 ANTI-TIGIT ANTIBODIES
专利摘要:
The present invention provides anti-TIGIT antibodies and antigen-binding fragments thereof that inhibit signaling via TIGIT. 公开号:BE1025333B1 申请号:E2017/5535 申请日:2017-07-31 公开日:2019-01-23 发明作者:Anthony Cooper；Christophe Quéva；Sofie Denies；Catherine Hoofd；Julia Cuende；Gregory Driessens 申请人:Iteos Therapeutics S.A.；Adimab Llc； IPC主号:
专利说明:
Anti-cancer immunotherapy relies on the modulation of the immune system to increase recognition and the response directed against tumor cells. Such modulation can be obtained by multiple mechanisms including the activation of costimulatory molecules present on immune cells or through the inhibition of coinhibitor receptors. Activation of an immune response is a complex mechanism involving many cell populations, such as antigen-presenting cells which are important for the initiation of an antigen-specific response and the effector cells responsible for cell destruction tumor. The mechanisms for modulating the activity of effector cells, such as cytotoxic T lymphocytes, are numerous and represent a target of choice in the context of anti-cancer immunotherapy TIGIT (T cell immunoreceptor with Ig and ITIM domains), also called WUCAM, VSIG9 or Vstm3, is a co-inhibitory receptor preferentially expressed on NK cells, CD8 + and CD4 + lymphocytes as well as on regulatory T lymphocytes (cells Treg, or simply "Tregs"). TIGIT is a transmembrane protein containing an ITIM domain in its intracellular part, a transmembrane domain and a variable domain of immunoglobulin on the extracellular part of the receptor. Several ligands of the TIGIT receptor have been described with CD155 / PVR demonstrating the best affinity followed by CD113 / PVRL3 and CD112 / PVRL2 (Yu et al. (2009) Nat. Immunol. 10:48.). DNAM / CD226, a co-stimulatory receptor, also expressed on NK cells and T lymphocytes, competes with TIGIT for binding with CD155 and CD112 but with a lower affinity, which suggests close control of activation of effector cells to avoid uncontrolled cytotoxicity against normal cells expressing the CD155 ligand. The expression of TIGIT is increased on tumor infiltrating lymphocytes (TIL) and in contexts of pathology, such as HIV infection. TIGIT expression marks depleted T cells which have weaker effector function compared to their TIGIT negative counterparts (Kurtulus et al. (2015) J. Clin. Invest. 276: 112; Chew et al. (2016) Plos Pathogens. 12). Conversely, Treg cells which express TIGIT demonstrate superior immunosuppressive activity compared to the TIGIT negative Treg cell population (Jolleretal. (2014) Immunity. 40: 569). Like other co-inhibitory receptors (PD1 or CTLA4) expressed on T lymphocytes which have been confirmed as relevant targets for immunotherapy and for which antagonistic antibodies have been approved for the treatment of cancer in humans, the development of an antagonistic anti-TIGIT antibody can help activate the immune system and BE2017 / 5535 better fight against cancer cells. It has been suggested that antagonistic anti-TIGIT antibodies as monotherapy, or in combination with an a-PD1 antibody, improve antitumor efficacy in preclinical models (Johnston et al. (2014) Cancer Cell 26: 1; WO2016 / 028656; US2016 / 0176963; US2016 / 0376365, the whole being incorporated here for reference). Thus, TIGIT-specific antagonist antibodies which inhibit the activity of the TIGIT receptor represent an opportunity to decrease the immunosuppressive effect associated with tumor microenvironments and, consequently, increase the anti-tumor immune response against tumor cells. SUMMARY OF THE INVENTION The present invention describes anti-TIGIT antibodies which can decrease the immunosuppressive effect of TIGIT receptor signaling. In particular, the antibodies of the invention can inhibit the immunosuppression due to TIGIT, by preventing the binding of the ligand on NK cells and T lymphocytes and / or the exhaustion of Treg cells positive for TIGIT, and / or by inducing internalization of the TIGIT receptor. In one aspect, the present invention describes an isolated antibody or an antigen binding fragment thereof which binds to human TIGIT which comprises a heavy chain variable domain comprising a heavy chain CDR1 (HCDR1), a chain heavy CDR2 (HCDR2) and a heavy chain CDR3 (HCDR3) chosen from the sequences HCDR1, HCDR2 and HCDR3 illustrated in FIG. 1 and which also comprises a variable domain of the light chain comprising a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2) and a light chain CDR3 (LCDR3) chosen from the sequences LCDR1, LCDR2 and LCDR3 illustrated in Figure 2. In some embodiments, the antibody or antigen binding fragment comprises a combination of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the combination is selected from the group of combinations formed by HCDRs from of each antibody in Figure 1 taken with the LCDRs of the corresponding antibody in Figure 2. In certain embodiments, an antibody or an antigen-binding fragment according to the invention can comprise a variable heavy chain domain having an amino acid sequence chosen from the group consisting of: SEQ ID NO: 211, 213 , 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 327, 329 and 331 and amino acid sequences demonstrating a sequence identity of at least 90% , 95%, 97%, 98% or 99% to them; and, optionally, comprises a variable domain of the light chain having an amino acid sequence chosen from the group composed of the amino acid sequences of SEQ ID NO: 212, 214, 216, 218, 220, 222, 224, 226 , 228, 230, 232, 234, 236, 238, 240, 328, 330 and 332 and amino acid sequences demonstrating a sequence identity of at least 90%, 95%, 97%, 98% or 99% to these. BE2017 / 5535 In some embodiments, the antibody or antigen binding fragment comprises a combination of a heavy chain variable domain and a light chain variable domain, in which the combination is selected from the group combinations formed by VH from each antibody in Figure 5, or from an amino acid sequence demonstrating a sequence identity of at least 90%, 95%, 97%, 98% or 99% to this ci, taken with the VL of the same antibody in Figure 5, or of an amino acid sequence demonstrating a sequence identity of at least 90%, 95%, 97%, 98% or 99% thereto . In certain embodiments, an antibody or an antigen binding fragment according to the invention comprises HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 in which HCDR1 comprises or is composed of SEQ ID NO: 16, HCDR2 comprises or is made up of SEQ ID NO: 17, HCDR3 includes or is made up of SEQ ID NO: 18, and LCDR1 understands or is made up of SEQ ID NO: 61, LCDR2 understands or is made up of SEQ ID NO: 62 , and LCDR3 includes or is composed of SEQ ID NO: 63. In certain embodiments, the variable domain of the heavy chain comprises or is composed of an amino acid sequence according to SEQ ID NO: 221 or an amino acid sequence demonstrating a sequence identity of at least 90% , 95%, 97%, 98% or 99% thereof, and the variable domain of the light chain comprises or is composed of an amino acid sequence according to SEQ ID NO: 222 or an acid sequence amines demonstrating a sequence identity of at least 90%, 95%, 97%, 98% or 99% thereof. In another aspect, the invention describes an isolated antibody or an antigen binding fragment thereof which binds to human TIGIT and which does not compete with CD155 for binding with TIGIT. In certain embodiments, the antibody or antigen binding fragment comprises HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 in which HCDR1 comprises or is composed of SEQ ID NO: 280, HCDR2 comprises or is composed of SEQ ID NO: 281, HCDR3 includes or is composed of SEQ ID NO: 282, and LCDR1 includes or is composed of SEQ ID NO: 292, LCDR2 includes or is composed of SEQ ID NO: 293, and LCDR3 includes or is composed of SEQ ID NO: 294. In certain embodiments, the variable domain of the heavy chain comprises or is composed of the amino acid sequence illustrated as SEQ ID NO: 333 or an amino acid sequence demonstrating a sequence identity of at least 90%, 95%, 97%, 98% or 99% to it, and the variable domain of the light chain comprises or is composed of the amino acid sequence illustrated as SEQ ID NO: 334 or an amino acid sequence demonstrating a sequence identity of at least 90%, 95%, 97%, 98% or 99% thereto. In certain embodiments of this type, the antibody or an antigen binding fragment thereof binds to human TIGIT and does not compete with CD155 for binding with TIGIT. In another aspect the invention describes an isolated antibody or an antigen binding fragment thereof, which cross-competes for binding with human TIGIT with an antibody according to the first aspect of the invention, by e.g. an antibody given as an example here. BE2017 / 5535 In another aspect, the invention describes an isolated antibody or an antigen-binding fragment thereof, which binds to the same epitope as an antibody according to the first aspect of the invention, e.g. antibody given as an example here. In another aspect, the invention describes an isolated anti-TIGIT antibody or an antigen binding fragment thereof which preferentially decreases TIGIT-expressing Treg cells, optionally in which the antibody or the binding fragment the antigen thereof is an antibody or an antigen binding fragment according to the first aspect of the invention, e.g., an antibody exemplified here. In another aspect, the invention describes a variant of the germ line or an affinity variant of an antibody according to the other aspects of the invention, eg, an antibody exemplified here. In another aspect, the invention describes an isolated polynucleotide encoding an antibody or antigen binding fragment, according to any other aspect of the invention, e.g., an antibody exemplified here. In another aspect, the invention describes an isolated polynucleotide encoding a VH and / or VL domain of an anti-TIGIT antibody, in which the polynucleotide comprises one or more sequences chosen from the group consisting of SEQ ID NO: 241- 270 and 335-342. In another aspect, the invention describes an expression vector comprising a polynucleotide according to the invention linked in operation to regulatory sequences which allow the expression of the antigen-binding polypeptide in a host cell or in a system. cell-free expression. In another aspect, the invention describes a host cell or a cell-free expression system containing an expression vector according to the invention. In another aspect, the invention describes a method of producing a recombinant antibody or an antigen binding fragment thereof which comprises culturing a host cell or an expression system free of cells according to the invention under conditions which allow the expression of the antibody or of the antigen-binding fragment and the recovery of the antibody or of the expressed antigen-binding fragment. In another aspect, the invention describes a pharmaceutical composition comprising an antibody or an antigen binding fragment according to the invention, e.g., an antibody exemplified herein, and at least one pharmaceutically acceptable carrier or excipient. In another aspect, the invention describes an antibody or antigen binding fragment according to the invention or a pharmaceutical composition according to the invention for use in therapy. BE2017 / 5535 In another aspect, the invention describes an antibody or antigen-binding fragment according to the invention (eg, an antibody given as an example here) or a pharmaceutical composition according to the invention for use in a method of treating cancer. In another aspect, the invention describes a method of treating cancer in a subject by administering an effective amount of an antibody or antigen-binding fragment according to the invention (e.g. , an antibody given as an example here) or a pharmaceutical composition according to the invention to a subject, thus treating cancer. In some embodiments, there is described an antibody or antigen binding fragment or a pharmaceutical composition for use in a method according to the invention, or a method of treating cancer according to the invention, wherein the method also includes administering an additional therapeutic agent. BRIEF DESCRIPTION OF THE FIGURES Figure 1 Table showing the sequences of the region determining the complementarity (CDR) of the variable domain of the heavy chain (VH) of the antibodies of the invention. Figure 2 Table presenting the CDR sequences of the variable domain of the light chain (VL) of the antibodies of the invention. Figure 3 Table showing the framework sequences (FR) of the variable domain of the heavy chain (VH) of the antibodies of the invention. Figure 4 Table showing the framework sequences (FR) of the variable domain of the light chain (VL) of the antibodies of the invention. Figure 5 Table showing the amino acid sequences of the variable domain of the heavy chain (VH) and the variable domain of the light chain (VL) of the antibodies of the invention. Figure 6 Table showing the sequences of the polynucleotides coding for the VH and VL domains of the antibodies according to the invention. Figure 7 Graph illustrating the results of a competition trial between hCD155 and the anti-TIGIT antibody for binding to Jurkat-hTIGIT cells. Figure 8 (Plate A) Graph illustrating the proportion of TIGIT positive cells among T cell subpopulations on PBMC cells from 7 healthy human donors. (Plate B) Graph illustrating the proportion of TIGIT positive cells among different immune subpopulations of PBMC from 7 healthy human donors. Figure 9 Graph illustrating the results of an anti-TIGIT antibody binding assay on Jurkat-hTIGIT cells. Figure 10 (Plates A and B) Graphs illustrating the results of an anti-TIGIT antibody binding assay on primary CD8 + T lymphocytes BE2017 / 5535 from healthy human PBMCs. (Plate C) Graph illustrating the results of an anti-TIGIT antibody binding test on primary memory CD8 + T lymphocytes and Tregs from healthy human PBMCs. FigureU (Plates A and B) Graphs illustrating the results of an anti-TIGIT antibody binding assay on primary CD8 + T lymphocytes from PBMCs of healthy cynomolgus. Figure 12 (Plates A, B and C) Graphs illustrating the effect of anti-TIGIT antibodies in a CHO-TCR-CD155 and Jurkat-hTIGIT bioassay. Figure 13 (Plates A, B and C) Graphs illustrating the effect of anti-TIGIT antibodies to increase the secretion of IFNg in a functional test on human primary CD8 + T lymphocytes from healthy donors activated with CHO- cells TCR-CD155. Figure 14 Histogram illustrating the effect of the anti-TIGIT antibody to increase the secretion of IFNg in a functional test on primary human CD8 + TILs from ascites of activated ovarian cancer patients with CHO-TCR-CD155 cells. Figure 15 (Plate A) Graphic illustrating the results of a competition test between the mouse CD155 and the anti-TIGIT antibody for binding to Jurkat-mTIGIT cells. (Plate B) Graph illustrating the effect of the antiTIGIT antibody to increase the secretion of IFNg in a functional test on T lymphocytes of OT-1 mice. (Plate C) Graph illustrating the effect of the anti-TIGIT antibody to increase cytotoxicity in a functional test on T lymphocytes of OT-1 mice. Figure 16 (Plate A) Graphic illustrating the anti-tumor efficacy of the anti-TIGIT antibody as monotherapy in a CT26 tumor model. (Boards B and C) Graphs illustrating the anti-tumor efficacy of the anti-TIGIT antibody in combination with anti-PD 1 in a CT26 tumor model. Figure 17 (Plate A) Graph illustrating the antitumor efficacy as a function of the isotype of the anti-TIGIT antibody as monotherapy in a CT26 tumor model. (Plate B) Graph illustrating the anti-tumor efficacy as a function of the isotype of the anti-TIGIT antibody in combination with the anti-PD1 in a CT26 tumor model. Figure 18 (Boards A and G) Graphs illustrating the modulation of the proportion of Treg cells in the total CD4 + T lymphocyte population in the CT26 tumor treated with the anti-TIGIT antibody as monotherapy or in combination with anti-PD 1. (Boards B and H) Graphs illustrating the modulation of the proportion of CD8 + T lymphocytes in the total population of CD45 + in the CT26 tumor treated with the anti-TIGIT antibody as monotherapy or in combination with the antiiPD1. (Boards C and I) Graphs illustrating the modulation of the ratio of CD8 + T lymphocytes / Treg cells in the CT26 tumor treated with the anti-TIGIT antibody in BE2017 / 5535 monotherapy or in combination with anti-PD1. (Plates D and J) Graph illustrating the modulation of CD4 + T lymphocytes secreting IFNg in the tumor CT26 treated with the anti-TIGIT antibody as monotherapy or in combination with anti-PD1. (Plate E) Graphic illustrating the modulation of CD8 + T lymphocytes secreting IFNg in the tumor CT26 treated with the anti-TIGIT antibody. (Plates L and F) Graphs illustrating the ratio of CD4 + T lymphocytes secreting IFNg / IL10 in the tumor CT26 treated with the anti-TIGIT antibody as monotherapy or in combination with anti-PD1. (Plates K) Graph illustrating the modulation of CD4 + T lymphocytes secreting IL-10 in the tumor CT26 treated with the anti-TIGIT antibody in combination with the anti-PD1 antibody. Figure 19 (Plate A) Graphic illustrating the effect of treatment with the anti-TIGIT antibody to modulate gene expression in the CT26 tumor and measured by NanoString. (Plate B) Graphic illustrating the modulation of the cytotoxic score in the CT26 tumor treated with the anti-TIGIT antibody as monotherapy or in combination with anti-PD1. (Plate C) Graph illustrating the modulation of the CD8 + T lymphocyte score in the CT26 tumor treated with the anti-TIGIT antibody as monotherapy or in combination with anti-PD1. Figure 20 (Plate A) Histograms illustrating the proportion of TIGIT + CD4 + , CD8 + T lymphocyte populations and Treg cells in PBMCs from healthy human volunteers. (Plate B) Graph illustrating the cytotoxic effect, in vitro, of the anti-TIGIT antibody on conventional populations of CD4 + , CD8 + T lymphocytes and Treg cells in PBMCs from healthy human volunteers. Figure 21 Graph illustrating the cytotoxic effect, ex vivo, of the anti-TIGIT antibody on conventional populations of CD4 + , CD8 + T lymphocytes and Treg cells in the CT26 tumor, in mice. DETAILED DESCRIPTION OF THE INVENTION In the present context, the term "immunoglobulin" includes a polypeptide having a combination of two heavy chains and two light chains, whether or not it has any relevant specific immunoreactivity. The term "antibody" describes such arrays that have known significant specific immunoreactive activity against an antigen of interest (eg, TIGIT). The terms "TIGIT antibody" or "anti-TIGIT antibody" are used herein to describe antibodies which demonstrate immunological specificity towards the TIGIT protein. Antibodies and immunoglobulins include light and heavy chains, in the presence or absence of an interchain covalent bond between the two. The basic structures of immunoglobulins in vertebrate systems are relatively well understood. BE2017 / 5535 The generic term "immunoglobulin" includes five distinct classes of antibodies that can be biochemically differentiated. Although all five classes of antibodies are within the scope of the present invention, the present discussion will generally focus on the IgG class of immunoglobulin molecules. With regard to IgG, the immunoglobulins comprise two identical light polypeptide chains with a molecular weight of approximately 23,000 Daltons, and two identical heavy chains with a molecular weight between 53,000 and 70,000. The four chains are linked by disulfide bridges in a "Y" configuration in which the light chains surround the heavy chains starting at the mouth of the "Y" and continuing through the variable region. The light chains of an antibody are classified as kappa or lambda (κ, λ). Each heavy chain class can be linked either with a kappa light chain or with a lambda light chain. Typically, the heavy and light chains are covalently linked to each other, and the "tail" portions of the two heavy chains are linked to each other by covalent disulfide bonds or bonds non-covalent when immunoglobulins are produced by B lymphocytes or genetically modified host cells. In the heavy chain, the amino acid sequences go from an N-terminal end at the split ends of the Y configuration to the C-terminal end at the bottom of the chain. Specialists in the field will understand that heavy chains are classified as gamma, mu, alpha, delta or epsilon, (γ, μ, α, δ, ε) with a few subclasses among them (e.g., γ 1-γ4) . It is the nature of this chain that determines the "class" of the antibody like IgG, IgM, IgA, IgD or IgE, respectively. Immunoglobulin subclasses (isotypes), e.g., lgG1, lgG2, lgG3, lgG4, lgA1, etc., are well characterized and are known to confer functional specialization. Modified versions of each of these classes and isotypes are readily distinguishable by a person skilled in the art in light of the present disclosure and, therefore, are within the scope of the present invention. As previously indicated, the variable region of an antibody allows the antibody to selectively recognize and specifically bind to epitopes on antigens. That is, the VL domain and the VH domain of an antibody combine to form the variable region which defines a 3D antigen binding site. This quaternary antibody structure forms the antigen binding site present at the end of each ΙΎ arm. More specifically, the antigen binding site is defined by three regions determining complementarity (CDR) on each of the VH and VL chains. In the present context, the terms “TIGIT protein” or “TIGIT antigen” or “TIGIT” are used interchangeably and describe the human T lymphocyte immunoreceptor (GenBank access number: NM_173799) which binds to the receptor / ligand poliovirus (PVR also called CD155). TIGIT is also called VSIG9, VSTM3 or WUCAM. The reference to TIGIT includes the native human TIGIT protein expressed naturally in the human host and / or BE2017 / 5535 on the surface of cultured human cell lines, as well as recombinant forms and fragments thereof, and also natural mutant forms. In the present context, the term "binding site" includes a region of a polypeptide which is responsible for selective binding to a target antigen of interest (eg, TIGIT). The binding domains include at least one binding site. Examples of binding domains include a variable antibody domain. The antibody molecules of the invention may include a single binding site or multiple (eg, two, three or four) binding sites. In the present context, the term "derivative" of a designated protein (eg, a TIGIT antibody or an antibody-binding fragment thereof) describes the origin of the polypeptide. In one embodiment, the polypeptide or amino acid sequence which is derived from a given starting polypeptide is a CDR sequence or a sequence related thereto. In one embodiment, the amino acid sequence which is derived from a given starting polypeptide is not contiguous. For example, in one embodiment, one, two, three, four, five, or six CDRs are derived from a starting antibody. In one embodiment, the polypeptide or amino acid sequence which is derived from a given starting polypeptide or from an amino acid sequence has an amino acid sequence which is essentially identical to that of the sequence starting material, or a part thereof, in which the part consists of at least 3 to 5 amino acids, at least 5 to 10 amino acids, at least 10 to 20 amino acids, d '' at least 20 to 30 amino acids or at least 30 to 50 amino acids, or which is identifiable by a person skilled in the art as having its origin in the starting sequence. In one embodiment, one or more CDR sequences derived from the starting antibody are modified to produce variant CDR sequences, e.g., affinity variants, in which the variant CDR sequences maintain the activity of link to TIGIT. In the present context, a "conservative amino acid substitution" is a substitution in which the amino acid residue is replaced by an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art, including basic side chains (eg, lysine, arginine, histidine), acid side chains (eg, aspartic acid, acid glutamic), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), non-polar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine , methionine, tryptophan), beta branching side chains (e.g. threonine, valine, isoleucine) and aromatic side chains (e.g. tyrosine, phenylalanine, tryptophan, histidine). Thus, a non-essential amino acid residue in an immunoglobulin polypeptide can be replaced by another amino acid residue belonging to the same side chain family. In another embodiment, an amino acid chain can be replaced with a chain having a similar structure which differs in order and / or composition from the members of the side chain family. BE2017 / 5535 In the present context, the term "heavy chain part" includes amino acid sequences derived from the constant domains of an immunoglobulin heavy chain. A polypeptide comprising a heavy chain part comprises at least: a CH1 domain, a hinge domain (e.g., the upper, middle and / or lower hinge region), a CH2 domain, a CH3 domain or a variant or fragment of this one. In one embodiment, an antibody or antigen binding fragment of the invention may comprise the Fc portion of an immunoglobulin heavy chain (e.g., a hinge portion, a CH2 domain and a CH3 domain ). In another embodiment, an antibody or an antigen binding fragment of the invention may lack part of a constant domain (eg, all or part of the CH2 domain). In some embodiments, at least one, and preferably all, the constant domains are derived from a heavy chain of human immunoglobulin. For example, in a preferred embodiment, the heavy chain portion comprises a fully human hinge domain. In other preferred embodiments, the heavy chain portion comprises a fully human Fc portion (eg, a hinge, CH2 and CH3 domain sequences from a human immunoglobulin). In certain embodiments, the constant domains constituting the heavy chain part come from different immunoglobulin molecules. For example, a heavy chain portion of a polypeptide may include a CH2 domain derived from an IgG 1 molecule and a hinge region derived from an IgG3 or IgG4 molecule. In other embodiments, the constant domains are chimeric domains comprising parts of different immunoglobulin molecules. For example, a hinge may include a first part from an IgGI molecule and a second part from an IgG3 or IgG4 molecule. As described above, a person skilled in the art will understand that the constant domains of the heavy chain part can be modified so that they vary in the amino acid sequence compared to the natural immunoglobulin molecule. (wild type). That is, the polypeptides of the invention disclosed herein may include alterations or modifications to one or more constant domains of the heavy chain (CH1, hinge, CH2 or CH3) and / or the domain of the constant region of the light chain (CL). Examples of modifications include additions, deletions or substitutions of one or more amino acids in one or more domains. In the present context, the terms “variable region” and “variable domain” are used interchangeably and are intended to have an equivalent meaning. The term "variable" describes the fact that certain parts of the VH and VL variable domains differ significantly in sequence among antibodies and are used in the binding and specificity of each given antibody for its target antigen. However, the variability is not evenly distributed across all of the variable domains of the antibodies. It is concentrated in three segments called the “hypervariable loops” in each of the VL and VH domains which are part of the antigen binding site. The first, second and third hypervariable loops of the chain domain BE2017 / 5535 slight V Lam bda are here called 1_1 (λ), Ι_2 (λ) and Ι_3 (λ) and can be defined as comprising residues 24-33 (L1 (À), composed of 9, 10 or 11 residues d amino acids), 49-53 (L2 (À), composed of 3 residues) and 90-96 (L3 (À), composed of 5 residues) in the VL domain (Morea et al., Methods 20, 267279, 2000 ). The first, second and third hypervariable loops of the V Ka ppa light chain domain are here called L1 (k), L2 (k) and L3 (k) and can be defined as comprising residues 25-33 (L1 ( k), composed of 6, 7, 8, 11, 12 or 13 amino acid residues), 49-53 (L2 (k), composed of 3 residues) and 90-97 (L3 (k), composed of 6 residues) in the VL field (Morea et al., Methods 20, 267-279, 2000). The first, second and third hypervariable loops of the VH domain are here called H1, H2 and H3 and can be defined as comprising residues 25-33 (H1, composed of 7, 8 or 9 residues), 52-56 (H2 , composed of 3 or 4 residues) and 91-105 (H3, of very variable length) in the VH domain (Morea et al., Methods 20, 267-279, 2000). Unless otherwise indicated, the terms L1, L2 and L3 respectively describe the first, the second and the third hypervariable loops of the VL domain, and include the hypervariable loops obtained from the two isotypes V Ka ppa and V Lam bda · The terms H1, H2 and H3 respectively describe the first, second and third hypervariable loops of the VH domain, and include hypervariable loops obtained from any of the known heavy chain isotypes, including y, ε, δ, a or μ. The hypervariable loops L1, L2, L3, H1, H2 and H3 may each include part of a "region determining complementarity" or "CDR", as defined below. The terms "hypervariable loop" and "region determining complementarity" are not strictly synonymous, since hypervariable loops (HV) are defined on the basis of a structure, while regions determining complementarity (CDR) are defined based on sequence variability (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991) and the HV and CDR limits may be different in certain areas VH and VL. The CDRs of the VL and VH domains can generally be defined as comprising the following amino acids: residues 24-34 (CDRL1), 50-56 (CDRL2) and 89-97 (CDRL3) in the variable domain of the light chain, and the residues 31-35 or 31-35b (CDRH1), 50-65 (CDRH2) and 95-102 (CDRH3) in the variable domain of the heavy chain; (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991). Thus, the HVs can be included in the corresponding CDRs and the references given here for the “hypervariable loops” of the VH and VL domains, should be interpreted as also encompassing the corresponding CDRs, and vice versa, unless otherwise indicated. The best-preserved parts of the variable domains are called the framework region (FR), as defined below. The variable domains of the native heavy and light chains each include four FRs (FR1, FR2, FR3 and FR4, respectively), largely adopting the configuration in β sheets, connected by the three hypervariable loops. The hypervariable loops in each chain BE2017 / 5535 are held together, close to each other, by the FRs, the hypervariable loops of the other chain contributing to the formation of the antigen binding site of the antibodies. A structural analysis of the antibodies revealed the relationship between the sequence and the shape of the binding site formed by the regions determining the complementarity (Chothia et al., J. Mol. Biol. 227, 799-817, 1992; Tramontano et al., J. Mol. Biol, 215, 175-182, 1990). Despite their high sequence variability, five of the six loops adopt only a small repertoire of main chain conformations, called "canonical structures". These conformations are firstly determined by the length of the loops and, secondly, by the presence of key residues at certain positions in the loops and in the frame regions which determine the conformation through their folding, the hydrogen bonds or the capacity to assume conformations. unusual main chain. In the present context, the term “CDR” or “complementarity determining region” describes the non-contiguous antigen combination sites found within the variable region of both the heavy chain and the light chain polypeptides. . These particular regions have been described by Kabat et al., J. Biol. Chem. 252, 6609-6616, 1977, by Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991, by Chothia et al., J. Mol. Biol. 196, 901-917, 1987, and by MacCallum et al., J. Mol. Biol. 262, 732-745, 1996, wherein the definitions include overlapping amino acid residues or subsets of amino acid residues when compared to each other. Amino acid residues which encompass the CDRs, as defined by each of the above references, are described for comparison. Preferably, the term "CDR" is a CDR as defined by Kabat based on the sequence comparisons. Table 1: Definitions of CDRs. CDR definitions Kabat ' Chothi Macca V H 31-35 26-32 30-35 V H 50-65 53-55 47-58 V H 95-102 96-101 93-101 V l 24-34 26-32 30-36 V l 50-56 50-52 46-55 V l 89-97 91-96 89-96 1 The numbering of residues follows the nomenclature of Kabat et al., Supra 2 The numbering of residues follows the nomenclature of Chothia et al., Supra 3 The numbering of residues follows the nomenclature of MacCallum et al., Supra In the present context, the term "framework region" or "FR region" includes amino acid residues that are part of the variable region, but which are not part of the CDRs (eg, in BE2017 / 5535 using the Kabat definition of CDR). Therefore, a variable region frame is about 100 to 120 amino acids in length but includes only amino acids outside of the CDRs. For the specific example of a heavy chain variable domain for the CDRs defined according to Kabat et al., The framework region 1 corresponds to the domain of the variable region encompassing amino acids 1 to 30; the framework region 2 corresponds to the domain of the variable region encompassing amino acids 36 to 49; the frame region 3 corresponds to the domain of the variable region including amino acids 66 to 94 and the frame region 4 corresponds to the domain of the variable region ranging from amino acids 103 to the end of the variable region. The framework regions for the light chain are similarly separated by each of the variable region of the light chain of the CDRs. Similarly, using the definition of CDR given by Chothia et al. or McCallum et al., the boundaries of the framework region are separated by their respective CDR ends as described above. In the preferred embodiments, the CDRs are as defined by Kabat. In natural antibodies, the six CDRs present on each monomeric antibody are short, non-contiguous amino acid sequences that are specifically positioned to form the antigen binding site when the antibody assumes its three-dimensional configuration in an aqueous environment . The rest of the heavy and light chain variable domains demonstrate less intermolecular variability in the amino acid sequence and are called the framework regions. The frame regions largely adopt a ß-sheet conformation and the CDRs form loops which connect, and in some cases are part of, the ß-sheet structure. Thus, these framework regions act to form a scaffold which allows the positioning of the six CDRs in a good orientation by interchain interactions, not covalent. The antigen binding site formed by the positioned CDRs defines a surface complementary to the epitope on the immunoreactive antigen. This complementary surface promotes the non-covalent binding of the antibody to the epitope of the immunoreactive antigen. The position of the CDRs can be easily identified by a person skilled in the art. In the present context, the term “fragment” describes a part or a portion of an antibody or of an antibody chain comprising less amino acid residues than an antibody or than an intact antibody chain or complete. The term "antigen binding fragment" describes a polypeptide fragment of an immunoglobulin or an antibody which binds to the antigen or which competes with the intact antibody (i.e. , with the intact antibody from which it is derived) for binding to the antigen (i.e., specific binding to TIGIT). In the present context, the term "fragment" of an antibody molecule includes fragments of the antibody antigen binding, eg, a variable domain of the light chain of an antibody (VL), a domain antibody heavy chain variable (VH), single chain antibody (scFv), F fragment (abj2, Fab fragment, Fd fragment, Fv fragment and single domain antibody fragment (DAb Fragments can be obtained, eg, by chemical or enzymatic treatment of an antibody or an intact or complete antibody chain by recombinant means. BE2017 / 5535 In the present context, the term "valence" describes the number of potential target binding sites in a polypeptide. Each target binding site specifically binds to a target molecule or to a specific site on a target molecule. When a polypeptide contains multiple target binding sites, each target binding site may specifically bind to the same molecule or to different molecules (e.g., may bind to different ligands or different antigens, or different epitopes on the same antigen). The binding molecules of the subject of the present invention have at least one specific binding site for TIGIT. In the present context, the term "specificity" describes the ability to bind (eg, to react immunologically with) a given target, eg, TIGIT. A polypeptide can be monospecific and contain one or more binding sites which specifically bind to a target or a polypeptide can be multispecific and contains two or more binding sites which specifically bind to the same target or to different targets. In one embodiment, an antibody of the invention is specific for several targets. For example, in this embodiment, a multispecies binding molecule of the invention binds to TIGIT and to a second target molecule. In this context, the second target molecule is a molecule other than TIGIT. In the present context, the term "synthetic" with respect to polypeptides includes polypeptides which include an unnatural amino acid sequence. For example, artificial polypeptides which are modified forms of natural polypeptides (eg, comprising a mutation such as addition, substitution or deletion) or which comprise a first amino acid sequence (which may be natural or artificial) which is linked in a linear amino acid sequence to a second amino acid sequence (which may be natural or artificial) to which it is not naturally linked in nature. In the present context, the term "modified" includes the manipulation of nucleic acid or polypeptide molecules by synthetic means (eg, recombinant techniques, in vitro peptide synthesis, enzymatic or chemical coupling of peptides, or certain combinations of these techniques). Preferably, the antibodies of the invention have been modified to improve one or more properties, such as antigen binding, stability / half-life or effector function. In the present context, the term "modified antibody" includes synthetic forms of the antibodies which are modified so that they are unnatural, eg, antibodies which comprise at least two parts of heavy chain but not two heavy chains complete (such as antibody or minibody with deleted domain); multispecific forms of antibodies (eg, bispecific, trispecific, etc.) modified to bind to two or more different antigens or to different epitopes on a single antigen; heavy chain molecules linked to scFv molecules, etc. ScFv molecules are known in the art and are described, e.g., in U.S. Patent 5,892,019. In addition, the term "modified antibody" includes multivalent forms of the antibodies (eg, trivalent, tetravalant, etc., antibodies which bind to two or more copies of the BE2017 / 5535 same antigen). In another embodiment, a modified antibody of the invention is a fusion protein comprising at least one heavy chain portion not containing the CH2 domain and comprising a binding domain of a polypeptide comprising the binding portion a member of a pair of receptor ligands. The term "modified antibody" can also be used herein to describe variants of the amino acid sequence of a TIGIT antibody of the invention. Those skilled in the art will understand that a TIGIT antibody of the invention can be modified to produce a variant TIGIT antibody which varies in the amino acid sequence compared to the TIGIT antibody from which it was derived. For example, nucleotide or amino acid substitutions leading to conservative substitutions or changes in "nonessential" amino acid residues can be made (eg, in CDR and / or frame). Amino acid substitutions may include the replacement of one or more amino acids with a natural or artificial amino acid. The term “variant of the germ line” is used here to specifically describe the variants in which the substitutions result in the replacement of one or more amino acid residues present at one or at given positions in the VH or VL d domain. 'A TIGIT antibody of the invention with an amino acid residue which is at an equivalent position in a reference human VH or VL domain encoded by the human germ line. Generally, for any "variant of the germ line", the substituted amino acid residues substituted in the variant of the germ line are taken exclusively, or essentially, from a single VH or VL domain encoded by the line human germ. In the present context, the term “affinity variant” describes a variant antibody which demonstrates one or more changes in the amino acid sequence in comparison with a reference TIGIT antibody of the invention, in which the variant d affinity demonstrates an altered affinity for TIGIT compared to the reference antibody. Preferably, the affinity variant will demonstrate an improved affinity for TIGIT, compared to the reference TIGIT antibody. The improvement may be apparent in the form of a lower KD for TIGIT, or a slower dissociation rate for TIGIT. Affinity variants generally demonstrate one or more changes in the amino acid sequence in CDRs, compared to the reference TIGIT antibody. Such substitutions may result in the replacement of the original amino acid present at a given position in the CDRs with a different amino acid residue, which may be a natural amino acid residue or an artificial amino acid residue. Amino acid substitutions can be conservative or non-conservative. In the present context, the term "affinity" or "binding affinity" should be understood based on the usual meaning in the field in the context of antibody binding, and reflects the strength and / or stability of the binding between an antigen and a binding site on an antibody or an antigen binding fragment thereof. BE2017 / 5535 The anti-TIGIT antibodies described here are characterized by a high affinity binding to human TIGIT. The binding affinity for TIGIT can be assessed using standard techniques known to those skilled in the art. Binding affinity can also be expressed as a dissociation constant for a given antibody, or K D. The lower the K D value, the stronger the binding interaction between an antibody and its target antigen. In one embodiment, the binding affinity of a Fab clone comprising a defined VH / VL pairing can be assessed using methods known in the art, e.g., by the ForteBio ™ system, by equilibration titration of the MSD solution (SET) or by surface plasmon resonance, eg using the Biacore ™ system as described in the examples attached. The Fab fragments of the antibodies according to the invention generally demonstrate a K D for the TIGIT measured by ForteBio ™ in the range going from 1 × 10 '10 to 5 × 10' 8 M, possibly 7 × 10 '10 to 4 × 10' 8 M. A KD in this range can be taken as an indication that the Fab, and a corresponding divalent mAb, demonstrate a high affinity binding for hTIGIT. Divalent mAbs comprising two Fabs (individually) demonstrating a KD for hTIGIT in the ranges mentioned are also taken to demonstrate a high affinity binding for hTIGIT. MSD a KD in the range of 1 x 10 '11 5 x 10' 9, possibly 2 x 10 '11 to 1 x 10' 9 can be taken as an indication of a high affinity binding to hTIGIT. The Fab fragments of the antibodies according to the invention generally demonstrate a KD for the TIGIT measured by Biacore ™ in the range going from 1 x 10 '10 M to 1 x 10' 10 M, possibly 2 x 10 ' 1 ° to 7 x 10 '10 M. A K D in this range can be taken as an indication that the Fab, and a corresponding divalent mAb, demonstrate a high affinity binding for hTIGIT. Binding affinity to human TIGIT can also be assessed using a cell-based system as described in the attached examples, in which the mAbs are tested for binding to mammalian cells (cell lines or cells ex vivo which express TIGIT), e.g. using ELISA or flow cytometry. A high affinity for TIGIT can be indicated, for example, by an EC 50 less than or equal to 0.5 nM by a flow cytometry analysis (for example, FACS) such as that described in Example 10. In certain embodiments, the antibodies of the invention demonstrate an EC 50 of binding to the cell less than or equal to 0.5 nM, possibly less than or equal to 0.2 nM. The determination of the affinity based on the cells expressed in the form of EC 50 is preferably carried out using Jurkat cells expressing hTIGIT or primary CD8 + T lymphocytes originating from human peripheral blood mononuclear cells (PBMC). In the present context, "Treg cells", or simply "Tregs", describe regulatory CD4 + T cells, ie, T cells which decrease the effector (s) of conventional T cells (T cells CD8 + or CD4 + ). Tregs can be identified by methods known in the art, e.g., using flow cytometry to identify CD4 cells expressing high levels of CD25 and low levels or the absence of CD127. BE2017 / 5535 As summarized above, the invention relates, at least in part, to the antibodies and antigen binding fragments thereof, which bind to TIGIT. The properties and characteristics of the TIGIT antibodies, and the antibody fragments according to the invention will now be described in more detail. In one aspect, the present invention describes an isolated antibody or an antigen binding fragment thereof which binds to human TIGIT which comprises a heavy chain variable domain comprising a heavy chain CDR1 (HCDR1), a chain heavy CDR2 (HCDR2) and a heavy chain CDR3 (HCDR3) chosen from the sequences HCDR1, HCDR2 and HCDR3 illustrated in FIG. 1 and which also comprises a variable domain of the light chain comprising a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2) and a CDR3 light chain (LCDR3) selected from the sequences LCDR1, LCDR2 and LCDR3 illustrated in Figure 2. That is, the invention describes an isolated antibody or a binding fragment antigen thereof which binds to human TIGIT and which comprises a heavy chain variable domain comprising a CDR1 heavy chain (HCDR1), a CDR2 heavy chain (HCDR2) and a CDR3 heavy chain (HCDR3), in which: (i) HCDR1 is chosen from the group composed of SEQ ID NOs: 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, 43, 271, 274 and 277; (ii) HCDR2 is chosen from the group composed of SEQ ID NOs: 2, 5, 8, 11, 14, 17, 20, 23, 26, 29, 32, 35, 38, 41.44, 272, 275 and 278; (iii) HCDR3 is chosen from the group composed of SEQ ID NOs: 3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 39, 42, 45, 273, 276 and 279; and which also comprises a light chain variable domain comprising a CDR1 light chain (LCDR1), a CDR2 light chain (LCDR2) and a CDR3 light chain (LCDR3), in which (iv) LCDR1 is chosen from the group consisting of SEQ IDs NO: 46, 49, 52, 55, 58, 61, 64, 67, 70, 73, 76, 79, 82, 85, 88, 283, 286 and 289; (v) LCDR2 is chosen from the group composed of SEQ ID NOs: 47, 50, 53, 56, 59, 62, 65, 68, 71, 74, 77, 80, 83, 86, 89, 284, 287 and 290; and (vi) LCDR3 is chosen from the group consisting of SEQ ID NOs: 48, 51, 54, 57, 60, 63, 66, 69, 72, 75, 78, 81, 84, 87, 90, 285, 288 and 291. Any anti-TIGIT antibody or an antigen binding fragment thereof comprising a VH domain paired with a VL domain to form an antigen binding site (human TIGIT) will comprise a combination of six CDRs: chain CDR3 variable heavy (HCDR3), CDR2 variable heavy chain (HCDR2), CDR1 variable heavy chain (HCDR1), CDR3 variable light chain (LCDR3), CDR2 variable light chain (LCDR2) and variable light chain BE2017 / 5535 CDR1 (LCDR1). Although many different combinations of the six CDRs selected from the groups of CDR sequences listed above are permitted, and within the scope of the invention, certain combinations of the six CDRs are particularly preferred; these being the “native” combinations within a single mAb demonstrating a high affinity for binding to human TIGIT. In some embodiments, the antibody or antigen binding fragment comprises a combination of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, in which the combination is selected from the group of combinations formed by HCDRs from of each antibody of Figure 1 taken with the LCDRs of the corresponding antibody of Figure 2. That is, in some embodiments, the antibody or the antigen binding fragment comprises a combination of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, in which the combination is chosen from the group consisting of: (i) HCDR1 comprising SEQ ID NO: 1, HCDR2 comprising SEQ ID NO: 2, HCDR3 comprising SEQ ID NO: 3, LCDR1 comprising SEQ ID NO: 46, LCDR2 comprising SEQ ID NO: 47 and LCDR3 comprising SEQ ID NO: 48; (ii) HCDR1 comprising SEQ ID NO: 4, HCDR2 comprising SEQ ID NO: 5, HCDR3 comprising SEQ ID NO: 6, LCDR1 comprising SEQ ID NO: 49, LCDR2 comprising SEQ ID NO: 50 and LCDR3 comprising SEQ ID NO: 51; (iii) HCDR1 comprising SEQ ID NO: 7, HCDR2 comprising SEQ ID NO: 8, HCDR3 comprising SEQ ID NO: 9, LCDR1 comprising SEQ ID NO: 52, LCDR2 comprising SEQ ID NO: 53 and LCDR3 comprising SEQ ID NO: 54; (iv) HCDR1 comprising SEQ ID NO: 10, HCDR2 comprising SEQ ID NO: 11, HCDR3 comprising SEQ ID NO: 12, LCDR1 comprising SEQ ID NO: 55, LCDR2 comprising SEQ ID NO: 56 and LCDR3 comprising SEQ ID NO: 57; (v) HCDR1 comprising SEQ ID NO: 13, HCDR2 comprising SEQ ID NO: 14, HCDR3 comprising SEQ ID NO: 15, LCDR1 comprising SEQ ID NO: 58, LCDR2 comprising SEQ ID NO: 59 and LCDR3 comprising SEQ ID NO: 60; (vi) HCDR1 comprising SEQ ID NO: 16, HCDR2 comprising SEQ ID NO: 17, HCDR3 comprising SEQ ID NO: 18, LCDR1 comprising SEQ ID NO: 61, LCDR2 comprising SEQ ID NO: 62 and LCDR3 comprising SEQ ID NO: 63; (vii) HCDR1 including SEQ ID NO: 19, HCDR2 including SEQ ID NO: 20, HCDR3 including SEQ ID NO: 21, LCDR1 including SEQ ID NO: 64, LCDR2 including SEQ ID NO: 65 and LCDR3 including SEQ ID NO: 66; BE2017 / 5535 (viii) HCDR1 including SEQ ID NO: 22, HCDR2 including SEQ ID NO: 23, HCDR3 including SEQ ID NO: 24, LCDR1 including SEQ ID NO: 67, LCDR2 including SEQ ID NO: 68 and LCDR3 comprising SEQ ID NO: 69; (ix) HCDR1 including SEQ ID NO: 25, HCDR2 including SEQ ID NO: 26, HCDR3 including SEQ ID NO: 27, LCDR1 including SEQ ID NO: 70, LCDR2 including SEQ ID NO: 71 and LCDR3 including SEQ ID NO: 72; (x) HCDR1 comprising SEQ ID NO: 28, HCDR2 comprising SEQ ID NO: 29, HCDR3 comprising SEQ ID NO: 30, LCDR1 comprising SEQ ID NO: 73, LCDR2 comprising SEQ ID NO: 74 and LCDR3 comprising SEQ ID NO: 75; (xi) HCDR1 including SEQ ID NO: 31, HCDR2 including SEQ ID NO: 32, HCDR3 including SEQ ID NO: 33, LCDR1 including SEQ ID NO: 76, LCDR2 including SEQ ID NO: 77 and LCDR3 including SEQ ID NO: 78; (xii) HCDR1 comprising SEQ ID NO: 34, HCDR2 comprising SEQ ID NO: 35, HCDR3 comprising SEQ ID NO: 36, LCDR1 comprising SEQ ID NO: 79, LCDR2 comprising SEQ ID NO: 80 and LCDR3 comprising SEQ ID NO: 81; (xiii) HCDR1 including SEQ ID NO: 37, HCDR2 including SEQ ID NO: 38, HCDR3 including SEQ ID NO: 39, LCDR1 including SEQ ID NO: 82, LCDR2 including SEQ ID NO: 83 and LCDR3 including SEQ ID NO: 84; (xiv) HCDR1 comprising SEQ ID NO: 40, HCDR2 comprising SEQ ID NO: 41, HCDR3 comprising SEQ ID NO: 42, LCDR1 comprising SEQ ID NO: 85, LCDR2 comprising SEQ ID NO: 86 and LCDR3 comprising SEQ ID NO: 87; (xv) HCDR1 including SEQ ID NO: 43, HCDR2 including SEQ ID NO: 44, HCDR3 including SEQ ID NO: 45, LCDR1 including SEQ ID NO: 88, LCDR2 including SEQ ID NO: 89 and LCDR3 including SEQ ID NO: 90; (xvi) HCDR1 including SEQ ID NO: 271, HCDR2 including SEQ ID NO: 272, HCDR3 including SEQ ID NO: 273, LCDR1 including SEQ ID NO: 283, LCDR2 including SEQ ID NO: 284 and LCDR3 including SEQ ID NO: 285; (xvii) HCDR1 comprising SEQ ID NO: 274, HCDR2 comprising SEQ ID NO: 275, HCDR3 comprising SEQ ID NO: 276, LCDR1 comprising SEQ ID NO: 286, LCDR2 comprising SEQ ID NO: 287 and LCDR3 comprising SEQ ID NO: 288; (xviii) HCDR1 comprising SEQ ID NO: 277, HCDR2 comprising SEQ ID NO: 278, HCDR3 including SEQ ID NO: 279, LCDR1 including SEQ ID NO: 289, LCDR2 including SEQ ID NQ: 290 and LCDR3 including SEQ ID NO: 291. BE2017 / 5535 In some embodiments, the antibody or antigen binding fragment comprises a heavy chain variable domain having an amino acid sequence selected from the group consisting of: SEQ ID NO: 211, 213, 215, 217 , 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 327, 329 and 331 and amino acid sequences demonstrating a sequence identity of at least 90%, 95%, 97%, 98% or 99% to them; and, optionally, comprises a variable domain of the light chain having an amino acid sequence chosen from the group composed of: amino acid sequences of SEQ ID NO: 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 328, 330 and 332 and amino acid sequences demonstrating a sequence identity of at least 90%, 95%, 97%, 98% or 99 % to these. Even if all the possible pairings of the VH domains and of the VL domains chosen from the VH and VL domain sequence groups listed above are permitted, and within the scope of the invention, certain combinations of VH and VL are particularly preferred; these being the “native” combinations within a single mAb demonstrating a high affinity for binding to human TIGIT. In some embodiments, the antibody or antigen binding fragment comprises a combination of a heavy chain variable domain and a light chain variable domain, in which the combination is selected from the group combinations formed by VH from each antibody in Figure 5, or from an amino acid sequence demonstrating a sequence identity of at least 90%, 95%, 97%, 98% or 99% to this ci, taken with the VL of the same antibody in Figure 5, or of an amino acid sequence demonstrating a sequence identity of at least 90%, 95%, 97%, 98% or 99% thereto . In certain embodiments, the antibody or the antigen-binding fragment comprises a combination of a heavy chain variable domain and a light chain variable domain, in which the combination is chosen from the group consisting of: (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 211 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 212; (ii) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 213 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 214; (iii) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 215 and a light chain variable domain comprising the amino acid sequence of SEQIDNO: 216; (iv) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 217 and a light chain variable domain comprising the amino acid sequence of SEQIDNO: 218; BE2017 / 5535 (v) of a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 219 and of a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 220; (vi) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 221 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 222; (vii) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 223 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 224; (viii) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 225 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 226; (ix) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 227 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 228; (x) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 229 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 230; (xi) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 231 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 232; (xii) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 233 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 234; (xiii) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 235 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 236; (xiv) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 237 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 238; (xv) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 239 and a light chain variable domain comprising the amino acid sequence of SEQ ID NQ: 240; BE2017 / 5535 (xvi) of a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 327 or an amino acid sequence identical to at least 90% thereof and a variable domain of light chain comprising the amino acid sequence of SEQ ID NO: 328 or an amino acid sequence identical to at least 90% thereof; (xvii) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 329 or an amino acid sequence identical to at least 90% thereof and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 330 or an amino acid sequence identical to at least 90% thereof; and (xviii) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 331 or an amino acid sequence identical to at least 90% thereof and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 332 or an amino acid sequence identical to at least 90% thereof. For each of the specific VH / VL combinations listed above, it is also permitted, and within the scope of the invention, to combine a VH domain having an amino acid sequence identical to at least 90%, 92%, 95 %, 97% or 99% to the VH domain sequence described with a VL domain having an amino acid sequence identical to at least 90%, 92%, 95%, 97% or 99% to the VL domain sequence described . The embodiments in which the amino acid sequence of the VH domain demonstrates a sequence identity of less than 100% with a given reference VH sequence, may nevertheless include heavy chain CDRs which are identical to HCDR1, HCDR2 and HCDR3 of the reference sequence while demonstrating variation in the amino acid sequence within the framework regions. Likewise, embodiments in which the amino acid sequence of the VL domain demonstrates a sequence identity of less than 100% with a given reference VL sequence, may nevertheless include light chain CDRs which are identical to LCDR1 , LCDR2 and LCDR3 of the reference sequence while demonstrating a variation in the amino acid sequence within the framework regions. In the previous paragraph, and elsewhere in this document, the structure of the antibody / antigen binding fragments is defined on the basis of a percentage of sequence identity with a reference sequence described (with an SEQ ID NO given). In this context, the percentage of sequence identity between two amino acid sequences can be determined by comparison of these two optimally aligned sequences in which the amino acid sequence to be compared can comprise additions or deletions with respect to to the reference sequence for optimal alignment between these two sequences. The percentage of identity is calculated by determining the number of identical positions for which the amino acid residue is identical between the two sequences, by dividing this number of identical positions by the total number of positions in the comparison window and by multiplying the result obtained by 100 to obtain the percentage of identity between these two sequences. Generally, the BE2017 / 5535 comparison will be the total length of the sequence being compared. For example, it is possible to use the BLAST program, "Blast 2 sequences" (Tatusova et al, "Blast 2 sequences a new tool for comparing protein and nucleotide sequences", FEMS Microbiol Lett. 174: 247-250) available on the site http://www.ncbi.nlm.nih.gov/gorf/bl2.html, the parameters used being those given by default (in particular, for the parameters of “penalty for open holes”: 5, and “penalty hole extension ”: 2; the chosen matrix being, for example, the“ BLOSUM 62 ”matrix proposed by the program), the percentage of identity between the two sequences which must be compared being calculated directly by the program. Determining the sequence identity of an interrogation sequence relative to a reference sequence is within the skill of the expert and can be performed using commercially available analysis software, such as BLAST ™ . In certain preferred embodiments, the antibody may comprise a heavy chain variable domain and a light chain variable domain in which HCDR1 comprises SEQ ID NO: 16, HCDR2 comprises SEQ ID NO: 17, HCDR3 comprises SEQ ID NO: 18, and the LCDR1 includes the SEQ ID NO: 61, LCDR2 includes the SEQ ID NO: 62, and the LCDR3 includes the SEQ ID NO: 63. In certain embodiments of this type, the variable domain of the heavy chain can comprise the amino acid sequence illustrated as SEQ ID NO: 221 or an amino acid sequence demonstrating a sequence identity of at least 90% , 95%, 97%, 98% or 99% thereof, and the variable domain of the light chain can comprise the amino acid sequence illustrated as SEQ ID NO: 222 or an amino acid sequence demonstrating sequence identity of at least 90%, 95%, 97%, 98% or 99% to it. The embodiments in which the amino acid sequence of the VH domain demonstrates a sequence identity of less than 100% with the sequence illustrated as SEQ ID NO: 221 may nevertheless include the heavy chain CDRs which are identical to HCDR1, HCDR2 and HCDR3 of SEQ ID NO: 221 (SEQ ID NO: 16, 17 and 18, respectively) while demonstrating variation in the amino acid sequence within the framework regions. Likewise, embodiments in which the amino acid sequence of the VL domain demonstrates a sequence identity of less than 100% with the sequence illustrated as SEQ ID NO: 222 may nevertheless include heavy chain CDRs which are identical to LCDR1, LCDR2 and LCDR3 of SEQ ID NO: 222 (SEQ ID NO: 61, 62 and 63, respectively) while demonstrating a variation in the amino acid sequence within the framework regions. TIGIT antibodies can take the form of various embodiments in which both a VH domain and a VL domain are present. The term "antibody" is used in the present context in its broadest sense and includes, without limitation, monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, multi-specific antibodies (eg. , bispecific antibodies), as long as they demonstrate the appropriate immunological specificity for a human TIGIT protein. The term "monoclonal antibody", as used herein, refers to an antibody obtained from a BE2017 / 5535 substantially homogeneous antibody population, that is to say that the individual antibodies forming the population are identical except for possible mutations of natural origin which may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. In addition, unlike conventional antibody preparations (polyclonal) which generally include different antibodies directed against different determinants (epitopes) on the antigen, each monoclonal antibody is directed against a single determinant or epitope on the antigen. "Antibody fragments" include part of a full-length antibody, usually the antigen binding region or the variable domain thereof. Examples of antibody fragments include Fab, Fab ', F (ab') 2, bispecific Fabs and Fv fragments, diabodies, linear antibodies, single chain antibody molecules, variable fragment single chain (scFv) and multispecific antibodies formed from antibody fragments (see Holliger and Hudson, Nature Biotechnol. 23: 1126-1136, 2005, the content of which is incorporated herein by reference). In nonlimiting embodiments, the TIGIT antibodies described here can comprise CH1 domains and / or CL domains, the amino acid sequence of which is completely or essentially human. If the TIGIT antibody is for therapeutic use in humans, it is usual for the entire constant region of the antibody, or at least part of it, to have an amino acid sequence totally or essentially human. Therefore, one or more or any combination of the CH1 domain, the hinge region, the CH2 domain, the CH3 domain and the CL domain (and the CH4 domain, if present) may be wholly or essentially human with respect to to its amino acid sequence. Such antibodies can be of any human isotype, human IgG4 and IgGI being particularly preferred. Advantageously, the CH1 domain, the hinge region, the CH2 domain, the CH3 domain and the CL domain (and the CH4 domain, if present) may all have a fully or essentially human amino acid sequence. In the context of the constant region of a humanized or chimeric antibody, or an antibody fragment, the term "essentially human" describes an amino acid sequence identity of at least 90%, or at least at least 92%, or at least 95%, or at least 97%, or at least 99% with a constant human region. The term "human amino acid sequence" in the present context describes an amino acid sequence which is encoded by a human immunoglobulin gene, which comprises mutated germ line genes, rearranged or somatic. Such antibodies can be of any human isotype, human IgG4 and IgGI being particularly preferred. Also described are TIGIT antibodies comprising constant domains of the “human” sequence which have been altered by one or more additions, deletions or substitutions of amino acids with respect to the human sequence. BE2017 / 5535 The TIGIT antibodies described here can be of any isotype. Antibodies for therapeutic use in humans will generally be of the IgA, IgD, IgE, IgG, IgM type, often of the IgG type, in which case, they may belong to any of the four subclasses lgG1, lgG2a and b , lgG3 or lgG4. Within each of these subclasses, it is permitted to make one or more substitutions, insertions or deletions of amino acids inside the Fc part, or to make other structural modifications, e.g. , to improve or reduce the Fc dependent functionality. In certain preferred embodiments, the TIGIT antibodies described herein are IgG antibodies. In certain embodiments, the antibodies according to the invention are IgG1 antibodies. In certain alternative embodiments, the antibodies according to the invention are IgG4 antibodies. IgG4 antibodies are known to undergo Fab arm exchange (FAE), which can give unpredictable pharmacodynamic properties of an IgG4 antibody. It has been shown that FAE can be prevented by an S228P mutation in the hinge region (Silva et al. J Biol Chem. 2015 Feb 27; 290 (9): 5462-5469). Consequently, in certain embodiments in which an antibody according to the invention is an IgG4 antibody, the antibody comprises the mutation S228P, i.e., a serine to proline mutation at position 228 (according to the numbering EU). In nonlimiting embodiments, it is envisaged that one or more substitutions, insertions or deletions of amino acids can be carried out inside the constant region of the heavy and / or light chain, particularly inside from the Fc region. Amino acid substitutions can result in the replacement of the substituted amino acid with a different natural amino acid, or with an artificial or modified amino acid. Other structural modifications are also permitted, such as, for example, changes in the glycosylation pattern (eg, by addition or deletion of glycosylation sites linked to N or O). Depending on the intended use of the TIGIT antibody, it may be desirable to modify the antibody of the invention with respect to its Fc receptor binding properties, eg, to modulate the effector function. In some embodiments, TIGIT antibodies may include an Fc region of a given antibody isotype, e.g., human IgGI, which is modified to reduce or substantially eliminate one or more antibody effector functions naturally associated with this antibody isotype. As demonstrated here, antibodies with effector lysis functions can be effective in reducing Treg cell populations but, surprisingly, without adversely affecting populations of conventional effector T cells. This selectivity allows a more powerful inhibition of the regulatory effect of Tregs while maintaining the anti-tumor effector T lymphocytes. Therefore, in some alternative embodiments, TIGIT antibodies retain one or more of the effector functions of the antibody naturally associated with this antibody isotype. For example, the TIGIT antibodies of the invention can be IgG1 antibodies which retain ADCC functionality. In other embodiments, TIGIT antibodies may include an Fc region of a given antibody isotype, e.g., human IgGI, which is modified to improve one or more effector functions of associated antibodies BE2017 / 5535 naturally to this antibody isotype. In this context, "antibody effector functions" include one or more of antibody-dependent cell cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC) and antibody-dependent cell phagocytosis (ADCP ), or all three. The monoclonal antibodies or antigen-binding fragments thereof which "cross-compete" with the TIGIT antibodies disclosed herein are those which bind to human TIGIT at the site (s) which are identical, overlap, or to the sites at which the TIGIT antibodies of the present invention bind. Competitive monoclonal antibodies or antigen-binding fragments thereof can be identified, e.g., by an antibody competition assay. For example, a sample of purified or partially purified human TIGIT can be attached to a solid support. Next, an antibody (or one of its antigen-binding fragments) among the present invention and a monoclonal antibody (or one of its antigen-binding fragments) which would be able to compete with an antibody of the invention are added. One of the two molecules is marked. If the labeled compound and the unlabeled compound bind to separate and independent sites on the TIGIT, the labeled compound will bind at the same level regardless of whether the supposedly competing compound is present or not. However, if the interaction sites are identical or overlap, the unlabeled compound will compete, and the amount of the labeled compound bound to the antigen will be decreased. If the unlabeled compound is present in excess, a very small amount, if present, of the labeled compound will bind. In the context of the present invention, the competing monoclonal antibodies (or antigen-binding fragments thereof) are those which reduce the binding of the antibody compounds of the present invention to the TIGIT by approximately 50%, approximately 60%, approximately 70%, approximately 80%, approximately 85%, approximately 90%, approximately 95% or approximately 99%. Details of the procedures for performing such competitive trials are well known in the art and can be found, e.g., in Harlow and Lane, Antibodies, A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1988, 567-569, 1988, ISBN 0-87969-314-2. Such tests can become quantitative by the use of purified antibodies. A standard curve is plotted by diluting an antibody against itself, i.e., the same antibody is used for both the marker and the competitor. The ability of an unlabelled competitive monoclonal antibody or an antigen binding fragment thereof to inhibit the binding of the labeled molecule to the plate is measured. The results are reported and the concentrations necessary to achieve the desired degree of binding inhibition are compared. Examples of TIGIT antibodies described here and having the sequences given in FIGS. 1 to 5 were developed from 5 clones of parent antibodies. Table 2 summarizes the antibody line described here. The human parent anti-TIGIT antibodies expressed in yeast and demonstrating a high functional activity against TIGIT were selected (gray lines, named 26 ...), underwent affinity maturation and were expressed in mammalian cells. to produce the second generation of antibodies (white lines below each parent, named 29 ...). In BE2017 / 5535 in addition, the antibody 31282 was produced from 29489 by an M-T substitution at amino acid 116 in the FR4 region of the VH. It is understood that this substitution removes a site of potential oxidation of the antibody and thus improves the stability without affecting the function. In addition, antibody 31288 was produced from 29494 by a V-L substitution at amino acid 2 in the FR1 region of VH and by an M-T substitution at amino acid 120 in the FR4 region of VH. It is understood that the V-L substitution restores the sequence of the VH4-39 germ line and the M-T substitution removes the potential oxidation site of the antibody and thus improves stability without affecting function. Table 2 Antibody clone VH CDR3 line Optimization process Germline ofVH 26518 26518 relative VH3-07 29478 26518 H1 / H2 / H3 VH3-30 26452 26452 relative VH 1-46 29487 26452 H1 / H2 / H3 VH 1-46 29489 26452 H1 / H2 / H3 VH 1-46 31282 29489 Amino acid mutationM116T VH 1-46 26486 26486 relative VH4-0B 29499 26486 H1 / H2 / H3 VH4-39 29494 26486 H1 / H2 / H3 VH4-39 31288 29494 Inversion of the germ line + mutation of amino acid M116T VH4-39 32919 31288 L1 / L2 / L3 VH4-39 32931 31288 L1 / L2 / L3 VH4-39 26521 26521 relative VH 1-69 29513 26521 H1 / H2 / H3 VH 1-69 26493 26493 relative VH3-09 29520 26493 H1 / H2 / H3 VH3-09 29523 26493 H1 / H2 / H3 VH3-33 29527 26493 H1 / H2 / H3 VH3-30 26432 26432 relative VH 1-69 The second generation of antibodies demonstrates higher affinity than the respective parent antibodies, as summarized in Table 3. BE2017 / 5535 Table 3 clone ForteBio Fab KD TIGIT Human Biotinylated TIGIT HIS (M) Monovalent ForteBio Fab KD TIGIT-Fc mouse(M) Monovale nt ForteBio Fab KD TIGIT-Fc cyno (M) Monovalent ForteBio IgG KD IgG KD TIGIT-Fc human (M) Avid MSD monovalent KD (M), TIGIT-His human Biovore monovale nt KD (M), human TIGIT-His Link to Human Jurkat TIGIT (Difference versus negative cells) Link to Jurkat TIGIT mice (Difference versus negative cells) 26518 1.24E-09 N.B. 4.47E-09 6.30E-10 154 233 29478 7.03E-10 9.18E-08 1.26E-09 5.27E-10 182 500 26452 5.08E-09 N.B. N.B. 4.74E-10 164 47 29487 2.08E-09 N.B. 1.55E-07 3.96E-10 161 95 29489 8.81E-10 N.B. 3.52E-08 3.53E-10 1.1E-10 2.48E-10 162 187 31282 1.34E-09 N.B. 3.77E-08 2.94E-10 26486 2.19E-08 N.B. N.B. 5.89E-10 143 199 29499 1.66E-09 2.55E-08 1.45E-08 3.19E-10 1.9E-11164 541 29494 1.66E-09 5.36E-08 1.86E-08 3.76E-10 7.0E-11 2.70E-10 164 511 31288 2.09E-092.51E-08 1.92E-10 32919 1.42E-096.57E-09 68032931 1.18E-091.97E-09 74129499 1.66E-09 2.55E-08 1.45E-08 3.19E-10 1.9E-11164 541 26521 9.87E-09 N.B. 1.49E-07 5.41E-10 146 218 29513 7.74E-10 8.55E-08 9.56E-09 3.92E-10 2.5E-11156 406 26493 4.06E-08 2.67E-08 N.B. 1.49E-09 80 463 29520 1.31E-09 1.95E-09 2.68E-09 3.84E-10 2.1E-10 7.16E-10 166 535 29523 3.84E-09 1.89E-08 2.79E-08 5.31E-10 1.7E-09150 502 29527 1.33E-09 2.02E-08 1.76E-08 3.50E-10 6.4E-10142 414 26432 1.31E-08 N.B. N.B. 4.62E-09 Therefore, in some embodiments, the antibodies or antigen binding fragments of the invention demonstrate a high affinity for human TIGIT. In certain embodiments, the Fab fragments of the antibodies according to the invention demonstrate a K D for TIGIT measured by ForteBio ™ in the range ranging from 1 × 10 10 to 5 × 10 8 M, possibly 7 × 10 10 at 4 x 10 ' 8 M. In certain embodiments, the antibodies according to the invention demonstrate a KD measured by MSD in the range from 1 x 10' 11 to 5 x 10 ' 9 M, possibly 2 x 10' 11 to 1 x 10 ' 9 . In certain embodiments, the Fab fragments of the antibodies according to the invention demonstrate a 10 KD for TIGIT measured by Biacore ™ in the range going from 1 × 10 −10 M to 1 × 10 −9 M, possibly 2 × 10 10 at 7 x 10 '10 M. In some embodiments, the antibodies or antigen binding fragments of the invention cross-react with TIGIT from mice and / or TIGIT from cynomolgus. Since the second generation antibodies "29 ..." are an affinity-matured progeny from the highly functional parent antibodies, it is expected that they will demonstrate functional properties at least similar or equivalent to the parent antibodies , and vice versa. As described herein, in some embodiments, an antibody or antigen binding fragment of the invention has equivalent affinity for TIGIT expressed on lymphocytes BE2017 / 5535 T CD8 and Treg cells. In the present context, an antibody or antigen binding fragment has "equivalent affinity" for CD8 T cells and Treg cells if the affinity for CD8 T cells is in the range of 0.5 to 1.5 times that of the affinity for Treg cells. For example, an antibody with equivalent affinity for CD8 T cells and Treg cells that demonstrates an affinity for Treg cells of 0.03 nM would demonstrate an affinity for CD8 T cells in the range of 0.015 to 0.045 nM. Table 3 presents a summary of the affinity properties of the anti-TIGIT antibodies of the invention, the gray cells indicating clones of the parent antibody, the second and third generation antibodies of each line being immediately illustrated in below the respective parent antibody (see also Table 2). As demonstrated in the attached examples, in some embodiments, antibodies of the invention compete with CD155 / PVR for binding with TIGIT. In some embodiments, the antibodies of the invention demonstrate competition with CD155 characterized by an IC 50 of 0.2 nM or less, preferably 0.1 nM or less. Without intending to be bound by theory, the competition of antibodies with CD155 for binding to TIGIT is expected to decrease the signaling levels induced by CD155 via TIGIT, thereby increasing lymphocyte activation levels. T effectors. As demonstrated in the attached examples, in certain embodiments, the antibodies of the invention demonstrate a high affinity for CD8 T lymphocytes expressing TIGIT and a high affinity for Treg cells expressing TIGIT. In certain embodiments, the antibodies of the invention demonstrate an affinity for CD8 T lymphocytes expressing TIGIT and Treg cells expressing TIGIT characterized by an EC 50 of less than 0.5 nM, preferably less than 0.3 nM, of preferably less than 0.2 nM. In certain embodiments, the antibodies of the invention demonstrate an equivalent affinity for CD8 T lymphocytes expressing TIGIT and for Treg cells expressing TIGIT. The antibodies according to the invention are also able to preferentially decrease the Treg cells expressing TIGIT. That is to say, the antibodies of the invention reduce the proportion of Treg cells expressing TIGIT relative to the total population of T lymphocytes by a magnitude greater than the reduction in the proportion of effector T lymphocytes (CD4 T lymphocytes). or CD8). This selective decrease in TIGIT-expressing Treg cells can be facilitated by selective lysis of TIGIT-expressing Treg cells (eg, by ADCC or CDC (see Figures 20 and 21). It is understood that TIGIT-expressing Treg cells are regulatory cells more potent than TregIT cells which do not express TIGIT. Without intending to be bound by theory, selective lysis depletion of TIGIT expressing Treg cells is expected to increase the function of effector T cells (by eg, T cell facilitated cytotoxicity, release of pro-inflammatory cytokines) by depleting the overall number of Treg cells and also depleting Treg cells demonstrating the most potent regulatory function. BE2017 / 5535 embodiments, the antibodies of the invention selectively lyse TIGIT-expressing Treg cells. Selective depletion of TIGIT-expressing Treg cells can also be facilitated by inducing the internalization of the TIGIT receptor so that it is no longer expressed at the cell membrane. Without intending to be bound by theory, by inducing the internalization of TIGIT so that Treg TIGIT + cells become Treg TIGIT- cells, it is expected that the regulatory function of these cells will become less powerful (given that TregIT TIGIT + cells are more powerful regulatory cells). Following the internalization of the receptor and the subsequent decrease in the regulatory power of these Treg cells, an increase in the effector function of T lymphocytes is expected. Consequently, in certain embodiments, the antibodies of l The invention demonstrates suppressive activity of TIGIT-expressing Treg cells, preferably by inducing TIGIT internalization by TIGIT-expressing Treg cells. It is particularly advantageous for the anti-TIGIT antibodies of the invention to demonstrate a high affinity for CD8 T lymphocytes and Treg cells and also to demonstrate a selective decrease in Treg cells, thereby promoting the effector function of T lymphocytes through two mechanisms. Maintaining the effector function of the antibody (eg, ADCC, CDC) results in an actual decrease in Treg cells and selectivity means that the effector function of the antibody does not cause an undesirable decrease in effector T cells . This selectivity is particularly surprising since previous attempts to produce an anti-TIGIT antibody have attempted to eliminate the effector function of the antibody in order to avoid lysis of the effector T lymphocytes expressing TIGIT. In addition, since the TIGIT antibodies of the invention demonstrate an affinity for effector T cells (eg, CD8 T cells), signaling via TIGIT in these cells can be inhibited by competition for binding to CD155 and / or the induction of TIGIT internalization on effector T lymphocytes. In combination, these effects of the antibodies of the invention can lead to significant upregulation of the effector function of T lymphocytes Other surprising advantageous properties demonstrated by the antibodies of the invention include increasing the effector function of T lymphocytes (eg, the release of proinflammatory cytokines) from tumor infiltrating lymphocytes (TIL). Exposure to tumor microenvironments can lead to the development of anergic, or so-called "depleted" phenotypes by TIL, possibly due to overexposure to the antigen and / or an immunosuppressive tumor microenvironment. Improving the effector function of TIL is desirable since it is these cells which infiltrate the tumor itself and are therefore positioned in the most suitable environment to reduce the size or growth of the tumor; however, due to the anergic or depleted phenotype of many TILs, it is expected to be difficult to potentiate their effector function. The increase in the pro-inflammatory response of TIL following exposure to the antibodies of the invention is therefore surprising and indicates that the antibodies can be particularly effective therapeutic agents. BE2017 / 5535 In certain embodiments, the invention describes anti-TIGIT antibodies or antigen binding fragments thereof in which the VH domain is derived from a sequence of the germ line of the human V region selected from : VH3-07, VH3-30, VH1-46, VH40B, VH4-39, VH 1-69, VH3-09, VH3-33, VH3-30. In certain preferred embodiments, the antibody or the antigen binding fragment thereof comprises a VH domain derived from a V1-46 germline from the human V region. A VH domain is "derived from" a particular V-region germ line sequence if the heavy chain variable region sequence is more likely to be derived from the given germ line than from any other line. As demonstrated in the attached examples, the present invention also describes antibodies which do not compete with CD155 / PVR for binding to TIGIT. Therefore, in another aspect, the invention describes an antibody or antigen binding fragment thereof which does not compete with CD155 / PVR for binding with human TIGIT. In certain embodiments of this type, the Fab fragments of the non-competitive CD155 antiTIGIT antibodies according to the invention demonstrate a K D for TIGIT measured by ForteBio ™ in the range from 5 × 10 ′ 9 to 5 × 10 ′ 8 M, possibly 1 x 10 ' 8 to 3 x 10' 8 M. In certain preferred embodiments, the antibody may comprise a heavy chain variable domain and a light chain variable domain in which HCDR1 comprises SEQ ID NO: 280, HCDR2 comprises SEQ ID NO: 281, HCDR3 comprises SEQ ID NO: 282, and LCDR1 includes SEQ ID NO: 292, LCDR2 includes SEQ ID NO: 293, and LCDR3 includes SEQ ID NO: 294. In certain embodiments of this type, the variable domain of the heavy chain can comprise the amino acid sequence illustrated as SEQ ID NO: 333 or an amino acid sequence demonstrating a sequence identity of at least 90% , 95%, 97%, 98% or 99% thereof, and the variable domain of the light chain may include the amino acid sequence illustrated as SEQ ID NO: 334 or an amino acid sequence demonstrating sequence identity of at least 90%, 95%, 97%, 98% or 99% to it. The embodiments in which the amino acid sequence of the VH domain demonstrates a sequence identity of less than 100% with the sequence illustrated as SEQ ID NO: 333 may nevertheless include the heavy chain CDRs which are identical to HCDR1, HCDR2 and HCDR3 of SEQ ID NO: 333 (SEQ ID NO: 280, 281 and 282, respectively) while demonstrating variation in the amino acid sequence within the framework regions. Likewise, embodiments in which the amino acid sequence of the VL domain demonstrates a sequence identity of less than 100% with the sequence illustrated as SEQ ID NO: 334 may nevertheless include light chain CDRs which are identical to LCDR1, LCDR2 and LCDR3 of SEQ ID NO: 334 (SEQ ID NO: 292, 293 and 294, respectively) while demonstrating a variation in the amino acid sequence within the framework regions. BE2017 / 5535 The invention also describes "germline variants" of the antibodies described herein. The invention further describes "affinity variants" of the antibodies described herein. The invention also describes an isolated antibody or antigen binding fragment thereof, which cross-competes for binding to human TIGIT with an antibody or antigen binding fragment described herein. The invention also describes an isolated antibody or an antigen binding fragment thereof, which binds to the same epitope as the antibody or the antigen binding fragment described herein. The invention also describes polynucleotide molecules coding for the TIGIT antibodies of the invention, and also expression vectors containing nucleotide sequences which code for the TIGIT antibodies of the invention linked in operation to regulatory sequences which allow the expression of the antigen binding polypeptide in a host cell or cell free expression system, and a host cell or cell free expression system containing this expression vector. Polynucleotide molecules encoding the TIGIT antibodies of the invention include, for example, recombinant DNA molecules. The terms "nucleic acid", "polynucleotide" or "polynucleotide molecule" as used in the present invention interchangeably refer to any single or double stranded DNA or RNA molecule and, in the case of a single-stranded molecule, the molecule of its complementary sequence. In the discussion of nucleic acid molecules, a sequence or structure of a given nucleic acid molecule can be described here according to the normal convention of description of the sequence in the 5 'to 3' direction. In some embodiments of the invention, the nucleic acids or polynucleotides are "isolated". This term, when applied to a nucleic acid molecule, describes a nucleic acid molecule which is separated from the sequences with which it is immediately contiguous in the natural genome of the organism from which it originates. For example, an "isolated nucleic acid" can comprise a DNA molecule inserted into a vector, such as a plasmid or a viral vector, or integrated into the genomic DNA of a prokaryotic or eukaryotic cell, or of a non-human host organism. When applied to RNA, the term "isolated polynucleotide" primarily describes an RNA molecule encoded by an isolated DNA molecule, as defined above. Furthermore, the term may describe an RNA molecule that has been purified / separated from other nucleic acids with which it would be associated in its natural state (i.e., in cells or tissues). An isolated polynucleotide (DNA or RNA) can also represent a molecule produced directly by biological or synthetic means and separated from the other components present during its production. For the recombinant production of a TIGIT antibody according to the invention, a recombinant polynucleotide encoding the antibody can be prepared (using standard molecular biology techniques) and inserted into a replication vector for expression in a chosen host cell, or BE2017 / 5535 a cell-free expression system. Suitable host cells can be prokaryotic, yeast, or higher eukaryotic cells, specifically mammalian cells. Examples of useful mammalian host cell lines are the SV1 transformed monkey CV1 renal line (COS-7, ATCC CRL 1651); the human embryonic renal line (cells 293 or 293 subcloned for growth in suspension culture, Graham et al., J. Gen. Virol. 36: 59-74, 1977); baby hamster kidney cells (BHK, ATCC CCL 10); Chinese hamster ovary cells / -DHFR (CHO, Urlaub et al., Proc. Natl. Acad. Sci. USA 77: 4216, 1980; or clones derived from CHO such as CHO-K1, ATCC CCL-61, Kao and Puck, Genetics of somatic mammalian cells, VII. Induction and isolation of nutritional mutants in Chinese hamster cells, Proc. Natl. Acad. Sci. 60: 1275-1281, 1968); mouse Sertoli cells (TM4; Mather, Biol. Reprod. 23: 243-252, 1980); mouse myeloma cells SP2 / 0-AG14 (ATCC CRL 1581; ATCC CRL 8287) or NS0 (HPA culture collections no. 85110503); monkey kidney cells (CV1 ATCC CCL 70); African green monkey kidney cells (VERO-76, ATCC CRL-1587); human cervical carcinoma cells (HELA, ATCC CCL 2); canine kidney cells (MDCK, ATCC CCL 34); Buffalo rat liver cells (BRL 3A, ATCC CRL 1442); human lung cells (W138, ATCC CCL 75); human liver cells (Hep G2, HB 8065); mouse mammary gland tumor (MMT 060562, ATCC CCL51); TRI cells (Mather et al., Annals N. Y. Acad. Sci. 383: 44-68, 1982); MRC 5 cells; FS4 cells and a human hepatoma line (Hep G2), as well as the DSM PERC-6 cell line. Expression vectors suitable for use in each of these host cells are also generally known in the art. It should be noted that the term "host cell" generally describes a cultured cell line. Whole human beings into which an expression vector coding for an antigen-binding polypeptide according to the invention has been introduced are explicitly excluded from the definition of a "host cell". In an important aspect, the invention also describes a method of producing a TIGIT antibody of the invention which comprises culturing a host cell (or a cell-free expression system) containing a polynucleotide (by eg, an expression vector) encoding the TIGIT antibody under conditions which allow expression of the TIGIT antibody, and recovery of the expressed TIGIT antibody. This recombinant expression method can be used for large-scale production of the TIGIT antibodies according to the invention, including monoclonal antibodies intended for therapeutic use in humans. Vectors, cell lines and methods of production suitable for the large scale manufacture of recombinant antibodies suitable for therapeutic use in vivo are generally available in the art and will be well known to those skilled in the art. Therefore, also in accordance with the invention, there is described an isolated polynucleotide encoding an antibody or an antigen binding fragment described herein. BE2017 / 5535 Also according to the invention, there is described an isolated polynucleotide encoding a VH and / or VL domain of an anti-TIGIT antibody, in which the polynucleotide comprises one or more sequences chosen from the group consisting of SEQ ID NO: 241 -270 and 335-342. Also, in accordance with the invention, there is described an expression vector comprising a polynucleotide, according to the invention, linked in operation to regulatory sequences which allow the expression of the antigen binding polypeptide in a host cell. or in a cell-free expression system. Also in accordance with the invention, there is described a host cell or a cell-free expression system containing an expression vector according to the invention. Also according to the invention there is described a method of producing a recombinant antibody or an antigen binding fragment thereof which comprises culturing the host cell or an expression system free of cells according to the invention under conditions which allow the expression of the antibody or of the antigen-binding fragment and the recovery of the antibody or of the expressed antigen-binding fragment. The invention also describes pharmaceutical compositions comprising an antibody or an antigen binding fragment according to the invention prepared with one or more pharmaceutically acceptable vehicles or excipients. Such compositions may include one or more of a combination (eg, two or more of two antibodies) of TIGIT antibodies. Techniques for the preparation of antibodies for therapeutic use in humans are well known in the art and are summarized, e.g., in Wang et al., Journal of Pharmaceutical Sciences, Vol. 96, pp1-26, 2007. The TIGIT antibodies and the pharmaceutical compositions described herein are useful in therapy, in particular in the therapeutic treatment of a disease, in particular pathologies which benefit from an inhibition of the function of TIGIT. The TIGIT antibodies, or the antigen binding fragments thereof, and the pharmaceutical compositions described herein can be used to inhibit the growth of cancer tumor cells in vivo and are therefore useful in the treatment of tumors. Therefore, other aspects of the invention relate to methods of inhibiting tumor cell growth in a human patient, and also methods of treating or preventing cancer, which include administering to a patient a therapeutically effective amount of a TIGIT antibody or antigen binding fragment or pharmaceutical compositions, as described herein. Another aspect of the invention describes a TIGIT antibody or an antigen binding fragment, as described herein, for use in inhibiting the growth of tumor cells in a BE2017 / 5535 human patient. Yet another aspect of the invention describes a TIGIT antibody or an antigen binding fragment, as described herein, for use in the treatment or prevention of cancer in a human patient. Preferred cancers whose growth can be inhibited by the use of the TIGIT antibodies described here include kidney cancer (e.g., renal cell carcinoma), breast cancer, brain tumors, acute or chronic leukemia, including acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, lymphomas (e.g., Hodgkin's and non-Hodgkin's lymphoma, lymphocytic lymphoma, primary CNS lymphoma, lymphoma lymphoma B, T-cell lymphoma), nasopharyngeal carcinoma, melanoma (e.g., metastatic malignant melanoma), prostate cancer, colon cancer, lung cancer, bone cancer, pancreatic cancer, skin cancer, cervico-facial cancer, malignant skin or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the an region ale, stomach cancer, testicular cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of endometriosis, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma soft tissue, cancer of the urethra, cancer of the penis, solid tumors of childhood, cancer of the bladder, cancer of the kidney or urethra, cancer of the renal pelvis, neoplasm of the system central nervous system (CNS), tumor angiogenesis, spinal axis tumor, brainstem glioma, pituitary adenoma, Kaposi's sarcoma, squamous cell cancer, squamous cell cancer, mesothelioma. In some embodiments, the method of treating cancer also includes administering an additional therapeutic agent, eg, a chemotherapeutic agent. As demonstrated here, the antibodies of the invention or the antigen binding fragments thereof are particularly effective when administered in combination with anti-PD-1 antibodies (i.e. ., specific antagonistic antibodies for the human immunoregulatory molecule PD-1). Administration of anti-TIGIT antibodies in combination with anti-PD-1 antibodies results in synergistic reduction in tumor growth compared to each antibody alone. Therefore, the invention also describes a method of treating cancer in a subject comprising administering to the subject an effective amount of an anti-TIGIT antibody or an antigen binding fragment thereof. according to the invention and also the administration of an effective amount of an anti-PD-1 antibody. Similar effects are expected to be observed using a combination of an anti-TIGIT antibody according to the invention and an anti-PD-L1 antibody. EXAMPLES BE2017 / 5535 EXAMPLE 1 Selection of TIGIT Antigen Binding Proteins The TIGIT ABPs were chosen from a synthetic library of human antibodies expressed and presented on the surface of yeast cells, generally in IgG format, as described, for example, in WO2009036379; W02010105256; W02012009568; and Xu et al., Protein Eng Des Sel., Vol. 26 (10), pp. 663-670 (2013)), and more specifically described above. The sequences and characteristics of the ABPs isolated from the recombinant libraries are given in Figures 1 to 6. Eight banks of naive synthetic human yeast each with a diversity of about 10 9 were propagated as previously described (see, e.g., Xu et al, 2013; W02009036379; W02010105256 and W02012009568). For the first two selection cycles, a magnetic bead sorting technique using the Miltenyi MACS system was performed, as described (see, e.g., Siegel et al., 2004). Briefly, yeast cells (approximately 10 10 cells / bank) were incubated with a biotinylated TIGIT-Fc antigen (Creative Biomart) in FACS wash buffer (phosphate buffered saline (PBS) / 0.1% serum - bovine albumin (BSA)). After washing with 50 ml of ice-cold washing buffer, the cell pellet was resuspended in 40 ml of washing buffer, and Streptavidin microbeads (500 µl) were added to the yeast and incubated for 15 min at 4 ° C . Then, the yeasts were pelletized, resuspended in 5 ml of washing buffer and loaded onto a Miltenyi LS column. After loading the 5 ml, the column was washed 3 times with 3 ml of FACS wash buffer. The column was then removed from the magnetic field, and the yeasts were eluted with 5 ml of culture medium and then cultured overnight. The following sorting cycles were carried out with flow cytometry. About 1x10 8 yeasts were pelletized, washed three times with washing buffer and incubated with a biotinylated TIGIT-Fc fusion antigen (10 nM) under equilibrium conditions at room temperature. The yeasts were washed twice and stained with LC-FITC (diluted 1: 100) and secondary reagents SA-633 (diluted 1: 500) or EA-PE (diluted 1:50) for 15 minutes at 4 ° C . After two washes with ice-cold wash buffer, the cell pellets were resuspended in 0.4 ml wash buffer and transferred to sorting tubes with filter caps. The sorting was carried out with a FACS ARIA sorter (BD Biosciences) and the sorting grids were assigned to select antibodies showing a specific binding compared to a reference control. Subsequent selection cycles have been used to reduce the number of non-specific antibodies by using soluble membrane proteins from CHO cells (see, e.g., WO2014179363 and Xu et al., Protein Eng Des Sel, Vol. 26 (10), pp. 663-670 (2013)), and to identify the binders which improved affinity for TIGIT using the TIGIT-Fc antigen. At the end of the last sorting cycle, the yeasts were seeded and individual colonies were selected for the characterization and the nomination of the clones for maturation by affinity. Functional activity was analyzed in 63 clones. Following the analysis, clones 26518, 26452, 26486, 26521 and 26493 showed the best functional activity and were selected for further optimization. BE2017 / 5535 Example 2 Optimization of the Antibody Optimization of naive clones was carried out using three maturation strategies: diversification of the light chain; diversification of CDRH1 and CDRH2; and diversification of CDRH3 within the diversity groups of CDRH1 and CDRH2 chosen. Light chain diversification: the heavy chain variable regions were extracted from the naive results (described above) and transformed into a light chain library with a diversity of 1 x 10 6 . Selections were made as described above with one MACS sorting cycle and three FACS sorting cycles using 10 nM or 1 nM biotinylated TIGIT-HIS antigen (Creative Biomart) for the respective cycles. Selection of CDRH1 and CDRH2: the CDRH3s from the clones selected from the light chain diversification procedure were recombined in a pre-established library with the variants CDRH1 and CDRH2 having a diversity of 1 × 10 8 and the selections were performed using the HIS-TIGIT monomeric antigen. The affinity pressures were applied using decreasing concentrations of the biotinylated HIS-TIGIT antigen (100 to 1 nM) under equilibrium conditions at room temperature. Selection of CDRH3 / CDRH1 / CDRH2: oligos have been ordered from. the IDT which included CDRH3 as well as the homologous flanking region on either side of CDRH3. A variety of amino acid positions in CDRH3 were tested by NNK variation at two positions per oligo across all of CHRH3. The CDRH3 oligos were double-stranded and used primers which paired with the flanking region of CDRH3. The remainder of the FWR1 to FWR3 of the heavy chain variable region was amplified from antibody groups which improved affinity and which were isolated from the diversity of CDRH1 and CDRH2 selected above. The library was then created by transforming the double-stranded CDRH3 oligo, the FWR1 to FWR3 fragments combined, and the heavy chain expression vector in yeast already containing the light chain of the original naive parent. The selections were made as in the previous cycles using sorting by FACS for four cycles. For each FACS cycle, the banks were evaluated for PSR binding, cross-reactivity between species and affinity pressure, and sorting was carried out to obtain populations with the desired characteristics. The affinity pressures for these selections were made as described above in the selection of CDRH1 and CDRH2. Example 3: Production and purification of antibodies A. Production in yeast In order to produce sufficient quantities of optimized and non-optimized antibodies selected for further characterization, the yeast clones were cultured to saturation and then incubated for 48 h at 30 ° C with shaking. After induction, the yeast cells were put in BE2017 / 5535 pellet and supernatants were collected for purification. The IgGs were purified using a protein A column and eluted with acetic acid, pH 2.0. The Fab fragments were produced by papain digestion and purified in a two-step process on protein A (GE LifeSciences) and KappaSelect (GE Healthcare LifeSciences). B. Production in mammalian cells In order to produce sufficient amounts of optimized and non-optimized antibodies selected for further characterization, a DNA vector encoding the specific antibody clones was produced and transduced in HEK cells. Synthetic DNA fragments of human codon optimized for the variable domains of the antibody were ordered from Geneart. The variable domain sequences were homogeneously ligated into the pUPE expression vectors containing the mouse IgKappa signal sequence and the constant regions of the respective antibody class. The expression vectors were verified by restriction analysis and DNA sequencing. For transient transfection, DNA maxipreps containing no endotoxin (Sigma) were produced and heavy and light chain vectors were co-transfected in HEK293EBNA1 cells, in Freestyle medium (ThermoFisherScientific), according to the protocols established. Primatone (final volume 0.55%) was added 24 h after transfection. The conditioned medium was collected 6 days after transfection. The antibodies were purified in batches by affinity chromatography on Mabselect sureLX (GE Healthcare). Bound antibodies were washed in two steps with PBS containing 1M NaCl and PBS. The antibodies were eluted with 20 mM citrate, 150 mM NaCl, pH3 and neutralized to a pH of about 7 with a volume of 1/6 of 1 M K2HPO4 / KH2PO4, pH 8. The antibodies were then further purified by gel filtration with a Superdex200 column, equilibrated with PBS. The fractions were analyzed by NuPAGE and the fractions containing the antibody were pooled. The final products were sterilized in a 0.22 μΜ syringe filter. The product was analyzed by NuPAGE and the endotoxin levels were measured by an LAL test. EXAMPLE 4 Determination of the Affinity for the Binding of Anti-TIGIT Antibodies to the Recombinant Human TIGIT Protein A. K D measurements on ForteBio The affinity measurements on ForteBio of the selected antibodies have generally been carried out as described previously (see, for example, Estep et al., Mabs, Vol. 5 (2), pp. 270-278 (2013)). Briefly, the affinity measurements on ForteBio were carried out by loading IgGs online on AHQ sensors. The sensors were equilibrated offline in a test buffer for 30 minutes and then monitored online for 60 seconds to establish reference values. The sensors loaded with IgG were exposed to 100 nM of antigen (human TIGIT-Fc, human TIGIT-His or TIGIT-Fc cyno) for 5 minutes, before being transferred BE2017 / 5535 in an analysis buffer for 5 minutes for a dissociation measurement. The kinetic parameters were analyzed using a 1: 1 binding model. More than 90 antibodies have been tested for affinity by ForteBio and Table 3 presents data for 15 anti-TIGIT antibodies demonstrating a strong binding to the recombinant TIGIT protein. B. Measurements of K D on MSD-SET The equilibrium affinity measurements of the selected antibodies were generally carried out as previously described (Estep et al., Mabs, Vol. 5 (2), pp. 270-278 (2013)). Briefly, equilibrium solution titers (SET) were performed in PBS + 0.1% BSA (PBSF) not containing IgG with the antigen (monomer TIGIT-His) maintained at a constant concentration of 10 to 100 μM and incubated with serial dilutions of 3 to 5 times Fab or mAbs, starting between 10 μM and 10 nM. The antibodies (20 nM in PBS) were deposited on standard MSD-ECL binding plates overnight at 4 ° C or at room temperature for 30 minutes. The plates were then blocked with BSA for 30 minutes with stirring at 700 rpm, followed by three washes with washing buffer (PBSF + 0.05% Tween 20). The SET samples were deposited and incubated on the plates for 150 seconds with shaking at 700 rpm followed by washing. The antigen captured on a plate was detected with 250 ng / ml of sulfotag-labeled streptavidin in PBSF incubated on the plate for 3 minutes. The plates were washed three times with washing buffer and then read on an MSD Sector Imager 2400 instrument, 1x reading buffer T with the supernatant. The percentage of free antigen has been reported in Prism as a function of the titrated antibody and fitted to a quadratic equation to extract the K D. In order to improve the yield, liquid handling robots were used in all MSD-SET experiments, including the preparation of the SET sample. The selected antibodies were tested for affinity by MSD and Table 4 presents data for 7 anti-TIGIT clones demonstrating a strong binding to the recombinant TIGIT protein. Table 4: MSD analysis of the affinity for the selected anti-TIGIT antibodies clone MSD affinityKD (M) ofTIGIT-His human monovalent 29489 1,1E-10 29494 7.0E-11 29499 1,9E-11 29513 2.5E-11 BE2017 / 5535 29520 2.1 E-10 29523 1.7E-09 29527 6.4E-10 C. Measurement on Biacore The biosensor analysis was carried out at 25 ° C in an HBS-EP buffer system (10 mM HEPES, pH 7.3, 150 mM NaCl, 3 mM EDTA, 0.05% P20 supernatant) in using a Biacore 8K optical biosensor fitted with a CM5 sensor chip (GE Healthcare, Marlboro, MA). The samples were kept at 8 ° C. An anti-human goat IgG capture antibody (specific fragment Fcy, Jackson ImmunoResearch Laboratories, Inc., West Grove, PA; 109-005-098) was immobilized (11,700 +/- 200 RU) on the two d cells. flow of the sensor chip using standard amine coupling chemistry. This type of surface provided a format for the reproducible capture of fresh test antibodies after each regeneration step. Flow cell 2 was used to analyze the captured antibody (60-90 UR) while flow cell 1 was used as the reference flow cell. Antigen concentrations ranging from 30 to 0.123 nM (3-fold dilution) were prepared in the circulating buffer. Each of the concentrations in the antigen sample was analyzed as a single replicate. Two control injections (buffer) were also analyzed and used to assess and remove the artifacts from the system. The association (300 s) and dissociation (600 s) phases for all the antigen concentrations were carried out at a flow rate of 30 μl / min. The surface was regenerated with three sequential injections (15 s, 15 s and 60 s) of 10 mM glycine, pH 1.5 at a flow rate of 30 μl / min. The data were aligned, doubly referenced and fitted to a 1: 1 linkage model using Biacore 8K evaluation software, version 1.0. The selected antibodies were tested for affinity by Biacore and Table 5 presents data for 5 anti-TIGIT clones demonstrating a strong binding to the recombinant TIGIT protein. Table 5: Analysis of the affinity by Biacore for the selected anti-TIGIT antibodies clone Biacore: monovalent KD (M) (IgG on CM5 chip, TIGIT-HIS solution (starting concentration dilution) human in25 nM, 3x 29489 2.48E-10 31282 2.94E-10 29494 2.70E-10 BE2017 / 5535 29520 7.16E-10 29527 1.20E-09 31288 1.92E-10 EXAMPLE 5 Competition Test Between Antagonistic Anti-TIGIT Antibodies and Natural TIGIT Ligands A. Epitope compartment Red384 byte / ligand blocking The compartmentalization of the epitopes / blocking by ligand of the selected antibodies was carried out with a cross-blocking test in standard sandwich format. Control anti-target IgGs were loaded onto AHQ sensors, and unoccupied Fc binding sites on the sensor were blocked with an irrelevant human IgG1 antibody. The sensors were then exposed to a target antigen at 100 nM (hTIGIT, Creative Biomart) followed by a second anti-target antigen or ligand (anti-TIGIT antibody and CD155 or CD113 or CD112). The data were processed with ForteBio 7.0 data analysis software. The additional binding by the second antibody or ligand after the antigen association indicates an unoccupied epitope (non-competitor), while an absence of binding indicates a blockage of the epitope (competitor or ligand blocking). Parent antibodies (before optimization) were tested for competition with natural ligands and Table 6 summarizes the data obtained for competition against CD155, CD112 and CD113. It was found that the parental clone 26432 does not compete with CD155 for binding to TIGIT. All other selected anti-TIGIT antibodies compete with the natural ligand for binding to the recombinant human TIGIT protein. Table 6: Analysis of compartmentalisation against natural ligands of TIGIT for non-optimized anti-TIGIT antibodies 26518 Yes Yes Yes26452 Yes Yes YesI ..................... ...................................... ............................................. î where26521 Yes Yes Yes BE2017 / 5535 26493 Yes YesYes 26432 No : B. Competition of anti-TIGIT antagonist antibodies with CD155 on JurkathTIGIT Jurkat cells overexpressing human TIGIT (Jurkat-hTIGIT) were collected and distributed to 10 5 cells / well and incubated with anti-human TIGIT antibodies at the following concentrations: 166.6; 53.24; 17.01; 5.43; 1.73; 0.55; 0.17; 0.05; 0.01; 5.78 x 10 '3; 1.85 x 10 '3; 5.9 x 10 ' 3 nM in complete medium for 45 minutes at 37 ° C. The excess antibody was washed, and then the cells were incubated with CD155-His at 5 pg / ml (Creative Biomart, PVR-3141H) for 45 minutes at 37 ° C. Then, bound CD155-His was detected using an anti-His coupled to PE (Biolegend, 362603, 2 µl per assay), incubated for 30 minutes at 4 ° C. The cells were analyzed by FACS with a BD LSRFortessa and the concentration (IC 50 ) which prevents half of the CD155 bonds was calculated on the basis of the mean geometric fluorescence. The results were as follows: 0.101 nM for clone 29489; 0.07 nM for clone 29494; 0.102 nM for clone 29520 and 0.078 nM for clone 29527, for the results illustrated in FIG. 7. The values of the other antibodies tested are summarized in Table 7 above. Overall, the results demonstrate strong competition by the antagonistic anti-TIGIT antibodies tested with CD155 for binding to TIGIT expressed at the membrane. Table 7: CI 50 data for the CD155 competition on human TIGIT 29489 0,101 29494 0,070 29499 0,103 29513 0.094 29520 0,102 29523 0.079 29527 0.078 Example 6: Characterization of hydrophobic interaction chromatography (MAbs. 2015 May-Jun; 7 (3): 553-561) Anti-TIGIT igG1 antibody samples were swapped in 1 M ammonium sulfate and 0.1 M sodium phosphate pH 6.5 on a Zeba centrifuge column BE2017 / 5535 kDa 0.5 ml (Thermo Pierce, catalog no. 87766). A sahn gradient was established on a Dionex ProPac HIC-10 column ranging from 1.8 M ammonium sulphate, 0.1 M sodium phosphate at pH 6.5 on the same condition as in the absence of sulphate ammonium. The gradient lasted 17 minutes at a flow rate of 0.75 ml / min. An acetonitrile washing step was added at the end of the cycle to remove any remaining protein and the column was rebalanced over 7 column volumes before the next injection cycle. The peaks of the retention times were measured at absorbance at A280 and the elution concentrations of ammonium sulfate were calculated based on the gradient and the flow rate. Table 8 summarizes the results obtained for 15 selected anti-TIGIT antibodies. Table 8: Analysis by hydrophobic interaction chromatography for the selected anti-TIGIT antibodies Cione Interaction chromatographyhydrophobic (HIC) Retention time (min) 26518 10.4 29478 iiiiiiiiiiliiiiiil 26452 9.3 29487 liiiiiiiiiiliiii 29489 10.6 26486 iiiiiiiiiiliiiiiil 29494 9.7 2949926521 12.4 29513 iiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiii 26493 8.8 2952029523 8.7 29527 iiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiii ^^ 26432 11.1 32919 liiiiiiiiililiiiiiiiiiiiiiiiiiiiiiiiiliiiiiiiiiiiiiiiiiiiiiii Example 7: Characterization of the preparation of the PSR polyspecificity reagent A. Preparation of the polyspecificity reagent: BE2017 / 5535 The polyspecificity reagent (PSR) was prepared according to Xu and. al., mAbs 2013. In short, 2.5 liters of CHO-S cells were used as starting material. The cells were pelletized at 2400 x g for 5 minutes in 500 ml centrifuge bottles filled to 400 ml. The cell pellets were combined and then resuspended in 25 ml of buffer B and pelletized at 2400 x g for 3 minutes. The buffer was then decanted and the washing repeated once. The cell pellets were resuspended in 3 times the pellet volume of buffer B containing 1 × protease inhibitor (Roche, cOmplete, EDTA-free) with a polytron homogenizer, the cells being kept on ice. The homogenate was then centrifuged at 2400 xg for 5 minutes and the supernatant retained and pelletized once again (2400 xg / 5 min) to ensure removal of unbroken cells, cell debris and nuclei; the supernatant thus obtained represents the total protein preparation. The supernatant was then transferred to two 45 ml Nalgene Oak Ridge centrifuge tubes and pelletized at 40,000 x g for 40 minutes at 4 ° C. The supernatants containing the separated systolic proteins (SCP) were then transferred to clean Oak Ridge tubes and centrifuged at 40,000 x g once more. In parallel, the pellets containing the membrane fraction (EMF) were retained and centrifuged at 40,000 g for 20 minutes to remove the residual supernatant. The EMF pellets were then rinsed with buffer B. 8 ml of buffer B were then added to the membrane pellets to dislodge the pellets and transfer them to a Dounce homogenizer. After homogenization of the pellets, they were transferred to a 50 ml conical tube and represented the final EMF preparation. One billion mammalian cells (eg, CHO, HEK293, Sf9) at approximately 10 6 to 10 7 cells / ml were transferred from the tissue culture environment into 4 conical tubes of 250 ml and pelletized at 550 xg for 3 minutes. All of the following steps were performed at 4 ° C or on ice with ice pads. The cells were washed with 100 ml of PBSF (1 x PBS + 1 mg / ml BSA) and pooled in a conical tube. After removal of the supernatant, the cell pellet was then resuspended in 30 ml of buffer B (50 mM HEPES, 0.15 M NaCl, mM CaCI2, 5 mM KCI, 5 mM MgCI2, 10% glycerol, pH 7.2) is pelletized at 550 xg for minutes. The buffer B supernatant was decanted and the cells suspended in 3 times the volume of the buffer B pellet plus 2.5 × the protease inhibitor (Roche, cOmplete, EDTA-free). Protease inhibitors in buffer B were systematically added from this step. The cells were homogenized four times by 30 second pulses (Polyton homogenizer, PT1200E) and the membrane fraction was pelletized at 40,000 xg for 1 hour at 4 ° C. The pellet was rinsed in 1 ml of buffer B; the supernatant retained and represents the "s". The pellet was transferred to a Dounce homogenizer with 3 ml of buffer B and resuspended by slowly moving the pestle from top to bottom for 30 to 35 strokes. The enriched membrane fraction (EMF) is put in a new collection tube, rinsing the pestle to collect all the potential proteins. The protein concentration of the purified EMF was determined using the Dc-protein test kit (BioRad). To dissolve the EMF, it was transferred to the buffer of BE2017 / 5535 solubilization (50 mM HEPES, 0.15 M NaCl, 2 mM CaCI2, 5 mM KCI, 5 mM MgCI2, 1% n-Dodecyl-bD-Maltopyranoside (DDM), 1 x protease inhibitor, pH 7.2 ) at a final concentration of 1 mg / ml. The mixture was rotated overnight at 4 ° C, followed by centrifugation in a 50 ml Oak Ridge tube (Fisher Scientific, 050529-ID) at 40,000 x g for 1 hour. The supernatant containing the soluble membrane proteins (SMP) was collected before the quantification of the protein yield as described above. For biotinylation, the NHS-LC-Biotin stock solution was prepared according to the manufacturer's protocol (Pierce, Thermo Fisher). Briefly, 20 µl of biotin reagent is added for each mg of EMF sample and incubated at 4 ° C for 3 hours with slow shaking. The volume is adjusted to 25 ml with buffer B and transferred to an Oak Ridge centrifuge tube. Then, pellet the biotinylated EMF (b-EMF) at 40,000 x g for one hour, and two rinses with 3 ml of buffer C (buffer B minus glycerol) without disturbing the pellet. The residual solution is removed and the residue resuspended with a Dounce homogenizer in 3 ml of buffer C as described above. The resuspended pellet now represents the biotinylated EMF (b-EMF). The b-SMPs are prepared by solubilizing as described above. B. PSR linkage analyzes PSR analyzes were carried out as described in patent WO2014 / 179363. Briefly, to characterize the PSR profile of the monoclonal antibodies presented on the yeast, two million yeasts presenting IgG were transferred to a 96-well test plate and pelletized at 3000 × g for 3 minutes to remove the supernatant. The pellet was resuspended in 50 µl of freshly prepared b-PSRs 1:10 dilution solution and incubated on ice for 20 minutes. The cells were washed twice with 200 μl of cold PBSF and the pellet resuspended in 50 μl of secondary labeling mixture (Extravidine-R-PE, anti-human LC-FITC and propidium iodide). The mixture was incubated on ice for 20 minutes followed by two washes with 200 µl ice cold PBSF. The cells were resuspended in 100 μl of ice-cold PBSF and the plate was analyzed in a FACS Canto (BD Biosciences) using an HTS sample injector. The flow cytometry data were analyzed for the average fluorescence intensity in the R-PE channel and normalized on appropriate controls to assess non-specific binding. Table 9 summarizes the polyspecific reagent binding results obtained for 15 selected anti-TIGIT antibodies, which is confirmed by the low scores for most of the clones. Table 9: Analysis of the polyspecificity reagent clone Polyspecificity reagent score (0-1) 26518 0.00 29478 llllllllllllllllllll ^ 26452 0.00 BE2017 / 5535 29487 | 1 | 29489 0.01 26486 lllllllllllillllllllllllll 29494 0.00 29499 lllllllllllillllllllllllll ^ 26521 0.00 29513 llllllllillllll 26493 0.00 2952029523 0.12 2952 / lllllllllllillllllllllllll 26432 0 00 31288 lllllllillllllllllllllllllll 3291932931 llllllllllllllll Example 8 Characterization of the Expression of TIGIT on Immune Populations Coming from Healthy Human PBMCs A. TIGIT expression profile on T lymphocyte subsets Flow cytometry analyzes were performed to assess the expression of TIGIT on immune cell subpopulations on freshly isolated PBMCs from healthy individuals. Conjugated antibodies were purchased from Ebioscience / Thermo Fisher Scientific, BioLegend or BD Biosciences. The cells were stained according to the manufacturer's instructions using the filtered FACS buffer (PBS + 2mM EDTA + 0.1% BSA) and the Brilliant Stain buffer (BD # 563794). Cells were blocked with the appropriate human FcBlock (BD # 564220) before staining and were fixed with Cl fixation buffer (eBioscience # 00-8222-49) before acquisition. The acquisition was carried out on a Fortessa FACS (BD Biosciences) and analyzed with FlowJo software (FlowJo, LLC). The viable cells were delimited thanks to their size and their granularity. Various subpopulations of immune cells have been defined as follows: CD19 + (B lymphocytes), CD3 'CD19' CD14 + (monocytes), CD3 + TCRab '(TCRgd T cells), CD3 + TCRab + (TCRab T lymphocytes), CD3 'CD19' CD14 'HLA-DR' CD56 low / high (NK cells), CD3 'CD19' CD14 'HLA-DR + (dendritic cells), CD3 + TCRab + CD4 + CD127 low CD25 + (regulatory T lymphocytes), CD3 + TCRab + CD4 + or CD8 + CD45RO 'CCR7 + (CD4 or CD8 naive T lymphocytes), CD3 + TCRab + CD4 + or CD8 + CD45RO + (memory T lymphocytes) and CD45ROCD62L' (effector T lymphocytes), As shown in Figure 8A and 8B, TIGIT is preferentially expressed on NK cells, regulatory T cells and memory CD8 T cells. The TIGIT protein is present at a lower level on other T lymphocyte subpopulations with a low BE2017 / 5535 proportion of naive T lymphocytes demonstrating TIGIT expression. In addition, TIGIT is not expressed on monocytes, dendritic cells and B lymphocytes (Fig. 8B). This dataset is in agreement with the published data (Yu et al. NI 2008 and Wang et al. EJI 2015). EXAMPLE 9 Cell Binding of Antagonistic Anti-TIGIT Antibodies A. Binding of anti-TIGIT antibodies to Jurkat-hTIGIT and Jurkat-mTIGIT The affinity of human anti-TIGIT antibodies was measured with Jurkat E6.1 cells transduced with a vector containing the human (Jurkat hTIGIT) or murine (JurkatmTIGIT) TIGIT sequence. In order to analyze the affinity of the antibodies selected for hTIGIT or mTIGIT, 10 5 cells were distributed per well and incubated with the anti-TIGIT antibody at a single dose of 100 nM (Table 3) or with a decreasing concentration (166 , 6; 53.24; 17.01; 5.43; 1.73; 0.55; 0.17; 0.05; 0.01; 5.78 x 10 '3; 1.85 x 10'3; 5.9 x 10 ' 3 nM) of the selected antibodies (Figure 9). The antibodies were incubated with the cells for 20 minutes at 4 ° C in FACS buffer. After washing, the cells were incubated with anti-human Ig (specific gamma Fc) -PE (eBioscience, 12-4998-82, at 2.5 μg / ml) for 20 minutes on ice and washed twice. The intensity of the geometric mean fluorescence was analyzed with LSR BD Fortessa. Cell binding was recorded as the average fluorescence intensity of PE on the transfected line compared to the non-transfected line for each antibody (Table 3). For the calculation of the CE50 binding, the semi-maximum binding concentration (CE50) to hTIGIT-Jurkat was calculated with a four-variable adjustment curve equation in Prism, and the values obtained were as follows: 0.082 nM for clone 29489; 0.07 nM for clone 29494; 0.119 nM for clone 29520 and 0.05 nM for clone 29527 for the data illustrated in FIG. 9. The results demonstrate a strong binding of human TIGIT expressed on the membrane for the antiTIGIT antibodies tested. B. Binding of anti-TIGIT antagonist antibodies to human primary T lymphocytes Isolated human PBMCs from healthy volunteers were analyzed for binding by anti-TIGIT agonist antibodies. The cells were distributed at 5 x 10 5 per well. The cells were incubated with anti-CD16 (Clone 3G8, BioLegend 302002), CD32 (Clone FLI8.26, BD Bioscience 557333) and CD64 (BD Bioscience 555525) at room temperature for 10 minutes, and anti-human antibodies TIGIT indicated were directly added to a final concentration of: 12.65; 4; 1.26; 0.40; 0.126; 0.040; 0.12 and 4x10 ' 3 nM in FACS buffer and incubated for 20 minutes at 4 ° C. After washing, the cells were incubated with anti-human Ig (specific gamma Fc) -PE (eBioscience, 12-4998-82, at 2.5 μg / ml) for 20 minutes at 4 ° C. Then, the cells were washed and incubated with the following antibodies and an LVD mixture for the results of FIGS. 10A and 10B: anti-CD4-PercP-Cy5.5 (clone A161A1, BioLegend 357414); anti-CD8BV510 (clone SK1, BD Bioscience 563919) and LVD efluor 520 (eBioscience 65-0867-14). For the BE2017 / 5535 Figure 10 C, the cells were washed and incubated with the following antibodies and the LVD mixture: LVD efluor 520 (eBioscience 65-0867-14), ant-TCRab-PercP-Cy5.5 (Clone IP26, Biolegend 306723 ), anti-CD4-BV510 (Clone SK3, BD Horizon 562970), anti-CD8-APC-Cy7 (Clone SK1, Biolegend 344714), anti-CD25-BV605 (Clone 2A3, Biolegend 562660), anti-CD 127-APC (A019D5, Biolegend 351316), anti-CCR7-BV421 (Clone G043H7, Biolegend 353207) and anti-CD45RO-PE-Cy7 (Clone UCHL1, Biolegend 304229). The EC 50 value for binding to human primary CD8 + T cells was calculated using the% of TIGIT positive cells positive on LVD'CD8 + T cells (Figures 10A and 10B). The EC 50 value for binding to CD8 + memory T cells or primary human Treg T cells was calculated using the% TIGIT cells positive on LVD TCRab + CD45RO + CD8 + polarized T cells (for memory T cells CD8 + ) or on LVD TCRab + T lymphocytes CD127 l0 CD25 hl CD4 + polarized (for Tregs) and is illustrated in Figure 10C. As shown in Figure 10A, the EC 50 value for binding to fully human CD8 + T cells is 0.123 nM for clone 29489; 0.181 nM, for clone 29520 and 0.253 nM for clone 29527. A direct comparison between 29489 and 31282 (mutant 29489 with an M to T mutation on residue 116) was carried out, and the EC50 value was 0.057 nM and 0.086 nM respectively, demonstrating a strong binding efficiency similar to the primary human CD8 + T lymphocytes for the 2 clones (FIG. 10B). The EC50 values obtained for binding to memory CD8 + T cells and to Tregs were 0.039 nM and 0.03 nM respectively, demonstrating a strong and similar affinity for the two populations (Figure 10C). C. Binding of anti-TIGIT antagonist antibodies to primary T lymphocytes cynomolgus PBMCs isolated from Macaca fascicularis were obtained from BioPRIM. The cells were thawed and stimulated with a T cell activation / propagation solution for non-human primates (Miltenyi Biotec) at 1: 2 (ball: cell ratio) according to the manufacturer's specifications. The next day, the cells were harvested, counted and distributed to 5 x 10 4 cells per well. The cells were incubated with anti-CD16 (Clone 3G8, BioLegend 302002), CD32 (Clone FLI8.26, BD Bioscience 557333) and CD64 (BD Bioscience 555525) at room temperature for 10 minutes, and anti-human antibodies TIGIT selected were directly added to a final concentration of: 12.65; 4; 1.26; 0.40; 0.126; 0.040; 0.12 and 4 x 10 ' 3 nM in FACS buffer and incubated for 20 minutes at 4 ° C. After washing, the cells were incubated with anti-human Ig (specific gamma Fc) -PE (eBioscience, 12-4998-82, at 2.5 μg / ml) for 20 minutes at 4 ° C. Then, the cells were washed and incubated with the following antibodies and an LVD mixture for the data illustrated in Figures 11A and 11B: anti-CD4-PercP-Cy5.5 (clone A161A1, BioLegend 357414); anti-CD8-BV510 (clone SK1, BD Bioscience 563919), CD69-APC-Cy7 (Clone FN50, BioLegend, 310914) and LVD efluor 520 (eBioscience 65-0867-14). The labeled cells have BE2017 / 5535 been analyzed by FACS using BD LSR Fortessa. The EC50 value for binding was calculated using the% of TIGIT positive labeled cells on LVD'CD69 + CD8 + polarized T cells. As shown in Figure 11, the EC 50 values for binding to cynomolgus CD8 + T lymphocytes were 0.487 nM for clone 29489, 1.73 nM for clone 29520 and 0.378 nM for clone 29527. Clones 29489 and 31282 (mutant 29489 with an M to T mutation on residue 116) were also compared, and the EC 50 values were 0.25 nM and 0.26 nM respectively for the example illustrated in Figure 11B , demonstrating a strong and similar affinity for the primary CD8 + T cells of cynomolgus for the two clones. BE2017 / 5535 Example 10: In vitro functional characterization of the antagonistic anti-TIGIT activity A. TIGIT bioassay on a CHO-TCR-CD155 and Jurkat-hTIGIT co-culture To characterize the functional consequence of blocking the human TIGIT receptor, we co-cultivated Jurkat cells, which express hTIGIT and a luciferase reporter activated during contact with TCRs (TIGIT effector cells "Thaw-and-Use" (Thaw and use ) from Promega), with the CHO-K1 cell line modified to express human CD155 and the TCR activator (CD155 aAPC / CHO-K1 "Thaw-and-Use" from Promega). Activation of Jurkat cells overexpressing TIGIT can be induced by contact with CHO-K1 cells expressing CD155 during contact of TCR with Jurkat cells and can be increased in the presence of an antagonistic anti-TIGIT antibody. In order to compare the power of the different antibodies to increase the activation of Jurkat cells, the experiment was carried out in the presence of increasing concentrations of antibodies and the EC50 values were calculated. The “Thaw-and-Use” CD155 aAPC / CHO-K1 cells (Promega, CS198811) were seeded according to the manufacturer's recommendations and incubated at 37 ° C, 5% CO2 in an O / N incubator. The following day, the TIGIT “Thaw-and-Use” effector cells (Promega, CS198811) were added according to the manufacturer's recommendations to the CD155 aAPC / CHO-K1 cell plates containing fresh complete medium with the anti-TIGIT antibody. at 133 nM (Figure 12A) or increasing concentrations (0.22; 0.54; 1.36; 3.41; 8.53; 21.3; 53.3; 133.33 and 333 nM) of antibodies anti-TIGIT (Figure 12B) and incubated at 37 ° C, 5% CO2 for 6 hours. After the 6 hours of incubation, the activation of the TIGIT effector cells was evaluated by measuring the luciferase activity using a Bio-GloTM Luciferase test system (Promega, G7941). As shown in Figure 12A, the addition of all selected clones (except 26493 which is specific for murine TIGIT) increased the luciferase signal compared to the control isotype, demonstrating the antagonistic activity of these antibodies which resulted in stronger activation of Jurkat-hTIGIT cells. Table 10 summarizes the multiplying factor of change in the induction of the expression of the luciferase obtained for the various anti-TIGIT antibodies in comparison with the isotype control clone (03847). Table 10: Multiplying factor of change in induction compared to the control isotype Clone name 1111¾ ^11¾¾¾multiplitteatif) 26518 2.89 BE2017 / 5535 As shown in Figure 12B, the activation of Jurkat-hTIGIT cells was evaluated with an anti-TIGIT antibody between 0.22 nM and 333 nM and gave an EC 50 value of 3.0 nM for the clone 29489; 4.4 nM for clone 29494; 2.3 nM for clone 29520 and 32 nM for clone 29527; 2.7 nM for clone 32919 and 3.2 nM for clone 32931 demonstrating a powerful functional activity following blockade of the inhibitory signal TIGIT. Clones 29489 and 31282 (mutant 29489 with an M to T mutation on residue 116) were also compared, and the EC 50 values were 4.3 nM and 8.1 nM respectively for the example illustrated in the Figure 12C, demonstrating similar functional activity for the two clones. B. Functional test based on human primary CD8 + T lymphocytes To characterize the functional consequence of blocking the human TIGIT receptor, we co-cultivated human CD8 + T lymphocytes from PBMCs of healthy human donors with the CHO-K1 cell line modified to express human CD155 and to activate human T lymphocytes . We observed that the release of IFNg by CD8 + T lymphocytes in the presence of CD155-expressing CHO-K1 cells could be increased by blocking hTIGIT with antagonistic anti-TIGIT antibodies. In order to compare the power of these antibodies to increase the release of IFNg, the experiment was carried out in the presence of increasing concentrations of antibodies and the EC 50 values were calculated. The “Thaw-and-Use” CD155 aAPC / CHO-K1 cells (Promega, CS198811) were seeded in 96-well plates with U-bottom according to the manufacturer's recommendations and incubated at 37 ° C, 5% CO2 in a O / N incubator. The following day, the CD8 + T lymphocytes were purified according to the manufacturer's recommendations using negative selection solutions (Stemcell Technologies, 17953) from mononuclear cells from human peripheral blood. BE2017 / 5535 frozen isolated from whole blood from healthy donors (Immunehealth). The purified CD8 T cells were then incubated at increasing concentrations (0.11 nM, 0.33 nM, 1.06 nM, 3.3 nM, 10.6 nM, 33.3 nM, 105.5 nM and 333 nM) of antibodies (100,000 CD8 T lymphocytes / 100 μl of complete medium containing the antibody) for 1 hour. Then, the antibody-CD8 mixture was added to the CD155 aAPC / CHO-K1 cell plates containing 50 µl of fresh complete medium and incubated at 37 ° C, 5% CO2 for 5 days. Finally, the concentrations of IFNg were evaluated in the cell supernatant with an ELISA test (Affymetrix eBioscience, 88-7316-86) which was carried out according to the manufacturer's recommendations. As shown in Figure 13A, all anti-TIGIT antibodies increased the secretion of IFNg compared to the control isotype. The highest increase was observed with clone 29489 (6.4 times) followed by 29494 (5.8 times), 29520 (5.4 times), 29499 (5.2 times), 29527 (4.5 times) and 29513 (3.2 times). A dose interval study (between 0.22 nM and 333 nM of anti-TIGIT antibody) was also carried out to assess the EC50 value for the increase in the secretion of IFNg by human primary CD8 T lymphocytes . As shown in Figure 13B, the antiTIGIT 29489 antibody demonstrated the best activity with an EC 50 of 3.5 nM followed by clone 29527 (EC 50 = 5.1 nM), clone 29494 (EC 50 = 6.1 nM) and the clone 29520 (EC 50 = 11.1 nM). Finally, clone 29489 and its mutant 31282 were tested in parallel and demonstrated a similar activity with an EC 50 value of 0.49 nM and 0.50 nM respectively (Figure 13C). Taken together, these data demonstrate potent functional activity of antagonistic anti-TIGIT antibodies to block the TIGIT inhibitory signal in human CD8 + T cells and to increase effector functions, characterized by a large increase in the production of IFNg. C. Functional test of human T IL To characterize the functional consequence of the blocking of the human TIGIT receptor on tumor infiltrating lymphocytes (TIL) of cancer patients, we co-cultivated human CD8 + T lymphocytes from the TIL of ovarian ascites of patient with CHO-K1 cell line modified to express human CD155 and to activate T lymphocytes. We observed that the release of IFNg by CD8 + T lymphocytes in the presence of modified CHO-K1 cells expressing CD155 can be increased by blocking hTIGIT with antagonistic antiTIGIT antibodies. The “Thaw-and-Use” CD155 aAPC / CHO-K1 cells (Promega, CS198811) were seeded in 96-well plates with U-bottom according to the manufacturer's recommendations and incubated at 37 ° C, 5% CO2 in a O / N incubator. The next day, CD8 + T cells were purified according to the manufacturer's recommendations using a negative selection column (Stemcell Technologies, 17953) from frozen human TIL isolated from ovarian ascites (Immunehealth). The purified CD8 + T cells were then incubated with clone 26452 of BE2017 / 5535 the anti-TIGIT antibody, the non-optimized parent of clones 29489 and 31282 (100,000 CD8 + T lymphocytes / 100 μl of complete medium containing the antibody) for 1 hour. Then, the antibody-CD8 mixture was added to the CD155 aAPC / CHO-K1 cell plates containing 50 µl of fresh complete medium and incubated at 37 ° C, 5% CO2 for 5 days. Finally, the concentrations of IFNg were evaluated in the cell supernatant with an ELISA test (Affymetrix eBioscience, 88-7316-86) which was carried out according to the manufacturer's recommendations. As shown in Figure 14, the secretion of IFNg was increased by almost 2-fold when the anti-TIGIT antibody was added to the coculture. These data demonstrate a powerful functional activity of antagonistic anti-TIGIT antibodies to block the TIGIT inhibitory signal in human CD8 + TILs and to increase the effector functions of T lymphocytes in a tumor environment. Example 11 Characterization of the Antagonistic Anti-TIGIT Antibody with Functional Activity in Mice A. CD155 competition test in mice for a substitute for the antagonistic anti-TIGIT antibody For this test, Jurkat cells (clone E6-1, ATCC TIB-152) modified to overexpress murine TIGIT (Jurkat-mTIGIT) were used. The cells were preincubated for 45 minutes at 37 ° C. with different concentrations of clone 26493 of the anti-TIGIT antibody (0.03 to 10 μg / ml) in 25 μl of complete medium (RPMI + 10% FCS). The cells were washed once and incubated with 4 μg / ml of the mouse CD155-His-Fc protein (Thermo Fisher, 50259M03H50) in 50 μl of complete medium for 45 minutes in an incubator. The cells were washed once, and stained with anti-His antibody coupled to PE (Biolegend, 362603) for 30 minutes at 4 ° C. The median fluorescent intensity (MFI) measured by FACS was used as a measure of the binding of CD155 to Jurkat-mTIGIT. FIG. 15A illustrates the dose-response curve for the anti-TIGIT clone 26493 for measuring the competition with CD155 with an IC 50 value calculated at 2.3 nM (the upper dotted line represents the signal from the isotype, the lower dashed line the signal from cells without CD155). These results demonstrate the functional efficacy of the anti-TIGIT antibody to compete with the ligand CD155 for murine TIGIT. B. In vitro functional test in mice: specific cytotoxicity of an antigen (OT-I) In order to evaluate the specific cytotoxic activity of an OT-I CD8 + T lymphocyte antigen on the target cells pulsed with OVA and to evaluate the effect of the anti-TIGIT antibody in this test, OT1 cells were were isolated from spleens of C57BL / 6-Tg (TcraTcrb) 1100Mjb / Crl (Charles River) mice by BE2017 / 5535 mechanical dissociation followed by negative selection for mouse T cells using an EasySep ™ mouse T cell isolation kit (Stemcell, catalog no. 19851). The PanO2 cancer cells which naturally express CD155, were treated with Mitomycin C (25pg / ml) and then pulsed with the OVA peptide (S7951-1MG, Sigma Aldrich, 1 pg / ml, 1 hour at 37 ° C) in order to be used as cells presenting the antigen ,. CD8 + T lymphocytes and PanO2 cells were co-cultured for 3 days in the presence of the clone 26493 antiTIGIT or of the control isotype at 133 nM. After 3 days, the supernatant was collected for the detection of IFN-gamma by ELISA (Figure 15B) and the T lymphocytes for the cytotoxicity test (Figure 15C). PanO2 OVA-pulsed cells were used as target cells. The target cells as well as non-pulsed Pan02 cells (internal controls), 1 × 10 6 each, were labeled with CFSE (C1157, ThermoFisher) and with the CelITrace ™ Far Red cell proliferation kit (C34564, ThermoFisher) respectively , according to the manufacturer's instructions. These cells were mixed (1: 1 ratio) and seeded at 2 x 10 4 cells per well. OT-1 CD8 + stimulated T cells were added to 1 x 10 5 cells / well (effector cells) giving an effector to target ratio of 10: 1 in the presence of the anti-TIGIT clone 26493 or of the isotype witness at 133 nM. After 24 hours, the cells were washed with PBS and detached by trypsinization. The cells were then stained with the "Live / dead fixable violet dead" cell staining kit (Molecular Probes, L34955). Cytotoxic destruction of target cells was then measured by monitoring the modulation of the ratio of living target cells to non-target cells by flow cytometry. Figure 15B demonstrates that the anti-TIGIT antibody increases the production of IFN-gamma by almost twice while Figure 15C demonstrates an increase in the cytotoxic activity of mouse CD8 + OT-I T lymphocytes by approximately 60%. Taken together, these results confirm the functional activity of the anti-TIGIT antibody to increase the effector function of mouse CD8 + T cells. Example 12 Antitumour Activity of the Antagonist Anti-TIGIT Antibody in Monotherapy and in Combination with the Anti-PD1 Antibodies in a Mouse Model A. In vivo anti-tumor activity of the antagonist anti-TIGIT antibody in monotherapy For this experiment, the anti-TIGIT clone 26493 was produced in mammalian cells on a mouse lgG2a isotype. 8 week old female Balb / c mice were inoculated with 500,000 CT26 colon cancer cells (ATCC® CRL-2638 ™) subcutaneously. On the 9th day after inoculation, when the tumor volumes were on average around 45 mm 3 , the mice were randomized into treatment groups with an equal tumor volume (n = 8 per group). The mice were treated with 200 μg of anti-TIGIT or of the control isotype (mlgG2a, Bioxcell) or with 200 μg of anti-PD-1 (RMP1-14, BioXcell) and 200 μg of the isotype control (mlgG2a, Bioxcell) or with 200 μg of anti-PD-1 (RMP1-14, BioXcell) and different concentrations of anti-TIGIT (200 μg, 60 μg, 20 μg) by intraperitoneal injections on day 9, day 12 and day 15. Tumor growth was monitored and tumor volumes were measured with a caliper BE2017 / 5535 electronic, three times a week, from day 9 to day 36. The mice were sacrificed when the tumor volume exceeded 2000 mm 3 . A statistical analysis of tumor growth curves was performed using a mixed linear model. The differences between the treatment groups were evaluated by testing the interaction of the time * treatment group. In order to test a synergistic effect between anti-TIGIT and anti-PD-1, the treatment groups were recorded by a combination of two variables; anti-TIGIT (yes / no) and anti-PD-1 (yes / no). A synergistic effect, in addition to the additive effect of each treatment (anti-TIGIT * time and anti-PD-1 * time) was evaluated by testing the anti-TIGIT * anti-PD-1 * time interaction. Figure 16A illustrates the mean tumor growth curves by group as well as individual growth curves for mice treated with the anti-TIGIT antibody as monotherapy. While in the control group, no mouse demonstrated regression of the tumor, 2 out of 8 mice treated with anti-TIGIT demonstrated a complete response. In the remaining mice, significant tumor growth retardation was present. In the control group, no mouse survived beyond 30 days, while in the treated group, 7 out of 8 mice survived beyond 30 days. Figure 16B illustrates the mean tumor growth curves by group as well as individual growth curves for mice treated with anti-PD1 as monotherapy or in combination with anti-TIGIT. A significant delay in tumor growth was observed in mice treated with anti-TIGIT + anti-PD-1 compared to anti-PD-1 monotherapy (p <0.0001). The combination of anti-TIGIT + anti-PD-1 made it possible to obtain a synergistic anti-tumor efficacy which was greater than the additive effect of the two monotherapy treatments (p = 0.02). The combination of anti-TIGIT (200 µg) and anti-PD1 antibodies resulted in a complete response in 7 out of 8 mice. Antitumor efficacy was maintained with the combination of anti-PD1 and lower doses of anti-TIGIT which gave a complete response in 8 out of 8 mice when the anti-TIGIT antibody was reduced to 60 pg and 5 out of 8 mice when the anti-TIGIT antibody was further reduced to 20 pg (Figure 16C). These data demonstrate the significant anti-tumor efficacy of anti-TIGIT therapy as monotherapy (p <0.0001) or in combination (p <0.0001) for the treatment of pre-established tumors. EXAMPLE 13 Antitumor Activity Dependent on the Isotype of the Antagonist Anti-TIGIT Antibody in Monotherapy and in Association with the Anti-PD1 Antibodies in a Mouse Model For this experiment, the anti-TIGIT clone 26493 was produced in mammalian cells on a mouse IgG2a and IgG 1 isotype. 8 week old female Balb / c mice were inoculated with 500,000 CT26 colon cancer cells (ATCC® CRL-2638 ™) subcutaneously. On the 10th day after inoculation, when the tumor volumes were on average around 100 mm 3 , BE2017 / 5535 the mice were randomized into treatment groups with an equal tumor volume (n = 10 per group). For the evaluation of monotherapy, the mice were treated with 200 μg of anti-TIGIT or of the control isotype (mlgG2a, Bioxcell) by intraperitoneal injections on day 10, day 13 and day 16. For evaluation of the combination with anti-PD-1, the mice were treated with 200 μg of antiiPD-1 (RMP1-14, BioXcell) and 200 μg of the control isotype (mlgG2a, Bioxcell) or with a combination of 200 µg of anti-PD-1 (RMP1-14, BioXcell) and 200 µg of anti-TIGIT by intraperitoneal injections on day 10, day 13 and day 16. Tumor growth was monitored and tumor volumes were were measured with an electronic caliper, three times a week, from day 10 to day 33. The mice were sacrificed when the tumor volume a exceeded 2000 mm 3 . Figure 17A illustrates the mean tumor growth curves by group as well as individual growth curves for monotherapy with anti-TIGIT antibody and Figure 17B for combination therapy between anti-TIGIT and anti-PD1 antibodies. Both as monotherapy and in combination with anti-PD-1, treatment with the anti-TIGIT antibody resulted in significant anti-tumor efficacy when administered in the form of the lgG2a mouse isotype (p = 0.0001 and p = 0.009, respectively). However, no anti-tumor efficacy could be observed with the anti-TIGIT in the form of the mouse lgG1 isotype, which suggests that the interaction of the Fc receptor with mlgG2a is important for the anti-tumor activity of the anti antibodies. -TIGIT antagonists in the CT26 model in mice. These results demonstrate an anti-tumor efficacy, in monotherapy or in combination, which depends on the anti-TIGIT isotype for the treatment of pre-established tumors. Example 14 Characterization of the Mechanism of Action of the In Vivo Antitumor Activity of the Antagonistic Anti-TIGIT Antibody A. Analysis by flow cytometry of the spleen and tumor In order to study the in vivo mode of action of the antagonistic anti-TIGIT antibody, tumors were analyzed by flow cytometry for the infiltrate of immune cells following treatment with the anti-TIGIT antibody, as monotherapy and in combination with anti-PD-1. The mice were inoculated and treated as described in Example 12. Two days after the second treatment, the mice (8 mice per group) were sacrificed and the tumors removed. The tumors were dissociated with a tumor dissociation kit (Miltenyi Biotec). For direct ex vivo staining, the cells were stained with anti-CD45, anti-CD4, anti-CD8 and anti-FoxP3 (all from eBioscience) after staining with a viability dye (Molecular Probes, L34955) and Fc-block. For ex vivo stimulation, the cells were incubated with a cell stimulation cocktail (eBioscience) and a protein transport inhibitor (eBioscience) for 3 hours. This was followed by staining with anti-CD4 and anti-CD8 and Fc-block antibodies. After fixing and permeabilization with commercial buffers (Cl fixing buffer and BE2017 / 5535 permeabilization), the cells were labeled with anti-IL-10 and anti-IFN-gamma (all from eBioscience). In all the figures, the percentage change compared to the relevant control group (control isotype for monotherapy, anti-PD-1 for the combination) is illustrated, with a negative value representing a decrease and a positive value representing an increase compared to the control group. FIG. 18A shows that the in vivo treatment of the tumor with the anti-TIGIT antibody mlgG2a results in a decrease in the proportion of regulatory T lymphocytes in the CD4 + TIL population of 28% compared to the control group, and which does not is not observed after treatment with anti-TIGIT mlgG1. This demonstrates that there is a decrease in TregIT TIGIT + cells, possibly explaining the differential efficiency of these two isotypes, as it is presented in example 14. FIG. 18B demonstrates that there is no decrease of CD8 + TILs, but rather, a small increase that is observed for the two isotypes (an increase of 17% compared to the controls for mlgG1 and 16% for mlgG2a). These combined results show an increase of more than 50% in the CD8 / Treg ratio in tumors treated with the anti-TIGIT mlgG2a (Figure 18C). The functionality of intratumoral T lymphocytes is also improved for the group treated with the anti-TIGIT antibody mlgG2a, with a large increase in the production of IFN-gamma for both CD4 + TILs (FIG. 18D) and TILs CD8 + (Figure 18E). This resulted in a large increase in the ratio of IFN-g producing cells to IL-10 producing cells after ex vivo stimulation in the CD4 + / CD8 + TIL population (Figure 18F). Figure 18G shows that the combination of anti-TIGIT mlgG2a with anti-PD-1 results in a 33% decrease in regulatory T cells compared to anti-PD1 monotherapy. Again, for CD8 + T cells the opposite is true, with an increase of 22% and 28% in the infiltration of CD8 + T cells, respectively for the isotypes mlgG1 and mlgG2a, in comparison to monotherapy PD-1 (Figure 18H). Together, this results in an increase of more than twice in the TIL CD8 + / Treg ratio in the tumor for the combination with the anti-TIGIT mlgG2a (Figure 181). In addition, treatment with the anti-TIGIT antibody mlgG2a in combination with the anti-PD-1 shows an enrichment of the Th1 versus Th2 phenotype for intratumoral CD4 + T lymphocytes, with a marked increase in CD4 cells producing IFN. -gamma (Figure 18J) and a decrease in CD4 cells producing IL-10 (Figure 18K). This leads to a large increase in the ratio of cells producing IFN-gamma versus IL-10 after ex vivo stimulation in the population of CD4 + TIL compared to mice treated with anti-PD-1 monotherapy (Figure 18L) . Table 11: Differential gene expression between mice treated with anti-TIGIT mlgG2a and mice treated with the support Gene symbol Log2 change factor Corrected p-value BE2017 / 5535 CCR2 -1.29 0.0000668 PRF1 1.79 0.0000668 CTSG 2.13 0.0000668 CTLA4 1.72 0.00309 GZMB 1.51 0.00309 CCL2 0.56 0.0174 IL2RA 1.61 0.0174 CD55 1.64 0.0213 IL2RB 0.872 0.0379 Cd274 0.982 0.0385 Klrgl 1.3 0.0402 Icos 1.26 0.0402 111 rn 0.87 0.0402 Cx3cr1 -0.82 .0428 Cira 0.896 .0428 CD33 -0.906 0.0479 CCl4 0.886 .0518 Table 12: Differential gene expression between mice treated with anti-TIGIT mlgG2a + anti-PD-1 and mice treated with anti-PD-1 Gene symbol Log2 change factor Corrected p-value CTSG 2.34 0.0000375 PRF1 1.69 0.000255 GZMB 1.71 0.000766 CD55 2.08 0.00131 EntpcH 0.839 0.00131 Klrgl 1.76 0.00132 ITGAL 0.874 0.0017 CTLA4 1.72 0.00173 IL2RA 1.82 0.00237 ITGB3 0.863 0.00237 SLC11A1 0.849 0.00329 CD36 1.44 0.0049 CD 180 0.899 0.00602 Icaml 0.893 0.00802 Cd274 1.06 0.00993 CD40 0.926 0.0113 BE2017 / 5535 Eomes 1.28 0.0113 Abcgl 0.869 0.0113 CCR2 -0.781 0.0122 Thy1 0.868 0.0165 CCL2 0.501 0.0203 GBP5 1.12 0.0216 Icos 1.24 0.0263 TGFBR2 0.458 0.0278 H2 K1 0.292 0.0307 SH2D1A 0,999 0.0307 IL2RB 0.808 0.0307 SELPLG 0.64 0.031 BST1 0.702 0.0317 Cd247 1 0,032 IRF8 0.699 0.0365 1121 r 0.899 0.0392 Gbp2b 1.11 0.0392 Statl 0.865 0.0427 C4b 0.922 .0428 ABCAL 0.537 0.044 TREM2 0.482 0.0454 B. Transcriptomic analysis of the tumor by NanoString In order to study the in vivo mode of action of the anti-TIGIT antibody, the infiltrate of immune cells from tumors treated with anti-TIGIT, as monotherapy and in combination with anti-PD5 1, was analyzed by transcriptomic analysis (Nanostring). The mice were inoculated and treated as described in Example 12. Two days after the third treatment with antiTIGIT and / or anti-PD1 antibodies, the mice were sacrificed and the tumors removed. The RNA was extracted and the expression of a selection of 770 genes involved in cancer immunology was directly quantified with nCounter technology (PanCancer Immune Profiling panel, Nanostring; carried out by 10 VIB Nucleomics Core). The data were analyzed with nSolver software (Nanostring). Figure 19A illustrates a volcano graph of the genes which are differentially regulated between the mice treated with the diluent and the mice treated with the anti-TIGIT mlgG2a. Genes with high statistical significance are found at the top of the graph, and genes expressed in very differentiated ways are found on both sides (left = negative regulation in mice treated with anti-TIGIT, right = regulation positive in mice treated with anti-TIGIT). Examples of genes exhibiting high upregulation include perforin, granzyme B and CTLA-4. The solid line represents an uncorrected p value of 0.01, the dotted line represents a corrected p value of 0.05 (Benjamini-Hochberg correction). Table 11 and Table 12 BE2017 / 5535 show the genes which are significantly differentially expressed for the anti-TIGIT mlgG2a in comparison with the diluent and the anti-PD-1 + anti-TIGIT mlgG2a versus the anti-PD1 respectively. When multiple genes have been summarized in the scores for functional immune cell subpopulations, the most significant result is an increase in the score for cytotoxic cells and CD8 + T lymphocytes (Figure 19B). The same changes are present in mice treated with anti-PD-1 + anti-TIGIT mlgG2a in comparison with anti-PD-1 alone. No change was observed in mice treated with anti-TIGIT mlgG1, as monotherapy or in combination with anti-PD-1. Taken together, these results demonstrate that the anti-tumor efficacy observed after in vivo treatment with the anti-TIGIT antibody is facilitated by a decrease in the Treg infiltrate in the tumor while the population of CD8 + effector T lymphocytes is increased. In addition, the effector function of TIL CD4 + and CD8 + is increased as shown by the higher proportion of cells producing IFNg, the increase in TH1 response and the increase in expression of genes important for cytotoxic functions of T lymphocytes Example 15 Activity of Antibody Dependent Cell Toxicity Induced by Antagonistic Anti-TIGIT Antibodies A. In vitro ADCC on human PBMCs from healthy donors PBMCs isolated from healthy human donors were resuspended in complete RPMI medium (supplemented with 10% heat inactivated SVF + 50 U Peniciline + 50 U Streptomycin, and supplemented with 200 IU IL-2 / ml) . 2.5x10 5 of human PBMC were distributed per well in a 96 U well plate. Clone 26452 of the anti-human TIGIT antibody produced in mammalian cells or the control IgG1 isotype (Biolegend, 403102) was added to a final concentration of 66.6; 0.66 and 0.006 nM for each corresponding well. The cells are incubated for 20 hours at 37 ° C with 5% CO2. The cells were then harvested and stained with the following antibodies: LVD efluor 520 (eBioscience 65-0867-14), ant-TCRab-PercPCy5.5 (Clone IP26, Biolegend 306723), anti-CD4-BV510 (Clone SK3, BD Horizon 562970), anti-CD8APC-Cy7 (Clone SK1, Biolegend 344714), anti-CD25-BV605 (Clone 2A3, Biolegend 562660), antiCD127-APC (A019D5, Biolegend 351316), anti-CCR7-BV421 (Clone G043H7 353207) and anti-CD45RO-PE-Cy7 (Clone UCHL1, Biolegend 304229). The results are presented for subpopulations of living cells. CD45 + CD4 + or CD45 + CD8 + represent the total CD4 + or CD8 + T lymphocytes. The CD45 + RO + CD4 + or CD45 + RO + CD8 + represent CD4 + T cells or CD8 + memory while the low CD4 + CD25 HIGH CD127 represent the Treg cells. The proportion of TIGIT + cells on Treg cells taken apart is higher than for CD8 + T cells and CD4 + memory polarized T cells, as shown in Figure 20A. BE2017 / 5535 Absolute quantification is carried out with AccuCheck counting balls (Life technologies) according to the manufacturer's specifications. After calculating the absolute number of cells per μl, the% of specific lysis is calculated using the following formula = (1- (absolute number of cells per μl on the sample treated with the TIGIT 26452 antibody / mean of triplicate without antibody treatment)) x100. As shown in Figure 20B, the anti-TIGIT 26452 hlgG1 antibody triggers higher specific lysis on Tregs (62.22%) than on total CD8 + T T cells (12.2%) or CD4 T cells + totals (16.36%). B. Ex-vivo ADCC on the tumor in mice To confirm that the mouse anti-TIGIT lgG2a antibody can decrease TIGIT + regulatory T cells, an ex-vivo ADCC assay has been developed. 8 week old female Balb / c mice were inoculated with 500,000 CT26 colon cancer cells (ATCC® CRL-2638 ™) subcutaneously. Three weeks after inoculation, the tumors were removed and dissociated with a tumor dissociation kit (Miltenyi Biotec). The single cell suspension was incubated with 133 nM anti-TIGIT antibody (mlgG1 or mlgG2a) for 20 hours (1 million cells / 200 µl in RPMI + 10% FCS). After 20 hours, the cells were labeled with anti-CD4, antiTIGIT, anti-CD8 and anti-FoxP3 (all from eBioscience) after staining with a viability dye (Molecular Probes, L34955) and Fc-block. Figure 21 illustrates the% decrease in the absolute number of TIGIT + cells compared to treatment with the control isotype for the various TIGIT + immune subpopulations. The largest decrease after treatment with the anti-TIGIT antibody mlgG2a is evident for regulatory T cells (a decrease around 40%), suggesting that these cells are more susceptible to ADCC than CD4 T cells + or classic CD8 + . Overall, these results demonstrate the effectiveness of the anti-TIGIT hlgG1 or mlgG2a to decrease the TIGIT + immune cells with preferential activity on the Treg population. Example 16: Prediction of immunogenicity using computer analysis (in silico) The immunogenic potential of clones 29494 and 29489 as well as for its variant 31282 was evaluated by computer prediction using the protein score EpiMatrix (De Groot et al. (2009) Clinical Immunol. 131: 189). To complete the analysis, the input sequences were analyzed in overlapping 9-mer frames and each frame was evaluated against a set of eight common HLA Class II alleles. These alleles are "super types". Each is functionally equivalent to, or almost equivalent to, several other "family member" alleles. Taken together, these eight super-type alleles, accompanied by members of their respective families, "cover" well over 95% of the human population (Southwood et al. (1998) J. Immunol 160: 3363). Each frame-by-allele "assessment" is a statement regarding the predicted HLA binding affinity. EpiMatrix assessment scores range from approximately -3 to +3 and are normally BE2017 / 5535 distributed. EpiMatrix assessment scores above 1.64 are defined as "keys"; i.e., they are potentially immunogenic and should be considered further. All other factors being equal, the more HLA ligands (i.e., EpiMatrix keys) present in a given protein, the higher the probability that this protein will induce an immune response. The EpiMatrix protein score represents the difference between the number of predicted T cell epitopes expected to be found in a protein of a given size, and the number of putative epitopes predicted by the EpiMatrix system. The EpiMatrix protein score is correlated with the observed immunogenicity. Scores of EpiMatrix protein scores are "normalized" and can be reported on a standardized scale. The “EpiMatrix Protein Score” for an “average” protein is zero. EpiMatrix protein scores greater than zero indicate the presence of excess MHC ligands and indicate higher potential for immunogenicity while scores below zero indicate the presence of a smaller amount of potential MHC ligands than expected and a lower potential for immunogenicity. Proteins with a score greater than +20 are considered to have significant immunogenicity potential. Adjustment for the presence of regulatory T cell epitopes. Antibodies are unique proteins because the amino acid sequence of their variable domain, particularly their complementarity determining region (CDR), can vary to an extraordinary extent. It is this variability that allows antibodies to recognize a wide variety of antigens. However, the recombination and mutation events that control the maturation of the antibody can also produce new epitopes or T-cell neo-epitopes. These neo-epitopes may appear to be "foreign" to circulating T cells. The presence of neo-epitopes in antibody sequences can result in the formation of a human-anti-human antibody response, also called a HAHA or ADA (anti-drug antibody) response. Regulatory T cells play an important role in suppressing immune responses against completely human proteins at the periphery, including those containing mutated sequences and / or highly variable sequences such as antibody CDRs. Regulatory T cells come into contact and are activated by epitopes of regulatory T cells. The inherent risk associated with the presence of neo-epitopes in antibody sequences appears to be balanced by the presence of epitopes of natural regulatory T cells. By screening the sequences of many isolated human antibodies, EpiVax identified several highly conserved HLA ligands that have regulatory potential. Experimental data suggests that many of these peptides are, in fact, actively tolerogenic in most subjects. These highly conserved, ubiquitous and regulatory T cell epitopes are called Tregitopes (De Groot et al. (2008) Blood 112: 3303) BE2017 / 5535 In many cases, the immunogenic potential of the neo-epitopes contained in humanized antibodies can effectively be controlled by the presence of a large number of Tregitopes. As part of the analysis of the immunogenicity of the antibodies, EpiVax developed an EpiMatrix score adjusted for the Tregitope and the corresponding prediction of an anti-therapeutic antibody response. In order to calculate the adjusted EpiMatrix score for the Tregitope, the Tregitope scores were deduced from the protein score EpiMatrix. Adjusted scores for Tregitope have been shown to correlate well with clinical immune responses observed for a set of 23 commercial antibodies (De Groot et al. (2009) Clinical Immunol. 131: 189). The antibody sequence score for clones 29489, 29494 and 31282 is found at the bottom of the EpiMatrix scale, indicating limited potential for immunogenicity. Regression analysis of the sublicensed monoclonal antibodies predicts an ADA response in approximately 0% of patients exposed for clones 29489 and 31282 of the antibody. For clone 29494, the analysis predicts an ADA response in 2.78% of the patients exposed for the reference VH sequence, and 2.88% for the variant VH sequence. The data is summarized in Table 13 below. Table 13: EpiMatrix scores adjusted by EpiMatrix and Tregitope Entry sequence Length Ratings EpiMatrix keys EpiMatrix Score EpiMatrix score adjusted for Tregitopeliilliiiii IIIIIIIIIII iiiiiiiiiiiiiiiiiiiiiiiiiin IIIIIIIIIII iiiiiililliiiiiii 29489_VL 108 800 39 -17.58 -51.75 31282_VH 121 904 40 -19.41 -47.26 liiiiiiiii iiiiiiiiii IIIIIIIIIII IIIIIIIIIII -17.58 IIIIIIIIIIIIIIIII iiiliiliili liiiiiiiii iiiilliiiiii IIIIIIIIIII IIIIIIIIIII -7.18 29494 VL 107 792 40 -12.2 -38.83 BE2017 / 5535 Legend of figures: Fig. 7: Competition with the human CD155 ligand Fig. 8: Expression of TIGIT on different immune populations originating from PBMCs of healthy donors Fig. 9: Liaison with Jurkat-hTIGIT Fig. 10: Binding to human primary CD8 + T cells Fig. 11: Binding to primary CD8 + T cells from Cynomolgus Fig. 12: Effect of anti-TIGIT antibodies in a CHO-TCR-CD155 and Jurkat-hTIGIT bioassay Fig. 13: Effect of anti-TIGIT anticoprs on human CD8 + T lymphocytes based on a functional test Fig. 14: Effect of anti-TIGIT anticoprs on human TIL based on a functional test Fig. 15: Characterization of the mouse anti-TIGIT replacement anticoprs which demonstrates functional activity in mice Fig. 16: Anti-tumor efficacy of the antagonistic anti-TIGIT antibody Fig. 17: Anti-tumor efficacy dependent on the isotype of the antagonistic anti-TIGIT antibody Fig. 18: Mechanism of the anti-tumor efficacy of the antagonistic anti-TIGIT anticoprs Fig. 19: Mechanism of the anti-tumor efficacy of the antagonist anti-TIGIT anticoprs (transcriptional analysis) Fig. 20: ADCC activity on healthy human PBMCs Fig. 21: Preferential decrease in Treg cells in a tumor suspension in mice
权利要求:
Claims (20) [1] 1. An isolated antibody or antigen binding fragment thereof which binds to human TIGIT and which comprises a heavy chain variable domain in which the antibody or antigen binding fragment comprises a combination of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, in which: HCDR1 includes SEQ ID NO: 16 (YTFTSYYMH), HCDR2 includes SEQ ID NO: 17 (VIGPSGASTSYAQKFQG), HCDR3 includes SEQ ID NO: 18 (ARDHSDYWSGIMEV), LCDR1 includes SEQ ID NO: 61 (RASQSVRSSYLA), LCDR2 includes SEQ ID NO: 62 (GASSRAT), and LCDR2 includes SEQ ID NO: 63 (QQYFSPPWT). [2] 2. The antibody or antigen binding fragment of claim 1, wherein the heavy chain variable domain comprises the amino acid sequence illustrated as SEQ ID NO: 221 or an amino acid sequence demonstrating sequence identity at least 90%, 95%, 97%, 98% or 99% thereof, and the variable domain of the light chain include the amino acid sequence illustrated as SEQ ID NO: 222 or a sequence d amino acids demonstrating a sequence identity of at least 90%, 95%, 97%, 98% or 99% thereto. [3] 3. The antibody or antigen binding fragment according to claim 5, wherein the heavy chain variable domain comprises the amino acid sequence illustrated as SEQ ID NO: 219 or an amino acid sequence demonstrating sequence identity at least 90%, 95%, 97%, 98% or 99% thereof, and the variable domain of the light chain include the amino acid sequence illustrated as SEQ ID NO: 220 or a sequence d amino acids demonstrating a sequence identity of at least 90%, 95%, 97%, 98% or 99% thereto. [4] 4. The antibody or antigen binding fragment according to any preceding claim, which is a human IgG antibody, preferably a human IgG1 antibody. [5] 5. Antibody or antigen-binding fragment according to any preceding claim, which demonstrates one or more effector functions chosen from cytotoxicity facilitated by the antibody-dependent cell (ADCC), complement-dependent cytotoxicity (CDC) and phagocytosis facilitated by the antibody-dependent cell (ADCP) against cells expressing TIGIT on the cell surface. BE2017 / 5535 [6] 6. Antibody or antigen binding fragment according to any preceding claim, which preferentially decreases Treg cells expressing TIGIT. [7] 7. Isolated anti-TIGIT antibody or antigen-binding fragment thereof which preferentially depletes TIGIT-expressing Treg cells, optionally in which the antibody or antigen-binding fragment is an antibody or a fragment of binding to the antigen according to any one of claims 1 to 3. [8] 8. Antibody or antigen binding fragment according to any preceding claim, which demonstrates an equivalent affinity for Treg cells expressing TIGIT and CD8 + T lymphocytes expressing TIGIT. [9] 9. Antibody or antigen binding fragment according to any preceding claim, which decreases the expression of TIGIT on CD8 + T lymphocytes and / or on Treg cells. [10] 10. An isolated polynucleotide encoding an antibody or an antigen binding fragment according to any preceding claim. [11] 11. The isolated polynucleotide according to claim 10, wherein the isolated polynucleotide comprises SEQ ID NO: 251 and SEQ ID NO: 252. [12] 12. Expression vector comprising the polynucleotide according to claim 10 or claim 11, operably linked to regulatory sequences which allow expression of the antigen binding polypeptide in a host cell or in a cell-free expression system. [13] 13. Host cell or expression system free of cells containing the expression vector according to claim 12. [14] 14. A method of producing a recombinant antibody or an antigen binding fragment thereof which comprises culturing the host cell or the cell-free expression system according to claim 13, under conditions which allow expression of the antibody or antigen binding fragment and recovery of the antibody or expressed antigen binding fragment. [15] 15. A pharmaceutical composition comprising an antibody or an antigen-binding fragment according to any one of claims 1 to 9, and at least one pharmaceutically acceptable carrier or excipient. [16] 16. The antibody or antigen binding fragment according to any of claims 1 to 9, or the pharmaceutical composition according to claim 15 for use in therapy. BE2017 / 5535 [17] 17. The antibody or antigen binding fragment according to any of claims 1 to 9, or the pharmaceutical composition according to claim 15 for use in a method of treating cancer. [18] 18. A method of treating cancer in a subject comprising administering an effective amount 5 of an antibody or antigen binding fragment according to any one of claims 1 to 9 or a pharmaceutical composition according to claim 15 to the subject, thereby treating cancer. [19] 19. An antibody or an antigen binding fragment or a pharmaceutical composition for use in a method according to claim 17, or a method of treating cancer according to claim 18, wherein the method also comprises administering an agent 10 additional therapy. [20] 20. Antibody or the antigen binding fragment or pharmaceutical composition for use in a method according to claim 19 or a method of treating cancer according to claim 19, wherein the therapeutic agent is selected from: an agent chemotherapeutic, anti-PD1 antibody, anti-PD-L1 antibody.
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同族专利:
公开号 | 公开日
引用文献:
公开号 | 申请日 | 公开日 | 申请人 | 专利标题 WO2016028656A1|2014-08-19|2016-02-25|Merck Sharp & Dohme Corp.|Anti-tigit antibodies| WO2016106302A1|2014-12-23|2016-06-30|Bristol-Myers Squibb Company|Antibodies to tigit| WO2016191643A2|2015-05-28|2016-12-01|Oncomed Pharmaceuticals, Inc.|Tigit-binding agents and uses thereof|
法律状态:
2019-02-25| FG| Patent granted|Effective date: 20190123 | 2020-12-04| HC| Change of name of the owners|Owner name: ITEOS BELGIUM SA; BE Free format text: DETAILS ASSIGNMENT: CHANGE OF OWNER(S), CHANGEMENT DE NOM DU PROPRIETAIRE; FORMER OWNER NAME: ITEOS THERAPEUTICS S.A. Effective date: 20201026 |
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申请号 | 申请日 | 专利标题 US201762606159P| true| 2017-07-27|2017-07-27| US62606159|2017-07-27|EP18750302.4A| EP3484925B1|2017-07-27|2018-07-26|Anti-tigit antibodies| CA3070791A| CA3070791A1|2017-07-27|2018-07-26|Anti-tigit antibodies| SG11202000660QA| SG11202000660QA|2017-07-27|2018-07-26|Anti-tigit antibodies| BR112020001499-0A| BR112020001499A2|2017-07-27|2018-07-26|anti-tigit antibodies| US16/634,506| US20200407445A1|2017-07-27|2018-07-26|Anti-tigit antibodies| EP20173561.0A| EP3719040A1|2017-07-27|2018-07-26|Anti-tigit antibodies| JP2020527852A| JP2020528768A|2017-07-27|2018-07-26|Anti-TIGIT antibody| KR1020207005664A| KR20200100589A|2017-07-27|2018-07-26|Anti-TIGIT antibody| AU2018306463A| AU2018306463A1|2017-07-27|2018-07-26|Anti-TIGIT antibodies| ES18750302T| ES2812236T3|2017-07-27|2018-07-26|Anti-TIGIT antibodies| CN201880048693.8A| CN110997720A|2017-07-27|2018-07-26|anti-TIGIT antibody| PCT/US2018/043968| WO2019023504A1|2017-07-27|2018-07-26|Anti-tigit antibodies| TW107126050A| TW201910351A|2017-07-27|2018-07-27|Anti-tigit antibodies| ARP180102114A| AR112768A1|2017-07-27|2018-07-27|ANTI-TIGIT ANTIBODIES| US16/159,506| US10329349B2|2017-07-27|2018-10-12|Anti-TIGIT antibodies| US16/413,557| US20190315867A1|2017-07-27|2019-05-15|Anti-tigit antibodies| IL272227A| IL272227D0|2017-07-27|2020-01-23|Anti-tigit antibodies| 相关专利
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