Novel anti-CTLA-4 antibodies and nucleotide sequences, and their uses

Abstract

Novel anti-CTLA-4 antibody molecules and nucleotide sequences and expression vectors, such as viruses, encoding such antibody molecules are described; the novel antibody molecules are antibody molecules that decrease Tregs and have an enhanced decrease effect on CTLA-4-positive cells, such as Tregs, compared to ipilimumab; the use of such antibody molecules or nucleotide sequences or viruses in medicine, such as in the treatment of cancer, such as solid tumors, is also described.

Description

DETAILED DESCRIPTION OF THE INVENTION CTLA-4-positive cells include regulatory T cells, Treg cells, Tregs, or Tregs (formerly known as suppressor T cells, sometimes also referred to as suppressor regulatory T cells), which are a subpopulation of T cells that can suppress other cells. immune systems in normal and pathological immune environments. Tregs are CD4-positive cells (CD4+ cells). There are other CD4+ T cells that are not Tregs; however, Tregs can be separated from CD4+ non-Treg cells because Tregs are also FOXP3 positive (FOXP3+) whereas CD4+ non-Treg cells are FOXP3 negative (FOXP3-). Similar to ipilimumab, the anti-CTLA-4 antibody molecules described herein act, at least in part, by depleting CTLA-4 positive cells, such as Tregs. Furthermore, similar to ipilimumab, the anti-CTLA-4 antibody molecules described herein block the interactions of CTLA-4 with B7.1 and B7.2. Thus, these antibodies may therefore help to overcome the suppressive effects induced by CTLA-4 on effector T cell proliferation. By Treg-lowering, or Treg-depleting, we mean herein a lowering, deletion, or removal of Tregs through physical clearance of cells. In particular, we refer to the decrease in intratumoral Tregs. Lowering of Tregs can be achieved by ADCC, ie, antibody-dependent cell-mediated cytotoxicity or antibody-dependent cellular cytotoxicity, and / or ADCP, ie, antibody-dependent cellular phagocytosis. This means that when an antibody molecule as described herein is administered to a subject, such as a human, it specifically binds to CTLA-4 expressed on the surface of Tregs, and this binding results in decreased of the Tregs. In some embodiments, CTLA-4 is preferentially expressed on tumor-infiltrating lymphocytes in the tumor microenvironment or on tumor cells. ADCC is an immune mechanism through which Fe receptor-bearing effector cells can recognize and kill antibody-coated target cells that express tumor-derived antigens, i.e., in the present case, CTLA-4, on their surface. . ADCP is a similar mechanism, although it results in the death of target cells through phagocytosis rather than cytotoxicity. Antibodies are well known to those skilled in the art of immunology and molecular biology. Typically, an antibody comprises two heavy (H) chains and two light (L) chains. In the present disclosure, we sometimes refer to this complete antibody molecule as a full-length or full-length antibody. The heavy chain of the antibody comprises a variable domain (VH) and three constant domains (CHI, CH2 and CH3), and the light chain of the antibody molecule comprises a variable domain (VL) and a constant domain (CL). The variable domains (sometimes collectively referred to as the Fv region) bind to the antibody target, or antigen. Each variable domain comprises three loops, called complementarity determining regions (CDRs), which are responsible for target binding. Constant domains are not directly involved in the binding of an antibody to an antigen, but they do exhibit various effector functions. Depending on the amino acid sequence of the constant region of their heavy chains, antibodies or immunoglobulins can be assigned to different classes. There are five main classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and in humans, several of these are divided into subclasses (isotypes), eg, IgG1, IgG2, IgG3, and IgG4; IgAl and IgA2. Another part of an antibody is the Fe domain (also known as the crystallizable fragment domain), which comprises two of the constant domains from each of the antibody's heavy chains. The Fe domain is responsible for the interactions between the antibody and the Fe receptor. Fe receptors are membrane proteins often found on the cell surface of immune system cells (ie, Fe receptors are found on the target cell membrane, also known as the plasma membrane or cytoplasmic membrane). The role of Fe receptors is to bind antibodies through the Fe domain and to internalize the antibody into the cell. In the immune system, this can result in antibody-mediated phagocytosis and antibody-dependent cell-mediated cytotoxicity. The term antibody molecule, as used herein, encompasses full-length or full-length antibodies, as well as functional fragments of full-length antibodies and derivatives of such antibody molecules. Functional fragments of a full-length antibody have the same antigen-binding characteristics as the corresponding full-length antibody and include the same variable domains (i.e., the VH and VL sequences) and / or the same CDR sequences as the full-length antibody. corresponding full size antibody. That the functional fragment has the same antigen-binding characteristics as the corresponding full-length antibody means that it binds to the same epitope on the target as the full-length antibody. Said functional fragment may correspond to the Fv part of a full-length antibody. Alternatively, said fragment may be a Fab, also called F(ab), which is a monovalent antigen-binding fragment that does not contain an Fe moiety, or an F(ab')2, which is a divalent antigen-binding fragment. containing two antigen-binding Fab moieties linked to each other by disulfide bonds, or an F(ab'), ie, a monovalent variant of an F(ab')2. Said fragment can also be a single chain variable fragment (scFv). A functional fragment does not always contain all six CDRs of a corresponding full-length antibody. It is appreciated that molecules containing three or fewer CDR regions (in some cases, even a single CDR or a portion thereof) may retain the antigen-binding activity of the antibody from which the CDR(s) is derived. For example, in Gao et al., 1994, J. Bioi. Chem., 269: 32389-93 it is described that a complete VL chain (including the three CDRs) has a high affinity for its substrate. Molecules containing two CDR regions have been described, for example, in Vaughan and Sollazzo 2001, Combinatorial! Chemistry & High Throughput Screening, 4: 417-430. On page 418 (right column - 3 Our Strategy for Design) a minibody is described that includes only the CDR H1 and H2 hypervariable regions interspersed within the framework regions. The minibody is described as being capable of attaching itself to a target. Pessi et al., 1993, Nature, 362: 367-9 and Bianchi et al., 1994, J. Mol. Biol., 236: 649-59 are referenced by Vaughan & Sollazzo and describe the H1 and H2 minibody and their properties in more detail. In Qiu et al., 2007, Nature Biotechnoiogy, 25: 921-9 it is shown that a molecule consisting of two linked CDRs is capable of binding antigen. Quiocho 1993, Nature, 362: 293-4 provides an overview of minibody technology. Ladner 2007, Nature Biotechnoiogy, 25: 875-7 comments that molecules containing two CDRs may retain antigen binding activity. Antibody molecules containing a single CDR region are described, for example, in Laune et al., 1997, JBC, 272: 30937-44, where a variety of hexapeptides derived from a CDR are shown to exhibit binding activity to the CDR. antigen and the / UUZDUU synthetic peptides of a complete and single CDR are reported to show strong binding activity. A variety of 12-mer peptides and associated framework regions are shown to have antigen-binding activity in Monnet et al., 1999, JBC, 274: 3789-96 and it is discussed that a CDR3-like peptide is only capable of binding antigen. . In Heap et al., 2005, J. Gen. Viro!., 86: 1791-1800, a microantibody (a molecule containing a single CDR) is reported to be capable of binding antigen and a cyclic peptide of a Anti-HIV antibody has antigen-binding activity and function. It is shown in Nicaise et al., 2004, Protein Science, 13: 1882-91 that a single CDR can confer antigen binding activity and affinity for its lysozyme antigen. Thus, antibody molecules having five, four, three, or fewer CDRs may retain the antigen-binding properties of the full-length antibodies from which they are derived. The antibody molecule can also be a derivative of a full length antibody or a fragment of such an antibody. The derivative has the same antigen-binding characteristics as the corresponding full-length antibody in that it binds to the same epitope on the target as the full-length antibody. Therefore, by the term antibody molecule, as used in the present description, we include all types of antibody molecules and functional fragments thereof and derivatives thereof, including: monoclonal antibodies, polyclonal antibodies, synthetic antibodies, antibodies produced recombinantly, multispecific antibodies, bispecific antibodies, human antibodies, humanized antibodies, chimeric antibodies, single chain antibodies, variable fragments (Fv), single chain variable fragments (scFv fragments), including divalent single chain variable fragments (di-scFv) and linked variable fragments by disulfide, Fab fragments, F(ab')z fragments, Fab1 fragments, antibody heavy chains, antibody light chains, antibody heavy chain homodimers, antibody light chain homodimers, antibody heavy chain heterodimers, antibody heterodimers antibody light chains, functional antigen-binding fragments of said homodimers and heterodimers. Furthermore, the term antibody molecule, as used herein, includes all classes of antibody molecules and functional fragments, including: IgG, IgGl, IgG2, IgG3, IgG4, IgA, IgM, IgD, and IgE. In some embodiments, the antibody is a human IgGl. The skilled person will know that mouse IgG2a and human IgGl interact productively with activating Fe gamma receptors and share the ability to activate deletion of target cells through activation of immune cells bearing activating Fe gamma receptors (eg, macrophages). and NK cells), for example, ADCP and ADCC. As such, while mouse IgG2a is the preferred isotype for deletion in mice, human IgG1 is a preferred isotype for deletion in humans. Conversely, optimal costimulation of TNFR superfamily agonist receptors, eg, 4-1BB, 0X40, TNFRII, CD40, is known to depend on FcyRII inhibitor antibody interaction. In the mouse, the IgGl isotype, which binds preferentially to inhibitory Fe receptor gamma (FcyRIIB) and only weakly to activating Fe gamma receptors, is known to be optimal for the costimulatory activity of the TNFR superfamily-targeted mAb. Although a direct equivalent of the mouse IgGl isotype has not been described in man, the antibodies can be engineered to show similarly enhanced binding to inhibitory over activating human Fe gamma receptors. Such genetically engineered TNFR superfamily-targeting antibodies also have enhanced costimulatory activity in vivo, in transgenic mice engineered to express human activating and inhibitory Fe gamma receptors (Dahan et al, 2016, Therapeutic Activity of Agonistic, Human Anti-CD40 Monoclonal Antibodies Requires Selective FcyR Engagement Cancer Ceii 29(6): 820-31). As described above, different types and forms of antibody molecules are included in the invention, and are known to the person skilled in immunology. It is known that antibodies used for therapeutic purposes are often modified with additional components that modify the properties of the antibody molecule. Accordingly, we include that an antibody molecule of the invention or an antibody molecule used according to the invention (for example, a monoclonal antibody molecule, and / or polyclonal antibody molecule, and / or bispecific antibody molecule) comprises a detectable moiety and / or a cytotoxic moiety. By detectable moiety, we include one or more from the group comprising: an enzyme; a radioactive atom; a fluorescent moiety; a chemiluminescent moiety; a bioluminescent residue. The detectable moiety allows the antibody molecule to be visualized in vitro and / or in vivo and / or ex vivo. By cytotoxic moiety, we include a radioactive moiety, and / or enzyme, eg, where the enzyme is a caspase, and / or toxin, eg, where the toxin is a bacterial toxin or a poison; wherein the cytotoxic moiety can induce cell lysis. We further include that the antibody molecule may be in isolated and / or purified form, and / or may be PEGylated. As discussed above, the CDRs of an antibody bind to the antibody target. The amino acid assignment to each CDR described herein is in accordance with the definitions according to Kabat EA et al., 1991, in Sequences of Proteins of Immunological Interest Fifth Edition, NIH Publication no. 91-3242, pp xv-xv¡¡. As the skilled person will know, there are also other methods for assigning / UUZDUU amino acids to each CDR. For example, the international information system ImMunoGeneTics (IMGT(R)) (http: / / www.imgt.org / and Lefranc and Lefranc The Immunoglobulin FactsBook published by Academic Press, 2001). In a further embodiment, the antibody molecule of the present invention or used according to the invention is an antibody molecule that is capable of competing with the specific antibodies described herein, such as the antibody molecules comprising SEQ ID NO: 15, 16, 17, 10, 18 and 19 or SEQ ID NO: 22, 23, 24, 10, 25 and 26. By "capable of competing" we mean that the competing antibody is capable of inhibiting or otherwise interfering, at least in part, with the binding of an antibody molecule as defined herein to the specific target. For example, said competitor antibody molecule may be capable of inhibiting the binding of an antibody molecule described herein by at least about 10%; for example at least about 20%, or at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90% , at least about 95%, about 100%, and / or inhibit the ability of the antibody described herein to prevent or reduce binding to the specific target by at least about 10%; for example at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 100%. Competitive binding can be determined by methods well known to those skilled in the art, such as enzyme-linked immunosorbent assay (ELISA). ELISA assays can be used to assess for antibodies that modify or block the epitope. Additional methods suitable for identifying competitive antibodies are described in Antibodies: A Laboratory Manual, Harlow & Lane, which is incorporated herein by reference (for example, see pages 567-569, 574-576, 583 and 590-612, 1988, CSHL, NY, ISBN 0-87969-314-2). It is well known that an antibody specifically binds to a defined target molecule or antigen, and this means that the antibody preferentially and selectively binds to its target and not to a non-target molecule. The CTLA-4 targets of the antibodies according to the present invention, or of the antibodies used according to the invention, are expressed on the surface of the cells, that is, they are cell surface antigens, which would include an epitope ( also known in this context as a cell surface epitope) for the antibody. Antigen and cell surface epitope are terms that an expert in immunology or cell biology will readily understand. By cell surface antigen, we include the cell surface antigen or at least the epitope thereof to which the antibody molecule described herein is exposed on the extracellular side of the cell membrane. Methods for assessing protein binding are known to those skilled in biochemistry and immunology. The skilled person will appreciate that these methods can be used to assess the binding of an antibody to a target and / or the binding of the Fe domain of an antibody to a Fe receptor; as well as the relative strength, or specificity, or inhibition, or prevention, or reduction in those interactions. Examples of methods that can be used to assess protein binding are, for example, immunoassays, BIAcore, Western blots, radioimmunoassay (RIA) and enzyme-linked immunosorbent assays (ELISA) (See Fundamental Immunology Second Edition, Raven Press, New York at pages 332-336 (1989) for a discussion on antibody specificity). Therefore, in the present description both an antibody molecule that specifically binds to CTLA-4 and an anti-CTLA-4 antibody molecule refers to an antibody molecule that specifically binds to the target CTLA-4 but does not bind to a non-target, or binds more loosely to a non-target (such as with lower affinity) than to the target. In some embodiments, the antibody molecule that specifically binds to CTLA-4 (or anti-CTLA-4 antibody molecule) refers to an antibody molecule that specifically binds to the extracellular domain of CTLA-4. In some embodiments, the antibody molecule that specifically binds to CTLA-4 (or the anti-CTLA-4 antibody molecule) does not cross-react with CD28. In some embodiments, the antibody molecule that specifically binds to CTLA-4 (or the anti-CTLA-4 antibody molecule) blocks the binding of CTLA-4 to CD80 and / or CD86, thereby inhibiting CLTA-signaling. 4. We further include the meaning that the antibody specifically binds to the target CTLA-4 at least twice as strongly, or at least five times as strongly, or at least 10 times as strongly, or at least 20 times as strongly, or at least 50 times stronger, or at least 100 times stronger, or at least 200 times stronger, or at least 500 times stronger, or at least approximately 1000 times stronger than a non-target. In addition, we include the meaning that the antibody specifically binds to the target CTLA-4 if it binds to the target with a Kd of at least about 101Kd, or at least / UUZDUU about 10'2Kd, or at least about 10'3Kd, or at least approximately 10'4Kd, or at least approximately 10'5Kd, or at least approximately 10'6Kd, or at least approximately 10-7Kd, or at least approximately 10-8Kd, or at least approximately 10-9Kd, or at least about 1010Kd, or at least about 1011Kd, or at least about 1012Kd, or at least about 1013Kd, or at least about 10'14Kd, or at least about 1015Kd. As mentioned above, the antibody molecules that specifically bind to CTLA-4 (or anti-CTLA-4 antibody molecules) described herein have an enhanced downregulating effect on CTLA-4 positive cells compared to with ipilimumab. The fact that antibody molecules have a downregulating effect on CTLA-4 positive cells means that upon administration to a subject, such as a human, said antibody specifically binds to CTLA-4 expressed on the surface of CTLA-4 positive cells, and this binding results in the depletion of such cells. In some embodiments, the CTLA-4-positive cells are CD4-positive (CD4+) cells, ie, cells that express CD4. In some embodiments, CTLA-4-positive cells are both CD4-positive and FOXP3-positive, ie, they express both CD4 and FOXP3. These cells are Treg. CD8-positive T cells also express CTLA-4, but Tregs express significantly higher levels of CTLA-4 than CD8-positive T cells. This makes Tregs more susceptible to downregulation compared to lower expressing CD8+ cells. In some situations, CTLA-4 is preferentially expressed on immune cells in the tumor microenvironment (tumor infiltrating cells, TILS). Therefore, in a tumor microenvironment, Tregs will be the cells with the highest expression of CTLA-4, resulting in antibody molecules that specifically bind to CTLA-4 (or anti-CTLA-4 antibody molecules). -CTLA-4) have a Treg-lowering effect. This is discussed in more detail below, for example, in Example 4 and in connection with Figure 13. In some embodiments, the CTLA-4 positive cells will be Tregs in a solid tumor. Such Tregs will have very high expression of CTLA-4, and therefore administration of antibody molecules that specifically bind to CTLA-4 will preferentially result in the decrease of such Tregs. As mentioned above, the anti-CTLA-4 antibody molecules described herein are Treg-lowering antibody molecules, which means that upon administration to a subject, such as a human, said antibody molecule binds / UUZDUU specifically to CTLA-4 expressed on the surface of Tregs, and this binding results in downregulation of Tregs. To decide whether an antibody molecule is an antibody molecule that has an enhanced lowering effect on CTLA-4 positive cells compared to ipilimumab as mentioned in the present disclosure, it is possible to use an antibody-dependent cellular cytotoxicity assay. (ADCC) in vitro or an in vivo assay in a PBMC-NOG / SCID model. The in vitro ADCC test which is performed by using an NK-92 cell line stably transfected to express the CD16-158V allele together with GFP, wherein the ADCC test comprises the following seven consecutive steps: 1) CTLA-4 positive cells, CD4 or Treg positive cells are isolated as target cells from the peripheral blood of healthy donors. This isolation can be performed using a CD4+ T cell isolation kit, such as a commercial kit from Miltenyi Biotec. 2) The target cells are then stimulated, eg for 48 hours, with CD3 / CD28, eg using CD3 / CD28 Dynabeads® and rhIL-2, such as 50 ng / ml rhIL-2. Stimulation can be performed at 37°C. 3) Then, the target cells are pre-incubated with the antibody molecule to be tested, for example, at 10 pg / ml for 30 min at 4 °C, and then mixed with NK cells. 4) The target cells are then incubated for an appropriate time, such as 4 hours, in RPMI 1640 + GlutaMAX medium containing HEPES buffer, sodium pyruvate and low IgG FBS. RPMI 1640 + GlutaMAX medium may contain 10 mM HEPES buffer, 1 mM sodium pyruvate, and 10% low IgG FBS, and effector-Target cell ratio may be 2:1. 5) Lysis is determined by flow cytometry. 6) Steps 1-5 are repeated, or performed in parallel, with the ipilimumab used in place of the antibody molecule tested in step 3. 7) The results of the lysis of the tested antibody molecule are compared with the results of the lysis of ipilimumab. Enhanced lysis for the tested antibody molecule compared to ipilimumab demonstrates that the tested antibody molecule has an enhanced knockdown effect on CTLA-4-positive cells, CD4-positive cells, or Treg-positive cells, respectively, depending on which cells are targeted. They were used. In some embodiments, the lowering effect improved in step 7) above is a significantly improved lowering effect. This test is demonstrated in more detail below in Example 4, in combination with Figure 12. The in vivo test is based on the combined use of PBMC mice and NOG / SCID mice, which in the present description is called the PBMC-NOG / SCID model. Both mice / UUZDUU PBMC like NOG / SCID mice are well known models. The in vivo test in the PBMC-NOG / SCID model comprises the following nine consecutive steps: 1) Human PBMC (peripheral blood mononuclear cells) are isolated, washed and resuspended in sterile PBS. In some embodiments, PBMCs were resuspended in PBS at 75 x 106 cells / ml. 2) NOG mice are injected iv (intravenously) with an appropriate amount, such as 200 μΙ, of the cell suspension from step 1). If 200 pl are injected, this corresponds to 15 x 10 6 cells / mouse. 3) A suitable time, such as 2 weeks, after injection, spleens from NOG mice are isolated and placed in single cell suspension. Optionally, a small sample of the single cell suspension is taken to determine CTLA-4 expression in human T cells by FACS, in order to confirm CTLA-4 expression. 4) The cell suspension from step 3) is resuspended in sterile PBS. In some embodiments, the cell suspension is resuspended in sterile PBS at 50 x 106 cells / ml. If determination of optional CTLA-4 expression is included in step 3, the remainder of the cell suspension is then resuspended in step 4. 5) SCID mice are injected i.p. (intraperitoneally) with a suitable amount, such as 200 μΙ, of the suspension from step 4. If 200 μΙ are injected, this corresponds to 10 x 10 6 cells / mouse. 6) A suitable time, such as 1 hour, after injection in step 5) SCID mice are treated with a suitable amount, such as 10 mg / kg, of the antibody molecule to be tested, ipilimumab or a isotype control monoclonal antibody. 7) Intraperitoneal fluid from treated SCID mice is collected at a suitable time, such as 24 hours, after treatment in step 6). 8) Human T cell subsets are identified and quantified by FACS using the following markers: CD45, CD4, CD8, CD25 and / or CD127. 9) The results of the identification and quantification of the T cell subsets of the mice treated with the tested antibody molecule are compared with the results of the identification and quantification of the T cell subsets of the mice treated with ipilimumab and with the Results of identification and quantification of T cell subsets from mice treated with the isotype control monoclonal antibody. A lower number of CTLA-4-positive cells in the intraperitoneal fluid of mice treated with the antibody molecule to be tested compared to the number of CTLA-4-positive cells in the intraperitoneal fluid of ipilimumab-treated mice demonstrates that the antibody molecule has enhanced downregulation effect on CTLA-4 positive cells compared to ipilimumab. A lower number of CD4-positive cells in the intraperitoneal fluid of mice treated with the antibody molecule to be tested compared to the number of CD4-positive cells in the intraperitoneal fluid of ipilimumab-treated mice demonstrates that the antibody molecule it has an enhanced downregulation effect on CD4-positive cells compared to ipilimumab. A lower number of Tregs in the intraperitoneal fluid of mice treated with the antibody molecule to be tested compared to the number of Tregs in the intraperitoneal fluid of ipilimumab-treated mice demonstrates that the antibody molecule has a knockdown effect. Improved decrease in Treg compared with ipilimumab. In this in vivo test, in some modalities it is of greater interest to observe the Treg decline at stage 7. This assay is demonstrated in more detail below in Example 4, in combination with Figures 14A-14B. The Treg decrease can also be assessed in an antibody-dependent cell phagocytosis (ADCP) assay, as is known to the skilled person. In some embodiments, the antibody molecules have a similar blocking effect on CTLA-4 interactions with B7.1 and B7.2 ligands as compared to Yervoy. This can be assessed by ELISA (as shown in Figures 10A and 10B) or in a more functional assay in which anti-CTLA-4 antibodies increase IL-2 production by T cells in response to PBMC stimulation. with SEB. In some embodiments, the anti-CTLA-4 antibody molecule is a human antibody molecule. In some embodiments, the anti-CTLA-4 antibody molecule is a humanized antibody molecule. In some embodiments, the anti-CTLA-4 antibody molecule is an antibody molecule of human origin, meaning that it originates from a human antibody molecule that has then been modified. In some embodiments, the anti-CTLA-4 antibody molecule is a human IgG1 antibody. In some embodiments, the anti-CTLA-4 antibody is an antibody in the form of a human IgGl antibody that exhibits enhanced binding to one or more activating Fc receptor(s) and / or is genetically engineered for enhanced binding to one or more activating Fc receptor(s). ; therefore, in some embodiments, the anti-CTLA-4 antibody is an Fe-engineered human IgGl antibody. In some embodiments, the anti-CTLA-4 antibody is a murine IgG2a antibody or IVIA / a / ZUZ I / UUZDUU humanized murine. In some embodiments, the anti-CTLA-4 antibody is a murine antibody that is cross-reactive with human CTLA-4. In some embodiments, the anti-CTLA-4 antibody is a monoclonal antibody. In some embodiments, the anti-CTLA-4 antibody is a polyclonal antibody. In some embodiments, the anti-CTLA-4 antibody molecule is an antibody molecule comprising one of three alternative VH-CDR1 sequences, one of three alternative VH-CDR2 sequences, one of two alternative VH-CDR2 sequences, Alternative CDR3, one of the two VL-CDR1 sequences, one of the two VL-CDR2 sequences and / or one of the two alternative VLCDR3 sequences presented in Table 1 below. In some embodiments, the anti-CTLA-4 antibody molecule is selected from the group consisting of antibody molecules comprising 1-6 of the CDRs selected from the group consisting of SEQ ID NO: 3, 6, 8, 10, 12 and 14. In some embodiments, the anti-CTLA-4 antibody molecule is selected from the group consisting of antibody molecules comprising the CDRs having SEQ ID NO: 3, 6, 8, 10, 12, and 14. In some embodiments, the anti-CTLA-4 antibody molecule is selected from the group consisting of antibody molecules comprising 1-6 of the CDRs, VH-CDR1, VHCDR2, VH-CDR3, VL-CDR1, and VL-CDR3, wherein VH-CDR1, if present, is selected from the group consisting of SEQ ID NO: 15, 22, 29, and 35; wherein VH-CDR2, if present, is selected from the group consisting of SEQ ID NO: 16, 23, 30, and 36; wherein VH-CDR3, if present, is selected from the group consisting of SEQ ID NO: 17, 24, 31, and 37; wherein VL-CDR1, if present, is selected from the group consisting of SEQ ID NO: 10 and 38. wherein VL-CDR2, if present, is selected from the group consisting of SEQ ID NO: 18, 25, 32, and 39; wherein VL-CDR3, if present, is selected from the group consisting of SEQ ID NO: 19, 26 and 40. In some embodiments, the anti-CTLA-4 antibody molecule is an antibody molecule selected from the group consisting of antibody molecules comprising 6 CDRs selected from the group consisting of: SEQ ID NO: 15, 16, 17, 10, 18 and 19; SEQ ID NO: 22, 23, 24, 10, 25 and 26; SEQ ID NO: 29, 30, 31, 10, 32 and 26; and SEQ ID NO: 35, 36, 37, 38, 39 and 40. In some embodiments, the anti-CTLA-4 antibody molecule is an antibody molecule comprising the 6 CDRs having SEQ ID NO: 15, 16, 17, 10, 18, and 19. In some embodiments, the anti-CTLA-4 antibody molecule is an antibody molecule comprising the 6 CDRs having SEQ ID NO: 22, 23, 24, 10, 25, and 26. In some embodiments, the anti-CTLA-4 antibody molecule is an antibody molecule selected from the group consisting of antibody molecules that comprise a VH selected from the group consisting of SEQ ID NO: 20, 27, 33, and 41. In some embodiments, the anti-CTLA-4 antibody molecule is an antibody molecule selected from the group consisting of antibody molecules comprising a VL selected from the group consisting of SEQ ID NO: 21, 28, 34, and 42. In some embodiments, the anti-CTLA-4 antibody molecule is an antibody molecule selected from the group consisting of antibody molecules comprising a VH and a VL selected from the group consisting of: SEQ ID NO: 20-21, 27 -28, 33-34 and 41-42. In some embodiments, the anti-CTLA-4 antibody molecule comprises a VH having the sequence SEQ ID NO: 20 and a VL having the sequence SEQ ID NO: 21. In some embodiments, the anti-CTLA-4 antibody molecule comprises a VH / UUZDUU having the sequence SEQ ID NO: TJ and a VL having the sequence SEQ ID NO: 28. IVIA / a / ZUZ I / UUZDUU Table 1: General CDR sequences of antibodies described in the present description Relevant part of the antibody Sequence Explanation of unidentified amino acid residues SEQ ID NO: VH-CDR1 alternative 1 FX1X2YX3MX4WX5R QAPG X1 = S 0 K; X2 = D, S 0 A; X3 = Y, S 0 A; X4 = S 0 N; and X5 = V 0 I 1 VH-CDR1 alternative 2 FSX1YX2MX3WVRQ OFF X1 = D 0 S; X2 = Y, S 0 A; and X3 = Y 0 N 2 VH-CDR1 alternative 3 FSX1YX2MX3WVRQ OFF X1 = D 0 Y; X2 = Y 0 S; and X3 = SoN 3 VH-CDR2, alternative 1 SX1ISX2X3X4X5X6X7 XsXgADSVKGR X1 = G 0 A; X2 = W, G 0 N; X3 = S 0 T; X4 = S 0 G; X5 = R 0 G; X6 = D, S 0 Y; X7 = K, T 0 I; Xs = G, Y, H 0 D; Xg = Y 0 F 4 VH-CDR2, alternative 2 SX1ISX2X3X4X5X6X7 XeYADSVKGR X1 = G 0 A; X2 = W, G 0 N; X3 = S 0 T; X4 = S 0 G; X5 = RoG;Xe=D, S 0 Y; X7 = K, To l;X8 = G, Y, Ho D 5 VH-CDR2, alternative 3 SX1ISX2SX3X4X5X6X7 YADSVKGR Xi = GoA;X2 = WoG;Xs = S 0 G; X4 = R 0 G; Xs = D oS;X6 = KoT;X7 = G oY, Ho D 6 VH-CDR3 alternative 1 X1X2X3X4X5X6X7X8X9X10 X11X12X13X14X15 X1 = T 0 A; X2 = T 0 R; X3 = D, I L ; X4 = L, R, SoG; Xs = A, V, S 0 Y; X6 = R, E, G 0 S; X7 = Y, M, L 0 G; Xs = N, H, Y 0 none; Xg = Q, D 0 none; X10 = W, A, D 0 none; Xn = L, F, R 0 none; X12 = A, D, G 0 none; Xi3 = D, I, M 0 none; X14 = D 0 none; and X15 = V 0 none 7 VH-CDR3 alternative 2 Xi X2DX3X4X5X6X7X8X9X10 X11X12X13 X1 = T 0 A; X2 = T 0 R; X3 = L 0 R; X4 = A 0V; Xs = R 0 E; Xe = Y 0 M; X? = N 0 none; Xs = Q 0 none; X9 = W 0 none; X10 = L, 0 none; Xn = A, 0 none; X12 = D 0 none 8 VL-CDR1 alternative 1 CX1GSSSNIGX2 X3YX4X5X6 X1 = T 0 S; X2 = A 0 S; X3 = G 0 N; X4 = D0V; X5 = V 0 Y; X6 = H 0 none 9 VL-CDR1 alternative 2 CTGSSSNIGAGYDVH 10 VL-CDR2 alternative 1 X1NX2X3RPS X1 = G, RoD;X2= D, N oS;yX3 = N,QoK 11 VL-CDR2 alternative 2 X1NX2X3RPS Xi = G 0 R; X2 = D 0 N; and X3 = N 0 Q 12 VL-CDR3 alternative 1 CX1X2X3DX4SLX5G X6VX7 X1 = A 0 Q; X2 = V, A 0 S; X3 = W 0 Y; X4 = D 0 S; Xs = N 0 S; Xs = V, W 0 P; and X7 = V 0 none 13 VL-CDR3 alternative 2 CAX1WDDSLNG X2V X1 = V 0 A; and X2 = V 0 W 14 Table 2: Specific anti-CTLA-4 antibody molecules; CDR sequences are marked in bold in full VH and VL sequences IVIA / a / ZUZ I / UUZDUU Antibody Clone Region Sequence SEQ ID NO: 4-E03 VH-CDR1 FSDYYMSWVR QAPG 15 VH-CDR2 SGISWSSRDK GYADSVKGR 16 VH-CDR3 TTDLARY 17 VL-CDR1 CTGSSSNIGAGYDVH 10 VL-CDR2 GNDNRPS 18 VL-CDR3 CAV WDDSLNG VV 19 VH EVQLLESGGG LVQPGGSLRL SCAASGFTFS DYYMSWVRQA PGKGLEWVSG ISWSSRDKGY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCTTDL ARYWGQGTLV TVSS 20 VL QSVLTQPPSA SGTPGQRVTI SCTGSSSNIG AGYDVHWYQQ LPGTAPKLLI YGNDNRPSGV PDRFSGSKSG TSASLAISGL RSEDEADYYC AVWDDSLNGV VFGGGTKLTV LG 21 human immunoglobulin G1 (lgG1) light chain (LC) MGWSCIILFLVATATGVHSQSVLTQPPSASGTPGQRVTISCT GSSSNIGAGYDVHWYQQLPGTAPKLLIYGNDNRPSGVPDRF SGSKSGTSASLAISGLRSEDEADYY CAVWDDSLNGVVFGGG TKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQ WKSHRSYSCQVTHEGSTVEKTVAPTECS 53 lgG1 human heavy chain (HC) MGWSCIILFLVATATGVHSEVQLLESGGGLVQPGGSLR LSCA ASGFTFSDYYMSWVRQAPGKGLEWVSGISWSSRDKGYADS VKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCTTDLARYWG QGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYF PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSS LGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEL LGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGK EYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTK NQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDG SFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSL SPGK 54 5-B07 VH-CDR1 FSSYSMNWVRQ APG 22 VH-CDR2 SAISGSGGST YYADSVKGR 23 VH-CDR3 ARDRVEMNQW LAD 24 VL-CDR1 CTGSSSNIGA GYDVH 10 VL-CDR2 RNNQRPS 25 VL-CDR3 CAAWDDSLNG WV 26 VH EVQLLESGGG LVQPGGSLRL SCAASGFTFS SYSMNWVR QA PGKGLEWVSA ISGSGGSTYY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARDR VEMNQWLADW GQGTLVTVSS 27 VL QSVLTQPPSA SGTPGQRVTI SCTGSSSNIG AGYDVHWYQQ LPGTAPKLLI YRNNQRPSGV PDRFSGSKSG TSASLAISGL RSEDEADYYC AAWDDSLNGW VFGGGTKLTV LG 28 murine immunoglobulin (lgG2a) light chain (LC) MGWSCIILFLVATATGVHSQSVLTQPPSASGTPGQRVTISCT GSSSNIGAGYDVHWYQQLPGTAPKLLIYRNNQRPSGVPDR FSGSKS GTSASLAISGLRSEDEADYYCAAWDDSLNGWVFG GGTKLTVLGQPKSSPSVTLFPPSSEELETNKATLVCTITDFY PGVVTVDWKVDGTPVTQGMETTQPSKQSNNKYMASSYLTL TARAWERHSSYSCQVTHEGHTVEKSLSRADCS 62 Murine immunoglobulin (lgG2a) heavy chain (HC) MGWSCIILFLVATATGVHSEVQLLESGGGLVQPGGSLRLSC AASGFTFSSYSMNWVRQAPGKGLEWVSAISGSGGSTYYA DSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRVE MNQWLADWGQGTLVTVSSAKTTAPSVYPLAPVCGDTTGSS VTLGCLVKGY FPEPVTLTWNSGSLSSGVHTFPAVLQSDLYT LSSSVTVTSSTWPSQSITCNVAHPASSTKVDKKIEPRGPTIK PCPPCKCPAPNLLGGPSVFIFPPKIKDVLMISLSSPIVTCVVVD VSEDDPDVQISWFVNNVEVHTAQTQTHREDYNSTLRVVSA LPIQHQDWMSGKEFKCKVNNKDLP APIERTISKPKGSVRAP QVYVLPPPEEEMTKKQVTLTCMVTDFMPEDIYVEWTNNGK TELNYKNTEPVLDSDGSYFMYSKLRVEKKNWVERNSYSCS VVHEGLHNHHTTKSFSRTPGK 63 2 -C06 VH-CDR1 FSSYAMSWVRQ APG 29 VH-CDR2 SGISGSGGYI HYADSVKGR 30 VH-CDR3 ATYSSGLHDA FDI 31 VL-CDR1 CTGSSSNIGA GYDVH 10 VL-CDR2 DNNKRPS 32 VL-CDR3 CAAWDDSLNG WV 26 VH EVQLLESGGG LVQPGGSLRL SCAASGFTFS SYAMSWVRQA PGKGLEWVSG ISGSGGYIHY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCATYS SGLHDAFDIW GQGTLVTVSS 33 VL EVQLLESGGG LVQPGGSLRL SCAASGFTFS SY AMSWVRQA PGKGLEWVSG ISGSGGYIHY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCATYS SGLHDAFDIW GQGTLVTVSS 34 2-F09 VH-CDR1 FKAYSMSWIR QAPG 35 VH-CDR2 SGISNTGGST DFADSVKGR 36 VH-CDR 3 ARLGYSGYDD RGMDV 37 VL -CDR1 CSGSSSNIGS NYVY 38 VL-CDR2 GNSNRPS 39 VL-CDR3 CQSYDSSLSG PVV 40 VH EVQLLESGGG LVQPGGSLRL SCAASGFTFK AYSMSWIRQA PGKGLEWVSG ISNTGGSTDF ADSVKGRFTI SRDNSKNTLY LQMNSLRAED T AMYYCARLG YSGYDDRGMD VWGQGTLVTV SS 41 VL QSVLTQPPSA SGTPGQRVTI SCSGSSSNIG SNYVYWYQQL PGTAPKLLIY GNSNRPSGVP DRFSGSKSGT SASLAISGLR SEDEADYYCQ SYDSSLSGPV VFGGGTKLTV LG 42 / UUZDUU In some embodiments, the anti-CTLA-4 antibody molecules described herein may also comprise one or both of the constant regions presented in Table 3 below. Table 3: Sequences of constant regions of antibodies described in the present description Region Sequence SEQ ID NO: CH ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKK VEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVV VDVSHEDPEVKFNW 43 CL QPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVK AGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEK TVAPTECS 44 In some embodiments, the anti-CTLA-4 antibody molecule is a molecule encoded by one of the nucleotide sequences presented in Table 4 below. Table 4: Specific Nucleotide Sequences Encoding Anti-CTLA-4 Antibody Molecules Clone Encoding Sequence SEQ ID NO: Heavy chain 4-E03 γ1 4-E03 VH GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCC TGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCA CCTTCAGTGACTACTACATGAGCTGGGTCCGCCAGGCTCCGGGGAAGGGG CTGGAGTGGGTCTCAGGCATTAGTTGGAGTAG TCGTGACAAAGGCTATGCGGACTCTGTGAAGGGCCGTTTCAC CATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAAT GAACAGCCTGAGAGCCGAGGACACTGCCGTGTATTACTGTA CCACAGATCTCGCTAGGTACTGGGGCCAGGGTACACTGGTC ACCGT GAGCTCAGCCTCCACCAAGGGCCCATCGGTCTTCCC CCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGG CCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTG ACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCA CACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCT CA GCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCC AGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCA AGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTC ACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGA CCGTCAGTCTTCCTCTTCCCCCAAAACCCAAGGACACCC TC ATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGA CGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACG TGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGG GAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCT CACCGTCCTGCACCAGGACTGGCTGAATG GCAAGGAGTACA AGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAG AAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACA GGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGA ACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCA GCGACATCGCCGTGGAGTGGGAGAGCA ATGGGCAGCCGGA GAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACG GCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCA GGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCAT GAGGCTCTGCACAACCACTACACGCAGAAGAGCCTTCTCCCT GTCTCCGGGTAAATGA 45 Light chain 4E03 λ 4-E03 VL CAGTCTGTGCTGACTCAGCCACCCTCAGCGTCTGGGACCCC CGGGCAGAGGGTCACCATCTCCTGCACTGGGAGCAGCTCCA ACATCGGGGCAGGTTATGATGTACACTGGTATCAGCAGCTCC CAGGAACGGCCCCCAAACTCCTCATCTATGGTAATGATAACC GG CCCTCAGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCT GGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGA GGATGAGGCTGATTATTACTGTGCAGTATGGGATGACAGCCT GAATGGTGTGGTATTCGGCGGAGGAACCAAGCTGACGGTCC TAGGTCAGCCCAAGGCTGCCCCCTCGGTC ACTCTGTTCCCG CCTCCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGT GTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGG CCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGT GGAAGTCC CACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCAC CGTGGAGAAGACAGTGGCCCCTACAGAATGTTCATGA 46 Heavy chain 5-B07 γ1 5-B07 VH GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCC TGGGGGGTCCCTGAGACTCTCCTGT GCAGCCTCTGGATTCA CCTTCAGTAGCTATAGCATGAACTGGGTCCGCCAGGCTCCAG GGAAGGGGCTGGAGTGGGTCTCAGCTATTAGTGGTAGTGGT GGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCAC CATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAAT GAACAGCCTGAGAGCC GAGGACACTGCCGTGTATTACTGTG CGAGAGATCGGGTAGAGATGAACCAGTGGCTGGCCGACTGGGGCCAGGGTACACTGGTCACCGTGAGCTCAGCCTCCACCAA GGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGA CTACT TCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCG CCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAG TCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCC CTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAA TCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGC CCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAG CACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCC CAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAG GTCA 47 CATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTC AAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGC CAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACC GTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTG AATGGCAAGGAGTACAAGTGCAAGGTCTCCA ACAAAGCCCTC CCAGCCCCCATCGAGAAAACCATTCCAAAGCCAAAGGGCA GCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGG ATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTC AAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAG CAATGGGCAGCCGGAGAACAACTACAAGACC ACGCCTCCCG TGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCA CCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCA TGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAG AAGAGCCTTCTCCCTGTCTCCGGGTAAATGA Light Chain 5B07 λ 5-B07 VL CAGTCT GTGCTGACTCAGCCACCCTCAGCGTCTGGGACCCC CGGGCAGAGGGTCACCATCTCCTGCACTGGGAGCAGCTCCA ACATCGGGGCAGGTTATGATGTACACTGGTATCAGCAGCTCC CAGGAACGGCCCCCAAACTCCTCATCTATAGGAATAATCAGC GGCCCTCAGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCT GGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGA GGATGAGGCTGATTATTACTGTGCAGCATGGGATGACAGCCT GAATGGTTGGGTGTTCGGCGGAGGAACCAAGCTGACGGTCC TAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCTCCTCTGAGGAGCTTCAAGCCAACAAGG CCACACTGGT GTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGG CCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCC CACAGAAGCTACAGCTGCCAGGTCACGCATGAAG GGAGCAC CGTGGAGAAGACAGTGGCCCCTACAGAATGTTCATGA 48 Heavy chain 2-C06 γ1 2-C06 VH GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCC TGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCA CCTTTAGCAGCTATGCCATGAGCTGGGTCCGCCAGGCTCCA GGGAAGGGGCTGGAGTGGGTCTCAGGTATTAGTGGCAGCG GTGGGTACATACACTATGCAGACTCCGTGAAGGGCCGGTTC ACCATCTCCAGAGACAATTCCAAGAACACG ST 49 GTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACTGC CGTGT ATTACTGTG CG ACCTATAG C AGTGG CCTG C ​​ATG AT GC1 1 1 1GATATCTGGGGCCAGGGTACACTGGTCACCGTGA GCTCAGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGG CACCCTCCTCCAAGAGCACCTCT GGGGGCACAGCGGCCC TGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGA CGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTG CACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACT CCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGG GCACCCAGACCTACATCTGCAACG TGAATCACAAGCCCAG CAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGT G AC AAAACTC AC AC ATG CCCACCGTG CCC AG CACCTG AA CTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAAC CCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCA CATGCGTGGTGGTGGACGTGAG CCACGAAGACCCTGAGG TCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATA ATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCA CGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGG ACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCA ACAAAGCCCTCCCAGCCCCCATCGAGAAA ACCATTCCCAA AGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCT GCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAG CCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATC GCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAA CTACAAGACCACGCCTCCCGTGCTGGACTCCGACG GCTC CTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGG TGGGAGCAGGGGAACGTGTTCTCATGCTCCGTGATGCAT GAGGCTCTGCACAACCACTACACGCAGAAGAGCCTTCCC CTGTCTCCGGGTAAATGA Light Chain 2C06 λ 2-C06 VL CAGTCTGTGCTGACTCAGCCACCCTCAGCGTCTGGGACCCC CGGGCAGAGGGTCACCATCTCCTGCACTGGGAGCAGCTCCA ACATCGGGGCAGGTTATGATGTACACTGGTATCAGCAGCTCC CAGGAACGGCCCCCAAACTCCTCAT CTATGACAATAATAAGC GACCCTCAGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCT GGCACCTCAGCTCTCCCTGGCCATCAGTGGGCTCCGGTCCGA GGATGAGGCTGATTATTACTGTGCAGCATGGGATGACAGCCT GAATGGTTGGGTGTTCGGCGGAGGAACCAAGCTGACGGTCC TAGGTCAGCCCAAG GCTGCCCCCT 50 CGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCC AACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCG GGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCG TCAAGGCGGGAGTGGAGACCACCACCCTCCAAACAAAGC AACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACG CC TGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCA CGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTAC AGAATGTTCATGA Heavy Chain 2-F09 and 1 2-F09 VH GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGC CTGGGGGGTCCCTGAGACTC TCCTGTGCAGCCTCTGGATTC ACCTTCAAAGCCTATAGCATGAGCTGGATCCGCCAGGCTCC AGGGAAGGGGCTGGAGTGGGTCTCAGGTATCAGTAACACG GGAGGTAGCACAGACTTCGCAGACTCCGTGAAGGGCCGGTT CACCATTCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCA AATGAACAGCCTGAGAG CCGAGGACACTGCCATGTATTACT GTGCGAGATTGGGATATAGTGGCTACGACGACCGTGGTATG GACGTCTGGGGCCAAGGTACACTGGTCACCGTGAGCTCAGC CTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCT CCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCT GGTC AAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGA ACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGG TGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATC TGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAA GAAAG TTGAGCCCAAATCTTGTGACAAAACTCACACATGCCC ACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCT TCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCC GGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCA CGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCG T GGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCA GTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCC TGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCAT CTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACA CC CTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTC AGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACAT CGCCGTGGAGTGGGAGAGCAATGGGCAGCCGG 51 AGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCG ACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAA GAGCAGGTGGCAGCAGGGGGAACGTCTTCTCATGCTCCGT GATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAG CCTTCCCCTGTCTCCGGGTAAATGA Light chain 2F09 λ 2-F09 VL CAGTCTGTGCTGACTCAGCCACCCTCAGCGTCTGGGACC CCCGGGCAGAGGGTCACCATCTCTTGTTCTGGAAGCAGC TCCAACATCGGAAGTAATTATGTGTACTGGTATCAGCAGC TCCCAGGAACGGCCCCCAAACTCCTCATCTATGGTAACAG CAATCGGCCCTCAGGGGTCCCTGACCGATTCTCT GGCTC CAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCT CCGGTCCGAGGATGAGGCTGATTATTACTGCCAGTCCTAT GACAGCAGCCTGAGTGGTCCTGTGGTATTCGGCGGAGGA ACCAAGCTGACGGTCCTAGGTCAGCCCAAGGCTGCCCCC TCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAG CCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTA CCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCA GCCCCGTCAAGGCGGGAGTGGAGACCACCACCACCCTCCA AACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGA GCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACA GCTGCCAGG TCACGCATGAAGGGAGCACCGTGGAGAAGA CAGTGGCCCCTACAGAATGTTCATGA 52 IVIA / a / ZUZ I / UUZDUU In some embodiments, it is advantageous if the antibody molecule binds to both human CTLA-4 (hCTLA-4) and cynomolgus monkey CTLA-4 (cmCTLA-4 or cyno CTLA-4). Cross-reactivity with CTLA-4 expressed in cells of the cynomolgus monkey, also called cynomolgus monkey or Macaca fascicularis, may be advantageous, as this allows the antibody molecule to be tested in the monkey without having to use a surrogate antibody, which targets in particular in tolerability. In some embodiments, it is advantageous if the antibody molecule binds to both human CTLA-4 (hCTLA-4) and murine CTLA-4 (mCTLA-4). This may be advantageous as it allows the antibody molecule to be tested in mice, with particular emphasis on effect and pharmacodynamics, without having to use a surrogate antibody. In some embodiments, the antibody molecule binds to all three of hCTLA-4, cmCTLA-4, and mCTLA-4. In some embodiments, it is necessary to use a surrogate antibody to test the functional activity of an antibody molecule in relevant in vivo mouse models. To ensure comparability between the effect of the antibody molecule in humans and the in vivo results for the surrogate antibody in mice, it is essential to select a functionally equivalent surrogate antibody that has the same in vitro characteristics as the human antibody molecule. In some embodiments, the antibody molecule does not bind to human CD28. The medically skilled person will know that drugs can be modified with different additives, for example, to change the rate at which the drug is absorbed by the body; and it can be modified in different ways, for example, to allow a particular route of administration to the body. Accordingly, we include that the antibody molecules, nucleotide sequences, plasmids, viruses and / or cells described in the present description can be combined with a pharmaceutically acceptable excipient, carrier, diluent, vehicle and / or adjuvant in a pharmaceutical composition. In this context, the term pharmaceutical composition can be used interchangeably with the terms pharmaceutical preparation, pharmaceutical formulation, therapeutic composition, therapeutic preparation, therapeutic formulation, and therapeutic entity. The pharmaceutical compositions described in the present description can comprise, or in some modalities consist of, antibody molecules, nucleotide sequences, plasmids, viruses or cells. The pharmaceutical compositions described herein may in some embodiments consist of or comprise plasmids comprising nucleotide sequences encoding the antibody molecules described above or comprising the nucleotide sequences described above. In some embodiments, pharmaceutical compositions may comprise nucleotide sequences encoding parts of or an entire antibody molecule described herein integrated into a viral cell or genome or viriome. The pharmaceutical composition may then comprise a cell or virus as a delivery vehicle for an antibody of the invention (or a delivery vehicle for a nucleotide sequence encoding an antibody of the invention). For example, in one embodiment, the virus may be in the form of a therapeutic oncolytic virus comprising nucleotide sequences encoding at least one of the antibody molecules described herein. In some embodiments, said oncolytic virus comprises nucleotide sequences encoding a full length human IgG antibody. In some embodiments, said oncolytic virus comprises nucleotide sequences encoding an scFv, Fab, or F(ab')2 antibody molecule. IVIA / a / ZUZ I / UUZDUU As described in the appended claims, in one embodiment the invention relates to a virus comprising a nucleotide sequence of the invention or a plasmid of the invention. Preferably, the virus is an oncolytic virus, such as a therapeutic oncolytic virus. As used herein, the term oncolytic refers to the ability of a virus to selectively replicate in dividing cells (for example, a proliferative cell such as a cancer cell) with the goal of slowing growth and / or using said dividing cell, either in vitro or in vivo, while showing no or minimal replication in non-dividing (eg normal or healthy) cells. Replication (or any form of replication such as replicating and replicating, etc.) means the duplication of a virus that can occur at the nucleic acid level or, preferably, at the infectious viral particle level. Said oncolytic virus can be obtained from any currently identified member of the virus. It may be a native virus that is naturally oncolytic, or it may be genetically modified by modifying one or more viral genes to increase tumor selectivity and / or replication preferentially in dividing cells, such as those involved in DNA replication, metabolism of nucleic acids, host tropism, surface adherence, virulence, lysis and dissemination (see, for example, Wong et al., 2010, Viruses 2: 78-106). Placing one or more viral gene(s) under the control of tissue-specific regulatory events or elements (eg, promoter) may also be contemplated. Illustrative oncolytic viruses include, without limitation, reovirus, Seneca Valley (SW) virus, vesicular stomatitis virus (VSV), Newcastle disease virus (NDV), herpes simplex virus (HSV), morbillivirus, adenovirus, poxvirus , retrovirus, measles virus, spumavirus, alpha virus, lentivirus, influenza virus, Sinbis virus, myxoma virus, rhabdovirus, picornavirus, coxsackievirus, parvovirus or the like. Such viruses are known to experts in medicine and virology. In some embodiments, said oncolytic virus is derived from a herpes virus. The Herpesviridae are a large family of DNA viruses that share a common structure and are composed of relatively large double-stranded, linear DNA genomes encoding 100-200 genes encapsulated within an icosahedral capsid that is wrapped in a lipid bilayer membrane. . Although the oncolytic herpes virus can be derived from different types of HSV, HSV1 and HSV2 are particularly preferred. The herpes virus can be genetically modified to restrict viral replication in tumors or reduce its cytotoxicity in non-dividing cells. For example, any viral gene involved in nucleic acid metabolism can be inactivated, such as thymidine kinase (Martuza et al., 1991, Science 252: 854-6), ribonucleotide reductase (RR) (Mineta et al., 1994 , Cancer Res. 54: 3363-66), or uracil-N-glycosylase (Pyles et al., 1994, 1 Viral. 68: 4963-72). Another aspect involves viral mutants with defects in the function of genes encoding virulence factors such as the gene ICP34.5 (Chambers et al., 1995, Proc. Nati. Acad. Sel. USA 92: 1411-5). Representative examples of Neolytic herpesviruses include NV1020 (eg, Geevarghese et al., 2010, Hum. Gene Ther. 21(9): 1119-28) and T-VEC (Harrington et al., 2015, Expert Rev. Anticancer Ther. 15(12): 1389-1403). In some embodiments, said oncolytic virus is derived from an adenovirus. Methods are available in the art for genetically modifying oncolytic adenoviruses. An advantageous strategy includes replacement of viral promoters with tumor-selective promoters or modifications of the adenoviral El gene product(s) to inactivate its function(s) of binding to p53 or retinoblastoma protein (Rb) that are altered in tumor cells. In the natural setting, the adenovirus ElB55kDa gene cooperates with another adenoviral product to inactivate p53 (p53 is frequently deregulated in cancer cells), thereby preventing apoptosis. Representative examples of oncolytic adenoviruses include ONYX-015 (for example, Khuri et al., 2000, Nat. Med 6(8): 879-85) and H101 also called Oncorine (Xia et al., 2004, Ai Zheng 23(12): 1666-70). In some embodiments, said oncolytic virus is a poxvirus. As used herein, the term "poxvirus" refers to a virus belonging to the family Poxviridae, with a specific preference for a poxvirus belonging to the subfamily Chordopoxviridae and more preferably to the genus Orthopoxvirus. Vaccinia virus, smallpox virus, canarypox virus, ectromelia virus, myxoma virus are particularly suitable in the context of the invention. The genomic sequences of such poxviruses are available in the art and in specialized databases (for example, Genbank with the accession number NC_006998, NC_003663 or AF482758.2, NC_005309, NC_004105, NC_001132 respectively). In specific and preferred embodiments, said oncolytic poxvirus is an oncolytic vaccinia virus. Vaccinia viruses are members of the poxvirus family characterized by a 200 kb double-stranded DNA genome that encodes numerous viral enzymes and factors that allow the virus to replicate independently of the host cell machinery. Most vaccinia virus particles are intracellular (IMV for intracellular mature virion) with a single lipid envelope and remain in the cytosol of infected cells until lysis. The other infectious form is a double-enveloped particle (EEV for extracellular enveloped virion) that buds from the infected cell without lysing it. Although it can be derived from any vaccinia virus strain, the Elstree, Wyeth, Copenhagen, Lister and Western Reserve strains are particularly preferred. The gene nomenclature used in the present description is that of the Copenhagen vaccinia strain, unless otherwise indicated. However, the correspondence between Copenhagen and other vaccinia strains is generally available in the literature. Preferably, said oncolytic vaccinia virus is modified by the / UUZDUU alteration of one or more viral genes. Said modification(s) lead(s) preferably to the absence of synthesis or the synthesis of a defective viral protein incapable of ensuring the activity of the protein produced under normal conditions by the unmodified gene. Illustrative modifications with the aim of altering viral genes involved in DNA metabolism, host virulence, IFN pathway are described in the literature (eg, Guse et al., 2011, Expert Opinion Biol. Ther. 11(5 ): 595-608) and the like. Modifications to alter a viral locus encompass the deletion, mutation, and / or substitution of one or more nucleotides (contiguous or not) within the viral gene or its regulatory elements. Modifications can be made by a number of routes known to those skilled in the art using standard recombinant techniques. More preferably, said oncolytic vaccinia viruses are modified by altering the gene encoding thymidine kinase (J2R locus). The enzyme thymidine kinase (TK) is involved in the synthesis of deoxyribonucleotides. TK is required for viral replication in normal cells since these cells generally have a low concentration of nucleotides, whereas it is dispensable in dividing cells that contain a high concentration of nucleotides. Alternatively, or in combination, said oncolytic vaccinia virus is modified by altering at least one or both genes encoding ribonucleotide reductase (RR). In the natural context, this enzyme catalyzes the reduction of ribonudeotides to deoxyribonucleotides, which represents a crucial step in DNA biosynthesis. The viral enzyme is similar in subunit structure to the mammalian enzyme, and is composed of two heterologous subunits, designated R1 and R2 encoded respectively by the I4L and F4L locus. In the context of the invention, either the I4L gene (encoding the R1 major subunit) or the F4L gene (encoding the R2 minor subunit) or both can be inactivated (eg as described in WO2009 / 065546 and Foloppe et al. others, 2008, Gene Ther., 15: 1361-71). The sequences of the J2R, I4L and F4L genes and their locations in the genome of various poxviruses are available in public databases. In some embodiments, said oncolytic virus comprises nucleotide sequences encoding the amino acid sequence having at least 80% identity to a sequence set forth in Table 2 above. In some embodiments, said oncolytic virus comprises an amino acid sequence having at least 85% identity to a sequence set forth in Table 2 above. In some embodiments, said oncolytic virus comprises an amino acid sequence having at least 90% identity to a sequence set forth in Table 2 above. In some embodiments, said oncolytic virus comprises an amino acid sequence having at least 95% identity to a sequence set forth in Table 2 above. In some embodiments, said oncolytic virus comprises nucleotide sequences encoding SEQ ID NO: 20 and ID NO: 21. In some embodiments, said oncolytic virus comprises nucleotide sequences encoding SEQ ID NO: 27 and ID NO: 28. In some embodiments, said oncolytic virus comprises nucleotide sequences encoding SEQ ID NO: 33 and ID NO: 34. In some embodiments, said oncolytic virus comprises nucleotide sequences encoding SEQ ID NO: 41 and ID NO: 42. In some embodiments, said oncolytic virus comprises nucleotide sequences having at least 80% identity to a sequence set forth in Table 4 above. In some embodiments, said oncolytic virus comprises nucleotide sequences having at least 85% identity to a sequence set forth in Table 4 above. In some embodiments, said oncolytic virus comprises nucleotide sequences having at least 90% identity to a sequence set forth in Table 4 above. In some embodiments, said oncolytic virus comprises nucleotide sequences having at least 95% identity to a sequence set forth in Table 4 above. In some embodiments, said oncolytic virus comprises SEQ ID NO: 45 and 46. In some embodiments, said oncolytic virus comprises SEQ ID NO: 47 and 48. In some embodiments, said oncolytic virus comprises SEQ ID NO: 49 and 50 In some embodiments, said oncolytic virus comprises SEQ ID NO: 51 and 52. Some oncolytic viruses have the capacity to harbor DNA inserts large enough to accommodate integration of full-length human antibody sequences. Attenuated vaccinia viruses and herpes simplex viruses are examples of therapeutic oncolytic viruses whose genome is large enough to allow integration of full-length IgG antibody sequences (Chan, W. M. et al. 2014 Annu Rev Virol 1(1): 119 -141; Bommareddy, P.K et al. 2018 Nat Rev Immunol 18(8): 498-513). Full-length IgG antibodies have been successfully integrated into oncolytic vaccinia virus, resulting in the expression and extracellular release (production) of full-length IgG antibodies upon infection of virus-susceptible host cells, e.g., cells cancerous (Kleinpeter, P., et al. 2016, Oncoimmunology 5 (10): el220467). Adenoviruses can also be genetically engineered to encode full-length IgG antibodies that are functionally produced and secreted upon cellular infection (Marino, N., et al. 2017 J Clin Invest 123(6): 2447-2463). In a preferred embodiment, said oncolytic virus is a poxvirus (for example, a vaccinia virus) defective for TK activity (resulting from disruption of the J2R locus) or defective for both TK and RR activities (resulting from disruption of both the J2R locus). J2R as from at least one of the I4L and / or F4L loci encoding RR) and comprising (a) nucleotide sequences encoding SEQ ID NO: 20 and ID NO: 21 or (b) nucleotide sequences encoding / UUZDUU encode SEQ ID NO: 27 and ID NO: 28 or (c) nucleotide sequences that encode SEQ ID NO: 33 and ID NO: 34 or (d) nucleotide sequences that encode SEQ ID NO: 41 and ID NO :42. Where appropriate, it may be advantageous that the nucleotide sequence(s) inserted into the oncolytic virus described herein include(s) additional regulatory elements to facilitate expression, trafficking, and biological activity. For example, a signal peptide can be included to facilitate secretion out of the producer cell (eg, an infected cell). The signal peptide is typically inserted at the N-terminus of the encoded polypeptide immediately after the Met primer. The choice of signal peptides is wide and accessible to those skilled in the art. For example, signal peptides originating from another immunoglobin (for example, an IgG heavy chain) can be used in the context of the invention to allow secretion of the anti-CTLA4 antibody described in the present description outside of the producer cell. For illustrative purposes, reference can be made to SEQ ID NO: 53 and SEQ ID NO: 54 comprising the light chain and the heavy chain of the 4-E03 antibody described herein equipped with IgG-originating peptide signals. A particularly preferred oncolytic virus is a vaccinia virus (for example, the Copenhagen strain) defective for both TK and RR activities (resulting from disruption of the J2R locus and I4L locus) and comprising nucleotide sequences encoding SEQ ID NO : 20 and SEQ ID NO: 21 or SEQ ID NO: 53 and SEQ ID NO: 54. In some embodiments, said oncolytic virus may additionally comprise nucleotide sequence(s) of therapeutic interest, such as the nucleotide sequence(s) encoding the immunomodulatory polypeptide(s) (i.e., a polypeptide involved in stimulating an immune response). immunity either directly or indirectly). Representative examples of suitable immunomodulatory polypeptides include, without limitation, cytokines and chemokines with a specific preference for granulocyte-macrophage colony-stimulating factor (GM-CSF) and particularly human, non-human primate or murine GM-CSF. Additional nucleotide sequence can be readily obtained by standard molecular biology techniques (eg, PCR amplification, cDNA cloning, chemical synthesis) using sequence data accessible in the art and the information provided herein. A particularly preferred oncolytic virus is a vaccinia virus (eg, Copenhagen strain) defective for both TK and RR activities (resulting from disruption of both the J2R locus and the I4L locus) and comprising nucleotide sequences encoding SEQ ID NO : 20 and ID NO: 21 or SEQ ID NO: 53 and SEQ ID NO: 54 and a nucleotide sequence encoding a GMCSF, with a specific preference for a human GM-CSF (eg, having SEQ ID NO: 55 or SEQ ID NO: 56) or a murine GM-CSF (eg, having SEQ ID NO: 57 or SEQ ID NO: 58). In addition, the nucleotide sequences that are inserted into said / UUZDUU oncolytic virus can be optimized to provide a high level of expression in a particular host cell or subject by modifying one or more codons. In addition to codon usage optimization, various modifications can also be contemplated to avoid clustering of rare, non-optimal codons present in concentrated areas and / or to delete or modify negative sequence elements that are expected to negatively influence codon levels. expression. Such negative sequence elements include without limitation regions having very high (>80%) or very low (<30%) GC content; AT-rich or GC-rich stretches of sequence; unstable direct or inverted repeat sequences; secondary structures R A; and / or internal cryptic regulatory elements such as internal TATA boxes, chi sites, ribosome entry sites, and / or splice donor / acceptor sites. In some embodiments, the nucleotide sequence(s) are placed under the control of appropriate regulatory elements for their proper expression in a host cell or subject. As used herein, the term "regulatory elements" refers to any element that enables, contributes, or modulates the expression of the coding nucleotide sequence(s) in a given host cell or subject, including its replication, duplication , transcription, splicing, translation, stability, and / or transport into or out of the expression cell. Those skilled in the art will appreciate that the choice of regulatory elements may depend on factors such as the nucleotide sequence itself, the virus into which it is inserted, the host cell or subject, the desired level of expression, and so on. The promoter is of special importance. In the context of the invention, this can be constitutive that directs the expression of the nucleotide sequence that it controls in many types of host cells or specific for certain host cells or regulated in response to specific events or exogenous factors (for example, by temperature , nutritional additive, hormone, etc.) or according to the phase of a viral cycle (for example, late or early). Promoters adapted for virus-mediated expression are known in the art. Representative examples of expression by an oncolytic poxvirus include, without limitation, the vaccinia p7.5K, pH5.R, pllK7.5, TK, p28, pll, pB2R, pA35R, K1L, and pSE / L promoters (Erbs et al., 2008, Cancer Gene Ther. 15(1): 18-28; Orubu et al. 2012, PloS One 7: e40167), chimeric early / late promoters and synthetic promoters (Chakrabarti et al., 1997, Biotechniques 23: 1094-7; Hammond et al., 1997, J. Viral Methods 66: 135-8 and Kumar and Boyle, 1990, Virology 179: 151-8). In preferred embodiments, the nucleotide sequences of the light and heavy chains of the antibody described herein are respectively placed under the control of promoters having the same transcriptional strength, and preferably under the control of the same promoter (for example, p7. 5K such as that described in SEQ ID NO: 59 or pH5.R such as that described in SEQ ID NO: 60) to obtain a similar expression level for both chains and therefore optimal assembly of the antibody as a protein heterotetrameric IVIA / a / ZUZ I / UUZDUU (ie, to avoid excess unassociated string). Additional nucleotide sequence (eg, encoding GM-CSF) can be placed under a different promoter (eg, pSE / L such as that described in SEQ ID NO: 61). The insertion of the nucleotide sequence(s) (possibly equipped with appropriate regulatory elements) into the genome of said Neolithic virus is performed by conventional means, either by use of appropriate restriction enzymes or, preferably, by homologous recombination. . The nucleotide sequence(s) can be independently inserted at any location in the viral genome. Various insertion sites can be considered, for example, in a non-essential viral gene, in an intergenic region or in a non-coding part of the genome of said oncolytic virus. The J2R locus and / or the I4L locus are particularly suitable for an oncolytic virus that is a poxvirus (eg, an oncolytic vaccinia virus). Upon insertion of the nucleotide sequence(s) into the viral genome, the viral locus at the insertion site can be at least partially removed. In one embodiment, this deletion or partial deletion may result in suppressed expression of the viral gene product encoded by the fully or partially deleted locus resulting in a virus defective for said viral function. A particularly preferred oncolytic virus is a TK and / or RR defective vaccinia virus comprising the cassette encoding the heavy chain inserted into the J2R locus and the cassette encoding the light chain inserted into the I4L locus. The cassette encoding the additional GM-CSF-encoding nucleotide sequence may be inserted at another location in the virus genome or at the J2R or I4L locus, with preference for insertion at the I4L locus. The present invention also provides a method for generating said oncolytic virus described in the present description, and particularly an oncolytic poxvirus, in a suitable host cell (producer cell). In some embodiments, said method comprises one or more step(s) of homologous recombination between a viral genome and a transfer plasmid comprising the nucleotide sequence(s) to be inserted (possibly with regulatory elements) flanking 5' and 3 ' with the viral sequences are presented respectively upstream and downstream of the insertion site. Said transfer plasmid can be generated and introduced into the host cell by routine techniques (eg by transfection). The virus genome can be introduced into the host cell by infection. The size of each flanking viral sequence can vary from at least 100 bp and at most 1500 bp on each side of the nucleotide sequence (preferably 200 to 550 bp and more preferably 250 to 500 bp). The homologous recombination that makes it possible to generate said oncolytic virus is preferably carried out in cultured cell lines (for example, HeLa, Vero) or in chicken embryonic fibroblast (CEF) cells obtained from embryonated eggs. In some embodiments, identification of the oncolytic virus that has incorporated the nucleotide sequences encoding anti-CTLA4 and possibly additional nucleotide sequence (eg, GM-CSF) may be facilitated by use of a screen and / or detectable gene. . In preferred embodiments, the transfer plasmid further comprises a selection marker with a specific preference for the GPT gene (encoding a guanine phosphoribosyl transferase) that allows growth in a selective medium (for example, in the presence of mycophenolic acid, xanthine and hypoxanthine) or a detectable gene encoding a detectable gene product such as GFP, e-GFP or mCherry. In addition, the use of an endonuclease capable of providing a di-strand break in said selectable or detectable gene may also be considered. Said endonuclease may be in the form of a protein or expressed by an expression vector. Once generated, such an oncolytic virus can be amplified in a suitable host cell using conventional techniques including culturing the transfected or infected host cell under suitable conditions to allow production and recovery of infectious particles. The present invention also relates to a method for producing the oncolytic virus described in the present description. Preferably, said method comprises the steps of a) preparing a producer cell line, b) transfecting or infecting the producer cell line prepared with the oncolytic virus, c) culturing the transfected or infected producer cell line under suitable conditions to allow virus production, d) recovering the virus produced from the culture of said producer cell line and optionally e) purifying said recovered virus. In some embodiments, the producer cell is selected from the group consisting of mammalian cells (eg, human or non-human) such as HeLa cells (eg, ATCC-CRM-CCL-2™ or ATCC-CCL-2.2™) , HER96, PER-C6 (Fallaux et al., 1998, Human Gene Ther. 9: 1909-17), hamster cell lines such as BHK-21 (ATCC CCL-10), etc., and bird cells such as those described in the WO2005 / 042728, WO2006 / 108846, WO2008 / 129058, WO2010 / 130756, WO2012 / 001075 as well as a primary chicken embryo fibroblast (CEF) prepared from chicken embryos obtained from fertilized eggs. Producer cells are preferably cultured in an appropriate medium which, if necessary, can be supplemented with suitable(or) non-suitable serum and / or growth factor(s) (for example, a chemically defined medium preferably free of animal-derived products or humans). Those skilled in the art can easily select an appropriate medium depending on the producing cells. Such means are commercially available. Producer cells are preferably cultured at +30°C to +38°C (more preferably about +37°C) for 1 to 8 days prior to infection. If necessary, multiple passages from 1 to 8 days can be performed to increase the total number of cells. In step b), the producer cells are infected with the oncolytic virus under / UUZDUU appropriate conditions by using an appropriate multiplicity of infection (MOI) to allow productive infection of the producer cells. For illustrative purposes, an appropriate MOI ranges from 10-3 to 20, with a specific preference for an MOI ranging from 0.01 to 5 and more preferably from 0.03 to 1. The infection step is carried out in a medium that may be the same or different from the medium used for the culture of producer cells. In step c), the infected producer cells are then grown under appropriate conditions well known to those skilled in the art until progeny virus particles are produced. The culture of infected producer cells is also preferably carried out in a medium that can be the same or different from the medium / medium used for the culture of producer cells and / or for the infection step, at a temperature between +32 °C and + 37 °C, for 1 to 5 days. In step d), the virus particles produced in step c) are collected from the culture supernatant and / or the producer cells. Recovery of the producer cells may require a step that allows disruption of the producer cell membrane to allow release of the virus. Disruption of the producer cell membrane can be induced by various techniques well known to those skilled in the art, including, but not limited to, freeze / thaw, hypotonic lysis, sonication, microfluidization, high shear homogenization (also called high speed) or high pressure homogenization. The recovered oncolytic virus can be at least partially purified before being dosed and used as described herein. A large number of purification steps and methods are available in the art, including, for example, clarification, enzymatic treatment (eg, endonuclease, protease, etc.), chromatographic, and filtration steps. Appropriate methods are described in the art (see, for example, WO2007 / 147528; WO2008 / 138533, WO2009 / 100521, WO2010 / 130753, WO2013 / 022764). In one embodiment, the present invention also provides a cell infected with the oncolytic virus described herein. The invention also includes pharmaceutical compositions comprising a virus, such as an oncolytic virus, as discussed above, and a pharmaceutically acceptable diluent, carrier, and / or adjuvant. In some embodiments, the pharmaceutical composition may be in the form of a CAR T cell, which carries parts or the complete antibody sequences described herein as part of the sequence encoding its chimeric antigen T cell receptor. The invention also encompasses pharmaceutical compositions comprising a / UUZDUU CAR T cell as discussed above and a pharmaceutically acceptable diluent, carrier and / or adjuvant. The invention also encompasses other therapeutic modalities or drug forms, such as antibody-drug conjugates, fusion proteins, etc., and a pharmaceutical composition comprising such therapeutic modalities. The antibody molecules, nucleotide sequences, plasmids, viruses, cells, and pharmaceutical compositions described herein may be suitable for parenteral administration including sterile aqueous and / or non-aqueous injection solutions that may contain antioxidants, and / or buffers. , and / or bacteriostatics and / or solutes that render the formulation isotonic with the blood of the intended recipient; and / or aqueous and / or non-aqueous sterile suspensions which may include suspending agents and / or thickening agents. The antibody molecules, nucleotide sequences, plasmids, cells, and pharmaceutical compositions described herein may be presented in unit-dose or multi-dose containers, for example, in sealed ampoules and vials, and may be stored under freeze-dried conditions ( ie, lyophilized) requiring only the addition of sterile liquid carrier, eg water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders and / or granules and / or tablets of the type described above. For parenteral administration to human patients, the daily dosage level of the anti-CTLA-4 antibody molecule will usually be from 1 mg / kg of the patient's body weight to 20 mg / kg, or in some cases even up to 100 mg / kg administered in single or divided doses. Lower doses may be used in special circumstances, for example in combination with prolonged administration. In any case, the actual dose that will be most suitable for any individual patient will be determined by the physician and will vary with the age, weight and response of the particular patient. The above dosages are illustrative of the average case. Of course, there may be individual cases where higher or lower dosage ranges are needed, and these are within the scope of this invention. Typically, a pharmaceutical composition (or medicament) described herein comprising an antibody molecule will contain the anti-CTLA-4 antibody molecule at a concentration of between about 2 mg / ml and 150 mg / ml or between about 2 mg / ml and 200mg / ml. In some embodiments, the pharmaceutical compositions will contain the anti-CTLA-4 antibody molecule at a concentration of 10 mg / ml. Typically, a pharmaceutical composition (or medicament) will contain the oncolytic virus described herein at a concentration of between approximately 103 to 1012vp (viral particles), iu (infectious unit) or pfu (plaque-forming units) depending on the virus and the quantitative technique. The amount of pfu present in a sample can be determined by counting the number of plaques after infection of permissive cells (eg, CEF or Vero cells) to obtain a plaque-forming unit (pfu) titer, the amount of vp after measuring the absorbance of A260, and the amount of iu by quantitative immunofluorescence, eg, by using anti-viral antibodies. As a general guide, individual doses that are suitable for a pharmaceutical composition comprising a variety of oncolytic poxviruses are from about 103 to about 1010 pfu, advantageously from about 103 pfu to about 109 pfu, preferably from about 104 pfu to about 108 pfu; and more preferably from about 104pfu to about 107pfu. Generally, in humans, oral or parenteral administration of the antibody molecules, nucleotide sequences, plasmids, viruses, cells and / or pharmaceutical compositions described in the present description is the preferred route, being the most convenient. For veterinary use, the antibody molecules, nucleotide sequences, plasmids, viruses, cells, and / or pharmaceutical compositions described herein are administered as a suitable acceptable formulation in accordance with normal veterinary practice and the regimen of administration will be determined by the veterinary surgeon. dosage and route of administration that will be most appropriate for a particular animal. Therefore, the present invention provides a pharmaceutical formulation comprising an amount of an antibody molecule, nucleotide sequence, plasmid, virus and / or cell of the invention effective to treat various conditions (as described above and below). Preferably, the antibody molecules, nucleotide sequences, plasmids, viruses, cells, and / or pharmaceutical compositions described herein are adapted for delivery by a route selected from the group comprising: intravenous; intratumoral; subcutaneous intramuscular. Administration can be in the form of a single injection or several repeated injections (eg, with the same or different doses, with the same or different routes, at the same or different sites of administration). For illustrative purposes, individual doses comprising approximately 104.5 x ΙΟ4, 105.5 x ΙΟ5, 106.5 x ΙΟ6, 107.5 x ΙΟ7, 108.5 x ΙΟ8, 109.5 x 109, or 1010pfu of an oncolytic poxvirus ( for example, the TK- and RR-defective vaccinia virus described herein) are particularly suitable for intratumoral administration. The present invention further includes antibody molecules, nucleotide sequences, plasmids, viruses, cells and / or pharmaceutical compositions described herein comprising pharmaceutically acceptable acid or base addition salts of the polypeptide binding moieties of the present invention . The acids which are used to prepare the pharmaceutically acceptable acid addition salts of the aforementioned basic compounds useful in this invention are those which form non-toxic acid addition salts, i.e., salts containing pharmacologically acceptable anions, such as salts of hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, acetate, lactate, citrate, acid citrate, tartrate, bitartrate, succinate, maleate, fumarate, gluconate, saccharate, benzoate, methanesulfonate, ethanesulfonate, benzenesulfonate, p -toluenesulfonate and pamoate [ie, l,r-methylene-bis-(2-hydroxy-3-naphthoate)], among others. Pharmaceutically acceptable basic addition salts may also be used to produce pharmaceutically acceptable salt forms of the agents according to the present invention. Chemical bases that can be used as reagents to prepare pharmaceutically acceptable base salts of the present agents that are acidic in nature are those that form non-toxic base salts with such compounds. Such non-toxic base salts include, but are not limited to, those derived from pharmacologically acceptable cations such as alkali metal cations (eg potassium and sodium) and alkaline earth metal cations (eg calcium and magnesium), ammonium or amine salts. water soluble such as N-methylglucamine-(meglumine) and lower alkanolammonium and other pharmaceutically acceptable basic organic amine salts, among others. The antibody molecules, nucleotide sequences, plasmids, viruses, and / or cells described herein may be lyophilized for storage and reconstituted in a suitable carrier prior to use. Any suitable lyophilization method (eg, spray drying, cake drying) and / or reconstitution techniques may be employed. Those skilled in the art will appreciate that lyophilization and reconstitution can lead to varying degrees of loss of antibody activity (for example, with conventional immunoglobulins, IgM antibodies tend to have greater loss of activity than IgG antibodies) and that usage levels may have to be adjusted incrementally to compensate. In one embodiment, the lyophilized (freeze dried) polypeptide binding moiety loses no more than about 20%, or no more than about 25%, or no more than about 30%, or no more than about 35%, or no more of about 40%, or not more than about 45%, or not more than about 50% of its activity (before lyophilization) when rehydrated. In some embodiments, the viral composition is suitably buffered at a physiological or slightly basic pH (eg, from about pH 7 to about pH 9 with a specific preference for a pH between 7 and 8.5 and more particularly near 8). It might be beneficial to further include a monovalent salt in the viral composition to ensure adequate osmotic pressure. Said monovalent salt may notably be selected from NaCI and KCI, preferably said monovalent salt is NaCI, preferably in a concentration of 10 to 500 mM (eg 50 mM). A suitable viral composition comprises 50 g / L sucrose / UUZDUU, 50 mM NaCI, 10 mM Tris-HCI, and 10 mM sodium glutamate, pH 8. The composition may also be formulated to include a cryoprotectant to protect the oncolytic virus at low temperature from storage. Suitable cryoprotectants include, without limitation, sucrose (or sucrose), trehalose, maltose, lactose, mannitol, sorbitol, and glycerol, preferably in a concentration of 0.5 to 20% (g weight / volume in I, referred to as w / v) as well. as well as high molecular weight polymers such as dextran or polyvinylpyrrolidone (PVP). The anti-CTLA-4 antibody molecules, nucleotide sequences, and pharmaceutical compositions described herein can be used in the treatment of cancer in a subject. We include that the subject could be mammalian or non-mammalian. Preferably not, the mammalian subject is a human or is a non-mammal, such as a horse, or a cow, or a sheep, or a pig, or a camel, or a dog or a cat. Most preferably, the mammalian subject is a human. By exhibit, we include that the subject exhibits a cancer symptom and / or a cancer diagnostic marker, and / or the cancer symptom and / or a cancer diagnostic marker can be measured and / or evaluated and / or quantified. It would be apparent to the medically skilled person what the cancer symptoms and diagnostic markers of cancer would be and how to measure and / or assess and / or quantify whether there is a reduction or increase in the severity of cancer symptoms, or a reduction o Increase in diagnostic markers of cancer; as well as how those cancer symptoms and / or cancer diagnostic markers could be used to form a prognosis for the cancer. Cancer treatments are often administered as a treatment cycle, that is, the therapeutic agent is administered over a period of time. The length of time of the course of treatment will depend on a number of factors, which may include the type of therapeutic agent being administered, the type of cancer being treated, the severity of the cancer being treated, and the age and health of the subject, among others. other reasons. By during treatment, we include that the subject is currently receiving a course of treatment, and / or receiving a therapeutic agent, and / or receiving a course of a therapeutic agent. In some embodiments, the cancer to be treated in accordance with the present invention is a solid tumor. In some modalities, the cancer is selected from the group consisting of advanced solid tumor, melanoma and other skin malignancies, synovial sarcoma, non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), cancer bladder cancer, prostate cancer, mesothelioma, ovarian cancer, breast cancer, renal cell cancer, hepatocellular carcinoma, head and neck cancer, and colorectal cancer. IVIA / a / ZUZ I / UUZDUU Each of the cancers described above is well known, and the symptoms and diagnostic markers of cancer are well described, as are the therapeutic agents used to treat those cancers. Accordingly, the symptoms, diagnostic markers of cancer, and therapeutic agents used to treat the above-mentioned types of cancer would be known to those skilled in medicine. Clinical definitions of the diagnosis, prognosis, and progression of a large number of cancers depend on certain classifications known as staging. Those staging systems operate to collate a number of different cancer diagnostic markers and cancer symptoms to provide a summary of the diagnosis and / or prognosis and / or progression of the cancer. The person skilled in oncology will know how to assess the diagnosis and / or prognosis and / or progression of cancer using a staging system, and what cancer diagnostic markers and cancer symptoms should be used to do so. By cancer staging, we include Raí's staging, which includes stage 0, stage I, stage II, stage III, and stage IV, and / or Binet's staging, which includes stage A, stage B, and stage C, and / or Ann Arbor staging, which includes stage I, stage II, stage III, and stage IV. It is known that cancer can cause abnormalities in cell morphology. These abnormalities often occur in a reproducible manner in certain types of cancer, which means that examination of these changes in morphology (also known as histological examination) can be used in the diagnosis or prognosis of cancer. Techniques for visualizing samples to examine cell morphology and prepare samples for visualization are well known in the art; for example, light microscopy or confocal microscopy. By histological examination, we include the presence of small, mature lymphocytes, and / or the presence of small, mature lymphocytes with a narrow border of cytoplasm, the presence of small, mature lymphocytes with a dense nucleus lacking discernible nucleoli, and / or the presence of small, mature lymphocytes with a narrow border of cytoplasm, and with a dense nucleus lacking discernible nucleoli, and / or the presence of atypical cells and / or lysed cells and / or prolymphocytes. Cancer is known to be the result of mutations in the cell's DNA, which can lead to the cell avoiding cell death or uncontrolled proliferation. Therefore, examining these mutations (also known as cytogenetic examination) can be a useful tool in evaluating the diagnosis and / or prognosis of a cancer. An example of this is the deletion of chromosome location 13ql4.1 that is characteristic of chronic lymphocytic leukemia. Techniques for examining mutations in cells are well known in the art; for example, fluorescent in situ hybridization (FISH). ΜΛ / a / ZUZ I / UUZDUU By cytogenetic examination, we include the examination of the DNA in a cell, and in particular the chromosomes. Cytogenetic examination can be used to identify DNA changes that may be associated with the presence of refractory cancer and / or recurrent cancer. This may include: deletions on the long arm of chromosome 13, and / or deletion of chromosome location 13ql4.1, and / or trisomy of chromosome 12, and / or deletions on the long arm of chromosome 12, and / or deletions on the long arm of chromosome 11, and / or the deletion of llq, and / or deletions on the long arm of chromosome 6, and / or the deletion of 6q, and / or deletions on the short arm of chromosome 17, and / or 17p deletion, and / or t(11:14) translocation, and / or (ql3:q32) translocation, and / or antigen receptor gene rearrangements, and / or BCL2 rearrangements, and / or BCL6 rearrangements, and / or t(14:18) translocations, and / or t(ll:14) translocations, and / or (ql3:q32) translocations, and / or (3:v) translocations, and / or translocations (8:14), and / or translocations (8:v), and / or translocations t(ll: 14) and (ql3:q32). Cancer subjects are known to exhibit certain physical symptoms, which are often the result of the cancer burden in the body. These symptoms often recur in the same cancer and may therefore be characteristic of the diagnosis and / or prognosis and / or progression of the disease. A medically skilled person will understand what physical symptoms are associated with what types of cancer, and how evaluating those physical systems can correlate with diagnosis and / or prognosis and / or disease progression. By physical symptoms, we include hepatomegaly and / or splenomegaly. BRIEF DESCRIPTION OF THE DRAWINGS In the examples below, reference is made to the following figures: Figures 1A to ID: Antibodies of the invention specifically bind to CTLA-4. ELISA showed that the antibodies bound human and cynomolgus CTLA-4 but not human CD28 protein. The binding of 2-C06 (Figure ΙΑ), 4-E-03 (Figure IB), 5-B07 (Figure 1C) was compared to Yervoy (Figure ID). Figures 2A to 2E: Dose-dependent binding of anti-CTLA-4 mAb to hCTLA-4-transfected cells. The anti-CTLA-4 mAb (Figure 2A to 2D) shows strong binding to 293T cells expressing Yervoy-like CTLA-4 (Figure 2E). Figure 3: Anti-CTLA-4 mAb binds to activated human CD4+ T cells in vitro. CD4+ T cells obtained from human peripheral blood were stimulated in vitro. Anti-CTLA-4 mAb binding (solid lines, top row) was analyzed by FACS and compared to Yervoy (dotted line, top row) and a commercial FACS antibody (bottom row). Figure 4: Binding block in activated CD4+ T cells in vitro. T cells were stained IVIA / a / ZUZ I / UUZDUU Human CD4+ activated in vitro with Alexa 647-labeled anti-CTLA-4 mAb (black line). Antibody binding was blocked by rhCTLA-4-Fc protein (grey line). Figure 5: Binding to activated cynomolgus CD4+ T cells in vitro. CD4+ T cells obtained from peripheral blood of cynomolgus were stimulated in vitro with CD3 / CD28 dynabeads. Anti-CTLA-4 mAb binding (solid lines, top row) was analyzed by FACS and compared to Yervoy (dotted line, top row) and a commercial FACS antibody (bottom row). Figure 6: Blockage of cell attachment by human and cynomolgus CTLA-4 protein. 293T-CTLA-4 cells were stained with Alexa 647-labeled anti-CTLA-4 mAb (black line). Antibody binding was blocked by rhCTLA-4-Fc protein (light gray line) and rcmCTLA-4-Fc protein (dark gray line). Figure 7: Binding to 293T cells expressing cynomolgus CTLA-4. 293T cells were transiently transfected with cynomolgus CTLA-4 and anti-CTLA4 mAb binding at different concentrations was analyzed by FACS. Figure 8: Expected lack of binding to resting cynomolgus / human PBMC. 4-E03, as well as 2-C06, 5-B07 and 2-F09 (data not shown), do not show any non-specific binding to different subsets of cells in human (top row) and cynomolgus (bottom row) PBMC as reported. analyzed by FACS. Figures 9A and 9B: Lack of expected direct agonist activity. CFSE-labeled CD4+ T cells from healthy donors were stimulated with coated anti-CD3 plus soluble anti-CTLA-4 mAb or anti-CD28. The % of dividing cells (CFSElow CD25+ cells) was determined after 3 days by FACS. (Figure 9A) FACS plots of a representative experiment (Figure 9B) summarizing the plot from 6 donors. Figures 10A and 10B: CD80 / CD86 blocking activity. The anti-CTLA-4 mAb blocks the binding of CD80 (Figure 10A) and CD86 (Figure 10B) to their CTLA-4 ligand as shown by ELISA. Figure 11: Functional ligand blockade in vitro. PBMC were stimulated with SEB plus titration doses of anti-CTLA-4 antibodies. The amount of IL-2 secreted in the supernatant was determined by MSD. In this figure, 1 representative donor out of 6 is shown. Figure 12: ADCC assay on activated CD4+ T cells in vitro. In vitro activated CD4+ T cells from healthy donors preopsonized with antiCTLA-4 mAb at 10 pg / ml were co-cultured with NK cells (NK-92 cell line) in a 2:1 ratio. ADCC activity was assessed by FACS as described below. The figure shows the mean + SD from 4-8 donors. / UUZDUU Figure 13: CTLA-4 is more expressed in tumor-resident Treg cells. Samples of freshly excised ovarian tumors and blood were obtained from patients during surgery. Ascites was collected from patients with different indications of cancer. CTLA expression in this patient material was compared to healthy PBMC. Tumor samples were minced and digested. Peripheral blood mononuclear cells were separated by centrifugation. CTLA-4 expression was assessed in CD4+ CD25+ CD127- Treg cells, CD4+ non-Treg cells, and CD8+ effector T cells by flow cytometry. Data represent individual patients / donors with n=12 for healthy PBMC, n=20 for ascites, n=9 for tumor, and n=5 for patient blood. CTLA-4 expression was also further assayed in human T cells that were activated in NOG mice in vivo and then isolated from the spleen of these mice (see Figure 14). Figure 14: Anti-CTLA-4 mAb mediates Treg lowering in vivo. Human PBMCs were injected iv into NOG mice. After approximately 2 weeks, spleens were removed and human Treg cells and CD8+ T cells were analyzed for CTLA-4 expression by FACS. Spleen cells isolated from NOG mice were transferred i.p. to SCID mice. lh later, the mice were treated i.p. with CTLA-4 hlgGl or control mAb. Intraperitoneal fluid was collected after 24 h and the frequency of human T cell subsets (14A: Treg and 14B: CD8+ T cells) was determined by flow cytometry. Figures 15A to 15D: Characterization of surrogate mouse anti-CTLA-4 mAb. Figures 15A and 15B: Blocking ELISAs were performed with m5-B07 to assess the blocking characteristics of the ligand. The antibody blocks the binding of Figure 15A CD80 and Figure 15B CD86 to their CTLA-4 ligand in a dose-dependent manner. Figures 15C and 15D: 5-B07 in mouse IgG2a format mediated Treg deletion in CT26 tumor model. Balb / c mice were injected subcutaneously with 1 x 10 6 CT26 cells and treatment started at the tumor size of approximately 7 x 7 mm. After 3 injections of 10 mg / kg antibody, single tumor cell suspensions were analyzed for immune cell content by FACS. Figure 15C: Ligand blockade of surrogate antibody 5-B07 causes decrease in T reg. This causes a change in the CD8+ T cell / Treg ratio as shown in Figure 15D. Figure 16: Generation of COPTG19384 and COPTG19385. Schematic representation of COPTG19384 and COPTG19385 used in this study. COPTG19385 contains a deletion of the J2R gene at the TK locus replaced by the anti-CTLA-4 heavy chain driven by p7.5K, and an I4L gene deletion at the RR locus replaced by the driven anti-CTLA-4 light chain by p7.5K. COPTG19384 contains a J2R gene deletion at the TK locus replaced by the anti-CTLA-4 heavy chain driven by p7.5K, and an I4L gene deletion at the RR locus replaced by the anti-CTLA-4 driven light chain. by p7.5K, and by GM-CSF driven by pSE / L. ΜΛ / a / ZUZ 1 / UUZDUU Figures 17A and 17B: Expression analysis of the monoclonal antibody 4-E03 in the supernatant of CEF cells infected with COPTG19384. Figure 17A) By Western blotting: CEF cells at MOI 0.05 were infected with COPTG19384 in triplicate. Cell supernatants were collected after 48 hours and analyzed by WB after electrophoresis in a non-reducing condition and using an anti-Ig (left blot) or anti-light chain (right blot) HRP-conjugated antibody. Figure 17B) by ELISA: CEF cells were infected at MOI 0.05 with COPTG19384 in triplicate or WTG17137. Cell supernatants were collected after 48 hours and analyzed by ELISA for the detection of the 4-E03 monoclonal antibody. Figure 18: Analysis of GM-CSF expression in the supernatant of CEF cells infected with COPTG19384 by ELISA. CEF cells at MOI 0.05 were infected with the primary research stock COPTG19384 in triplicate or WTG17137. Cell supernatants were collected after 48 hours and analyzed by ELISA for the detection of GM-CSF. Figures 19A to 19C: Replication studies of COP WT, COPTG19384 (two batches) and WTG17137 (two batches) in normal and tumor hepatocytes. Figure 19A) Replication rate in normal human hepatocytes. Figure 19B) Replication rate in malignant HepG2. Figure 19C) Therapeutic indices calculated from replication rates measured in HepG2 and hepatocytes. Figure 20: Replication of COPTG19384 and WTG17137 in reconstructed human skin. The replication of COPTG19384 and WTG17137 was evaluated after 7 days and with variable inocula from 10 to 105pfu. The results are the means and the SEM of three measurements. Figures 21A to 21C: Oeneolytic activities of COPTG19384 and WTG17137 in three human tumor cell lines: MIA PaCa-2 (Figure 21A), LoVo (Figure 21B) and HepG2 (Figure 21C). Figures 22A and 22B: Expression level of monoclonal antibody 4-E03 and GM-CSF in (Figure 22A) supernatants from infected HepG2 and LoVo and (Figure 22B) supernatants from 5 different infected human tumor cell lines. Figure 22A) The expression levels of 4-E03 and GM-CSF were evaluated after 5 days of incubation, at an MOI of 10.5 to 10.2 for COPTG19384 and at an MOI of 10.2 for WTG17137 used as negative control. Figure 22B) Expression levels of 4-E03 and GM-CSF were assessed 48 hours after infection by COPTG19384 at MOI 0.05. Figures 23A and 23B: Binding of different batches of 4-E03 to the CTLA-4 protein. The binding of recombinantly produced 4-E03 by CHO (research batch 4-E03) or HEK (toxin batch 4-E03) cells to recombinant human (Figure 23A) and cynomolgus (Figure 23B) protein was compared with binding of 4-E03 purified from the supernatant of infected MIA PaCa-2 tumor cells (4-E03 TG) by ELISA. / UUZDUU Figures 24A and 24B: Binding of different batches of 4-E03 to cells expressing CTLA4. The binding of recombinantly produced 4-E03 by CHO (research batch 4E03) or HEK (toxin batch 4-E03) cells to human (Figure 24A) and cynomolgus (Figure 24B) CTLA-4 expressing cells was compared. with the binding of 4-E03 purified from the supernatant of infected MIA PaCa-2 tumor cells (4-E03 TG) by flow cytometry. Figure 25: Kinetics of virus accumulation in the LoVo xenograft tumor. The kinetics of virus accumulation in LoVo xenografted tumor after i.t. single dose of COPTG19384 or WTG17137 at two different doses (ΙΟ4 or 105pfu). The solid or broken lines represent the median of the three values ​​determined at each moment. Figures 26A and 26B: Kinetics of mAb 4-E03 and GM-CSF accumulation in LoVo xenograft tumor Figure 26A) The kinetics of mAb 4-E03 accumulation in LoVo xenograft tumor was evaluated after i.t. single dose of COPTG19384 or WTG17137 at two different doses (ΙΟ4 or 105pfu) or after i.p. single 3 mg / kg monoclonal antibody 4-E03. Solid or dashed lines represent the median of the three values. Figure 26B) The kinetics of GM-CSF accumulation in LoVo xenografted tumor after i.t. single dose of COPTG19384 at two different doses (ΙΟ4o 105pfu) or WTG17137 (105pfu). The solid lines represent the median of the three values ​​determined at each moment. Figures 27A and 27B: Kinetics of mAb 4-E03 and GM-CSF concentrations in sera of LoVo xenografted mice. Figure 27A) The kinetics of mAb 4-E03 concentrations in serum were evaluated after i.t. single injection in LoVo xenograft tumor of COPTG19384 or WTG17137 at two different doses (ΙΟ4 or 105pfu) or after i.p. single 3 mg / kg monoclonal antibody 4-E03. The solid lines represent the median of the three values. Figure 27B) Kinetics of serum GM-CSF concentrations after i.t. unique in LoVo xenograft tumor of COPTG19384 at two different doses (ΙΟ4o 105pfu) or WTG17137 (105pfu). The dashed lines represent the median of the three values ​​determined at each moment. Figure 28: Kinetics of virus accumulation in CT26 tumors. Kinetics of virus accumulation in CT26 tumors after three injections (i.t. at DO, D2, and D4) of WTG18058, COPTG19421, or COPTG19407 at 107 pfu / injection. Figures 29A and 29B: Kinetics of accumulation of mAb m5-B07 and mGM-CSF in the CT26 tumor. Figure 29A) Kinetics of mAb m5-B07 concentrations in CT26 tumor during and after three i.t. of WTG18058, COPTG19421 or COPTG19407 (107 pfu / injection) or after an i.p. single 3 mg / kg monoclonal antibody m5-B07. The solid lines represent the median of the three values. Figure 29B) Kinetics of / UUZDUU mGM-CSF concentrations in CT26 tumor during and after three i.t. of WTG18058, COPTG19421 or COPTG19407 (107 pfu / injection) or after an i.p. single 3 mg / kg monoclonal antibody m5-B07. The solid lines represent the median of the three values ​​determined at each moment. Figure 30: Kinetics of mAb m5-B07 concentrations in CT26 pattern sera. Kinetics of mAb m5-B07 concentrations in serum after three i.t. in WTG18058, COPTG19421 or COPTG19407 tumor CT26 (107 pfu / injection) or after i.p. single 3 mg / kg monoclonal antibody m5-B07. The solid lines represent the median of the three values ​​determined at each moment. Figures 31A and 31B: Antitumor activity in the CT26 model: effect of COPTG19347 + / - anti-PD1 on the growth of the CT26 tumor (Figure 31A) and the survival of the mice (Figure 31B). CT26 cells were injected s.c. in BalB / c mice at D-7. COPTG19347 (107pfu), WTG18058 (107pfu), WTG18058 or buffer were injected i.t., at OD, D2 and D4, possibly followed by i.p. of 250 pg / mouse of anti-PD1 RMPI-14 on D7, DIO, D14, D17 and D21. Figure 32: Evaluation of the dose effect in the CT26 model (compilation of observed survival data after treatment of COPTG19407, COPTG19421 and WTG18058). Figures 33A to 33E: Antitumor activity of COPTG19407 compared to WTG18058 plus m5-B07 in the CT26 tumor model. CT26 cells were injected s.c. in Balb / c mice. Treatment of the mice was started when the tumors reached approximately 100 mm3. Mice were injected on DO, D2, and D5 with COPTG19407 (8.5 x 106 pfu i.t.), WTG18058 (8.5 x 106 pfu i.t.), m5-B07 (10 mg / kg i.p.), or the combination of WTG18058 (8.5 x 106 pfu i.t.) plus m5. -B07 (10 mg / kg i.p.). Figures 33A to 33D: Tumor growth was followed and Figure 33E: Survival over time. Figures 34A to 34E Individual tumor volume curves of BALB / c mice bearing subcutaneous A20 tumors. A20 cells were injected s.c. in Balb / c mice. Treatment of mice was started when tumors reached 80-100 mm3 Mice were injected at DO, D2 and D4 with vehicle (i.t.), COPTG19407 (4.75 x 106pfu i.t.), WTG18058 (4.75 x 106pfu i.t.), RMPI- 14 (anti-mPD-1) (250 pg / mouse i.p.) or the combination of COPTG19407 (4.75 x 106pfu i.t.) plus RMPI-14 (250 pg / mouse i.p.). Figure 34A) Animals from group 1 treated with vehicle. Figure 34B) Group 2 animals treated with WTG18058. Figure 34C) Group 3 animals treated with COPTG19407. Figure 34D) Group 4 animals treated with murine anti-PD1. Figure 34E) Group 5 animals treated with COPTG19407 and murine anti-PD1. Figure 35: Mean tumor volume curves of BALB / cN mice bearing subcutaneous A20 tumors. Each point represents the mean tumor volume recorded per group. Tumor volumes of all animals were monitored for 64 days. Mice were treated with vehicle (group 1), WTG18058 (group 2), COPTG19407 (group 3), the murine anti-PD1 antibody RMP1-14(BioXCell) (group 4), and COPTG19407 and RMP1-14 (group 5). Animals were randomized on D7 and treated for the period D7 to D31. The last mice were sacrificed on D61. Figures 36A and 36B: Anti-tumor activity in the A20 model: effect of COPTG19407 + / - anti-PD1 on the growth of the A20 tumor (Figure 36A) and the survival of the mice (Figure 36B). A20 cells were injected s.c. in Balb / c mice. Treatment of the mice was started when the tumors reached 80 to 100 mm3. Mice were treated with vehicle (i.t.) (group 1), anti-PD-1 (group 2), isotype (group 3), WTG18058 (105pfu i.t.) (group 4), WTG18058 (105pfu i.t.) + isotype ( group 5), WTG18058 (105pfu i.t.) + anti-PD-1 (group 6), WTG19407 (105pfu i.t.) (group 7), WTG19407 (105pfu i.t.) + isotype (group 8) and WTG19407 (105pfu i.t.) + antiPD -1 (group 9). Figures 37A to 37E: Individual tumor volume curves of C57BL / 6 mice bearing subcutaneous C38 tumors. C38 cells were injected s.c. in C57bl / 6 mice. Treatment of the mice was started when the tumors reached 80-100 mm3. Mice were injected on DO, D2 and D4 with vehicle (i.t.), COPTG19407 (4.75 x 106pfu i.t.), WTG18058 (4.75 x 106pfu i.t.), RMP1-14 (anti-mPD-1) (250 pg / mouse i.p.) or the combination of COPTG19407 (4.75 x 106 pfu i.t.) plus RMP1-14 (250 pg / mouse i.p.). Figure 37A) Animals from group 1 treated with vehicle. Figure 37B) Animals from group 2 treated with WTG18058. Figure 37C) Group 3 animals treated with COPTG19407. Figure 37D) Group 4 animals treated with murine anti-PD1. Figure 37E) Group 5 animals treated with COPTG19407 and murine anti-PD1. Figure 38: Mean tumor volume curves of C57BL / 6 mice bearing subcutaneous C38 tumors. Each point represents the mean tumor volume recorded per group. Tumor volumes of all animals were monitored for 61 days. Mice were treated with vehicle (group 1), WTG18058 (group 2), COPTG19407 (group 3), the murine anti-PD1 antibody RMP1-14 (BioXCell) (group 4), and COPTG19407 and RMP1-14 (group 5). Animals were randomized on D7 and treated for the period D7 to D31. The last mice were sacrificed on D61. Figures 39A to 39E: Individual tumor volume curves of BALB / c mice bearing subcutaneous EMT6 tumors. EMT6 cells were injected s.c. in Balb / C mice. Treatment of mice was started when tumors reached 80-100 mm3. Mice were injected at DO, D2 and D4 with vehicle (i.t.), COPTG19407 (4.75 x 106pfu i.t.), WTG18058 (4.75 x 106pfu i.t.), RMP1-14 ( anti-mPD-1) (250 pg / mouse i.p.) or the combination of COPTG19407 (4.75 x 106pfu i.t.) plus RMP1-14 (250 pg / mouse i.p.). Figure 39A) Animals from group 1 treated with vehicle. / UUZDUU Figure 39B) Group 2 animals treated with WTG18058. Figure 39C) Group 3 animals treated with COPTG19407. Figure 39D) Group 4 animals treated with murine anti-PD1. Figure 39E) Group 5 animals treated with COPTG19407 and murine anti-PD1. Figure 40: Mean tumor volume curves of BALB / c mice bearing tumors subcutaneous EMT6. Each point represents the mean tumor volume recorded per group. Tumor volumes of all animals were monitored for 61 days. Mice were treated with vehicle (group 1), WTG18058 (group 2), COPTG19407 (group 3), the murine anti-PD1 antibody RMP1-14 (BioXCell) (group 4), and COPTG19407 and RMP1-14 (group 5). Animals were randomized on D7 and treated for the period D7 to D31. The last mice were sacrificed in D56. The curves stopped after the death of more than 20% of the mice. TABLE A Summary of characteristics of anti-CTLA-4 antibodies ADCC (% CTLA4+) 25.6 LO s 42.8 15.1 46.3 Block mCD86 N / A N / A N / A N / A N / A mCD80 N / A N / A N / A N / A N / A mCD86 complete OR CD or complete complete complete hCD80 complete complete complete complete complete Protein binding mCD28 nonbinding r- nonbinding nonbinding nonbinding cmCTLA4 nonbinding in nonbinding 0.3 nonbinding cmCTLA4 0.2 0.2 OJ cz> 0.4 0.2 hCD28 nonbinding nonbinding nonbinding nonbinding nonbinding hCTLA4 0.2 0.3 0.3 0.5 0.2 Blocking protein binding cm 09 09 GO 09 09 Cell binding cyno weak cñ O c: c tñ Cell binding EC50 0.2 cS 610 0.15 0.13 c o o 002-C06 004-E03 002-F09 5-B07 Yervoy / UUZDUU EXAMPLES Specific, non-limiting examples embodying certain aspects of the invention will now be described. In the examples, the rh protein refers to a human recombinant protein (for example, rhIL-2 refers to the human recombinant IL-2 protein) and the rcm protein refers to a cynomolgus recombinant protein (for example, rcmCTLA4 refers to the cynomolgus protein). recombinant CTLA-4 from cynomolgus). In addition to the sequences mentioned above, some additional sequences are used in the examples, which are set out in Table 5 below. Table 5: Additional sequences used in the examples Sequence SEQ ID NO: Human GM-CSF without signal peptide APARSPSPSTQPWEHVNAIQEARRLLNLSRDTAAEMNETVEVISEMF DLQEPTCLQTRLELYKQGLRGSLTKLKGPLTMMASHYKQHCPPTPE TSCATQTITFESFKENLKDFLLVIPFDCWEPVQE 55 Human GM-CSF with signal peptide MWLQSLLLLGTVACS ISAPARSPSPSTQPWEHVNAIQEARRLLNLSR DTAAEMNETVEVISEMFDLQEPTCLQTRLELYKQGLRGSLTKLKGPL TMMASHYKQHCPPTPETSCATQTITFESFKENLKDFLLVIPFDCWEP VQE 56 Murine GM-CSF without signal peptide APTRSPITVTRPWKHVEAIKEALNLLDDMPVTLNEEVEVVSNEFS FKK LTCVQTRLKIFEQGLRGNFTKLKGALNMTASYYQTYCPPTPETDCET QVTTYADFIDSLKTFLTDIPFECKKPVQK 57 Murine GM-CSF with signal peptide MWLQNLLFLGIVVYSLSAPTRSPITVTRPWKHVEAIKEALNLLDDMPV TLNEEVEVVSNEFSFKKLTCVQTRLKIFEQGLRGN FTKLKGALNMTA SYYQTYCPPTPETDCETQVTTYADFIDSLKTFLTDIPFECKKPVQK 58 Promoter p7.5K CCACCCACI 1 1 1 IAIAGTAAGI 1 1 1 1CACCCATAAATAATAAATACA ATAATTAATTTCCGTAAAAGTAGAAAATATATTCTAATTTATTGCA CGGTAAGGAAGTAGAATCATAAAGAACAGT 59 Promoter pH5.R TTTATTCTATACTTAAAAAATGAAAATAAATACAAAGGTTCTTGAGG GTTGT GTTAAATTGAAAGCGAGAAATAATCATAAATTATTCATTAT CGCGATATCCGTTAAGTTTG 60 Promoter pSE / L AAAAATTGAAA1 1 1 1A1 1 1 1 1 1 1 1 1 1 1 1 GGAATATAAATA 61 Example 1: Generation of CTLA-4 Specific Antibodies Isolation of scFv Antibody Fragments The n-Coder® scFv library (Biolnvent; Sóderlind E, et al. Nat Biotechnol 2000; 18(8): 852-6) was used to isolate scFv antibody fragments that recognize human CTLA-4. The phage library was used in three consecutive tests against recombinant human protein. After phage incubation, cells were washed to remove unbound phage. Binding phage were eluted with trypsin and amplified in E. coli. The resulting phage pool was converted to scFv format. E. coli was transformed with scFv-carrying plasmids and individual scFv clones were expressed. Identification of unique scFv binding to CTLA-4 Converted scFvs from the third screen were assayed using a homogeneous FMAT assay (Applied Biosystems, Carlsbad, CA, USA) for binding to transfected 293 FT cells expressing human CTLA-4 or a non-target protein. Briefly, transfected cells were added to clear bottom plates, along with scFv-containing supernatant from expression plates (diluted 1:7), mouse His-tag antibody (0.4 Z^g / ml; R&D Systems) and an APC-conjugated goat anti-mouse antibody (0.2 pg / ml; cat. no. 115-136-146, Jackson Immunoresearch). FMAT plates were incubated at room temperature (approximately 20-25 °C) for 9 h before reading. Target-specific bacterial clones were scored as active and collected in a 96-well plate. IgG binding to CTLA-4 in ELISA 96-well plates (Lumitrac 600 LIA plate, Greiner) were coated overnight at 4°C with recombinant human CTLA-4-Fc protein (R&D Systems) at 1 pmol / well, recombinant human CTLA-4-Fc protein (R&D Systems) at 1 pmol / well, cynomolgos (cm) (R&D Systems) at 1 pmol / well, recombinant mouse CTLA-4-Fc protein (R&D Systems) at 0.3 pmol / well, recombinant human CD28-Fc protein (R&D Systems) at 1 pmol / well or recombinant mouse CD28-Fc protein (R&D Systems) at 0.3 pmol / well. After washing, titrated doses of anti-CTLA-4 mAb from 0 pg / ml to 0.06 ng / ml (66 nM to 0.3 pM) were allowed to bind for 1 hour. The plates were then washed again and bound antibodies were detected with an anti-human F(ab)HRP secondary antibody (Jackson ImmunoResearch) diluted to 50 ng / ml. Super Signal ELISA Pico (Thermo Scientific) was used as a substrate and plates were analyzed using the Tecan Ultra microplate reader. All antibodies were shown to bind to human and cynomolgus CTLA-4 protein but not to human CD28 protein as assayed by ELISA. In addition, / UUZDUU was shown to bind 5-B07 to mouse CTLA-4 but not to mouse CD28 (see Figures 1A to ID). IgG binding to CTLA-4 expressing 293T cells in flow cytometry Converted IgG clones were analyzed for binding to CTLA-4 expressing 293T cells (purchased from Crownbio). Cells were incubated with different concentrations (as indicated in Figures 2A to 2E) of anti-CTLA-4 mAb at 4 °C for 20 min before washing and staining with an APC-labeled goat-expressed anti-human secondary antibody (# 109-136-088, Jackson ImmunoResearch). Dead cells were excluded from analyzes by using Fixable Viability Dye eFluor780 (eBiosciences). Data acquisition was performed on a FACSVerse (BD Biosciences, Franklin Lakes, NI) and analyzed with the FlowJo program (Tree Star, Ashland, OR). Anti-CTLA-4 mAbs were shown to bind to 293T cells expressing human CTLA-4 in a dose-dependent manner with an EC50 value similar to that of Yervoy (Figures 2A to 2E). 293T cells stably transfected with human CTLA-4, 293T cells transiently transfected with cynomolgus CTLA-4, human or naïve cynomolgus PBMCs, in vitro activated human or cynomolgus CD4+ T cells, were incubated with the concentrations of indicated anti-CTLA-4 mAbs at 4 °C for 20 min before washing and staining with an APC-labeled anti-human secondary antibody (Jackson ImmunoResearch). Example 2: Anti-CTLA-4 mAb specifically binds to (primary) cells expressing human v cynomolgus CTLA-4 CTLA-4-specific mAb binds to primary human and cynomolgus in vitro activated CD4+ T cells, but not to naïve PBMC isolated from healthy donors PBMC were isolated from buffy coats. Briefly, leukocytes were diluted 1:3 in PBS and loaded onto Ficoll-Paque Plus pads (Amersham). The samples were centrifuged at 800 xg for 20 min at 20°C. The upper phase containing plasma was removed and the mononuclear cells of the distinct white band at the plasma / Ficoll interface were isolated. Human peripheral CD4+ T cells were purified by negative selection using the MACS CD4 T cell isolation kit (Miltenyi Biotec). CD4+ T cells were activated in vitro with CD3 / CD28 dynabeads (Life Technologies) plus 50 ng / ml rhIL-2 (R&D Systems) in RIO medium (RPMI containing 2 mM glutamine, 1 mM pyruvate, 100 ul penicillin and streptomycin / ml and 10% FBS (GIBCO from Life Technologies) for 3 days to upregulate CTLA-4 expression CD4+ T cells were isolated from cynomolgus using non-human CD4 microbeads (Miltenyi Biotec) and incubated with 50 ng / ml PMA (Sigma-Aldrich) IVIA / a / ZUZ I / UUZDUU and 100 ng / ml ionomycin (Sigma-Aldrich) for 3 days. Naive human or cynomolgus PBMC, in vitro activated human or cynomolgus CD4+ T cells were incubated with the indicated concentrations of anti-CTLA-4 mAb at 4 °C for 20 min before washing and staining with an APC-labeled anti-human secondary antibody. (Jackson ImmunoResearch). Anti-CTLA-4 mAb binding was analyzed by FACS using a BD FACS Verse. The antibodies were shown to bind to activated human (Figure 3) and cynomolgus (Figure 5) CD4+ T cells in vitro but not to resting PBMC (Figure 8). Binding to T cells endogenously expressing CTLA-4 is similar to Yervoy staining (Figure 3, top row, dotted line) and as a positive control a commercial anti-CTLA-4 FACS antibody from BD Biosciences (clone BNI3 ; Figure 3, bottom row). As shown in Figure 4, staining of in vitro activated human CD4+ T cells (black line) can be completely blocked by rhCTLA-4-Fc (grey line), demonstrating the specificity of the antibodies. In this competitive binding assay, 2 pg / ml Alexa 647-labeled anti-CTLA-4 mAb was mixed with recombinant human CTLA-4-Fc protein (50 pg / ml) prior to incubation with CTLA-4-expressing cells. . IgG binding was detected by FACS. Transfected 293T cells expressing human and cynomolgus CTLA-4 confirm cynomolgus cross-reactivity of the tested antibodies Cynomolgus cross-reactivity of the antibodies was further confirmed in transfected CTLA-4 expressing 293T cells. As demonstrated in Figure 6, the binding of CTLA-4-specific antibodies to transfected cells expressing human CTLA-4 can be inhibited by recombinant human and cynomolgus protein (both R&D Systems). The antibodies were also shown to bind to transfected cells expressing cynomolgus CTLA-4 (Figure 7, top row). This binding can again be blocked by a recombinant protein from cynomolgus (bottom row, gray line). Experiments were performed in a similar manner to the competitive assay described above in Example 2 in connection with Figure 4. Lack of expected direct agonistic activity In vitro proliferation assays were performed to exclude unanticipated direct agonist activity (eg, due to non-specific binding). Human peripheral CD4+ T cells were purified from healthy PBMC by negative selection using the MACS CD4 T cell isolation kit (Miltenyi Biotec) and subsequently labeled with CFSE (2 μΜ, Molecular Probes). Antibodies were cross-linked with goat anti-human IgG expressed F(ab')2, Fcy fragment specific or / UUZDUU goat expressed anti-mouse IgG F(abj2, Fcy fragment specific at a molar ratio IgG:F(ab')2 = 1.5:1 for 1 h at RT 1 x 105 purified human CD4+ T cells were stimulated with plaque-bound anti-CD3 (0.5 pg / ml; clone UCHT1, R&D Systems) and 4 pg / ml soluble anti-CTLA-4, crosslinked or anti-CD28 (clone CD28.2, BioLegend) crosslinked for 72 hours at 37 C. Cells were washed and stained with a BV421-conjugated anti-CD25 antibody (clone M-A251, BD Biosciences). of dividing CD25+ / CFSEIow cells was analyzed by FACS. Figures 9A and 9B demonstrate that none of the anti-CTLA-4 mAbs tested induce T cell proliferation in contrast to anti-CD28 stimulation. Example 3 - CD80 / CD86 anti-CTLA-4 mAb blocking ligand binding ligand blocking ELISA The anti-CTLA-4 IgG ligand blocking activity was assessed by ELISA. For this purpose, recombinant human CTLA-4-Fc protein (R&D Systems) was coated on 96-well plates (Lumitrac 600 LIA plate, Greiner) at 1 pmol / well. After washing, the titrated doses of anti-CTLA-4 mAb were allowed to bind for 1 hour. His-tagged ligands were added at 200 nM and 100 nM, respectively (rhCD80 and rhCD86; R&D Systems) and the plates were further incubated for 15 minutes. After washing, bound ligand was detected with an HRP-tagged anti-His antibody (R&D Systems). Super Signal ELISA Pico (Thermo Scientific) was used as a substrate and plates were analyzed using the Tecan Ultra microplate reader. As shown in Figures 10A and 10B, the anti-CTLA-4 antibodies tested show Yervoy-like ligand-blocking activity. Functional ligand blockade in vitro For the SEB PBMC assay, total PBMC from healthy donors were seeded in 96-well plates (1 x 105 cells / well) and stimulated with 1 pg / ml Staphylococcus enterotoxin B (SEB, Sigma Aldrich) in the presence of titrated doses of IgG anti CTLA-4. IL-2 secretion was measured by MSD (Mesoscale) on day 3 according to the manufacturer's instructions. Antibodies 4-E03 and 2-C06 were shown to increase IL-2 production and their potency was shown to be similar to Yervoy's. In Figure 11, a representative donor of 6 is shown. Example 4 - Anti-CTLA-4 mAb decreases cells expressing CTLA-4 in vitro and in vivo Antibody Dependent Cellular Cytotoxicity (ADCC) ADCC assays were performed by using an NK-92 cell line. ΜΛ / a / ZUZ 1 / UUZDUU stably transfected to express the CD16-158V allele together with GFP (purchased from Conkwest, San Diego, CA; Binyamin, L., et al., 2008, Blocking NK cell inhibitory self-recognition promotes antibody-dependent cellular cytotoxicity in a model of anti-lymphoma therapy.Journal of immunology 180, 6392-6401). Target CD4+ T cells were isolated from peripheral blood of healthy donors using a CD4+ T cell isolation kit (Miltenyi Biotec). Cells were stimulated for 48 hours with CD3 / CD28 dynabeads (Life Technologies, Thermo Fisher) and 50 ng / ml rhIL-2 (RyD Systems) at 37°C. Target cells were preincubated with mAb at 10 pg / ml for 30 min at 4 °C before mixing with NK cells. Cells were incubated for 4 h in RPMI 1640 + GlutaMAX medium (Invitrogen) containing 10 mM HEPES buffer, 1 mM sodium pyruvate and 10% low IgG FBS at a 2:1 target:effector cell ratio. Lysis was determined by flow cytometry. Briefly, at the end of the incubation, the cell suspension was stained with BV510-conjugated anti-CD4 (clone RPA-T4, BD Biosciences) along with 10 nM SYTOX Red (Invitrogen) or Fixable Viability Dye eFluor780 (eBioscience) killed cell stain. ) for 20 min in the dark at 4 °C and then cells were analyzed using a FACSVerse (BD Biosciences). 4-E03 showed significantly improved deletion of CTLA-4+ T cells in vitro compared to Yervoy (Figure 12). CTLA-4 expression in material from the primary patient In order to validate the translational potential of the above finding on the decreased activity of anti-CTLA-4 mAb, we examined the expression of CTLA-4 in the primary patient material. The Ethics Committee of Skane University Hospital obtained ethical approval for the use of clinical samples. Informed consent was provided in accordance with the Declaration of Helsinki. Samples were obtained through the Department of Gynecology and the Department of Oncology, Skánes University Hospital, Lund. The ascitic fluid was evaluated as the single cell suspension that had been isolated. Tumor material was cut into small pieces and incubated in RIO with DNase I (Sigma Aldrich) and Liberase™ (Roche Diagnostics) for 20 min at 37°C. The remaining tissue was mechanically disrupted and, together with the cell suspension, passed through a 70 pm cell filter. Cells isolated from ascitic fluid and tumors were stained. To identify different subsets of T cells, the following antibodies were used: CD4-BV510 (RPA-T4), CD25-BV421 (M-A251), anti-CD127-FITC (HIL-7R-M21), CTLA-4PE (BNI3), CD8-PeCy7 (RPA-T8), CD3-APC (UCHT1), CD45-PercP-Cy5.5 (HI30), mouse IgG2a isotype, κ PE control (G155-178; all from BD Biosciences). Data acquisition was performed using FACSVerse and data was analyzed with FlowJo. As shown in Figure 13, CTLA-4 is more expressed in Treg cells. IVIA / a / ZUZ I / UUZDUU intratumoral, making them a good target to decrease CTLA-4-specific antibodies. Model PBMC-NOG / SCID To confirm the in vitro findings on the lowering activity of CTLA-4-specific antibodies, we tested the lowering capacity of the anti-CTLA-4 mAb in a PBMC-NOG / SCID model in vivo. The model is based on the well-established hu-PBMC-NOG model (Sqndergaard H. et al., Clin Exp Immunol. 2013 May; 172(2): 300-10. doi: 10.1111 / cei.12051; Cox JH et al. , PLoS One. 2013 Dec 23;8(12):e82944.doi:10.1371 / journal.pone.0082944.eCollection 2013) and internally modified as described below. Mice were bred and maintained at local facilities in accordance with local office guidelines. Eight week old females C.B. 17 scid (Bosma GC et al., Nature. 10 Feb 1983; 301(5900): 527-30) and NOG (NOD / Shi-sc / í / / IL-2RYnul1; Ito M et al., 2002, NOD / Mice were supplied by Taconic (Bomholt, Denmark) and were maintained at local animal facilities. For the PBMC-NOG / SCID (primary human xenograft) model, human PBMCs were isolated using Ficoll Paque PLUS and after washing cells were resuspended in sterile PBS at 75 x 10 6 cells / ml. NOG mice were injected i.v. with 200 pl of cell suspension corresponding to 15 x 106 cells / mouse. 2 weeks after injection, spleens were isolated and placed in single cell suspension. Subsequently, a small sample was taken to determine the expression of CTLA-4 in human T cells by FACS. As indicated in Figure 13, CTLA-4 is more expressed on Treg cells compared to other T cells reflecting the situation in human patients. Most of the cells were resuspended in sterile PBS at 50 x 106 cells / ml. SCID mice were injected i.p. with 200 μΙ of the suspension corresponding to 10 x 106 cells / mouse. Mice were treated with 10 mg / kg anti-CTLA-4 hlgGl, Yervoy, or lh isotype control mAb later. The intraperitoneal fluid of the mice was collected after 24 h. Human T cell subsets were identified and quantified by FACS using the following markers: CD45, CD4, CD8, CD25, CD127 (all from BD Biosciences). All antibodies tested showed similar or better Treg-lowering activity than Yervoy. Other T cell populations, such as CD8 + effector T cells, were not affected (Figures 14A-14B). Example 5: The selected surrogate antibody m5-B07 shows the same functional characteristics as 4-E03 In some of the examples, particularly the in vivo examples, the antibody clone 5-B07 in mIgG2a format (also called m5-B07) is used. This is a mouse antibody, which is a surrogate antibody for the human antibodies described herein. It has been selected as a surrogate antibody since it binds to murine CTLA-4 and therefore blocks ligand binding (Figures 15A and 15B). In addition, it also shows Treg-lowering activity (Figures 15C and 15D). ligand blocking ELISA The ligand blocking activity of 5-B07 was assessed by ELISA. For this purpose, recombinant mouse CTLA-4-Fc protein (Sino Biological Inc.) was coated on 96-well plates (Lumitrac 600 LIA plate, Greiner) at 1 pmol / well. After washing, the titrated doses of anti-CTLA-4 mAb were allowed to bind for 1 hour. His-tagged ligands were added at 200 nM and 100 nM, respectively (rmCD80 and rmCD86; Sino Biological Inc.) and the plates were further incubated for 15 minutes. After washing, bound ligand was detected with an HRP-tagged anti-His antibody (R&D Systems). Super Signal ELISA Pico (Thermo Scientific) was used as a substrate and plates were analyzed using the Tecan Ultra microplate reader. As shown in Figures 15A to 15D, the antibody blocks the binding of (Figure 15A) CD80 and (Figure 15B) CD86 to their CTLA-4 ligand. Treg-lowering activity in vivo The effects of CTLA-4-specific antibodies on T cell subsets in the tumor in vivo were investigated in the CT26 tumor model as described below. Mice were bred and maintained at local facilities in accordance with local office guidelines. Taconic (Bomholt, Denmark) supplied six to eight week old Balb / C females and kept them in local animal facilities. CT26 cells (ATCC) were grown in glutamax-buffered RPMI, supplemented with 10% FCS. When the cells were semi-confluent they were detached with trypsin and resuspended in sterile PBS at 10 x 10 6 cells / ml. Mice were injected s.c. with 100 μΙ of the cell suspension corresponding to 1 x 106 cells / mouse. When the tumors reached approximately 7x7 mm, the mice were treated twice weekly i.p. with 10 mg / kg of the indicated antibodies as indicated in the figures. After the third administration, the tumors were dissected, mechanically divided into small pieces and digested 2 times x 5 min with vertexes between, using a mixture of 100 pg / ml liberase (Roche) and 100 µg / ml of DNase (Sigma) at 37 °C. The cell suspension (400 g for 10 min) was then washed with PBS containing 10% FBS. Subsequently, cells were resuspended in MACS buffer and stained with a panel of antibodies that stained CD45, CD3, CD8, CD4, and CD25 (all from BD Biosciences). Prior to staining, / UUZDUU cells were blocked against non-specific binding using 100 gg / ml IVIG. Cells were analyzed using a FACS Verse (BD Biosciences). Mouse Treg cells were identified as CD45+CD3+CD4+CD25+ cells. As shown in Figure 15C, 5-B07 in mouse IgG2a format mediates Treg deletion in the tumor associated with D) increased CD8 / Treg ratio compared to other CTLA-4-specific n-CoDeR antibodies and the commercially available well described clone 9H10. Example 6 - Generation of a virus that expresses mAb anti-CTLA4 (COPTG19385) or mAb anti-CTLA4 and GM-CSF (COPTG19384), expression of transqenes and characterization of genetic stabilities COPTG19384 and COPTG19385 are vaccinia viruses (Copenhagen strain) encoding the anti-CTLA4 monoclonal antibody (4-E03). COPTG19384 further encodes human GM-CSF. More particularly, COPTG19384 and COPTG19385 are defective for thymidine kinase (TK, J2R locus) and ribonucleotide reductase (RR, I4L locus) activities. As illustrated in Figure 16, the expression cassette encoding the heavy chain 4-E03 (HC; SEQ ID NO: 54) under the control of the p7.5K promoter (SEQ ID NO: 59) was inserted into the J2R locus. , and the expression cassette encoding the 4-E03 IgG light chain (LC, SEQ ID NO: 53) under the control of the p7.5K promoter SEQ ID NO: 59) was placed in the I4L locus. For COPTG19384, the expression cassette encoding human GM-CSF (SEQ ID NO: 56) under the control of the pSE / L promoter (SEQ ID NO: 61) was also placed in the I4L locus. The same promoter (p7.5K) was used to control the expression of HC and LC to obtain the same level of expression for both chains and therefore optimal assembly of the antibody as a heterotetrameric protein (i.e., to avoid excess of unassociated string). However, the same promoter for both antibody chains prevents insertion at the same locus (identical DNA sequences increase the risk of recombination and after transgene deletion). Thus, the cassette encoding 4-E03 HC was inserted into the J2R locus and the cassette encoding 4-E03 LC into the I4L locus. The cassette encoding the GM-CSF transgene but under a different promoter (pSE / L) was also inserted into the I4L locus, like the antibody light chain. Generation of COPTG19384 The vaccinia virus transfer plasmids, pTG19339 and pTG19341, were designed to allow insertion of nucleotide sequences by homologous recombination into the J2R and I4L loci of the vaccinia virus genome, respectively. They originate from plasmid pUC18 into which the flanking sequences (BRG and BRD) surrounding the J2R (pTG19339) or I4L (pTG19341) locus were cloned. Each plasmid also contains the p7.5K promoter. IVIA / a / ZUZ I / UUZDUU A synthetic fragment called the 1436 bp HC Fragment containing the HC gene of the 4-E03 antibody was produced. An LC fragment containing the 4-E03 antibody LC gene and hGM-CSF gene under the control of pSE / L was generated by a synthetic route and inserted into a plasmid vector. Coding sequences were optimized for human codon usage, a Kozak (ACC) sequence was added before the ATG start codon and a transcriptional terminator (T ι l l i NT) was added after the stop codon. In addition, some patterns were excluded: l l l l l NT, GGGGG, CCCCC which are deleterious for expression in poxviruses. The HC fragment was inserted into XVI / II restricted pTG19339 by homologous recombination, giving rise to pTG19367. The LC-bearing plasmid was restricted by SnaB1 and the resulting LC-GMCSF fragment was inserted by homologous recombination into ΡνιΛΊ-restricted pTG19341, giving rise to pTG19384. In this plasmid, the expression cassettes were inserted head to tail between the recombination arms allowing homologous recombination at the I4L locus of the vaccinia virus genome. COPTG19384 was generated in chicken embryo fibroblast (CEF) by two successive homologous recombinations for successive insertion into the I4L and J2R loci and by using COPTG19156 as parental virus and the two transfer plasmids pTG19367 and pTG19384. CEFs were isolated from 12-day-old embryonated SPF eggs (Charles River). Embryos were mechanically dislocated, solubilized in Tryple Select solution (Invitrogen) and cultured in an MBE (Eagle Based Medium; Gibco) supplemented with 5% FCS (Gibco) and 2 mM L-glutamine. Homologous recombination between the transfer plasmids and the parental vaccinia virus allows the generation of recombinant vaccinia viruses that have lost the GFP and mCherry expression cassettes and have gained the antibody and GM-CSF expression cassettes. COPTG19156 contains the mCherry gene expression cassette at its I4L locus and the GFP gene expression cassette at its J2R locus. Homologous recombination between the transfer plasmid pTG19367 and the parental COPTG19156 allows the generation of recombinant vaccinia viruses that have lost their GFP expression cassette and gained the 4-E03 heavy chain expression cassette and selection was performed by isolation of red fluorescent plates. This intermediate recombinant virus (COPTG19367) was used as the parent virus for a second round of homologous recombination with pTG19384 as the transfer plasmid for the generation of recombinant vaccinia viruses that have lost their mCherry expression cassette and gained the upstream expression cassettes. light 4-E03 and GM-CSF. Selection of COPTG19384 (Figure 16) was performed by isolating non-fluorescent white plaques. COPTG19384 viral stock was amplified in CEF in two F175 flasks to generate adequate virus stocks that can be aliquoted and stored at -80 °C until use. The viral stock was titrated in CEF cells and the infectious titers were expressed in pfu / ml and calculated with the following formula: number of lytic areas x dilution factor x 4. For illustrative purposes, the produced viral stock was titrated 6.8 x 106pfu / ml. This pool was analyzed by PCR to verify the integrity of the expression cassettes and recombination arms using the appropriate primer pairs. The pool was also analyzed by sequencing of both expression cassettes. Alignment of sequencing results showed 100% homology to the expected theoretical sequence. When necessary, viral preparations were purified using standard techniques (eg as described in WO2007 / 147528). Generation of COPTG19385 The vaccinia virus transfer plasmids, pTG19339 and pTG19341, were designed to allow insertion of nucleotide sequences by homologous recombination into the J2R and I4L loci of the vaccinia virus genome, respectively. They originate from plasmid pUC18 into which the flanking sequences (BRG and BRD) surrounding the J2R (pTG19339) or I4L (pTG19341) locus were cloned. Each plasmid also contains the p7.5K promoter. A synthetic fragment called the 1436 bp HC Fragment containing the HC gene of the 4-E03 antibody was produced. Coding sequences were optimized for human codon usage, a Kozak (ACC) sequence was added before the ATG start codon and a transcriptional terminator (ι l l l NT) was added after the stop codon. In addition, some patterns were excluded: l l l l l NT, GGGGG, CCCCC which are deleterious for expression in poxviruses. The HC fragment was inserted into PvuII restricted pTG19339 by homologous recombination, giving rise to pTG19367. The plasmid containing the expression cassette encoding only the 4E03 light chain was obtained by deletion of the cassette encoding the hGM-CSF gene under the control of pSE / L in plasmid pTG19384 (described above). pTG19384 was restricted with Nhel and Xbal (compatible sticky ends) and ligated again, resulting in pTG19385. COPTG19385 was generated in chicken embryo fibroblast (CEF) by two successive homologous recombinations for successive insertion into the J2R and I4L loci and by using COPTG19156 as parental virus and the two transfer plasmids pTG19367 and pTG19385. CEFs were isolated from 12-day-old embryonated SPF eggs (Charles River). Embryos were mechanically dislocated, solubilized in Tryple Select solution (Invitrogen) and cultured in an MBE (Eagle Based Medium; Gibco) supplemented with 5% FCS (Gibco) and 2 mM L-glutamine. Homologous recombination between the transfer plasmids and the parental vaccinia virus allows the generation of recombinant vaccinia viruses that have lost the / UUZDUU GFP and mCherry expression cassettes and have gained the antibody expression cassettes. COPTG19156 contains the mCherry gene expression cassette at its I4L locus and the GFP gene expression cassette at its J2R locus. Homologous recombination between the transfer plasmid pTG19367 and the parental COPTG19156 allows the generation of recombinant vaccinia viruses that have lost their GFP expression cassette and gained the 4-E03 heavy chain expression cassette and selection was performed by isolation of red fluorescent plates. This intermediate recombinant virus (COPTG19367) was used as the parent virus for a second round of homologous recombination with pTG19385 as the transfer plasmid for the generation of recombinant vaccinia viruses that have lost their mCherry expression cassette and gained the upstream expression cassette. light 4-E03. Selection of COPTG19385 was performed by isolating non-fluorescent white plaques. COPTG19385 viral stock was amplified in CEFs in two F175 flasks to generate appropriate virus stocks that can be aliquoted and stored at -80 °C until use. The viral stock was titrated in CEF cells and the infectious titers were expressed in pfu / ml and calculated with the following formula: number of lytic areas x dilution factor x 4. For illustrative purposes, the produced viral stock titrated 1.04 x 107pfu / ml. This pool was analyzed by PCR to verify the integrity of the expression cassettes and recombination arms using the appropriate primer pairs. The pool was also analyzed by sequencing of both expression cassettes. Alignment of sequencing results showed 100% homology to the expected theoretical sequence. When necessary, viral preparations were purified using standard techniques (eg as described in WO2007 / 147528). expression of transactions Supernatants of COPTG19384-infected CEF cells were assessed for virus-mediated expression of the 4-E03 monoclonal antibody by Western blotting (WB) and compared to the recombinantly produced antibody (40 ng of 4-E03). WB allows visualization of the presence of non-functional molecules that do not bind to CTLA4 (eg, molecules with incomplete chain assembly, aggregates). CEF cells were infected at MOI 0.05 with COPTG19384 viral stock in triplicate. Cell supernatants were collected after 48 hours and analyzed by WB after electrophoresis in a non-reducing condition and using an anti-Ig (left blot) or anti-light chain (right blot) HRP-conjugated antibody. The results illustrated in Figure 17A indicate that the WB profile in non-reducing condition of the mAb produced by infected CEFs is close to that of purified 4-E03 with a similar apparent size between 100 and 150 kDa, indicating correct folding and chain assembly. / UUZDUU Quantification in supernatants of the secreted functional 4-E03 antibodies and GM-CSF was performed with ELISA. The ELISA allowed quantitative measurement of the amount of functional polypeptide produced in cell supernatants. WTG17137 was used as a negative control. This is a vaccinia virus (Copenhagen strain) deleted at the J2R and I4L loci encoding the FCU1 suicide gene (described in WO2009 / 065546). For 4-E03 antibody estimation, microplates were coated by overnight incubation at 4 °C with 100 µl per well of CTLA4-Fc at 0.25 pg / ml. After incubation, the coating solution was discarded, the blocking solution was added, and the plates were incubated for 1-2 hours at RT before washing. Calibration standards 4-E03 (0.097 to 100 ng / mL), or samples (in triplicate) diluted in the blocking solution, were added to the wells and the plates were incubated for 2 hours at 37°C before washing. HRP-conjugated antibody diluted in the blocking solution was added to each well and the plates were incubated for 1 hour at 37°C before washing. After incubation with TMB solution 30 min at RT in the dark, 2 M H2SO4 (stop solution) was added to stop the enzymatic reaction. The absorbances were read at 450 nm in a microplate reader. Absorbances were plotted against the antibody concentration of the calibration standard. As illustrated in Figure 17B, functional mAb 4-E03 was produced in COPTG19384-infected cells, reaching concentrations close to 1 pg / ml. Virus-mediated expression of GM-CSF in the same supernatants was also assessed using the Quantikine® ELISA (R&D Systems Ref SGM00). Briefly, this assay uses two anti-hGM-CSF antibodies. The first antibody used to capture the hGM-CSF in the samples was coated onto the surface of the wells of a 96-well plate. The second is conjugated and added to the plate in solution to detect the captured hGM-CSF. The concentration of hGM-CSF in the sample is then calculated by interpolation from a calibration curve established with some purified hGM-CSF provided by the kit. As illustrated in Figure 18, the expression level of GM-CSF was approximately 6 pg / ml. In conclusion, concentrations equal to or greater than 1 pg / ml were detected for both transgenes, indicating a satisfactory level of expression. Genetic stability: Genetic stability tests were performed after five passages of the virus in CEF in serum-free medium with a multiplicity of infection (MOI) of 10-4. Passage P5 was diluted and inoculated into CEF cells in 60 mm culture dishes to obtain 20 to 40 viral plaques per dish. One hundred viral plaques were isolated and subcultured. After one cycle of amplification, isolated viral plaques were inoculated into CEF cells and tested by PCR and ELISA. PCR analysis and transgene expression showed that more than 90% of / UUZDUU clones have a correct profile. Since the acceptance criterion for clinical development of a product is genetic stability greater than or equal to 90%, COPTG19384 was considered genetically stable. Example 7 - In vitro characterization of COPTG19384 Hepatocyte replication studies: tumor selectivity of a virus expressing anti-CTLA4 mAb and GM-CSF COPTG19384 carries two deletions of genes encoding viral enzymes (TK and RR) involved in nucleotide metabolism. When functional, these enzymes allow the virus to replicate in the cytoplasm of most cells, including those in a quiescent state (ie, with a low available nucleotide background). A host range study was performed in primary and malignant cells to verify that insertion of the different transgenes into the J2R and I4L loci does not modify host range selectivity. The replication of COPTG19384 was evaluated in normal primary human cells (hepatocytes, prepared by Biopredic) and in tumor cells from the same organ (HepG2 from hepatocellular carcinoma, ATCC® HB-8065™). Replication rates and therapeutic indices were calculated and compared to those of wild-type Copenhagen vaccinia (COP WT, non-deletion virus) as references for non-selective vaccinia virus and recombinant double-deletion WTG17137 virus (deleted in the 1990s). J2R and I4L genes with the FCU1 suicide gene inserted in place of J2R), respectively. Two batches were tested, a research batch (Batch 1) and a GMP-produced batch (Batch 2). The replication rate was determined as the ratio of total infectious particles at the end of incubation / initial infectious particles (inoculum). The therapeutic index of each virus was determined as the ratio: replication rate in HepG2 cells / replication rate in hepatocytes. The higher the ratio, the better the selectivity of the virus towards tumor cells. Primary hepatocytes were cultured in hepatic basal cell medium supplemented with 1.6% additives for hepatocyte culture medium. HepG2 were seeded in 12-well plates at 4 E+05 cells / well and incubated for 24 h at 37 °C with 25% CO. Prior to infection, the culture medium was removed and 70 pfu / well of virus in PBS for hepatocytes or FCS-supplemented PBS for HepG2 was added to each well. Infected cells were incubated for 30 min at 37 °C with 5% CO2 and then 1.5 ml / well of culture medium were added. The plates were incubated at 37 °C with 5% CO2 for 3 days and then stored at -80 °C. The plates were then thawed and the wells sonicated for 30 seconds at 40% amplitude before titration in Vero cells. Replication rate in normal human hepatocytes: Normal hepatocytes were chosen to monitor the capacity of COPTG19384 in normal human cells, since these primary cells can regularly be obtained / UUZPUU directly from donors. In those cells, the WT COP spread well with a replication rate of more than 50,000 (Figure 19A). In other words, each initial infection virus particle produced approximately 50,000 new viruses. In the case of the two double deletion recombinant viruses (ie, WTG17137 and COPTG19384), this replication rate was dramatically reduced from 5 to 15 depending on the virus or virus batch ( Figure 19A ). This latter result indicates that the attenuated replication towards normal cells caused by the two deletions was conserved between WTG17137 and COPTG19384. Replication rate in HepG2 tumor cells: HepG2 cells were chosen to monitor the ability of COPTG19384 to replicate in human tumor cells since these cells are a malignant counterpart to normal hepatocytes. In those cells, all five viruses tested had a fairly similar replication rate, reaching approximately 100,000 new viruses, whichever was the initial virus tested (Figure 19B). Therefore, double deletion in WTG17137 and COPTG19384 and vectoring of the transgenes did not impair their ability to replicate in malignant cells. therapeutic index: As illustrated in Figure 19C, the calculated index is only two for COP WT, indicating poor selectivity of COP WT for tumor vs. normal cells. In contrast, for both WTG17137 and COPTG19384 (and for both virus batches tested) this index ranges from 8.2 E+03 to 1.8 E+04. This confirms that the two recombinant viruses have the same good selectivity towards tumor cells versus normal cells. These results demonstrated that COPTG19384 and WTG17137 have very similar replicative properties in tumor and healthy cells. Compared to COP WT, its replication in tumor Hep G2 is similar while it is highly altered in healthy hepatocytes. Thus, deletion of the two genes (J2R and I4L) restricts replication of the deleted virus to multiplying cells (ie, with a large nucleotide background), including tumor cells. Since the expression and replication of the transgenes are tightly coupled, COPTG19384 is an efficient vector for selective delivery of therapeutic proteins into the tumor. Replication assays in CEF v LoVo: The replication of COPTG19384 was assessed in CEF (producer cells) isolated from 11- or 12-day-old embryonated specific pathogen-free eggs (Charles Rivers) and in a human tumor cell line (LoVo; ATCC® CCL-229™). CEF and LoVo cells in suspension were prepared and infected at MOIs of 10'3 for CEF and 10'2 for LoVo (three wells per cell type per time point). After different incubation times, viral titration was performed in Vero cells (CCL-81™). The replication of COPTG19384 was compared to that of WTG17137 as a reference. / UUZDUU The results show that the replication of COPTG19384 and WTG17137 was similar in both CEF and LoVo (data not shown). Replication assays on reconstructed human skin: The replication of COPTG19384 in reconstructed human skin (TSkin™ / Full Thickness Human Skin Model) was also evaluated. Thirty-six T-Skin™ samples obtained from (EPISKIN SA) were cultured in 6-well plates and kept in fresh culture medium. WTG17137 and COPTG19384 were distributed in each well (in triplicate) to obtain the final concentration attended (ie, 101 to 105 pfu / well). A corresponding negative control of medium without virus (mock) was also tested. The plates were incubated at 37°C with 5% CO2 for 7 days and the T-Skin™ samples were collected and cut into two pieces. The infectious titer was determined in one of the two pieces by using Vero cells for virus titration. Figure 20 shows that COPTG19384 replicates in reconstructed skin to the same extent as control WTG17137, supporting the fact that vectoring of both GM-CSF and mAb 4-E03 did not change the replication behavior of the virus. vaccinia in reconstructed human skin. oncolytic assay The oncolytic activity is representative of the lytic activity of the viral samples tested in tumor cells. It was evaluated by quantifying cell viability after 5 days of incubation in different tumor cell lines: the LoVo human colorectal adenocarcinoma cell line (ATCC® CCL-229™), the MIA PaCa-2 human pancreatic tumor cell line ( ATCC® CCL-1420) and the human hepatocarcinoma cell line HepG2 (ATCC® HB8065™). The oncolytic activities of COPTG19384 were compared with those of WTG17137 as a reference. A negative control corresponding to uninfected cells (Mock infected cells) was also plated. Cells were prepared, distributed in Eppendorf tubes (1.2 x 106 cells / tube) before infecting them with the virus at an MOI of 10.5 to 10.2 and incubated for 30 min at 37 °C. Appropriate complete medium was added to the Eppendorf tube and an aliquot of this suspension was added to each well (in triplicate) in a 6-well plate containing 2 ml of the appropriate complete medium. The plates were incubated at 37 °C with 5% CO2 for 5 days and cell viability was determined in the Vi-Cell counter. Results were expressed as a percentage of the cell viability of Mock infected cells. Cell supernatants were also recovered for determination of the concentration of mAb 4-E03 and GM-CSF. Figures 21A to 21C show that the oncolytic activities of COPTG19384 and WTG17137 are similar in the three tumor cell lines tested. Expression level of transgenes IVIA / a / ZUZ I / UUZDUU Expression levels of both the 4-E03 monoclonal antibody and GM-CSF were measured by ELISA (as described in Example 6) in culture supernatants of HepG2 and LoVo cells recovered after determination of oncolytic activity (5 days of infection at variable MOI). The expression levels of 4-E03 and GM-CSF were further measured by ELISA (see example 6) in supernatants of 5 cell lines grown under the following conditions, respectively the human gastric carcinoma cell line Hs-746 T (ATCC® HTB-135™), the SK-OV-3 human ovarian tumor cell line (ATCC® HTB-77™), the MIA PaCa-2 human pancreatic tumor cell line (ATCC® CCL-1420), the LoVo human colorectal adenocarcinoma cell line (ATCC® CCL-229™) and the human colorectal carcinoma cell line HCT 116 (ATCC® CCL-247™). Each cell line was cultured (in triplicate) in 6-well plates (106 cells / well ) and incubated at 37 °C with 5 % CO2 for 24 h before being infected at MOI 0.05. Cell supernatants were recovered 48 h after infection for determination of the concentration of mAb 4-E03 and GM-CSF. As expected, MOI, time post infection, and cell line are important parameters influencing the expression level of transgenes in supernatants from infected cells. Figure 22A shows that a fairly replication-permissive (HepG2) and a fairly resistant (LoVo) tumor cell line, when infected with COPTG19384, can produce approximately the same amount of mAb 4-E03 and GM- in their culture supernatants. CSF. However, the maximum expression for HepG2 is reached at a 10-fold lower MOI than for LoVo. Furthermore, for all 5 tumor cell lines tested, transgene expression was greater than 0.1 and greater than 1 pg / ml for mAb 4-E03 and GMCSF respectively (Figure 22B). It should be noted that the ELISA assay used to measure the concentration of mAb 4-E03 uses the CTLA4 antigen to capture the antibody. In other words, the antibodies measured by this assay are at least partially functional (i.e., they recognize their antigen, the other functions of the antibody are carried by the Fe part), Purification of mAb 4-E03 v analysis of allycosylation profile In order to produce a fairly large amount of mAb 4-E03 from infected cells, 15 F175 flasks containing ~4.7 107 Mia-PACA cells / flask were infected at MOI of 0.01 with COPTG19384 and incubated 72 h. Culture supernatant of MIA Paca-2 cells (approximately 450 mL containing 670 pg mAb 4-E03 as determined by ELISA) was collected, mixed, and clarified by centrifugation to remove most cell debris. The clarified supernatants were filtered on 0.2 pm filters and 2 mM EDTA (to inhibit putative metal proteases) and 20 mM Tris pH 7.5 (to raise the pH) were added. The filtered supernatant was then passed through a ProtA Hitrap column (GE healthcare, ref. 17-5079-01). The column was transferred and connected to a Purifier FPLC (GE Healthcare) and IVIA / a / ZUZ I / UUZDUU purification program (THM / ProtA 1 ml frac bieu injection loop) was applied. The eluted fractions containing the mAb were loaded on NuPage Bis-Tris 4-12% gels (Thermo NP0323) after the addition of Laemlli buffer (Biorad) containing or not containing beta-mercaptoethanol for the reduction or not of the disulfide bonds of the mAb. mAb. The gel was stained with InstantBIue (Expedeon, ISB1L). Three fractions corresponding to the main elution peak were pooled and, after dialysis against the formulation buffer, the antibody concentration was determined by absorbance at 280 nm. The final concentration of the purified mAb was 0.29 mg / ml. The first characterization was an evaluation of chain assembly by electrophoresis under reducing and non-reducing conditions. Under non-reducing conditions, the 2 light and 2 heavy chains assemble to form the native and functional antibody. It appeared that 4-E03 purified from infected MIA PaCa-2 and the recombinantly produced 4-E03 have indistinguishable electrophoresis profiles under both reducing and non-reducing conditions. In other words, 4-E03 purified from infected MIA PaCa-2 has the expected ratio of light to heavy chains and is correctly assembled into 2 light and 2 heavy chain heterotetramers. The presence of this heterotetramer was also confirmed by mass spectrometry. The purified mAb was subjected to mass spectrometry for glycosylation analysis. Briefly, the mAb was either digested or not digested with the IdeS protease which specifically cleaves IgG at the hinge (resulting in F(ab')2 and Fe parts). The masses of the whole antibody or portions of Fe bearing N-glycosylation were determined and each mass was fitted with a theoretical mass calculated from the Fe primary sequence and a glycosylation pattern. The glycosylation profile of the purified mAb 4E03 from infected MIA PaCa-2 was compared to that of recombinantly produced and purified compounds 4-E03 and MabThera as a benchmark for clinically used human IgGl. The results show that the glycosylation profiles of 4-E03 produced from infected PaCa2 MIA had a different glycosylation profile from the two antibody controls with a majority of G0F (88%) whereas both recombinant 4-E03 and MabThera have a similar glycosylation profile with the typical distribution of G0F, GIF and G2F. However, MIA PaCa-2 was suspected to be the cause of the low GIF and G2F species in the purified mAb 4-E03 due to a low level of Beta-1,4-galactosyltransferase 1 transcript which could be the cause of the lack of galactosyl residue (and thus a lack of Glf and G2F) of the 4-E03 expressed in MIA PaCa-2. The same type of purification was further performed followed by mass analysis of the permeate recovered during the purification of the produced COPTG19384 in CEF. The results show that the glycosylation profiles of 4-E03 from infected CEFs were very similar to those of MabThera or recombinant 4-E03. This last result suggests that the glycosylation profile of the antibody is affected more by the cell line used than by the infection itself. / UUZDUU 4-E03 purified from the supernatant of infected MIA PaCa-2 cells (4E03 TG) further exhibits the same binding characteristics as 4-E03 produced recombinantly by CHO (research batch) or HEK (toxic batch) cells. (Figures 23A, 23B and 24A, 24B). This was demonstrated by ELISA (described in Example 1) to test for binding to recombinant human CTLA-4 (Figure 23A) and cynomolgus (Figure 23B) protein. A FACS analysis testing for binding to cells expressing human (Figure 24A) and cynomolgus (Figure 24B) CTLA-4 (see Examples 1 and 2) confirmed similar cross-reactivity and binding affinities for the different batches of CTLA-4. 4-E03. Pattern of disulfide bonds and glycosylation of GM-CSF To investigate the glycosylation pattern and the presence of some disulfide bonds, different human tumor cell lines were infected with COPTG19384 and their supernatants were analyzed by the same WB method. MIA-Paca-2, LoVo, HepG2 and HCT116 cells were infected at MOI 0.01 and incubated 72 h in culture medium without serum. Culture supernatants were collected, clarified by centrifugation, and then filtered on a 0.2 pm filter. Supernatants were stored at -20 °C until analysis. They were treated by addition of 8 µL of 5X Rapid PNGase F buffer followed by incubation at 75°C for 5 minutes. One μΙ of PNGase F (to remove N-glycans from the glycoprotein) was then added and the mixture was incubated for 30 minutes at 50°C. Twenty-five μΙ samples were prepared by adding 5 μΙ 4x Laemmli buffer with or without beta-mercaptoethanol (reducing and non-reducing conditions) prior to Western blotting. Immune complexes were detected using the Amersham ECL Prime Western blot and chemiluminescence was recorded with a Molecular Imager ChemiDOC XRS (Biorad). GM-CSF from infected HCT116, LoVo and MIA PaCa-2 showed the same glycosylation pattern whereas GM-CSF produced by infected HepG2 migrated as a non-N-glycosylated molecule. These results indicate that the GM-CSF produced by COPTG19384-infected human tumor cells has the expected post-translational modifications (ie, disulfide bonds and N-glycosylation). However, these modifications are likely to vary depending on the tumor cell lines used for infection and their specific metabolic state. Example 8: Pharmacokinetics after intratumoral injections of COPTG19384 Expression kinetics in tumor and bloodstream of anti-CTLA4, GM-CSF and virus antibodies after intratumoral invection fi.t.) of vaccinia virus in a LoVo xenoinhibitor model. Protocol LoVo cells 5 x 10 6 cells were implanted in the right flank of Swiss nude / UUZDUU mice (Charles River, France). After approximately two weeks, when the volume of the tumors reached ~120 mm3, the mice were randomized and divided into 6 groups of 15 animals. • Group 1 mice received an i.t. of COPTG19384 at a dose of 1 x 10 4 pfu / mouse in DO (first day of treatment). • Group 2 mice received an i.t. of COPTG19384 at a dose of 1 x 10 5 pfu / mouse in OD. • Group 3 mice received an i.t. of WTG17137 at a dose of 1 x 10 4 pfu / mouse in DO. • Group 4 mice received an i.t. of WTG17137 at a dose of 1 x 10 5 pfu / mouse in DO. • Group 5 mice received an intraperitoneal (i.p.) administration of 4-E03 at a dose of 3 mg / kg in DO. • Group 6 mice received an i.p. of ipilimumab (Yervoy) at a dose of 3 mg / kg in DO. Tumor and blood were collected from 3 animals on days 1, 3, 6, 10, and 20. Tumors were weighed and homogenized for immediate processing. A quarter of the homogenized tumors were collected for virus titer and the remaining suspension was centrifuged and the supernatants were stored at -20 °C until use. The blood was divided into two parts: one was added to a heparin tube (25 IU / 100 μΙ blood) for the titration assay and frozen at -80 °C until analysis. Cleared sera were produced from the other part and stored at -20 °C until use. Virus titer was determined in tumor and blood samples by titration in Vero cells. Virus replication kinetics in the LoVo model: In the LoVo model, in which COPTG19384 was injected once at two doses (1 x 104 or 1 x 105 pfu) virus replication was monitored and compared with that of WTG17137 injected under the same conditions. The results shown in Figure 25 show a significant dispersion of the three measured virus titer values ​​for each time point. However, the results further show that both viruses and at both doses replicate in the tumor and maintain a fairly high titer / g tumor from day 3 to 20 days post-injection. There is no obvious difference in virus titer, at any given time, between the two doses of virus or between the two viruses used. Interestingly, all blood samples were negative for virus detection except one sample (WTG17137, dose: 1 x 107 pfu on day 10) for which only 13 pfu / ml was detected (data not shown). Together these results indicate that after an injection of 1 x 104 or 1 x 105 IVIA / a / ZUZ I / UU4DUU pfu, virus replication was maintained in LoVo tumors for at least 20 days with barely detectable presence in the bloodstream. It should be noted that the LoVo xenograft model is highly favorable for virus replication, as it uses permissive human tumor cells and Swiss nude mice that have severely impaired immune systems with therefore limited antiviral activity. Kinetics of transgene expression in the LoVo model: As expected, the kinetics of transgene expression in the tumor followed the kinetics of virus replication with a maximum concentration (Cmax) at days 6 or 10 for mAb 4-E03 (Figure 26A) and GM-CSF ( Figure 26B). For the single injection of mAb 4-E03 (or ipilimumab), Cmaxes in tumor and blood were observed at the first time point (day 1) and mAb concentrations measured subsequently were consistent with the pharmacokinetics of a single injection. Human IgGl in mouse (Figures 27A and 27B). Furthermore, 4-E03 concentrations in the tumor at Cmax and subsequently (i.e., 6-10-20 days post-injection) were approximately 10-fold higher after COPTG19384 treatment (for both doses) than after from a single injection of 4-E03 mAb at a therapeutic dose of 3 mg / kg (Figure 26A). In contrast, the blood concentration of the mAb after COPTG19384 treatment was always lower than those measured after i.p. injection. of 3 mg / kg of 4-E03 (Figure 27A). This result indicates that mAb vectoring allows to achieve a high concentration in the tumor without exceeding or even reaching the blood concentration obtained with therapeutic mAb dosing. The kinetics of GM-CSF expression after COPTG19384 treatment follows that observed with 4-E03 (Figure 26B). Interestingly, the GM-CSF levels measured in the tumor are below the level of 4-E03 for the same samples, although LoVo infected in vitro by COPTG19384 express approximately twice as much GM-CSF as 4-E03. Blood concentrations of GM-CSF were also very low compared to 4-E03 (Figure 27B). This result is in agreement with the in vivo half-life of GM-CSF which is very short compared to that of a human IgGl. These results indicate that the vectored antibodies and GM-CSF are mainly expressed in the tumor after i.t. of COPTG19384 with minimal systemic exposure. These results confirm that vectoring is particularly suitable for transgenes with toxicological (eg anti-CTLA4) or pharmacokinetic (eg GM-CSF) concerns. Tumor and bloodstream expression kinetics of antiCTLA4, GM-CSF, and virus antibodies after intratumoral injection of vaccinia virus in a syngeneic CT26 model ΜΛ / a / zuz i / uuzouu Evaluation of viral activities in the immunocompetent CT26 murine model requires the generation of several surrogate viruses encoding a murine anti-mCTLA4 with or without murine GM-CSF: • COPTG19407 is a Vaccinia virus (Copenhagen strain) containing an expression cassette encoding the murine IgG2 m5-B07 heavy chain (SEQ ID NO: 63) under the p7.5 promoter at the J2R locus, and an expression cassette encoding expression encoding the m5B07 light chain (SEQ ID NO: 62) under the p7.5 promoter and murine GM-CSF (SEQ ID NO: 58) under the pSE / L promoter at the I4L locus. • COPTG19421 is a Vaccinia virus (Copenhagen strain) containing an expression cassette encoding the m5-B07 heavy chain under the p7.5 promoter at the J2R locus, and an expression cassette encoding the m5-B07 light chain. under the p7.5 promoter at the I4L locus. • WTG18058, used as reference, is a Vaccinia virus (Copenhagen strain) deleted in the J2R and I4L genes, without any transgene (empty virus). These vaccinia viruses were generated as the human counterparts by two successive homologous reassortments at the J2R (TK) and then I4L (RR) loci following the process described in Example 6. The ELISA method to quantify m5-B07 and mGM-CSF antibodies was similar to that described above (example 6, transgene expression for 4-E03 and GM-CSF) except that murine CTLA4-Fc antigen was used to capture the murine antibody and the Quantikine ELISA Mouse GM-CSF kit (R&D Systems) was used. to quantify mGM-CSF. The oncolytic activity of these viruses was further evaluated in several cell lines (one sarcoma: MCA205 and two colon carcinomas CT26 and MC38) and found to be similar to that of WTG18058, showing that murine antibody vectoring with or without mGM-CSF had no impact on the oncolytic abilities of vaccinia virus (data not shown). Protocol: CT26 cells (2 x 105 cells) were implanted in the right flank of Balb / c mice (Charles River, France). After approximately one week when the volume of the tumors reached 25-50 mm3 the mice were randomized and divided into 3 groups of 20 animals (groups 1 to 3) and a group 4 of 10 animals. Tumor and blood were collected and treated as described for the LoVo model, except that they were collected on days 1, 4, 8, and 10 for the first 3 groups and on day 1 for group 4. • Group 1 mice received an i.t. of WTG18058 at a dose of 1 x 107 pfu / mouse on OD, D2 and D4. • Group 2 mice received an i.t. of COPTG19407 at a dose of 1 x 107 pfu / mouse on OD, D2 and D4. IVIA / a / ZUZ I / UUZDUU • Group 3 mice received i.t. of COPTG19421 at a dose of 1 x 107 pfu / mouse on OD, D2 and D4. • Group 4 mice received i.p. of m5-B07 at a dose of 3 mg / kg in DO. Virus replication kinetics in the CT26 model: In the CT26 model, where two surrogate viruses were injected three times (1 x 107 pfu / injection), virus replication was monitored and compared with that of WTG18058 injected under the same conditions. The results shown in Figure 28 show, as for the LoVo model, a significant dispersion of the three values ​​of virus titers measured for each moment. However, the titers for all three viruses were maintained over time and up to 10 days, indicating that the two transgenes did not affect virus clearance or replication, at least in this time window. Infectious virus particles were not detected in any of the blood samples (data not shown). Kinetics of transgene expression in the CT26 model As in the LoVo model, expression of transgenes in the tumor reflected virus replication. In other words, m5-B07 antibody (Figure 29A) and mGM-CSF (Figure 29B) were detected in the tumor at a fairly constant level during the 10-day follow-up. In the case of the monoclonal antibody, the Cmax achieved after injections of COPTG19421 or COPTG19407 was approximately 10-fold lower than the Cmax observed with i.p. injection. single m5-B07 antibody at 3 mg / ml (Figure 29A). In serum, the difference was even more pronounced with ~100-fold lower circulating m5-B07 concentration after virus treatments versus 3 mg / kg m5-B07 injection (Figure 30). For GM-CSF, only COPTG19407 treatment produced a measurable concentration of mGM-CSF in CT26 tumors, indicating that the measured cytokine was of recombinant, rather than endogenous, origin. As in the LoVo model, the mGM-CSF concentrations measured in the tumor were lower than those of m5-B07 (Figure 29B). Furthermore, mGMCSF produced by the tumor was not detectable in any serum samples, probably due to a short half-life of the molecule that prevents any systemic accumulation. Example 9: Studies of antitumor activity COPTG19347 is a Vaccinia virus (Copenhagen strain) deleted in the J2R and I4L genes and encoding a complete murine antibody (namely m5-B07, heavy and light chain) that recognizes the murine CTLA4 antigen. COPTG19421 vs. COPTG19347 both expressed m5-B07 but under different promoters: namely p7.5K and pH5.R respectively. Quantitation of m5-B07 in infected cell supernatants was assessed ranging from approximately 1 pg / ml in CT26 infected at MOI 101 to approximately 4 pg / ml in / UUZDUU cells. MCA205 infected at MOI 10'2. The highest expression in MCA205 vs. CT26 was also observed for mGM-CSF in the culture supernatant of COPTG19407-infected cells (data not shown). Antitumor activity in mice carrying the CT26 model in combination with anti-PD1 Protocol: CT26 cells (2 x 105 cells) were implanted in the right flank of Balb / c mice (Charles River, France). When the tumors reached a volume of 25-50 mm3, the mice were randomized into five groups of ten animals. Briefly, mice were treated by three i.t. administrations, 2 days apart, of virus followed by i.p. of murine anti-PD1 (RMP1-14 BioXcell) twice a week for three weeks. More specifically, • Group 1 mice received the vehicle; • Group 2 mice received an i.t. from 1 x 107 pfu of COPTG19347 in OD, D2 and D4; • Group 3 mice received an i.t. of 1 x 107 pfu of COPTG19347 on DO, D2 and D4 and intraperitoneal administration i.p. from 250 pg / mouse of RMP1-14, to D7, Dll, D14, D18 and D22; • Group 4 mice received i.p. 250 pg / mouse of RMP1-14, on D7, Dll, D14, D18 and D22; • Group 5 mice received an i.t. of 1 x 107 pfu of WTG18058 in OD, D2 and D4. The dimensions of the tumors were measured twice a week with calipers and their volumes were calculated by using the formula (n / 6) (length x width 2 ). The animals were sacrificed when their tumor volume reached 2000 mm3. Antitumor activities of COPTG19347 in the CT26 model: As illustrated in Figures 31A and 31B, treatment with COPTG19347 produced not only inhibition of tumor growth (Figure 31A) but also tumor regression which ultimately results in tumor-free mice surviving up to 100 days (Figure 31B). . Treatment with COPTG19347 produced 60% tumor-free mice at day 100. Co-treatment with anti-PD-1 antibody did not significantly improve tumor growth inhibition or the percentage of mice surviving for a long time (approximately 70% from tumor-free mice at day 100). In comparison, RPMI-14 treatment did not provide any antitumor effect (the same behavior as untreated mice (all dead in the first 40 days) while WTG18058 had a Π poor activity (approximately 10% tumor-free mice at day 100). Dose-effect evaluation in the CT26 model: The three surrogate viruses (different promoter to drive m5-B07 and with or without m-GM-CSF) were compared and a dose increase of COPTG19407 versus WTG18058 (7.5 x 104, 7.5 x 105 or 7.5 x 106 pfu) was performed. Experimental conditions were exactly as described above, except that co-treatment with anti-PD1 was omitted. The results of two independent experiments clearly demonstrated that the three tested viruses COPTG19407, COPTG19421 and COPTG19347 had strong antitumor activity at the dose of 7.5 x 106 pfu. This confirms that neither the mGM-CSF encoded in COPTG19407 nor the use of a weaker promoter in COPTG19421 and COPTG19407 impaired the antitumor activity of the weaponized viruses. At the highest dose tested (ie, 7.5 x 106pfu), the number of tumor-free mice at 80 days was between 5 / 10 and 7 / 10 depending on the virus and experiment versus 0 / 10 mice. treated with empty virus as summarized in the following table. / UUZDUU Table 6: Effect of surrogate viruses COPTG19407, COPTG19421 and COPTG19347 on tumor growth after i.t. Virus name TK locus RR locus Dose (pfu) Number of tumor-free mice on D100 COPTG19407 p7.5K-HC* p7.5K-LC*; pSE / L-GM-CSF 7.5 x 106 7 / 10 COPTG19421 p7.5K -HC* p7.5K -LC* 7.5 x 106 5 / 10 COPTG19347 pH5.R-HC* pH5.R-LC* 7.5 x 106 7 / 10 VVTG 18058 - - 7.5 x 106 0 / 10 Simulated - 0 / 10 *HC and LC represent the heavy and light chains of the murine anti-mCTLA4 antibody m5-B07 respectively Furthermore, dose escalation performed with the empty virus (VTGV18058) and with the surrogate COPTG19384 (COPTG19407) demonstrated that even at a relative low dose (7.5 x 104pfu) the antibody-expressing virus still had some clear antitumor activities with 4 / 10 and 2 / 10 tumor-free mice at 80-98 days vs. 0 / 10 for WTG18058 treatment at the same low dose as shown in the Table below. Table 7: Dose effect of COPTG19407 or WTG18058 on tumor growth after i.t. IVIA / a / ZUZ I / UUZDUU Virus name TK locus RR locus Dose (pfu) Number of tumor-free mice on D100 COPTG19407 p7.5K-HC* p7.5K-LO*; pSE / L-GM-CSF 7.5 x 106 7 / 10 7.5 x 105 8 / 10 7.5 x 104 4 / 10 VVTG18058 - - 7.5 x 106 0 / 10 7.5 x 105 0 / 10 7.5 x 104 0 / 10 A summary of survival data is presented in Figure 32 (overall survival graph compiled from two independent studies). Statistical analysis, using a log-rank test, was performed to determine if there were significant differences between the survival of each group. Antitumor activity of COPTG19407 compared to the combination of WTG18058 plus m5-B07 CT26 tumor-bearing mice were prepared as previously described (Example 5). Briefly, CT26 cells were injected s.c. in BALB / c mice. Treatment of the mice was started when the tumors reached approx. 100mm3. Mice were then injected on DO, D2, and D5 with COPTG19407 (8.5 x 106 pfu i.t.), WTG18058 (8.5 x 106 pfu i.t.), m5B07 (10 mg / kg i.p.), or the combination of WTG18058 (8.5 x 106 pfu i.t.) plus m5. -B07 (10 mg / kg i.p.). Then, the dimensions of the tumors were measured twice a week and the mice were sacrificed when the tumors reached 2000 mm3. As shown in Figures 33A to 33E, tumor growth was significantly inhibited when mice were treated with anti-CTLA-4 expressing COPTG19407 virus and GM-CSF, while the combination of knockdown virus plus anti-CTLA4 m5- B07 did not result in improved therapy compared to the use of a single agent. In the groups treated with m5-B07 alone, WTG18058 virus alone, or the combination of m5-B07 and WTG18058, only 20% of the mice survived past day 70 (Figure 33E). In contrast, 90% of the mice survived more than 100 days after COPTG19407 administration, demonstrating the power of the vectoring strategy. Antitumor activity of anti-CTLA-4 and GM-CSF encoding VVTK-RR- in mice bearing A20 subcutaneous murine B-cell lymphoma The A20 cell line is a BALB / c B lymphoma cell line derived from a spontaneous reticulum cell neoplasm found in an aged BALB / cAnN mouse (ATCC TIB208™). Protocol (1): Tumors were induced by subcutaneous injection of 5 x 10 6 A20 cells into the right flank of female Balb / cN mice (Charles River, France). When the tumors reached a mean volume of 95 mm3, 50 mice were randomized into 5 groups of ten animals. • Group 1 mice received vehicle administration on OD, D2 and D4, • Group 2 mice received i.t. of WTG18058 at a dose of 4.75 x 106pfu on OD, D2 and D4, • Group 3 mice received an i.t. of COPTG19407 at a dose of 4.75 x 106 pfu on OD, D2 and D4, • Group 4 mice received i.p. of antiPD-1 antibody at a dose of 250 pg on D7, DIO, D14, D17, D21 and D24, • Group 5 mice received i.t. of COPTG19407 at a dose of 4.75 x 106 pfu on DO, D2 and D4 combined with i.p. of the anti-PD-1 antibody at a dose of 250 pg on D7, DIO, D14, D17, D21 and D24. Antitumor activity: Tumor volumes of all animals were monitored throughout the study. The antitumor activity of the treatments is based on the evaluation of the criteria of tumor doubling time, tumor growth retardation and tumor growth inhibition (% T / C). Tumor doubling time was similar for Groups 1, 2 and 4, ranging from 5.14 days (Group 1) to 6.37 days (Group 2). For Group 3, the tumor doubling time could not be accurately calculated as the tumors did not grow exponentially, indicating greater treatment efficacy, compared to Groups 1, 2 and 4. Similarly, it was calculated the tumor doubling time using only one animal in Group 5. 9 out of 10 mice in Group 3 had tumors that regressed on D15 and did not grow substantially in volume from D25 to end of study on D64. At D64, the tumor volumes of these 9 mice ranged from 4 mm3 (technical limit of tumor detection) to 59.77 mm3. Similarly, in Group 5, tumors regressed in 9 mice after initiation of treatment, to reach values ​​ranging from 7.24 to 63.21 mm3 at end of study / UUZDUU on D64. As can be seen in Figures 34A to 34E presenting the individual tumor volume curves for each treatment group (Groups 1 to 5 corresponding to Figures 33A to 33E), tumors did not grow in animals in groups 3 and 5 that received COPTG19407 confirming the strong antitumor activity of the virus expressing antibodies with or without anti-PD1. Tumor growth delay was calculated by estimating the time required for tumors to reach a mean target volume of 300 mm3. The results were similar to those obtained with tumor doubling time, as this parameter could only be calculated for tumors that reached the target volume of 300 mm3, which was not the case for most of the animals in Groups 3. and 5. Groups 1, 2, and 4 had mean tumor growth delays of 16, 21, and 17 days, respectively, which were not significantly different from each other. In addition, in group 3 (n = 2) they had a mean tumor delay of 14 days, which indicated that the tumors that did grow in this group grew at the same rate as groups 1, 2 and 4, however, these tumors actually regressed in both animals. In comparison, the single tumor that grew in Group 5 (n = 1) had a significantly longer (p < 0.0026) tumor growth delay than all other groups, of 43 days. Tumor growth inhibition (% T / C) was calculated after comparing the mean tumor volume of vehicle-treated Group 1 with the other treatment groups. Group 2 had an optimal %T / C of 34% on D22, indicating transient marginal antitumor activity, but this value increased to 71% on D31. Moderate antitumor activity (10-30%, %T / C) was observed in Group 4. In comparison, Groups 3 and 5 showed marked antitumor activity (%T / C less than 10%) from D27 to D31 (last calculable value of % T / C). Figure 35 illustrates mean tumor volume curves in BALB / cN mice bearing subcutaneous A20 tumors demonstrating the dramatic effect of COPTG19407 virus expressing anti-CTLA4 and Gm-CSF with or without anti-PD1 on tumor growth. Protocol (2): Tumors were induced by subcutaneous injection of 5 x 106 A20 cells into the right flank of BALB / cN female mice. When the tumors reached a mean volume of 80-100 mm3, 90 animals were randomized into 9 groups of 10 animals. • Group 1 mice received an i.t. of vehicle on OD, D2 and D4 • Group 2 mice received i.p. of antiPD-1 antibody at a dose of 250 pg / mouse / injection on DO, D4, D7, DIO, D14 and D17 • Group 3 mice received i.p. of the isotype at a dose of 250 pg / mouse / injection on DO, D4, D7, DIO, D14 and D17 • Group 4 mice received an i.t. from WTG18058 to IVIA / a / ZUZ I / UUZDUU a dose of 1 χ 105pfu on DO, D2 and D4 • Group 5 mice received i.t. of WTG18058 at a dose of 1 χ 105pfu on DO, D2 and D4, and an i.p. isotype at a dose of 250 pg / mouse / injection on DO, D4, D7, DIO, D14 and D17 • Group 6 mice received i.t. of WTG18058 at a dose of 1 χ 105pfu on DO, D2 and D4 and an i.p. of anti-PD-1 antibody at a dose of 250 pg / mouse / injection on DO, D4, D7, DIO, D14 and D17 • Group 7 mice were administered COPTG19407 at a dose of 1 χ 105pfu on DO, D2 and D4 • Group 8 mice received an i.t. of COPTG19407 at a dose of 1 χ 105pfu on DO, D2 and D4, combined with i.p. isotype at a dose of 250 pg / mouse / injection on DO, D4, D7, DIO, D14, D17 and D24. • Group 9 mice received an administration of COPTG19407 at a dose of 1 χ 105pfu on DO, D2 and D4, combined with i.p. of anti-PD-1 antibody at a dose of 250 pg / mouse / injection on OD, D4, D7, DIO, D14 and D17. Antitumor activity: The dose of COPTG19407 is suboptimal and demonstrated mild antitumor activity similar to that of anti-PD-1 treatment in terms of tumor volume and mouse survival. In contrast, the combination of COPTG19407 with anti-PD-1 showed strong antitumor activity resulting in a small tumor volume compared to the other groups as shown in Figure 36A (approximately 290 mm3 at day 36 compared to approximately 630 mm3 and 750 mm3 at day 24 for mice receiving anti-PD-1 alone or COPTG19407 alone respectively), and much better survival of the animals as presented in Figure 36B (7 animals still alive at day 57 in the group 9 versus only two or one in groups 2 or 7 respectively). Anti-tumor activity study of anti-CTLA-4 v GM-CSF encoding WTK-RR in mice bearing C38 subcutaneous colon tumor cells C38 is a murine colon adenocarcinoma originating from the American Type Culture Collection (ATCC CRL-2779™). Protocol: Tumor fragments (30-50 mg) were implanted subcutaneously in the right flank of female C57BL / 6J mice (Janvier, France). When the tumors reached a mean volume of approximately 60 mm3, 50 animals were randomized into five groups of ten animals. / UUZDUU • Group 1 mice received vehicle administration on OD, D2 and D4, • Group 2 mice received i.t. of a WTG18058 at a dose of 4.75 χ 106pfu on DO, D2 and D4, • Group 3 mice received an i.t. of COPTG19407 at a dose of 4.75 χ 106pfu on DO, D2 and D4, • Group 4 mice received i.p. of the murine anti-PD-1 antibody at a dose of 250 pg on D7, DIO, D14, D17, D21 and D24, • Group 5 mice received an administration of COPTG19407 at a dose of 4.75 χ 106pfu on DO, D2 and D4 combined with an i.p. of antiPD-1 antibody at a dose of 250 pg on D7, DIO, D14, D17, D21 and D24. Antitumor activity: As before, the tumor volumes of all animals were monitored throughout the study. Tumor doubling time was similar for Groups 1 and 2 at approximately 6.7 days. Group 4 (n = 5) had a longer tumor doubling time (10.4 days) but there were no significant differences between groups. For Groups 3 and 5, the tumor doubling time was calculable but with fewer animals (n = 2) since most tumors in mice in these groups did not grow exponentially, indicating greater treatment efficacy, in compared to Groups 1, 2, and 4. As can be seen in Figures 37A to 37E, 8 out of 10 mice in Group 3 had tumors that regressed from D15 and did not grow substantially in volume, and five mice had no detectable tumors at end of study on D61. Similarly, in Group 5, tumors regressed in 8 mice after the start of treatment, to reach values ​​ranging from 0 (n = 2) to 47.82 mm3 at the end of the study on D61. Tumor growth delay was calculated by estimating the time required for tumors to reach a mean target volume of 300 mm3. The results were similar to those obtained with tumor doubling time, as this parameter could only be calculated for tumors that reached the target volume of 300 mm3, which was not the case for most of the animals in Groups 3. and 5. There were no significant differences between the groups. Groups 1, 2, and 4 (n=5) had mean delays in tumor growth of 23 to 27 days, respectively. Furthermore, groups 3 (n = 2) and 5 (n = 3) had mean growth delays of 18 and 24 days, respectively. This indicated that the tumors that did grow in these two groups grew at rates similar to those in groups 1, 2, and 4. Tumor growth inhibition (% T / C) was calculated after comparing the mean tumor volume of vehicle-treated Group 1 with the other treatment groups. Group 2 did not show any inhibition of tumor growth, as %T / C remained above 100% for the duration of the study. In comparison, groups 3, 4 and 5 all showed marked antitumor activity (%T / C less than 10%) from D31 (group 3 only) to D42 (last calculable %T / C value). Figure 38 illustrates mean tumor volume curves from C57BL / 6 mice bearing subcutaneous C38 tumors demonstrating the dramatic effect of anti-CTLA4 / GM-CSF expressing COPTG19407 virus with or without anti-PD1 on tumor growth. Study of the antitumor activity of anti-CTLA-4 and GM-CSF that it encodes WTK-RR- in mice bearing EMT6 subcutaneous breast tumor cells EMT6 is a murine breast carcinoma originating from the ATTC (ATCC CRL2755™). Protocol: Tumors were induced by subcutaneous injection of 1 x 10 6 EMT6 cells in female BALB / cByJ mice (Charles River, France). When the tumors reached a mean volume of approximately 51 mm3, fifty mice per individual tumor volume were randomized into five groups of ten animals. • Group 1 mice received vehicle administration on OD, D2 and D4, • Group 2 mice received i.t. of WTG18058 at a dose of 4.75 x 106pfu on OD, D2 and D4, • Group 3 mice received an i.t. of COPTG19407 at a dose of 4.75 x 106 pfu on OD, D2 and D4, • Group 4 mice received i.p. of antiPD-1 antibody at a dose of 250 pg on D7, DIO, D14, D17, D21 and D24, • Group 5 mice received an administration of COPTG19407 at a dose of 4.75 x 106pfu on DO, D2 and D4 combined with an i.p. administration of antiPD-1 antibody at a dose of 250 pg on D7, DIO, D14, D17, D21 and D24. Tumor volumes of all animals were monitored throughout the study by evaluating the criteria of tumor doubling time, tumor growth retardation, and tumor growth inhibition (%T / C). Antitumor activity: Tumor doubling time was similar for Groups 1, 2, and 4 at approximately 5.4 days. For Group 3, the tumor doubling time could not be calculated as most of the tumors did not grow exponentially, indicating higher treatment efficacy, compared to Groups 1, 2 and 4. A similar effect was observed in Group 5, ΜλΌΖϋΖ 1 / UUZDUU where only one animal was used for calculation of tumor doubling time. As can be seen in the individual tumor volume plots (Figures 39A to 39E), 8 out of 10 mice in Group 3 had tumors that regressed from D15 and did not grow substantially in volume, and seven mice had no detectable tumors at the end of the day. of the study in D61. Similarly, in Group 5, tumors regressed in 9 mice after the start of treatment, to reach values ​​ranging from 0 (n = 8) to 13.24 mm3 at the end of the study on D56. Tumor growth delay was calculated after estimating the time required for tumors to reach a mean target volume of 200 mm3. The results were similar to those obtained with tumor doubling time, as this parameter could only be calculated for tumors that reached the target volume of 200 mm3, which was not the case for most of the animals in Groups 3. and 5. Groups 1, 2, and 4 had median delays in tumor growth of approximately 19 days. In addition, groups 3 and 5 (n = 2 for both groups) had mean growth delays of 24 and 12 days, respectively. This indicated that the tumors that did grow in these two groups grew at rates similar to those in groups 1, 2, and 4. There were no significant differences between the groups. Tumor growth inhibition (% T / C) was calculated after comparing the mean tumor volume of vehicle-treated Group 1 with the other treatment groups. Group 2 showed transient marginal tumor growth inhibition on D28 but increased to 79% on D31. Group 4 showed no antitumor activity, with a %T / C > 60% for the duration of the study. In comparison, groups 3 and 5 showed marked antitumor activity (%T / C less than 10%) from D24 to D31 (last calculable %T / C value). Figure 40 illustrates mean tumor volume curves in BALB / cByJ mice bearing subcutaneous EMT6 tumors demonstrating the dramatic effect of anti-CTLA4 / GM-CSF expressing COPTG19407 virus with or without anti-PD1 on tumor growth. Reexposure to CT26 Balb / c mice exposed to CT26 tumor cells that survived after treatment with 104, 105, or 106 pfu of COPTG19421 or 106 pfu of COPTG19407 were re-exposed to CT26 tumor cells or were exposed to Renca cells (renal adenocarcinoma cells: control), to study whether a specific antitumor immune response was generated. / UUZDUU Table 8: Effect of CT26 tumor cell rechallenge or RenCa tumor cell challenge on the number of tumor-free mice IVIA / a / ZUZ I / UUZDUU Group Tumor-free / total mice Control Naive CT26 0 / 5 VVTG18058 105 + CT26 rechallenge 0 / 1 COPTG19421 106 + CT26 rechallenge 3 / 3 COPTG19421 106 + RenCa 0 / 2 challenge COPTG194 07 1 04 + CT26 rechallenge 1 / 1 COPTG194 07 1 04 + exposure to RenCa 0 / 1 COPTG194 07 1 05 + re-exposure to CT26 1 / 2 COPTG194 07 1 05 + exposure to RenCa 0 / 2 COPTG194 07 1 06 + re-exposure to CT26 2 / 4 COPTG194 07 1 06 + exposure to RenCa 0 / 3 The results presented in Table 8 show that 0 / 8 mice that had received COPTG19421 or COPTG19407 were tumor-free after RenCa challenge, whereas 7 / 10 mice that had received COPTG19421 or COPTG19407 were tumor-free after rechallenge. to CT26. This indicates that COPTG19421 and COPTG19407 generated a specific immune memory against CT26 cells.

Claims

1. An antibody molecule that specifically binds to CTLA-4 and has an improved downregulating effect on CTLA-4 positive cells compared to ipilimumab.

2. The antibody molecule according to claim 1, further characterized in that it has an improved downregulating effect on CD4-positive cells compared to ipilimumab.

3. The antibody molecule according to claim 1 or 2, further characterized in that it has an improved downreg effect compared to ipilimumab.

4. The antibody molecule according to any of claims 1 to 3, further characterized in that it is deemed to have an improved downregulating effect on CTLA-4-positive cells, CD4-positive cells and / or Treg cells compared to ipilimumab if it provides an improved downregulation in: (i) an in vitro ADCC assay performed using an NK-92 cell line stably transfected to express the CD16-158V allele together with GFP, wherein the ADCC assay comprises the following consecutive steps: 1) CTLA-4-positive cells, CD4-positive cells or Treg cells are isolated as target cells from peripheral blood of healthy donors; 2) the target cells are then stimulated with CD3 / CD28 and rhIL-2; 3) the target cells are then pre-incubated with the antibody molecule and subsequently mixed with NK cells;4) the target cells are then incubated in RPMI 1640 + GlutaMAX medium containing HEPES buffer 1, sodium pyruvate and low IgG FBS; 5) lysis is determined by flow cytometry; 6) steps 1 to 5 are repeated, or performed in parallel, using ipilimumab instead of the antibody molecule in step 3; and 7) the antibody molecule lysis results are compared with the ipilimumab lysis results, and improved antibody molecule lysis compared with ipilimumab demonstrates that the antibody molecule has an enhanced downregulating effect on CTLA-4 positive cells, CD4 positive cells and / or Treg cells; and / or (i) an in vivo test in a PBIMC-NOG / SCID model, wherein the in vivo test comprises the following consecutive steps: 1) human PBMCs are isolated, washed and resuspended in sterile PBS; 2) NOG mice are injected iv with the cell suspension from step 1);3) The spleens of NOG mice are isolated and placed in a single-cell suspension; 4) the cell suspension from step 3) is resuspended in sterile PBS; 5) the suspension from step 4 is injected into SCID mice; 6) the SCID mice are then treated with the antibody molecule, ipilimumab, or an isotype control monoclonal antibody; 7) intraperitoneal fluid is collected from the treated SCID mice; 8) human T-cell subsets are identified and quantified by FACS using the following markers: CD45, CD4, CD8, CD25, CD127;9) The results of the identification and quantification of T cell subsets from mice treated with the antibody molecule are compared with the results of the identification and quantification of T cell subsets from mice treated with ipilimumab and with the results of the identification and quantification of T cell subsets from mice treated with the isotype control monoclonal antibody, and a lower number of CTLA-4 positive cells, CD4 positive cells and / or Treg cells in the intraperitoneal fluid of mice treated with the antibody molecule to be tested compared with the number of CTLA-4 positive cells, CD4 positive cells and / or Treg cells in the intraperitoneal fluid of mice treated with ipilimumab demonstrates that the antibody molecule has an improved decreasing effect on CTLA-4 positive cells, CD4 positive cells and / or Treg cells compared with ipilimumab.

5. The antibody molecule according to any of claims 1 to 4, further characterized in that the antibody molecule is selected from the group consisting of antibody molecules comprising 1-6 of the CDRs selected from the group consisting of SEQ ID NO: 3, 6, 8, 10, 12 and 14. 6 - The antibody molecule according to any one of claims 1 to 5, further characterized in that the antibody molecule is selected from the group consisting of antibody molecules comprising 1-6 of the CDRs VH-CDR1, VH-CDR2, VH-CDR3, VLCDR1 and VL-CDR3, wherein VH-CDR1, if present, is selected from the group consisting of SEQ ID NO: 15, 22, 29 and 35; wherein VH-CDR2, if present, is selected from the group consisting of SEQ ID NO: 16, 23, 30 and 36; wherein VH-CDR3, if present, is selected from the group consisting of SEQ ID NO: 17, 24, 31 and 37; where VL-CDR1, if present, is selected from the group consisting of SEQ ID NO: 10 and 38. where VL-CDR2, if present, is selected from the group consisting of SEQ ID NO: 18, 25, 32 and 39; where VL-CDR3, if present, is selected from the group consisting of SEQ ID NO: 19, 26 and 40.

7. The antibody molecule according to any of claims 1-6, further characterized in that the antibody molecule comprises the 6 CDRs having SEQ ID NO: 15, 16, 17, 10, 18 and 19 or the 6 CDRs having SEQ ID NO: 22, 23, 24, 10, 25 and 26.

8. An antibody molecule that binds specifically to CTLA-4, wherein the antibody molecules comprise either the 6 CDRs having SEQ ID NO: 15, 16, 17, 10, 18 and 19, or the 6 CDRs having SEQ ID NO: 22, 23, 24, 10, 25 and 26.

9. The antibody molecule according to any of claims 1 to 8, further characterized in that the antibody molecule comprises a variable heavy chain selected from the group consisting of SEQ ID NO: 20 and 27 and / or a variable light chain selected from the group consisting of SEQ ID NO: 21 and 28.

10. The antibody molecule according to any of claims 1 to 9, further characterized in that the antibody molecule comprises the heavy chain constant region SEQ ID NO: 43 and / or the light chain constant region SEQ ID NO:

44. 11 - The antibody molecule according to any of claims 1 to 6, further characterized in that the antibody molecule is an antibody molecule that is capable of competing for binding to CTLA-4 with an antibody molecule as defined in any of claims 7 to 10.

12. The antibody molecule according to any of claims 1 to 11, further characterized in that the antibody molecule is selected from the group consisting of a full-size antibody, a chimeric antibody, a single-chain antibody, a Fab, an Fv, an scFv, a Fab' and a (Fabjz).

13. The antibody molecule according to any of claims 1 to 12, further characterized in that it binds to human CTLA-4 (hCTLA-4) and / or to monkey cynomolgus CTLA-4 (cmCTLA-4) and / or to murine CTLA-4 (mCTLA-4). 14 - The antibody molecule according to any of claims 1 to 13, further characterized in that it does not bind to human CD28.

15. The antibody molecule according to any of claims 1 to 14, further characterized in that the antibody molecule is selected from the group consisting of a human IgG antibody, a humanized IgG antibody, and an IgG antibody of human origin. 16.- The antibody molecule according to claim 15, further characterized in that the antibody molecule is a human IgG1 antibody. 17 - The antibody molecule according to any of claims 1 to 16, further characterized in that the antibody molecule is a monoclonal antibody.

18. An isolated nucleotide sequence encoding an antibody molecule as defined in any of claims 1 to 17.

19. The isolated nucleotide sequence according to claim 18, further characterized in that it comprises or consists of a sequence selected from the group consisting of SEQ ID NO: 45 to 52.

20. A plasmid comprising a nucleotide sequence as defined in claim 18 or 19.

21. A virus comprising a nucleotide sequence as defined in claim 18 or 19 or a plasmid as defined in claim 20.

22. The virus according to claim 21, further characterized in that it is an oncolytic virus and preferably an oncolytic poxvirus.

23. The virus according to claim 22, further characterized in that said poxvirus belongs to the subfamily Chordopoxviridae, more preferably to the genus Orthopoxvirus preferably selected from the group consisting of Vaccinia virus, smallpox virus, canary pox virus, ectromelia virus and myxoma virus.

24. The virus according to claim 23, further characterized in that said oncolytic virus is a vaccinia virus defective for thymidine kinase (TK) and / or ribonucleotide reductase (RR) activities and comprising nucleotide sequences encoding SEQ ID NO: 20 and ID NO: 21 or SEQ ID NO: 53 and ID NO:

54.

25. The virus according to claim 24, further characterized in that said vaccinia oncolytic virus further comprises a nucleotide sequence encoding a GM-CSF, having a specific preference for a human GM-CSF (e.g., having SEQ ID NO: 55 or SEQ ID NO: 56) or a murine GM-CSF (e.g., having SEQ ID NO: 57 or SEQ ID NO: 58).

26. The virus according to any of claims 21 to 25, further characterized in that the cassette encoding the heavy chain is inserted into locus J2R and the cassette encoding the light chain is inserted into locus I4L.

27. A cell comprising a nucleotide sequence as defined in claim 18 or 19, or a plasmid as defined in claim 20, or a virus as defined in any of claims 21 to 26. 28 - An antibody molecule as defined in any of claims 1 to 17, a nucleotide sequence according to claim 18 or 19, a plasmid according to claim 20, a virus according to any of claims 21 to 26 or a cell according to claim 27 for use in medicine.

29. An antibody molecule as defined in any of claims 1 to 17, a nucleotide sequence according to claim 18 or 19, a plasmid according to claim 20, a virus according to any of claims 21 to 26 or a cell according to claim 27 for use in the treatment of cancer.

30. The use of an antibody molecule as claimed in any of claims 1 to 17, a nucleotide sequence as claimed in claim 18 or 19, a plasmid as claimed in claim 20, a virus as claimed in any of claims 21 to 26 or a cell as claimed in claim 27 for the manufacture of a pharmaceutical composition for the treatment of cancer.

31. A pharmaceutical composition comprising or consisting of an antibody molecule as defined in any of claims 1 to 17, a nucleotide sequence as claimed in claim 18 or 19, a plasmid as claimed in claim 20, a virus as claimed in any of claims 21 to 26 or a cell as claimed in claim 27, and optionally a pharmaceutically acceptable diluent, carrier, vehicle and / or excipient.

32. The pharmaceutical composition according to claim 31, for use in the treatment of cancer.

33. The use of a pharmaceutical composition as claimed in claim 31, for the manufacture of a medicament for the treatment of cancer.

34. The antibody molecule for use according to claim 29, the nucleotide sequence for use according to claim 29, the plasmid for use according to claim 29, the virus for use according to claim 29, the cell for use according to claim 29, the use as claimed in claim 30, the pharmaceutical composition for use according to claim 32, or the use as claimed in claim 34, wherein the cancer is a solid cancer.