IL-18BP antagonist antibodies and their use in monotherapy and combination therapy in the treatment of cancer

Abstract

IL-18BP antagonist antibodies and their use in monotherapy and combination therapy in the treatment of cancer are provided. [Solution] The present invention relates to anti-IL18-BP antibodies and uses thereof. The present invention relates to, for example, monotherapy and combination therapy using the immune checkpoint inhibitor antibodies described herein. An object of the present invention is to provide anti-IL18-BP antibodies or uses in disease treatment. The present invention fulfills this need by providing anti-IL18-BP antibodies (including antigen-binding fragments), particularly anti-IL18-BP antibodies that block IL18-BP, which can be used to treat diseases such as cancer.

Description

[Technical Field]

[0001] This application claims priority to U.S. Patent Application Nos. 63 / 320,202, filed March 15, 2022, 63 / 351,242, filed June 10, 2022, and 63 / 478,898, filed January 6, 2023, which are incorporated by reference in their entireties. [Background technology]

[0002] Interleukin 18 (IL-18) stimulates T cells, NK cells, and IL-18 is a proinflammatory cytokine that can stimulate myeloid cells. IL-18 has been proposed as an immunotherapeutic agent for the treatment of cancer, given its ability to stimulate anti-tumor immune cells. However, the clinical effectiveness of IL-18 is limited. Therefore, there is a need for compositions and methods that provide effective IL-18 signaling activity for treating and preventing cancer and other diseases and disorders.

[0003] Interleukin 18 binding protein (IL1 IL18-BP) binds to IL18 and prevents IL18 from binding to the IL18 receptor, thus functioning as an inhibitor of the proinflammatory cytokine IL18. IL18-BP inhibits IL18-induced activation and proliferation of T cells and NK cells, as well as proinflammatory cytokine production, resulting in reduced T cell and NK cell activity and T helper type 1 immune responses.

[0004] It is an object of the present invention to provide anti-IL18-BP antibodies or uses in treating diseases. The present invention fulfills this need by providing anti-IL18-BP antibodies (including antigen-binding fragments), particularly anti-IL18-BP antibodies that block IL18-BP, and can be used to treat diseases such as cancer. Summary of the Invention

[0005] The present invention provides compositions and methods related to anti-IL18-BP antibodies.

[0006] In some embodiments, the invention provides compositions comprising an anti-IL18-BP (interleukin-18 binding protein) antibody for use in treating cancer, for activating T cells, NK cells, NKT cells, dendritic cells, MAIT T cells, γδ T cells and / or innate lymphoid cells (ILCs), and / or for modulating myeloid cells, wherein the antibody antagonizes at least one immunoinhibitory effect of IL18-BP, optionally wherein the anti-IL18-BP antibody blocks the IL18:IL18-BP binding interaction, optionally wherein the anti-IL18-BP antibody exhibits a binding affinity of less than 1 pM.

[0007] In some embodiments, the composition comprises an anti-IL18-BP antibody that competes for binding with an antibody that binds to the secretory chain of human IL18-BP of SEQ ID NO: 254 and / or human IL18-BP of SEQ ID NO: 255 and / or an antibody that competes for binding to IL18.

[0008] In some embodiments, the composition comprises an anti-IL18-BP antibody, wherein the anti-IL18-BP competes for binding with antibodies described in U.S. Patent No. 8,436,148, WO 2019213686, WO 200107480, WO 2019051015, WO 2014126277(A1), WO 2012177595, U.S. Patent Application Publication No. 20140364341, and WO 2018060447.

[0009] In some embodiments, the composition comprises an anti-IL18-BP antibody, wherein the antibody i. The sequences of vhCDR1 (SEQ ID NO: 1), vhCDR2 (SEQ ID NO: 32), vhCDR3 (SEQ ID NO: 3), vlCDR1 (SEQ ID NO: 4), vlCDR2 (SEQ ID NO: 5), and vlCDR3 (SEQ ID NO: 6) of Figure 1A (66650); ii. The sequences of vhCDR1 (SEQ ID NO: 7), vhCDR2 (SEQ ID NO: 8), vhCDR3 (SEQ ID NO: 9), vlCDR1 (SEQ ID NO: 10), vlCDR2 (SEQ ID NO: 11), and vlCDR3 (SEQ ID NO: 12) of Figure 1B (66670); iii. The sequences of vhCDR1 (SEQ ID NO: 13), vhCDR2 (SEQ ID NO: 14), vhCDR3 (SEQ ID NO: 15), vlCDR1 (SEQ ID NO: 16), vlCDR2 (SEQ ID NO: 17), and vlCDR3 (SEQ ID NO: 18) of Figure 1C (66692); iv. The sequences of vhCDR1 (SEQ ID NO: 19), vhCDR2 (SEQ ID NO: 20), vhCDR3 (SEQ ID NO: 21), vlCDR1 (SEQ ID NO: 22), vlCDR2 (SEQ ID NO: 23), and vlCDR3 (SEQ ID NO: 24) of Figure 1D (66716); v. The sequences of vhCDR1 (SEQ ID NO: 25), vhCDR2 (SEQ ID NO: 26), vhCDR3 (SEQ ID NO: 27), vlCDR1 (SEQ ID NO: 28), vlCDR2 (SEQ ID NO: 29), and vlCDR3 (SEQ ID NO: 30) of Figure 1E (66650); vi. The sequences of vhCDR1 (SEQ ID NO: 31), vhCDR2 (SEQ ID NO: 32), vhCDR3 (SEQ ID NO: 33), vlCDR1 (SEQ ID NO: 34), vlCDR2 (SEQ ID NO: 35), and vlCDR3 (SEQ ID NO: 36) of Figure 1F (66670); vii. The sequences of vhCDR1 (SEQ ID NO: 37), vhCDR2 (SEQ ID NO: 38), vhCDR3 (SEQ ID NO: 39), vlCDR1 (SEQ ID NO: 40), vlCDR2 (SEQ ID NO: 41), and vlCDR3 (SEQ ID NO: 42) of Figure 1G (66692); viii. The sequences of vhCDR1 (SEQ ID NO: 43), vhCDR2 (SEQ ID NO: 44), vhCDR3 (SEQ ID NO: 45), vlCDR1 (SEQ ID NO: 46), vlCDR2 (SEQ ID NO: 47), and vlCDR3 (SEQ ID NO: 48) of Figure 1H (66716); ix. The sequences of vhCDR1 (SEQ ID NO: 43), vhCDR2 (SEQ ID NO: 44), vhCDR3 (SEQ ID NO: 45), vlCDR1 (SEQ ID NO: 46), vlCDR2 (SEQ ID NO: 47), and vlCDR3 (SEQ ID NO: 48) of Figure 1H (66716); x. The sequences of vhCDR1 (SEQ ID NO: 844), vhCDR2 (SEQ ID NO: 845), vhCDR3 (SEQ ID NO: 846), vlCDR1 (SEQ ID NO: 847), vlCDR2 (SEQ ID NO: 848), and vlCDR3 (SEQ ID NO: 849) of Figure 1I (66650); xi. The sequences of vhCDR1 (SEQ ID NO: 850), vhCDR2 (SEQ ID NO: 851), vhCDR3 (SEQ ID NO: 852), vlCDR1 (SEQ ID NO: 853), vlCDR2 (SEQ ID NO: 854) and vlCDR3 (SEQ ID NO: 855) of Figure 1J (66670); xii. The sequences of vhCDR1 (SEQ ID NO: 856), vhCDR2 (SEQ ID NO: 857), vhCDR3 (SEQ ID NO: 858), vlCDR1 (SEQ ID NO: 859), vlCDR2 (SEQ ID NO: 860), and vlCDR3 (SEQ ID NO: 861) of Figure 1K (66692); xiii. The sequences of vhCDR1 (SEQ ID NO: 862), vhCDR2 (SEQ ID NO: 863), vhCDR3 (SEQ ID NO: 864), vlCDR1 (SEQ ID NO: 865), vlCDR2 (SEQ ID NO: 866), and vlCDR3 (SEQ ID NO: 867) of Figure 1L (66716); xiv. The sequences of vhCDR1 (SEQ ID NO: 55), vhCDR2 (SEQ ID NO: 56), vhCDR3 (SEQ ID NO: 57), vlCDR1 (SEQ ID NO: 60), vlCDR2 (SEQ ID NO: 61), and vlCDR3 (SEQ ID NO: 62) of Figure 2A (71709); xv. The sequences of vhCDR1 (SEQ ID NO: 65), vhCDR2 (SEQ ID NO: 66), vhCDR3 (SEQ ID NO: 67), vlCDR1 (SEQ ID NO: 70), vlCDR2 (SEQ ID NO: 71), and vlCDR3 (SEQ ID NO: 72) of Figure 2B (71719); xvi. The sequences of vhCDR1 (SEQ ID NO: 75), vhCDR2 (SEQ ID NO: 76), vhCDR3 (SEQ ID NO: 77), vlCDR1 (SEQ ID NO: 80), vlCDR2 (SEQ ID NO: 81), and vlCDR3 (SEQ ID NO: 82) of Figure 2C (71720); xvii. The sequences of vhCDR1 (SEQ ID NO: 85), vhCDR2 (SEQ ID NO: 86), vhCDR3 (SEQ ID NO: 87), vlCDR1 (SEQ ID NO: 90), vlCDR2 (SEQ ID NO: 91) and vlCDR3 (SEQ ID NO: 92) of Figure 2D (71722); xviii. The sequences of vhCDR1 (SEQ ID NO: 95), vhCDR2 (SEQ ID NO: 96), vhCDR3 (SEQ ID NO: 97), vlCDR1 (SEQ ID NO: 100), vlCDR2 (SEQ ID NO: 101) and vlCDR3 (SEQ ID NO: 102) of Figure 2E (71701); xix. The sequences of vhCDR1 (SEQ ID NO: 105), vhCDR2 (SEQ ID NO: 106), vhCDR3 (SEQ ID NO: 107), vlCDR1 (SEQ ID NO: 110), vlCDR2 (SEQ ID NO: 111) and vlCDR3 (SEQ ID NO: 112) of Figure 2F (71663); xx. The sequences of vhCDR1 (SEQ ID NO: 115), vhCDR2 (SEQ ID NO: 116), vhCDR3 (SEQ ID NO: 117), vlCDR1 (SEQ ID NO: 120), vlCDR2 (SEQ ID NO: 121) and vlCDR3 (SEQ ID NO: 122) of Figure 2G (71662); xxi. The sequences of vhCDR1 (SEQ ID NO: 125), vhCDR2 (SEQ ID NO: 126), vhCDR3 (SEQ ID NO: 127), vlCDR1 (SEQ ID NO: 130), vlCDR2 (SEQ ID NO: 131) and vlCDR3 (SEQ ID NO: 132) of Figure 2H (66692); xxii. The sequences of vhCDR1 (SEQ ID NO: 135), vhCDR2 (SEQ ID NO: 136), vhCDR3 (SEQ ID NO: 137), vlCDR1 (SEQ ID NO: 140), vlCDR2 (SEQ ID NO: 141) and vlCDR3 (SEQ ID NO: 142) of Figure 2I (71710); xxiii. The sequences of vhCDR1 (SEQ ID NO: 145), vhCDR2 (SEQ ID NO: 146), vhCDR3 (SEQ ID NO: 147), vlCDR1 (SEQ ID NO: 150), vlCDR2 (SEQ ID NO: 151) and vlCDR3 (SEQ ID NO: 152) of Figure 2J (71717); xxiv. The sequences of vhCDR1 (SEQ ID NO: 155), vhCDR2 (SEQ ID NO: 156), vhCDR3 (SEQ ID NO: 157), vlCDR1 (SEQ ID NO: 160), vlCDR2 (SEQ ID NO: 161) and vlCDR3 (SEQ ID NO: 162) of Figure 2K (71739); xxv. The sequences of vhCDR1 (SEQ ID NO: 165), vhCDR2 (SEQ ID NO: 166), vhCDR3 (SEQ ID NO: 167), vlCDR1 (SEQ ID NO: 170), vlCDR2 (SEQ ID NO: 171) and vlCDR3 (SEQ ID NO: 172) of Figure 2L (71736); xxvi. The sequences of vhCDR1 (SEQ ID NO: 175), vhCDR2 (SEQ ID NO: 176), vhCDR3 (SEQ ID NO: 177), vlCDR1 (SEQ ID NO: 180), vlCDR2 (SEQ ID NO: 181) and vlCDR3 (SEQ ID NO: 182) of Figure 2M (71707); xxvii. The sequences of vhCDR1 (SEQ ID NO: 185), vhCDR2 (SEQ ID NO: 186), vhCDR3 (SEQ ID NO: 187), vlCDR1 (SEQ ID NO: 190), vlCDR2 (SEQ ID NO: 191) and vlCDR3 (SEQ ID NO: 192) of Figure 2N (66716); xxviii. The sequences of vhCDR1 (SEQ ID NO: 195), vhCDR2 (SEQ ID NO: 196), vhCDR3 (SEQ ID NO: 197), vlCDR1 (SEQ ID NO: 200), vlCDR2 (SEQ ID NO: 201) and vlCDR3 (SEQ ID NO: 202) of Figure 2O (71728); xxix. The sequences of vhCDR1 (SEQ ID NO: 205), vhCDR2 (SEQ ID NO: 206), vhCDR3 (SEQ ID NO: 207), vlCDR1 (SEQ ID NO: 210), vlCDR2 (SEQ ID NO: 211) and vlCDR3 (SEQ ID NO: 212) of Figure 2P (71741); xxx. The sequences of vhCDR1 (SEQ ID NO: 215), vhCDR2 (SEQ ID NO: 216), vhCDR3 (SEQ ID NO: 217), vlCDR1 (SEQ ID NO: 220), vlCDR2 (SEQ ID NO: 221) and vlCDR3 (SEQ ID NO: 222) of Figure 2Q (71742); xxxi. The sequences of vhCDR1 (SEQ ID NO: 225), vhCDR2 (SEQ ID NO: 226), vhCDR3 (SEQ ID NO: 227), vlCDR1 (SEQ ID NO: 230), vlCDR2 (SEQ ID NO: 231) and vlCDR3 (SEQ ID NO: 232) of Figure 2R (71744); xxxii. The sequences of vhCDR1 (SEQ ID NO: 235), vhCDR2 (SEQ ID NO: 236), vhCDR3 (SEQ ID NO: 237), vlCDR1 (SEQ ID NO: 240), vlCDR2 (SEQ ID NO: 241) and vlCDR3 (SEQ ID NO: 242) of Figure 2S (71753), and xxxiii. Comprises vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences selected from the group consisting of vhCDR1 (SEQ ID NO: 245), vhCDR2 (SEQ ID NO: 246), vhCDR3 (SEQ ID NO: 247), vlCDR1 (SEQ ID NO: 250), vlCDR2 (SEQ ID NO: 251), and vlCDR3 (SEQ ID NO: 252) sequences of Figure 2T (71755).

[0010] In some embodiments, the composition comprises an antibody, wherein the antibody i. The heavy chain variable domain (SEQ ID NO: 54) and light chain variable domain (SEQ ID NO: 59) of Figure 2A (71709); ii. The heavy chain variable domain (SEQ ID NO: 64) and light chain variable domain (SEQ ID NO: 69) of Figure 2B (71719); iii. The heavy chain variable domain (SEQ ID NO: 74) and light chain variable domain (SEQ ID NO: 79) of Figure 2C (71720); iv. The heavy chain variable domain (SEQ ID NO: 84) and light chain variable domain (SEQ ID NO: 89) of Figure 2D (71722); v. The heavy chain variable domain (SEQ ID NO: 94) and light chain variable domain (SEQ ID NO: 99) of Figure 2E (71701); vi. The heavy chain variable domain (SEQ ID NO: 104) and light chain variable domain (SEQ ID NO: 109) of Figure 2F (71663); vii. The heavy chain variable domain (SEQ ID NO: 114) and light chain variable domain (SEQ ID NO: 119) of Figure 2G (71662); viii. The heavy chain variable domain (SEQ ID NO: 124) and light chain variable domain (SEQ ID NO: 129) of Figure 2H (66692); ix. The heavy chain variable domain (SEQ ID NO: 134) and light chain variable domain (SEQ ID NO: 139) of Figure 2I (71710); x. The heavy chain variable domain (SEQ ID NO: 144) and light chain variable domain (SEQ ID NO: 149) of Figure 2J (71717); xi. The heavy chain variable domain (SEQ ID NO: 154) and light chain variable domain (SEQ ID NO: 159) of Figure 2K (71739); xii. The heavy chain variable domain (SEQ ID NO: 164) and light chain variable domain (SEQ ID NO: 169) of Figure 2L (71736); xiii. The heavy chain variable domain (SEQ ID NO: 174) and light chain variable domain (SEQ ID NO: 179) of Figure 2M (71707); xiv. The heavy chain variable domain (SEQ ID NO: 184) and light chain variable domain (SEQ ID NO: 189) of Figure 2N (66716); xv. The heavy chain variable domain (SEQ ID NO: 194) and light chain variable domain (SEQ ID NO: 199) of Figure 2O (71728); xvi. The heavy chain variable domain (SEQ ID NO: 204) and light chain variable domain (SEQ ID NO: 209) of Figure 2P (71741); xvii. The heavy chain variable domain (SEQ ID NO: 214) and light chain variable domain (SEQ ID NO: 219) of Figure 2Q (71742); xviii. The heavy chain variable domain (SEQ ID NO: 224) and light chain variable domain (SEQ ID NO: 229) of Figure 2R (71744); xix. The heavy chain variable domain (SEQ ID NO: 234) and light chain variable domain (SEQ ID NO: 239) of Figure 2S (71753), and xx. Comprises an antibody heavy chain variable domain and a light chain variable domain selected from the group consisting of the heavy chain variable domain (SEQ ID NO: 244) and the light chain variable domain (SEQ ID NO: 249) of Figure 2T (71755).

[0011] In some embodiments, the antibody comprises a CH1-hinge-CH2-CH3 region derived from human IgG1, IgG2, IgG3, or IgG4, where the hinge region optionally comprises a mutation.

[0012] In some embodiments, the antibody comprises a CH1-hinge-CH2-CH3 region derived from human IgG4.

[0013] In some embodiments, the hinge region comprises a mutation.

[0014] In some embodiments, the antibody comprises the CL region of a human κ2 light chain.

[0015] In some embodiments, the antibody comprises the CL region of a human λ2 light chain.

[0016] In some embodiments, the antibody i. a heavy chain variable domain, a) CDR-H1 having the sequence YTFX-X2-YA-X3-H, where X is N, R, D, G or K, X2 is S, H, I or Q, and X3 is M or V; and b) a CDR-H2 having the sequence WIHAGTGXT-X2-YSQKFQG, where X is N, A, or V and X2 is K or LW-IH; and c) a heavy chain variable domain comprising a CDR-H3 having the sequence ARGLGXVGPTGTSWFDP (wherein X is S or E); ii. a light chain variable domain, a) a CDR-L1 having the sequence RASQGISSWLA; b) a CDR-L2 having the sequence EASSLES; c) a light chain variable domain comprising a CDR-L3 having the sequence QQYRX-X2-PFT (wherein X is S, V, Y, L, or Q and X2 is F, S, or G).

[0017] In some embodiments, the antibody i. a heavy chain variable domain, a) a CDR-H1 having the sequence GTFX-X2-Y-X3-IS, where X is S or N, X2 is E or S, and X3 is V or P; b) a CDR-H2 having the sequence GIIPGAGTAXYAQKFQG (wherein X is N) or IG-IIPX-X2-G-X3-A-X4-YAQKFQG (wherein X is G or I, X2 is S or A; X3 is T or S, and X4 is N or I); and c) a heavy chain variable domain comprising a CDR-H3 having the sequence ARGRHXHET (wherein X is S or F); ii. a light chain variable domain, a) a CDR-L1 having the sequence RASQGISSWLA; b) a CDR-L2 having the sequence AASSLQS; c) a light chain variable domain comprising a CDR-L3 having the sequence QQVYX-X2-PWT (wherein X is S or R and X2 is L or FQ-).

[0018] In some embodiments, the antibody i. a heavy chain variable domain, a) a CDR-H1 having the sequence FTFXN-X2-AM-SF-TFXN-X2-X3-MS, where X is G or D or S and X2 is T or V or Y; b) a CDR-H2 having the sequence AISX-X1-X2-GSTYYADSVK-GA-ISANAGSTYYADSVKG, where X is G or A, X2 is N or S, and X3 is A or G; and c) a heavy chain variable domain comprising a CDR-H3 having the sequence AKGPDRQVFDY; ii. a light chain variable domain, a) a CDR-L1 having the sequence RASQGIXSWLA (wherein X is S or D); b) a CDR-L2 having the sequence AASSLQS; c) a light chain variable domain comprising a CDR-L3 having the sequence QHAX-X1-FPY-TQ-HALXFPYT (wherein X is Y or L and X1 is S or F).

[0019] In some embodiments, the antibody i. a heavy chain variable domain, a) a CDR-H1 having the sequence GSISSX-X2-Y-X3-WG, where X is S or P, X2 is E or D, and X3 is G, Y, or P; and b) a CDR-H2 having the sequence SIX-X2-X3-G-X4-TYYNPSLKS, where X is Y or V, X2 is Y or N, X3 is Q or S, and X4 is S or A; and c) a heavy chain variable domain comprising a CDR-H3 having the sequence ARGPXRQ-X2-FDY (wherein X is Y or H and X2 is V or L); ii. a light chain variable domain, a) a CDR-L1 having the sequence RASQGISSWLA; b) a CDR-L2 having the sequence AASSLQS; c) a light chain variable domain comprising a CDR-L3 having the sequence QQGX-X2-FPYT (wherein X is S or F and X2 is S or V).

[0020] In some embodiments, the antibody i. a heavy chain variable domain, a) a CDR-H1 having the sequence YTFX-X2-YA-X3-H, where X is any amino acid, X2 is any amino acid, and X3 is any amino acid; b) a CDR-H2 having the sequence WIHAGTGXT-X2-YSQKFQG, where X is any amino acid and X2 is any amino acid; and c) a heavy chain variable domain comprising a CDR-H3 having the sequence ARGLGXVGPTGTSWFDP (wherein X is any amino acid); ii. a light chain variable domain, a) a CDR-L1 having the sequence RASQGISSWLA; b) a CDR-L2 having the sequence EASSLES; c) a light chain variable domain comprising a CDR-L3 having the sequence QQYRX-X2-PFT (wherein X is any amino acid and X2 is any amino acid).

[0021] In some embodiments, the antibody i. a heavy chain variable domain, a) a CDR-H1 having the sequence GTFX-X2-Y-X3-IS, where X is any amino acid, X2 is any amino acid, and X3 is any amino acid; b) a CDR-H2 having the sequence GIIPG-X2-GTA-X3-YAQKFQG, where X is G or Y, X2 is A or S, and X3 is N, I, or V; and c) a heavy chain variable domain comprising a CDR-H3 having the sequence ARGRHXHET (wherein X is any amino acid); ii. a light chain variable domain, a) a CDR-L1 having the sequence RASQGISSWLA; b) a CDR-L2 having the sequence AASSLQS; c) a light chain variable domain comprising a CDR-L3 having the sequence QQVYX-X2-PWT (wherein X is any amino acid and X2 is any amino acid).

[0022] In some embodiments, the antibody i. a heavy chain variable domain, a) the sequence FTFXN-X2-AMS, where X is any amino acid and X2 is any amino acid; b) a CDR-H2 having the sequence AISX-X1-X2-GSTYYADSVKG, where X is any amino acid, X2 is any amino acid, and X3 is any amino acid; and c) a heavy chain variable domain comprising a CDR-H3 having the sequence AKGPDRQVFDY; ii. a light chain variable domain, a) a CDR-L1 having the sequence RASQGIXSWLA (wherein X is any amino acid); b) a CDR-L2 having the sequence AASSLQS; c) a light chain variable domain comprising a CDR-L3 having the sequence QHAX-X1-FPYT (wherein X is any amino acid and X2 is any amino acid).

[0023] In some embodiments, the antibody i. a heavy chain variable domain, a) a CDR-H1 having the sequence GSISSX-X2-Y-X3-WG, where X is any amino acid, X2 is any amino acid, and X3 is any amino acid; b) a CDR-H2 having the sequence SIX-X2-X3-G-X4-TYYNPSLKS, where X is any amino acid, X2 is any amino acid, X3 is any amino acid, and X4 is any amino acid; and c) a heavy chain variable domain and a CDR-H3 having the sequence ARGPXRQ-X2-FDY (wherein X is any amino acid and X2 is any amino acid); ii. a light chain variable domain, a) a CDR-L1 having the sequence RASQGISSWLA; b) a CDR-L2 having the sequence AASSLQS; c) a light chain variable domain comprising a CDR-L3 having the sequence QQGX-X2-FPYT (wherein X is any amino acid and X2 is any amino acid).

[0024] In some embodiments, the antibody i. a heavy chain variable domain, a) CDR-H1 having the sequence YTFX-X2-YA-X3-H, where X is N, R, D, G, T, Q, S, A, or K, X2 is S, H, I, N, L, Y, or Q, and X3 is M or V; and b) a CDR-H2 having the sequence XI-X2-AG-X3-X4-X5-T-X6-YSQKFQG, where X is W or Y, X2 is H or N, X3 is S, T or A, X4 is G or A, X5 is N, A, T or V, and X6 is E, K or L; and c) a heavy chain variable domain comprising a CDR-H3 having the sequence ARGLGXVGPTGTSWFDP (wherein X is S, L, A, K, or E); ii. a light chain variable domain, a) a CDR-L1 having the sequence RASQGISSWLA; b) a CDR-L2 having the sequence EASSES (wherein X is L or S); c) a light chain variable domain comprising a CDR-L3 having the sequence QQYRX-X2-PFT (wherein X is S, V, Y, L, T, or Q and X2 is F, S, Y, or G).

[0025] In some embodiments, the antibody i. a heavy chain variable domain, a) a CDR-H1 having the sequence GTFX-X2-Y-X3-IS, where X is S or N, X2 is E or S, and X3 is V or P; b) a CDR-H2 having the sequence GIIPX-X2-GTA-X3-YAQKFQG, where X is G, S, I or Y, X2 is A, V or S, and X3 is N, I or V; and c) a heavy chain variable domain comprising a CDR-H3 having the sequence ARGRHXHET (wherein X is S, G, or F); ii. a light chain variable domain, a) a CDR-L1 having the sequence RASQGISSWLA; b) a CDR-L2 having the sequence AASSLQS; c) a light chain variable domain comprising a CDR-L3 having the sequence QQXY-X2-X3-PWT (wherein X is V or L, X2 is S or R, and X3 is L, I, or F).

[0026] In some embodiments, the antibody i. a heavy chain variable domain, a) CDR-H1 having the sequence FTFX-X2-X3-X4-MS, where X is G, S, P or D or S, X2 is N, S or P, X3 is T, V or Y, and X4 is A, H or I; b) a CDR-H2 having the sequence AISX-X2-X3-X4-X5-T-X6-YADSVKG, where X is G or A, X2 is N, T, E or S, X3 is A or G, X4 is A or G, X5 is S or G, and X6 is Y or F; and c) a heavy chain variable domain comprising a CDR-H3 having the sequence AKGPDRQVFDY; ii. a light chain variable domain, a) a CDR-L1 having the sequence RASQGIXSWLA (wherein X is S or D); b) a CDR-L2 having the sequence AASSLQS; c) a CDR-L3 having the sequence QHX-X2-X3-FPYT (wherein X is A or G, X2 is Y, R or L, and X3 is S, R, L or F); and and a light chain variable domain comprising:

[0027] In some embodiments, the antibody i. a heavy chain variable domain, a) a CDR-H1 having the sequence GSIXS-X2-X3-Y-X4-W-X5, where X is S or F, X2 is S or P, X3 is E or D, X4 is G, P or Y, and X5 is G or S; and b) a CDR-H2 having the sequence XI-X2-X3-X4-G-X5-TYYNPSLKS, where X is S or V, X2 is Y, V, F or A, X3 is Y, F or N, X4 is Q, A or S, and X5 is S, A or N; and c) a heavy chain variable domain comprising a CDR-H3 having the sequence ARGPXRQ-X2-FDY (wherein X is Y, H, or F, and X2 is V or L); ii. a light chain variable domain, a) a CDR-L1 having the sequence RASQGISSWLA; b) a CDR-L2 having the sequence AASSLQS; c) a light chain variable domain comprising a CDR-L3 having the sequence QQGX-X2-FPYT (wherein X is S, N, W, or F and X2 is S or V).

[0028] In some embodiments, the antibody i) vhCDR1, vhCDR2, and vhCDR3 from VH1-03.66650, VH1-69.66670, VH3-23.66692, or VH1-39.66716; ii) vlCDR1, vlCDR2, and vlCDR3 from VH1-03.66650, VH1-69.66670, VH3-23.66692, or VH1-39.66716. vlCDR1, vlCDR2, and vlCDR3 from VH1-03.66650, VH1-69.66670, VH3-23.66692, or VH1-39.66716.

[0029] In some embodiments, the antibody i)VH1-03.66650, VH1-69.66670, VH3-23.66692, VH1-39.66716, ADI-71663, ADI-71 662, ADI-66692, ADI-71701, ADI-71709, ADI-71710, ADI-71707, ADI-71717, ADI-717 19, ADI-71220, ADI-71722, ADI-71736, ADI-71739, ADI-71728, ADI-66716, ADI-717 41, vhCDR1, vhCDR2, and vhCDR3 derived from ADI-71742, ADI-71744, ADI-71753, or ADI-71755, ii)VL-κ-1-5, VL-κ-1-12, ADI-71663, ADI-71662, ADI-66692, ADI-71701, ADI-71709, ADI-71710, ADI-71707, ADI-71717, ADI-71719, ADI-71220, ADI-7 1722, ADI-71736, ADI-71739, ADI-71728, ADI-66716, ADI-71741, ADI-71742, ADI-71744, ADI-71753, or vlCDR3 derived from ADI-71755, Optionally, the CDRs comprise 0 to 4 substitutions, with no individual CDR comprising more than one substitution, and vhCDR3 and vlCDR3 comprising no substitutions.

[0030] In some embodiments, the anti-IL18-BP antibody i) a heavy chain variable domain comprising a sequence exhibiting at least 90%, at least 95%, or at least 98% identity to a heavy chain variable domain derived from ADI-71663, ADI-71662, ADI-66692, ADI-71701, ADI-71709, ADI-71710, ADI-71707, ADI-71717, ADI-71719, ADI-71220, ADI-71722, ADI-71736, ADI-71739, ADI-71728, ADI-66716, ADI-71741, ADI-71742, ADI-71744, ADI-71753, or ADI-71755, wherein each individual vhCDR comprises no more than one substitution and vhCDR3 comprises no substitutions; ii)ADI-71663, ADI-71662, ADI-66692, ADI-71701, ADI-71709, ADI-71710, ADI-71707, ADI-7171 7, ADI-71719, ADI-71220, ADI-71722, ADI-71736, ADI-71739, ADI-71728, ADI-66716, ADI-71741 a light chain variable domain comprising a sequence exhibiting at least 90%, at least 95%, or at least 98% identity to a light chain variable domain derived from ADI-71742, ADI-71744, ADI-71753, or ADI-71755, wherein each individual vlCDR comprises no more than one substitution, and vlCDR3 comprises no substitutions.

[0031] In some embodiments, the anti-IL18-BP antibody i) a heavy chain variable domain comprising vhCDR1, vhCDR2, and vhCDR3 derived from ADI-71663, ADI-71662, ADI-66692, ADI-71701, ADI-71709, ADI-71710, ADI-71707, ADI-71717, ADI-71719, ADI-71220, ADI-71722, ADI-71736, ADI-71739, ADI-71728, ADI-66716, ADI-71741, ADI-71742, ADI-71744, ADI-71753, or ADI-71755, wherein the heavy chain variable domain is derived from ADI-71663, ADI- a heavy chain variable domain comprising a sequence exhibiting at least 90% identity to a heavy chain variable domain from ADI-71662, ADI-66692, ADI-71701, ADI-71709, ADI-71710, ADI-71707, ADI-71717, ADI-71719, ADI-71220, ADI-71722, ADI-71736, ADI-71739, ADI-71728, ADI-66716, ADI-71741, ADI-71742, ADI-71744, ADI-71753, or ADI-71755, wherein each individual vhCDR comprises no more than one substitution and vhCDR3 comprises no substitutions; ii) a light chain variable domain comprising vlCDR1, vlCDR2, and vlCDR3 derived from ADI-71663, ADI-71662, ADI-66692, ADI-71701, ADI-71709, ADI-71710, ADI-71707, ADI-71717, ADI-71719, ADI-71220, ADI-71722, ADI-71736, ADI-71739, ADI-71728, ADI-66716, ADI-71741, ADI-71742, ADI-71744, ADI-71753, or ADI-71755, wherein the light chain variable domain is and a light chain variable domain comprising a sequence exhibiting at least 90% identity to a light chain variable domain derived from ADI-662, ADI-66692, ADI-71701, ADI-71709, ADI-71710, ADI-71707, ADI-71717, ADI-71719, ADI-71220, ADI-71722, ADI-71736, ADI-71739, ADI-71728, ADI-66716, ADI-71741, ADI-71742, ADI-71744, ADI-71753, or ADI-71755, wherein each individual vlCDR comprises no more than one substitution and vlCDR3 does not contain a substitution.

[0032] In some embodiments, the antibody comprises a heavy chain variable region derived from ADI-71663, ADI-71662, ADI-66692, ADI-71701, ADI-71709, ADI-71710, ADI-71707, ADI-71717, ADI-71719, ADI-71220, ADI-71722, ADI-71736, ADI-71739, ADI-71728, ADI-66716, ADI-71741, ADI-71742, ADI-71744, ADI-71753, or ADI-71755. and a light chain variable domain derived from ADI-71663, ADI-71662, ADI-66692, ADI-71701, ADI-71709, ADI-71710, ADI-71707, ADI-71717, ADI-71719, ADI-71220, ADI-71722, ADI-71736, ADI-71739, ADI-71728, ADI-66716, ADI-71741, ADI-71742, ADI-71744, ADI-71753, or ADI-71755.

[0033] In some embodiments, the antibody comprises a CH1-hinge-CH2-CH3 region derived from human IgG4.

[0034] In some embodiments, the hinge region comprises a mutation.

[0035] In some embodiments, the antibody comprises the CL region of a human κ2 light chain.

[0036] In some embodiments, the antibody comprises the CL region of a human λ2 light chain.

[0037] In some embodiments, the antibody a)VH1-03.66650, VH1-69.66670, VH3-23.66692, VH1-39.66716, ADI-71663, ADI-71662, AD I-66692, ADI-71701, ADI-71709, ADI-71710, ADI-71707, ADI-71717, ADI-71719, ADI-71220 a heavy chain variable domain comprising vhCDR1, vhCDR2, and vhCDR3 from an antibody selected from the group consisting of ADI-71722, ADI-71736, ADI-71739, ADI-71728, ADI-66716, ADI-71741, ADI-71742, ADI-71744, ADI-71753, or ADI-71755; b) a light chain variable domain comprising vlCDR1, vlCDR2, and vlCDR3 from an antibody selected from the group consisting of VL-κ-1-5, VL-κ-1-12, ADI-71663, ADI-71662, ADI-66692, ADI-71701, ADI-71709, ADI-71710, ADI-71707, ADI-71717, ADI-71719, ADI-71220, ADI-71722, ADI-71736, ADI-71739, ADI-71728, ADI-66716, ADI-71741, ADI-71742, ADI-71744, ADI-71753, or ADI-71755.

[0038] In some embodiments, the antibody a)VH1-03.66650, VH1-69.66670, VH3-23.66692, VH1-39.66716, ADI-71663, ADI-71662, AD I-66692, ADI-71701, ADI-71709, ADI-71710, ADI-71707, ADI-71717, ADI-71719, ADI-71220 a heavy chain variable domain comprising vhCDR1, vhCDR2, and vhCDR3 from an antibody selected from the group consisting of ADI-71722, ADI-71736, ADI-71739, ADI-71728, ADI-66716, ADI-71741, ADI-71742, ADI-71744, ADI-71753, or ADI-71755; b) a light chain variable domain comprising a vlCDR1, vlCDR2, and vlCDR3 derived from an antibody selected from the group consisting of VL-κ-1-5, VL-κ-1-12, ADI-71663, ADI-71662, ADI-66692, ADI-71701, ADI-71709, ADI-71710, ADI-71707, ADI-71717, ADI-71719, ADI-71220, ADI-71722, ADI-71736, ADI-71739, ADI-71728, ADI-66716, ADI-71741, ADI-71742, ADI-71744, ADI-71753, or ADI-71755; Optionally, 1) each CDR individually contains 0 to 4 substitutions, no individual CDR contains more than one substitution, and vhCDR3 and vlCDR3 contain no substitutions; 2) each CDR individually contains one substitution; or 3) each individual vhCDR contains no more than one substitution, and vhCDR3 contains no substitutions.

[0039] In some embodiments, the antibody comprises a CH1-hinge-CH2-CH3 region derived from human IgG1, IgG2, IgG3, or IgG4, where the hinge region optionally comprises a mutation.

[0040] In some embodiments, the antibody comprises a CH1-hinge-CH2-CH3 region derived from human IgG4.

[0041] In some embodiments, the hinge region comprises a mutation.

[0042] In some embodiments, the antibody comprises the CL region of a human κ2 light chain.

[0043] In some embodiments, the antibody comprises the CL region of a human λ2 light chain.

[0044] In some embodiments, the antibody competes for binding with an antibody of any one of the preceding claims.

[0045] The present invention also provides a method of treating cancer in a patient, comprising administering an anti-IL18-BP antibody according to any of the preceding claims, wherein the cancer is treated.

[0046] The present invention also provides a method of treating cancer in a patient, comprising administering an anti-IL18-BP antibody, wherein the anti-IL18-BP antibody activates T cells, NK cells, NKT cells, dendritic cells, MAIT T cells, γδT cells, and / or innate lymphoid cells (ILCs) and / or regulates myeloid cells, thereby treating the cancer.

[0047] The present invention also provides a method for activating T cells in a patient, the method comprising administering an anti-IL18-BP antibody according to any of the preceding claims, whereby the T cells are activated.

[0048] The present invention also provides a method for activating NK cells in a patient, the method comprising administering an anti-IL18-BP antibody according to any of the preceding claims, whereby the NK cells are activated.

[0049] The present invention also provides a method for activating NKT cells in a patient, the method comprising administering an anti-IL18-BP antibody according to any of the preceding claims, thereby activating the NKT cells.

[0050] The present invention also provides a method for modulating bone marrow cells in a patient, the method comprising administering an anti-IL18-BP antibody according to any of the preceding claims, wherein said bone marrow cells are modulated.

[0051] The present invention also provides a method for activating dendritic cells in a patient, the method comprising administering an anti-IL18-BP antibody according to any of the preceding claims, whereby the dendritic cells are activated.

[0052] The present invention also provides a method for activating dendritic cells in a patient, the method comprising administering an anti-IL18-BP antibody according to any of the preceding claims, thereby activating the MAIT T cells.

[0053] The present invention also provides a method for activating dendritic cells in a patient, the method comprising administering an anti-IL18-BP antibody according to any of the preceding claims, thereby activating the γδ T cells.

[0054] The present invention also provides a method for activating ILC cells in a patient, the method comprising administering an anti-IL18-BP antibody according to any of the preceding claims, whereby the ILC cells are activated.

[0055] The present invention also relates to tumor microenvironment (TME) and / or lymphatic and a method for increasing IL-18-mediated immunostimulatory activity in TME and / or lymph nodes, the method comprising administering an anti-IL18-BP antibody, wherein the anti-IL18-BP antibody increases IL-18-mediated immunostimulatory activity in TME and / or lymph nodes.

[0056] The present invention also provides a method for restoring IL-18 activity to T cells, NK cells, NKT cells, myeloid cells, dendritic cells, MAIT T cells, γδ T cells, and / or innate lymphoid cells (ILCs), comprising administering an anti-IL18-BP antibody, wherein the anti-IL18-BP antibody restores activity to T cells, NK cells, NKT cells, myeloid cells, dendritic cells, MAIT T cells, γδ T cells, and / or innate lymphoid cells (ILCs).

[0057] In some embodiments, the anti-IL18-BP antibody is administered as a stable liquid pharmaceutical formulation.

[0058] In some embodiments, the T cells are cytotoxic T cells (CTLs). )

[0059] The present invention also relates to a method for treating T cells comprising administering to a subject a therapeutic agent selected from the group consisting of: + T cells and CD8 + 48. The method of claim 47, wherein the cell is selected from the group consisting of T cells.

[0060] In some embodiments, the treatment objectives include an increase in tumor growth inhibition of at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, 225%, 250%, 275%, 300%, 325%, 350%, 375%, 400%, 425%, 450%, 475%, 500%, 525%, 550%, 575%, 600%, 625%, 650%, 675%, 700%, 725%, 750%, 775%, 800%, 825%, 850%, 875%, 900%, 925%, 950%, 975%, or 1000% compared to a control or untreated patient.

[0061] Treated subjects experience a decrease in serotonin levels by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, 225%, 250%, 275%, 300%, 325%, 350%, 375%, 400%, 425%, 450%, 475% compared to controls or untreated patients 55. The method of claim 40, wherein the patient exhibits a reduction in tumor growth of 500%, 525%, 550%, 575%, 600%, 625%, 650%, 675%, 700%, 725%, 750%, 775%, 800%, 825%, 850%, 875%, 900%, 925%, 950%, 975%, or 1000%.

[0062] The present invention also provides the methods described herein, wherein the NK cells are CD16+ lymphocytes.

[0063] The present invention also provides the methods described herein, wherein the NK cells are CD56+ NK cells.

[0064] The present invention also provides methods described herein, wherein activation is measured as an increase in expression of one or more activation markers.

[0065] The present invention also provides a method as described herein, wherein the activation marker is selected from the group consisting of CD107a, CD137, CD69, granzymes, and perforin.

[0066] The present invention also provides a method as described herein, wherein activation is measured as an increase in proliferation of said NK cells.

[0067] The invention also provides the methods described herein, wherein activation is measured as an increase in the secretion of one or more cytokines.

[0068] The present invention also provides the methods described herein, wherein the one or more cytokines are selected from the group consisting of IFNγ, TNF, GMCSF, MIG (CXCL9), IP-10 (CXCL10) and MCP1 (CCL2).

[0069] The present invention also provides methods described herein, wherein activation is measured as an increase in direct killing of target cells.

[0070] In some embodiments, the method further comprises administering a second antibody.

[0071] In some embodiments, the second antibody is an antibody that binds to and / or inhibits a human checkpoint receptor protein.

[0072] In some embodiments, the second antibody is selected from the group consisting of an anti-PVRIG antibody, an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-TIGIT antibody, an anti-CTLA-4 antibody, an anti-PD-L2 antibody, an anti-B7-H3 antibody, an anti-B7-H4 antibody, an anti-CEACAM-1 antibody, an anti-PVR antibody, an anti-LAG3 antibody, an anti-CD112 antibody, an anti-CD96 antibody, an anti-TIM3 antibody, an anti-BTLA antibody, an anti-ICOS antibody, an anti-OX40 antibody, or an anti-41BB antibody, an anti-CD27 antibody, or an anti-GITR antibody.

[0073] In some embodiments, the PVRIG antibody is selected from the group consisting of CHA.7.518.1.H4(S241P) and CHA.7.538.1.2.H4(S241P).

[0074] In some embodiments, the anti-PVRIG antibody comprises i) a heavy chain variable domain comprising vhCDR1, vhCDR2, and vhCDR3 derived from CHA.7.518.1.H4(S241P) (sequence number 260), and ii) a light chain variable domain comprising vlCDR1, vlCDR2, and vlCDR3 derived from CHA.7.518.1.H4(S241P) (sequence number 265).

[0075] In some embodiments, the anti-PVRIG antibody comprises i) a heavy chain variable domain comprising vhCDR1, vhCDR2, and vhCDR3 derived from CHA.7.538.1.2.H4(S241P) (sequence number 270), and ii) a light chain variable domain comprising vlCDR1, vlCDR2, and vlCDR3 derived from CHA.7.538.1.2.H4(S241P) (sequence number 275).

[0076] In some embodiments, the anti-PVRIG antibody comprises i) a heavy chain variable domain comprising vhCDR1, vhCDR2, and vhCDR3 from CHA.7.518.4 (sequence number 1453, Figure 36AG), and ii) a light chain variable domain comprising vlCDR1, vlCDR2, and vlCDR3 from CHA.7.518.4 (sequence number 1457, Figure 36AG).

[0077] In some embodiments, the anti-PVRIG antibody is selected from the group consisting of GSK4381562 / SRF816 (GSK / Surface), NTX2R13 (Nectin Therapeutics), anti-PVRIG antibodies described in WO 2017 / 041004, anti-PVRIG antibodies described in WO 2001 / 008879, anti-PVRIG antibodies described in WO 2018 / 017864, and anti-PVRIG antibodies described in WO 2118 / 000205.

[0078] In some embodiments, the anti-TIGIT antibody is selected from the group consisting of CPA.9.083.H4(S241P) and CPA.9.086.H4(S241P).

[0079] In some embodiments, the anti-TIGIT antibody comprises i) a heavy chain variable domain comprising vhCDR1, vhCDR2, and vhCDR3 derived from CPA.9.083.H4(S241P) (SEQ ID NO: 350), and ii) a light chain variable domain comprising vlCDR1, vlCDR2, and vlCDR3 derived from CPA.9.083.H4(S241P) (SEQ ID NO: 355).

[0080] In some embodiments, the anti-TIGIT antibody comprises i) a heavy chain variable domain comprising vhCDR1, vhCDR2, and vhCDR3 derived from CPA.9.086.H4(S241P) (SEQ ID NO: 360), and ii) a light chain variable domain comprising vlCDR1, vlCDR2, and vlCDR3 derived from CPA.9.086.H4(S241P) (SEQ ID NO: 365).

[0081] In some embodiments, the anti-TIGIT antibody comprises i) a heavy chain variable domain comprising vhCDR1, vhCDR2, and vhCDR3 from CHA.9.547.18 (sequence number 1177, Figure 34QQQQ), and ii) a light chain variable domain comprising vlCDR1, vlCDR2, and vlCDR3 from CHA.9.547.18 (sequence number 1181, Figure 34QQQQ).

[0082] In some embodiments, the anti-TIGIT antibody is selected from the group consisting of EOS-448 (GlaxoSmithKline, iTeos Therapeutics), BMS-986207, domvanalimab (AB154, Arcus Biosciences, Inc.), AB308 (Arcus Bioscience), osipellimab (aBGB-A1217, BeiGene), tiragolumab (MTIG7192A, RocheGenentech), BAT6021 (Bio-Thera Solutions), BAT6005 (Bio-Thera Solutions), IBI939 (Innovent Biologics, U.S. Patent Application Publication No. 2021 / 00040201), JS006 (Junshi Bioscience / COHERUS), ASP8374 (Astellas Pharma Inc), vibostolimab (MK-7684, Merck Sharp & Co., Inc.), and / or EGFR-1000 (European Biosciences). Dohme), M6332 (Merck KGAA), etigilimab (OMP-313M32, Mereo BioPharma), SEA-TGT (Seagen), HB0030 (Huabo Biopharma), AK127 (AKESO), IBI939 (Innovent Biologics), and anti-TIGIT antibodies include Genentech antibody (MTIG7192A).

[0083] In some embodiments, the anti-PD-1 antibody is selected from the group consisting of nivolumab (Opdivo®; BMS; CheckMate078), pembrolizumab (KEYTRUDA®; Merck), TSR-042 (Tesaro), cemiplimab (REGN2810; Regeneron Pharmaceuticals, see U.S. Patent Application Publication No. 20170174779), BMS-936559, spartalizumab (PDR001, Novartis), pidilizumab (CT-011; Pfizer Inc), tislelizumab (BGB-A317, BeiGene), camrelizumab (SHR-1210, Incyte and Jiangsu HengRui), SHR-1210 (CTR20170299 and CTR20170322), SHR-1210 (CTR20160175 and CTR20170090), sintilimab (Tyvyt®; Eli lily and Innovent Biologics), toripalimab (JS001, Shanghai Junshi Bioscience), JS-001 (CTR20160274), IBI308 (CTR20160735), BGB-A317 (CTR20160872), penprimab (AK105, Akeso Biopharma), dimvelelimab (Arcus), BAT1306 (Bio-Thera Solutions Ltd), sasanlimab (PF-06801591, Pfizer), dostallimab-gxly (GlaxoSmithKline LLC), prorugolimab (Biocad), kadonilimab (Akeso Inc), geptanolimab (Genor BioPharma Co Ltd), selplulimab (Shanghai Henlius Biotech Inc), balstilimab (Agenus Inc), retifanlimab (Incyte Corp), cetrelimab (Johnson & Johnson), CS-1003 (EQRx Inc), IBI-318 (Innovent Biologics Inc), ivonesimab (Akeso Inc), pucotenlimab (Lepu Biopharma Co Ltd), QL-1604 (Qilu Pharmaceutical Co Ltd), SCTI-10A (SinoCelltech Group Ltd), tebotelimab (MacroGenics Inc), AZD-7789 (AstraZeneca Plc), budigalimab (AbbVie Inc), EMB-02 (EpimAb Biotherapeutics Inc), ezabenlimab (Boehringer Ingelheim International GmbH), F-520 (Shandong New Time Pharmaceutical Co Ltd), HX-009 (Waterstone Hanxbio Pty Ltd), zelvalimab (Amgen), peresolimab (Eli Lilly and Co), rosnilimab (AnaptysBio Inc), budalimab (Xencor), izularimab (Xencor), lorigellimab (MacroGenics Inc), YBL-006 (Y-Biologics Inc), and ONO-4685 (Ono Pharmaceutical Co Ltd), LY-3434172 (Eli Lilly and Co).

[0084] In some embodiments, the anti-PD-L1 antibody is selected from the group consisting of atezolizumab (TECENTRIQ®; MPDL3280A; IMpower110; Roche / Genentech), avelumab (BAVENCIO®, MSB001071 8C, EMD Serono & Pfizer), and durvalumab (MEDI4736, IMFINZI®, AstraZeneca). Other antibodies under development include lodapolimab (LY3300054, Eli Lily), pimivalimab (Jounce Therapeutics Inc), SHR-1316 (Jiangsu Hengrui Medicine Co Ltd), emvafolimab (Jiangsu Simcere Pharmaceutical Co Ltd), sugemalimab (CStone Pharmaceuticals Co Ltd), cosibelimab (Checkpoint Therapeutics Inc), pakmilimab (CytomX Therapeutics Inc), IBI-318, IBI-322, IBI-323 (Innovent Biologics Inc), INBRX-105 (Inhibrx Inc), KN-046 (Alphamab Oncology), 6MW-3211 (Mabwell Shanghai Bioscience Co Ltd), BNT-311 (BioNTech SE), and FS-118 (F-star Therapeutics Inc), GNC-038 (Systimmune Inc), GR-1405 (Genrix (Shanghai) Biopharmaceutical Co Ltd), HS-636 (Zhejiang Hisun Pharmaceutical Co Ltd), LP-002 (Lepu Biopharma Co Ltd), PM-1003 (Biotheus Inc), PM-8001 (Biotheus Inc), STIA-1015 (ImmuneOncia Therapeutics LLC), ATG-101 (Antengene Corp Ltd), BJ-005 (BJ Bioscience Inc), CDX-527 (Celldex Therapeutics Inc), GNC-035 (Systimmune Inc), GNC-039 (Systimmune Inc), HLX-20 (Shanghai Henlius Biotech Inc), JS-003 (Shanghai Junshi Bioscience Co Ltd), LY-3434172 (Eli Lilly and Co), MCLA-145 (Merus NV), MSB-2311 (Transcenta Holding Ltd), PF-07257876 (Pfizer Inc), Q-1802 (QureBio Ltd), QL-301 (QLSF Biotherapeutics Inc), QLF-31907 (Qilu Pharmaceutical Co Ltd), RC-98(RemeGen Co. Ltd), TST-005 (Transcenta Holding Ltd), atezolizumab (IMpower133), BMS-936559 / MDX-1105, and / or RG-7446 / MPDL3280A, and YW243.55.S70.

[0085] In some embodiments, the anti-IL18-BP antibody and the second antibody are administered sequentially or simultaneously, in any order, in one or more formulations.

[0086] In some embodiments, the anti-IL18-BP antibody is for use in combination with an immunostimulatory antibody, cytokine therapy, or immunomodulatory agent, cytotoxic agent, chemotherapeutic agent, growth inhibitory agent, antihormonal agent, kinase inhibitor, anti-angiogenic agent, cardioprotective agent, immunosuppressant, agent that promotes blood cell proliferation, angiogenesis inhibitor, protein tyrosine kinase (PTK) inhibitor, or other therapeutic agent.

[0087] In some embodiments, the method further comprises administering one or more inflammasome activators.

[0088] In some embodiments, the inflammasome activator is a chemotherapeutic agent.

[0089] In some embodiments, the chemotherapeutic agent is selected from the group consisting of platinum, paclitaxel (taxol), sorafenib, doxorubicin, sorafenib, 5-FU, gemcitabine, and irinotecan (CPT-11).

[0090] In some embodiments, the platinum chemotherapeutic agent is oxaliplatin or cisplatin.

[0091] In some embodiments, the inflammasome activator is a CD39 inhibitor.

[0092] In some embodiments, the CD39 inhibitor is an anti-CD39 antibody.

[0093] In some embodiments, the cancer is angiogenic tumors, melanoma, non-melanoma skin cancer (squamous cell carcinoma and basal cell carcinoma), mesothelioma, squamous cell carcinoma, lung cancer, small cell lung cancer, non-small cell lung cancer, neuroendocrine lung cancer (including pleural mesothelioma, neuroendocrine lung carcinoma), NSCL (large cell), NSCLC large cell adenocarcinoma, non-small cell lung cancer (NSCLC), NSCLC squamous cell, soft tissue sarcoma, Kaposi's sarcoma, adenocarcinoma of the lung, squamous cell carcinoma of the lung, NSCLC with a TPS of PDL1≧50%, neuroendocrine lung carcinoma, atypical carcinoid lung cancer, cancer of the peritoneum, esophageal cancer, hepatocellular carcinoma, liver cancer (including HCC), gastric cancer, cancer) (including gastrointestinal cancer), pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, urothelial carcinoma, bladder cancer, liver cancer, glioma, brain cancer (and edema such as that associated with brain tumors), breast cancer (including, for example, triple-negative breast cancer), testicular cancer, testicular germ cell tumor, colon cancer, colorectal cancer (CRC), colorectal cancer MSS (MSS-CRC); refractory MSS colorectal;MSS (microsatellite stable), primary peritoneal cancer, primary peritoneal ovarian cancer, microsatellite stable primary peritoneal cancer, platinum-resistant microsatellite stable primary peritoneal cancer, CRC (MSS unknown), rectal cancer, endometrial cancer (including endometrioid carcinoma), uterine cancer, salivary gland cancer, kidney cancer, renal cell carcinoma (RCC), renal cell carcinoma (RCC), gastroesophageal junction cancer, prostate cancer, vulvar cancer, thyroid cancer, liver cancer, carcinoid cancer, head and neck cancer, B-cell lymphoma (non-Hodgkin's lymphoma) Hodgkin's lymphoma (NHL), including low-grade / follicular non-Hodgkin's lymphoma (NHL), small lymphocytic (SL) NHL, intermediate-grade / follicular NHL, intermediate-grade diffuse NHL, diffuse large B-cell lymphoma, high-grade immunoblastic NHL, high-grade lymphoblastic NHL, high-grade small non-cleaved cell NHL, bulky mass disease NHL, mantle cell lymphoma, AIDS-related lymphoma, and Waldenstrom's macroglobulinemia, Hodgkin's lymphoma (HLL) D), chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), T-cell acute lymphoblastic leukemia (T-ALL), acute myeloid leukemia (AML), hairy cell leukemia, chronic myeloblastic leukemia, multiple myeloma, post-transplant lymphoproliferative disorder (PTLD), abnormal vascular proliferation associated with phacomatosis, Meigs syndrome, Merkel cell carcinoma, MSI-high cancer, KRAS-mutated tumors, adult T-cell leukemia / lymphoma, adenoid cystic carcinoma (including adenoid cystic carcinoma) , melanoma, malignant melanoma, metastatic melanoma, pancreatic cancer, pancreatic adenocarcinoma, ovarian cancer (including ovarian carcinoma), pleural mesothelioma, cervical squamous cell carcinoma (cervical SCC), anal squamous cell carcinoma (anal SCC), carcinoma of unknown primary, gallbladder carcinoma, pleural mesothelioma, chordoma, endometrial sarcoma, chondrosarcoma, uterine sarcoma, uveal melanoma, amyloidosis, AL-amyloidosis, astrocytoma, and / or myelodysplastic syndrome (MDS);

[0094] In some embodiments, the cancer is selected from the group consisting of renal clear cell carcinoma (RCC), lung cancer, NSCLC, lung adenocarcinoma, lung squamous cell carcinoma, gastric adenocarcinoma, ovarian cancer, endometrial cancer, breast cancer, triple-negative breast cancer (TNBC), head and neck tumors, colorectal adenocarcinoma, melanoma, and metastatic melanoma.

[0095] The present invention also provides an anti-IL18BP antibody as described herein for use in the treatment of cancer by activating T cells, NK cells, NKT cells, dendritic cells, MAIT T cells, γδT cells, and / or innate lymphoid cells (ILCs) in a patient, and / or modulating myeloid cells.

[0096] The present invention also provides an anti-IL18BP antibody as described herein for use in increasing IL-18-mediated immunostimulatory activity in the tumor microenvironment (TME) and / or lymph nodes.

[0097] The present invention also provides an anti-IL18BP antibody as described herein for treating cancer in a recipient patient.

[0098] The present invention also provides an anti-IL18BP antibody as described herein for use according to any of the preceding claims.

[0099] The present invention also provides an anti-IL18BP antibody described herein for use in combination with a second antibody, in some embodiments, the second antibody is selected from the group consisting of an anti-PVRIG antibody, an anti-PD-1 antibody, an anti-PD-L1 antibody, and an anti-TIGIT antibody.

[0100] The present invention also relates to an anti-IL18-BP antibody having a IL-18 activity of less than 0.005 pM, less than 0.01 pM, less than 0.02 pM, less than 0.03 pM, less than 0.04 pM, less than 0.05 pM, less than 0.06 pM, less than 0.07 pM, less than 0.08 pM, less than 0.09 pM, less than 0.10 pM, less than 0.15 pM, less than 0.20 pM, less than 0.25 pM, less than 0.30 pM, less than 0.3 Provided are anti-IL18BP antibodies as described herein that exhibit a binding affinity or KD of less than 5 pM, less than 0.40 pM, less than 0.45 pM, less than 0.50 pM, less than 0.55 pM, less than 0.60 pM, less than 0.65 pM, less than 0.70 pM, less than 0.75 pM, less than 0.80 pM, less than 0.85 pM, less than 0.90 pM, less than 0.95 pM, or less than 1 pM. [Brief explanation of the drawings]

[0101] [Figure 1A] FIG. 1 shows the sequences of vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 of antibody 66650. [Figure 1B] FIG. 1 shows the sequences of vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 of antibody 66670. [Figure 1C] FIG. 1 shows the sequences of vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 of antibody 66692. [Figure 1D] FIG. 1 shows the sequences of vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 of antibody 66716. [Figure 1E] FIG. 1 shows the sequences of vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 of antibody 66650. [Figure 1F] FIG. 1 shows the sequences of vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 of antibody 66670. [Figure 1G] FIG. 1 shows the sequences of vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 of antibody 66692. [Figure 1H] FIG. 1 shows the sequences of vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 of antibody 66716. [Figure 1I] FIG. 1 shows the sequences of vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 of antibody 66650. [Figure 1J] FIG. 1 shows the sequences of vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 of antibody 66670. [Figure 1K] FIG. 1 shows the sequences of vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 of antibody 66692. [Figure 1L]FIG. 1 shows the sequences of vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 of antibody 66716. [Figure 1M] FIG. 1 is a diagram of IgG sequences, including IgG1, IgG2, IgG3, and IgG4.

[0102] [Figure 2A] FIG. 1 shows the sequences of the variable heavy and light chains, vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, vlCDR3, and full length of antibody ADI-71709. [Figure 2B] FIG. 1 shows the sequences of the variable heavy and light chains, vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, vlCDR3, and full length of ADI-71719. [Figure 2C] FIG. 1 shows the sequences of the variable heavy and light chains, vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, vlCDR3, and full length of ADI-71720. [Figure 2D] FIG. 1 shows the sequences of the variable heavy and light chains, vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, vlCDR3, and full length of ADI-71722. [Figure 2E] FIG. 1 shows the sequences of the variable heavy and light chains, vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, vlCDR3, and full length of ADI-71701. [Figure 2F] FIG. 1 shows the sequences of the variable heavy and light chains, vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, vlCDR3, and full length of ADI-71663. [Figure 2G] FIG. 1 shows the sequences of the variable heavy and light chains, vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, vlCDR3, and full length of ADI-71662. [Figure 2H] FIG. 1 shows the sequences of the variable heavy and light chains, vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, vlCDR3, and full length of ADI-66692. [Figure 2I] FIG. 1 shows the sequences of the variable heavy and light chains, vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, vlCDR3, and full length of ADI-71710. [Figure 2J] FIG. 1 shows the sequences of the variable heavy and light chains, vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, vlCDR3, and full length of ADI-71717. [Figure 2K] FIG. 1 shows the sequences of the variable heavy and light chains, vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, vlCDR3, and full length of ADI-71739. [Figure 2L] FIG. 1 shows the sequences of the variable heavy and light chains, vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, vlCDR3, and full length of ADI-71736. [Figure 2M] FIG. 1 shows the sequences of the variable heavy and light chains, vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, vlCDR3, and full length of ADI-71707. [Figure 2N] FIG. 1 shows the sequences of the variable heavy and light chains, vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, vlCDR3, and full length of ADI-66716. [Figure 2O] FIG. 1 shows the sequences of the variable heavy and light chains, vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, vlCDR3, and full length of ADI-71728. [Figure 2P] FIG. 1 shows the sequences of the variable heavy and light chains, vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, vlCDR3, and full length of ADI-71741. [Figure 2Q] FIG. 1 shows the sequences of the variable heavy and light chains, vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, vlCDR3, and full length of ADI-71742. [Figure 2R]FIG. 1 shows the sequences of the variable heavy and light chains, vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, vlCDR3, and full length of ADI-71744. [Figure 2S] FIG. 1 shows the sequences of the variable heavy and light chains, vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, vlCDR3, and full length of ADI-71753. [Figure 2T] FIG. 1 shows the sequences of the variable heavy and light chains, vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, vlCDR3, and full length of ADI-71755. [Figure 2U] FIG. 1 shows the sequences of the variable heavy and light chains, vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, vlCDR3, and full length of AB-837 (also referred to as "AbD35328," "837," or "Ab837").

[0103] [Figure 3A] FIG. 1 shows the alignment of CDRH and CDRL sequences between VH3-23 and VL-κ-1-12 germline sequences 71663, 71662 and 66692. [Figure 3B] FIG. 1 shows the alignment of CDRH and CDRL sequences between VH1-03 and VL-κ-1-5 germline sequences 71701, 71707, 71709, 71710, and 71717. [Figure 3C] FIG. 1 shows the alignment of CDRH and CDRL sequences between VH1-69 and VL-κ-1-2 germline sequences 71719, 71720, 71722 and 71728. [Figure 3D] FIG. 1 shows the alignment of CDRH and CDRL sequences between VH4-39 and VL-κ-1-12 germline sequences 71736, 71739 and 66716. [Figure 3E] FIG. 1 shows the alignment of CDRH and CDRL sequences between VH4-39 and VL-κ-1-12 germline sequences 71736, 71739, 66716, 71742, 71744, 71741, 71753 and 71755.

[0104] [Figure 4A] FIG. 1 shows the expression of IL18 across all TCGA tumors. [Figure 4B] Figure 1 shows the expression of IL18-BP across all TCGA tumors.Box plot of log10 RPKM for each TCGA tumor, with a baseline of 1 RPKM.

[0105] [Figure 5A-1] FIG. 1 shows IL18 stratified by IFNγ expression by tumor type in TCGA. [Figure 5A-2] FIG. 1 shows IL18 stratified by IFNγ expression by tumor type in TCGA. [Figure 5A-3] FIG. 1 shows IL18 stratified by IFNγ expression by tumor type in TCGA. [Figure 5B-1] Figure 1 shows IL18-BP stratified by IFNγ expression by tumor type in TCGA. Boxplot of log10 RPKM for each TCGA tumor, with a baseline of 1 RPKM. For tumor abbreviations, see Table 1. IFNγ-high represents the upper quartile and IFNγ-low represents the lower quartile. FC - fold change, P - p-value of Student's T-test between IFNγ-high and IFNγ-low. Fraction represents the number of samples in IFNγ-high / IFNγ-low. [Figure 5B-2] Figure 1 shows IL18-BP stratified by IFNγ expression by tumor type in TCGA. Boxplot of log10 RPKM for each TCGA tumor, with a baseline of 1 RPKM. For tumor abbreviations, see Table 1. IFNγ-high represents the upper quartile and IFNγ-low represents the lower quartile. FC - fold change, P - p-value of Student's T-test between IFNγ-high and IFNγ-low. Fraction represents the number of samples in IFNγ-high / IFNγ-low. [Figure 5B-3]Figure 1 shows IL18-BP stratified by IFNγ expression by tumor type in TCGA. Boxplot of log10 RPKM for each TCGA tumor, with a baseline of 1 RPKM. For tumor abbreviations, see Table 1. IFNγ-high represents the upper quartile and IFNγ-low represents the lower quartile. FC - fold change, P - p-value of Student's T-test between IFNγ-high and IFNγ-low. Fraction represents the number of samples in IFNγ-high / IFNγ-low.

[0106] [Figure 6A-1] Figure 1 shows core inflammasome signatures stratified by IFNγ expression by tumor type in TCGA. Boxplot of log10 RPKM for each TCGA tumor, with a baseline of 1 RPKM. For tumor abbreviations, see Table 1. IFNγ-high represents the upper quartile, and IFNγ-low represents the lower quartile. FC - fold change, P - Student's T-test p-value between IFNγ-high and IFNγ-low. Fraction represents the number of samples in IFNγ-high / IFNγ-low. [Figure 6A-2] Figure 1 shows core inflammasome signatures stratified by IFNγ expression by tumor type in TCGA. Boxplot of log10 RPKM for each TCGA tumor, with a baseline of 1 RPKM. For tumor abbreviations, see Table 1. IFNγ-high represents the upper quartile, and IFNγ-low represents the lower quartile. FC - fold change, P - Student's T-test p-value between IFNγ-high and IFNγ-low. Fraction represents the number of samples in IFNγ-high / IFNγ-low. [Figure 6A-3] Figure 1 shows core inflammasome signatures stratified by IFNγ expression by tumor type in TCGA. Boxplot of log10 RPKM for each TCGA tumor, with a baseline of 1 RPKM. For tumor abbreviations, see Table 1. IFNγ-high represents the upper quartile, and IFNγ-low represents the lower quartile. FC - fold change, P - Student's T-test p-value between IFNγ-high and IFNγ-low. Fraction represents the number of samples in IFNγ-high / IFNγ-low. [Figure 6B]FIG. 1 shows cosine similarity heatmaps and dandograms between core inflammasome genes, IL18, IL18-BP, IL18R, and additional upstream inflammasome genes.

[0107] [Figure 7] Figure 7A shows dot plots of IL18 and IL18-BP, the expression of two genes in breast cancer subtypes before and after treatment in TNBC, and Figure 7B shows dot plots of IL18 and IL18-BP, the expression of two genes before and after treatment in TNBC, also divided by expanding TCR clones (_E) and non-expanding TCR clones (_NE).

[0108] [Figure 8] FIG. 10 shows an affinity matrix of mAbs to human IL18-BP versus human and cynomolgus monkey ("cyno") IL18-BP by Biacore.

[0109] [Figure 9] Competition with human IL18 for binding of IL18-BP-Fc performed in an AlfaLISA assay using 15 nM purified Ab with a hIgG1 backbone.

[0110] [Figure 10] FIG. 10: Blocking activity of parental mAbs against human IL18-BP analyzed by ELISA.

[0111] [Figure 11] FIG. 10: Blocking activity of parental mAbs against cynomolgus IL18-BP analyzed by ELISA.

[0112] [Figure 12] FIG. 10. IC50 values ​​of anti-human IL18-BP Abs measured by ELISA.

[0113] [Figure 13] FIG. 10: Ability of mAbs against human IL18-BP to rescue human IL18 bound by IL18-BP-Fc protein demonstrated using IL18 HEK293 reporter cells.

[0114] [Figure 14A] 1 shows that anti-IL-18BP antibodies completely restored IL-18 activity on NK cells. Schematic diagram of the assay setup. [Figure 14B] FIG. 1 shows that anti-IL-18BP antibody completely restored IL-18 activity on NK cells. [Figure 14C] FIG. 1 shows that anti-IL-18BP antibody completely restored IL-18 activity on NK cells. [Figure 14D] FIG. 1 shows that anti-IL-18BP antibody completely restored IL-18 activity on NK cells. [Figure 14E] FIG. 1 shows that anti-IL-18BP antibody completely restored IL-18 activity on NK cells. [Figure 14F] FIG. 1 shows that anti-IL-18BP antibody completely restored IL-18 activity on NK cells. [Figure 14G] FIG. 1 shows that anti-IL-18BP antibody completely restored IL-18 activity on NK cells. [Figure 14H]Schematic diagram of the assay setup showing that anti-IL-18BP antibodies fully restored IL-18 activity on NK cells. Thawed NK cells from four donors were cultured with rhIL-18 (3 or 10 ng / mL) and rhIL-18BP (1 μg / mL) in the presence of rhIL-12 (10 ng / mL) for 30 minutes to allow IL-18-IL-18BP complex formation. After 30 minutes of incubation, cells were treated with increasing doses of anti-IL-18BP antibodies (20 μg / mL to 0.25 μg / mL; dilution factor of 1:3 (FIGS. 14A-G); or 10 μg / mL to 0.325 μg / mL; dilution factor of 1:2 (FIGS. 14H-N)) or isotype control (20 μg / mL (FIGS. 14A-G) or 10 μg / mL (FIGS. 14A-G)). [Figure 14I] Figure 14 shows that anti-IL-18BP antibodies were able to fully restore IFNγ secretion (Figures 14B-14D, 14I-14N) and CD69 expression (Figures 14E-14G) in a dose-dependent manner. The isotype control was unable to restore IL-18 activity. [Figure 14J] Figure 14 shows that anti-IL-18BP antibodies were able to fully restore IFNγ secretion (Figures 14B-14D, 14I-14N) and CD69 expression (Figures 14E-14G) in a dose-dependent manner. The isotype control was unable to restore IL-18 activity. [Figure 14K] Figure 14 shows that anti-IL-18BP antibodies were able to fully restore IFNγ secretion (Figures 14B-14D, 14I-14N) and CD69 expression (Figures 14E-14G) in a dose-dependent manner. The isotype control was unable to restore IL-18 activity. [Figure 14L] Figure 14 shows that anti-IL-18BP antibodies were able to fully restore IFNγ secretion (Figures 14B-14D, 14I-14N) and CD69 expression (Figures 14E-14G) in a dose-dependent manner. The isotype control was unable to restore IL-18 activity. [Figure 14M]Figure 14 shows that anti-IL-18BP antibodies were able to fully restore IFNγ secretion (Figures 14B-14D, 14I-14N) and CD69 expression (Figures 14E-14G) in a dose-dependent manner. The isotype control was unable to restore IL-18 activity. [Figure 14N] Figure 14 shows that anti-IL-18BP antibodies were able to fully restore IFNγ secretion (Figures 14B-14D, 14I-14N) and CD69 expression (Figures 14E-14G) in a dose-dependent manner. Isotype controls were unable to restore IL-18 activity. Figure 14 shows dose-response curves and calculated EC50s for % rescue by anti-IL-18BP antibodies. Representative data are from one donor. Rescue by anti-IL-18BP Ab is calculated as [(IL-12 + IL-18 + IL-18BP + anti-IL-18BP Ab) - (IL-12 + IL-18 + IL-18BP + isotype)] / [(IL-12 + IL-18) - (IL-12 + IL-18 + IL-18BP + isotype)].

[0115] [Figure 15A] 1 shows that anti-IL-18BP antibodies blocked IL-18BP secreted from PBMCs. 2 shows a schematic diagram of the assay setup. [Figure 15B]

[0023] Figure 1 shows that anti-IL-18BP antibodies blocked IL-18BP secreted from PBMCs. Figure 1 shows that anti-IL-18BP antibodies were able to induce dose-dependent IFNγ secretion above IL-12 + IL-18 control levels, suggesting that the antibodies can block endogenous IL-18BP activity. Representative data are from one donor. [Figure 15C]

[0023] Figure 1 shows that anti-IL-18BP antibodies blocked IL-18BP secreted from PBMCs. Figure 1 shows that anti-IL-18BP antibodies were able to induce dose-dependent IFNγ secretion above IL-12 + IL-18 control levels, suggesting that the antibodies can block endogenous IL-18BP activity. Representative data are from one donor. [Figure 15D] Figure 15B shows that anti-IL-18BP antibody blocked IL-18BP secreted from PBMCs. Schematic diagram of the assay setup. Thawed PBMCs from two donors were incubated with rhIL-12 (10 ng / mL), rhIL-18 (33.3 ng / mL) and a dose titration of anti-IL-18BP antibody (Figure 15B: 20 ​​μg / mL to 0.625 μg / mL; dilution factor of 1:2). [Figure 15E] Figures 15E-15J show that anti-IL-18BP antibodies blocked IL-18BP secreted from PBMCs. Cells were cultured for 24 hours with IL-12 (6 μg / mL - 0.002 μg / mL; 1:3 dilution factor) or isotype control (20 μg / mL). Anti-IL-18BP antibodies were able to induce dose-dependent IFNγ secretion above IL-12 + IL-18 control levels, suggesting that the antibodies can block endogenous IL-18BP activity. Representative data are from one donor. [Figure 15F] Figures 15E-15J show that anti-IL-18BP antibodies blocked IL-18BP secreted from PBMCs. Cells were cultured for 24 hours with IL-12 (6 μg / mL - 0.002 μg / mL; 1:3 dilution factor) or isotype control (20 μg / mL). Anti-IL-18BP antibodies were able to induce dose-dependent IFNγ secretion above IL-12 + IL-18 control levels, suggesting that the antibodies can block endogenous IL-18BP activity. Representative data are from one donor. [Figure 15G]Figures 15E-15J show that anti-IL-18BP antibodies blocked IL-18BP secreted from PBMCs. Cells were cultured for 24 hours with IL-12 (6 μg / mL - 0.002 μg / mL; 1:3 dilution factor) or isotype control (20 μg / mL). Anti-IL-18BP antibodies were able to induce dose-dependent IFNγ secretion above IL-12 + IL-18 control levels, suggesting that the antibodies can block endogenous IL-18BP activity. Representative data are from one donor. [Figure 15H] Figures 15E-15J show that anti-IL-18BP antibodies blocked IL-18BP secreted from PBMCs. Cells were cultured for 24 hours with IL-12 (6 μg / mL - 0.002 μg / mL; 1:3 dilution factor) or isotype control (20 μg / mL). Anti-IL-18BP antibodies were able to induce dose-dependent IFNγ secretion above IL-12 + IL-18 control levels, suggesting that the antibodies can block endogenous IL-18BP activity. Representative data are from one donor. [Figure 15I] Figures 15E-15J show that anti-IL-18BP antibodies blocked IL-18BP secreted from PBMCs. Cells were cultured for 24 hours with IL-12 (6 μg / mL - 0.002 μg / mL; 1:3 dilution factor) or isotype control (20 μg / mL). Anti-IL-18BP antibodies were able to induce dose-dependent IFNγ secretion above IL-12 + IL-18 control levels, suggesting that the antibodies can block endogenous IL-18BP activity. Representative data are from one donor. [Figure 15J]Figures 15E-15J show that anti-IL-18BP antibodies blocked IL-18BP secreted from PBMCs. Cells were cultured for 24 hours with IL-12 (6 μg / mL - 0.002 μg / mL; 1:3 dilution factor) or isotype control (20 μg / mL). Anti-IL-18BP antibodies were able to induce dose-dependent IFNγ secretion above IL-12 + IL-18 control levels, suggesting that the antibodies can block endogenous IL-18BP activity. Representative data are from one donor.

[0116] [Figure 16] FIG. 1 shows the affinity measurement of anti-mouse mIL18BP Ab to mouse IL18-BP protein by ELISA.

[0117] [Figure 17] FIG. 1 shows SPR kinetic measurements of anti-mouse IL18-BP (AbD35328 (also called "837," "Ab837," or "AB-837")).

[0118] [Figure 18] FIG. 1 shows analysis of mAb performance in functional blocking of mIL18-BP-mIL-18 interaction by ELISA.

[0119] [Figure 19] FIG. 1 shows IC50 analysis of anti-mouse IL18-BP (AbD35328).

[0120] [Figure 20] FIG. 1 shows the functional blocking activity of purified mAbs against mouse IL18-BP through IFNγ secretion.

[0121] [Figure 21] FIG. 1 shows EC50 analysis for anti-mouse IL18-BP.

[0122] [Figure 22A]22A to 22L show the evaluation of anti-IL18-BP monotherapy or combination therapy with anti-PD-L1 Ab in a mouse syngeneic CT26 tumor model. (Figure 22A) Tumor growth measurements for each group in monotherapy. [Figure 22B] FIG. 1 shows an analysis of survival percentages for each group in monotherapy. [Figure 22C] FIG. 11 is a summary of tumor growth measurements for individual mice in each monotherapy group. [Figure 22D] FIG. 11 is a summary of tumor growth measurements for individual mice in each monotherapy group. [Figure 22E] FIG. 11 is a summary of tumor growth measurements for individual mice in each monotherapy group. [Figure 22F] FIG. 11 is a summary of tumor growth measurements for individual mice in each monotherapy group. [Figure 22G] FIG. 1 shows tumor growth measurements for each group in combination therapy. [Figure 22H] FIG. 1. Survival analysis of combination therapy. [Figure 22I] FIG. 14. Summary of tumor growth measurements for individual mice in each combination therapy group. [Figure 22J] FIG. 14. Summary of tumor growth measurements for individual mice in each combination therapy group. [Figure 22K] FIG. 14. Summary of tumor growth measurements for individual mice in each combination therapy group. [Figure 22L] FIG. 10: Statistical analysis of the effect of combination therapy.

[0123] [Figure 23A] Figures 23A to 23L show the evaluation of anti-IL18-BP monotherapy or combination therapy with anti-PD-L1 Ab in the syngeneic B16 / Db-hmgp100 mouse tumor model. (Figure 23A) Tumor growth measurements for each group in monotherapy. [Figure 23B] FIG. 1 shows an analysis of survival percentages for each group in monotherapy. [Figure 23C] FIG. 11 is a summary of tumor growth measurements for individual mice in each monotherapy group. [Figure 23D] FIG. 11 is a summary of tumor growth measurements for individual mice in each monotherapy group. [Figure 23E] FIG. 11 is a summary of tumor growth measurements for individual mice in each monotherapy group. [Figure 23F] FIG. 11 is a summary of tumor growth measurements for individual mice in each monotherapy group. [Figure 23G] FIG. 1 shows tumor growth measurements for each group in combination therapy. [Figure 23H] FIG. 1 shows an analysis of survival percentages for each group in the combined therapy. [Figure 23I] FIG. 14. Summary of tumor growth measurements for individual mice in each combination therapy group. [Figure 23J] FIG. 14. Summary of tumor growth measurements for individual mice in each combination therapy group. [Figure 23K] FIG. 14. Summary of tumor growth measurements for individual mice in each combination therapy group. [Figure 23L] FIG. 10: Statistical analysis of the effect of combination therapy.

[0124] [Figure 24A] 24A to 24G show the activity of the combination of anti-IL18-BP and anti-TIGIT in the B16 / Db-hmgp100 syngeneic mouse tumor model. (FIG. 24A) Tumor growth measurements for each group in the combined therapy. [Figure 24B] FIG. 1 shows an analysis of the percentage survival of each combination therapy group. [Figure 24C] FIG. 14. Summary of tumor growth measurements for individual mice in each combination therapy group. [Figure 24D] FIG. 14. Summary of tumor growth measurements for individual mice in each combination therapy group. [Figure 24E] FIG. 14. Summary of tumor growth measurements for individual mice in each combination therapy group. [Figure 24F] FIG. 14. Summary of tumor growth measurements for individual mice in each combination therapy group. [Figure 24G] FIG. 10: Statistical analysis of the effect of combination therapy.

[0125] [Figure 25A] 25A to 25G show the activity of the combination of anti-IL18-BP and anti-PVRIG in the B16 / Db-hmgp100 syngeneic mouse tumor model. (FIG. 25A) Tumor growth measurements for each group in the combination therapy. [Figure 25B] FIG. 1 shows an analysis of survival percentages for each group in the combined therapy. [Figure 25C] FIG. 14. Summary of tumor growth measurements for individual mice in each combination therapy group. [Figure 25D] FIG. 14. Summary of tumor growth measurements for individual mice in each combination therapy group. [Figure 25E] FIG. 14. Summary of tumor growth measurements for individual mice in each combination therapy group. [Figure 25F] FIG. 14. Summary of tumor growth measurements for individual mice in each combination therapy group. [Figure 25G] FIG. 10: Statistical analysis of the effect of combination therapy.

[0126] [Figure 26A] Figures 26A-G show the monotherapy activity of anti-IL18-BP and anti-mPD-L1 in the syngeneic E0771 orthotopic mouse tumor model. (Figure 26A) Tumor growth measurements for each group in monotherapy. [Figure 26B] FIG. 11 is a summary of tumor growth measurements for individual mice in each monotherapy group. [Figure 26C] FIG. 11 is a summary of tumor growth measurements for individual mice in each monotherapy group. [Figure 26D] FIG. 11 is a summary of tumor growth measurements for individual mice in each monotherapy group. [Figure 26E] FIG. 11 is a summary of tumor growth measurements for individual mice in each monotherapy group. [Figure 26F] FIG. 1 shows an analysis of survival percentages for each group in monotherapy. [Figure 26G] FIG. Statistical analysis of the effect of monotherapy.

[0127] [Figure 27A] Figure 1 shows a tumor re-challenge experiment in the E0771 TNBC model. Groups of 5-10 C57BL / 6 tumor-naive, age-matched mice were orthotopically inoculated with E0771 (0.5 x 10 cells). When tumors reached a volume of 250 mm, mice were treated with the indicated mAb: AB-837 mIgG1-D265A or isotype control, followed by five booster doses. Two months later, tumor-free, age-matched mice were orthotopically re-inoculated with E0771. Tumor volumes are expressed as mean volume ± SEM. [Figure 27B] Figure 1 shows a tumor rechallenge experiment in the E0771 TNBC model. Groups of 5-10 C57BL / 6 tumor-naive, age-matched mice were orthotopically inoculated with E0771 (0.5 x 10 cells). When tumors reached a volume of 250 mm, mice were treated with the indicated mAb: AB-837 mIgG1-D265A or isotype control, followed by five booster doses. Two months later, tumor-free, age-matched mice were orthotopically re-inoculated with E0771. Individual tumor measurements for each mouse are shown. CR - complete responder; PR - partial responder (TV ≤ 500 mm). [Figure 27C] Figure 1 shows a tumor re-challenge experiment in the E0771 TNBC model. Groups of 5-10 C57BL / 6 tumor-naive, age-matched mice were orthotopically inoculated with E0771 (0.5 x 10 cells). When tumors reached a volume of 250 mm, mice were treated with the indicated mAb: AB-837 mIgG1-D265A or isotype control, followed by five booster doses. Two months later, tumor-free, age-matched mice were orthotopically re-inoculated with E0771. Figure 2 shows Kaplan-Meier survival curves for each group. [Figure 27D]Figure 1 shows a tumor re-challenge experiment in the E0771 TNBC model. Groups of 5-10 C57BL / 6 tumor-naive, age-matched mice were orthotopically inoculated with E0771 (0.5 x 10 cells). When tumors reached a volume of 250 mm, mice were treated with the indicated mAb: AB-837 mIgG1-D265A or isotype control, followed by five booster doses. Two months later, tumor-free, age-matched mice were orthotopically re-inoculated with E0771. Figure 2 shows spleen weight / body weight ratio. [Figure 27E] Figure 1 shows a tumor re-challenge experiment in the E0771 TNBC model. Groups of 5-10 C57BL / 6 tumor-naive, age-matched mice were orthotopically inoculated with E0771 (0.5 x 10 cells). When tumors reached a volume of 250 mm, mice were treated with the indicated mAb: AB-837 mIgG1-D265A or isotype control, followed by five booster doses. Two months later, tumor-free, age-matched mice were orthotopically re-inoculated with E0771. Figure 2 shows the percentage of CD44+CD62L-CD8+ effector T cells. [Figure 27F] Figure 1 shows a tumor re-challenge experiment in the E0771 TNBC model. Groups of 5-10 C57BL / 6 tumor-naive, age-matched mice were orthotopically inoculated with E0771 (0.5 x 10 cells). When tumors reached a volume of 250 mm, mice were treated with the indicated mAb: AB-837 mIgG1-D265A or isotype control, followed by five booster doses. Two months later, tumor-free, age-matched mice were orthotopically re-inoculated with E0771. Figure 2 shows the number of CD19+ cells per mg of spleen.

[0128] [Figure 28A] FIG. 1 shows the amino acid sequence of the human IL18-BP protein. [Figure 28B] FIG. 1 shows secreted human IL18-BP protein chains. [Figure 28C] 1 shows the amino acid sequence of the mouse IL18-BP protein, with the signal peptide sequence highlighted. [Figure 28D] FIG. 1 shows secreted mouse IL18-BP protein chains. [Figure 28E] FIG. 1 shows the amino acid sequence of human IL18 protein. [Figure 28F] FIG. 1 shows the amino acid sequence of mouse IL18 protein.

[0129] [Figure 29A] FIG. 1 shows the sequences of the variable heavy and light chains and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2 and vlCDR3 of CHA.7.518.1.H4(S241P). [Figure 29B] FIG. 1 shows the sequences of the variable heavy and light chains and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2 and vlCDR3 of CHA.7.538.1.2.H4(S241P).

[0130] [Figure 30A] FIG. 1 shows the sequences of the variable heavy and light chains and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2 and vlCDR3 of CPA.9.083.H4(S241P). [Figure 30B] FIG. 1 shows the sequences of the variable heavy and light chains and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2 and vlCDR3 of CPA.9.086.H4(S241P).

[0131] [Figure 31] FIG. 1 shows the ability of mAbs against human IL18-BP to rescue human IL18 bound by IL18-BP in human serum, as demonstrated by ELISA.

[0132] [Figure 32] FIG. 1 shows the ability of mAbs against human IL18-BP to rescue cyno IL18 bound by cyno IL18-BP, as demonstrated using ELISA.

[0133] [Figure 33] FIG. 1 shows that TIGIT and IL18Rα are co-expressed in the TME.

[0134] [Figure 34A] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34B] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34C] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34D] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34E]Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34F] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34G] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34H] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34I] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34J]Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34K] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34L] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34M] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34N] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34O]Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34P] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34Q] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34R] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34S] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34T]Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34U] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34V] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34W] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34X] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34Y]Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34Z] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34AA] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34BB] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34CC] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34DD]Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34EE] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34FF] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34GG] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34HH] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34II]Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34JJ] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34KK] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34LL] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34MM] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34NN]Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34OO] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34PP] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34QQ] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34RR] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34SS]Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34TT] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34UU] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34VV] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34WW] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34XX]Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34YY] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34ZZ] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34AAA] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Fig. 34BBB] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34CCC]Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Fig.34DDD] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34EEE] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34FFF] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34GGG] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34HHH]Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34III] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34JJJ] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34KKK] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34LLL] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34MMM]Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34NNN] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34OOO] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34PPP] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34QQQ] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34RRR]Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34SSS] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34TTT] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34UUU] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34VVV] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34WWW]Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34XXX] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34YYY] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34ZZZ] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34AAAA] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34BBBB]Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34CCCC] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Fig.34DDDD] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34EEEE] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34FFFF] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34GGGG]Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34HHHH] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34III] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34JJJJ] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34KKKK] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34LLLL]Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34MMMM] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34NNNN] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34OOOO] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34PPPP] Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies. [Figure 34QQQQ]Figure 1 shows the sequences of four anti-TIGIT antibodies that block the interaction of TIGIT with PVR: CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), and CHA.9.547.13.H4(S241P), as well as benchmark antibodies BM26 and BM29, and numerous other anti-TIGIT antibodies.

[0135] [Figure 35A] FIG. 1 shows the amino acid sequences of the constant domains of human IgG1 (with some useful amino acid substitutions), IgG2, and IgG3. [Figure 35B] FIG. 1 shows the amino acid sequences of the human IgG4 constant domain, the IgG4 constant domain with hinge variants that find particular use in the present invention, and the constant domains of the kappa and lambda light chains.

[0136] [Figure 36A] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36B] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36C] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36D] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36E] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36F]FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36G] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36H] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36I] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36J] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36K] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36L] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36M] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36N] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36O]FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36P] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36Q] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36R] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36S] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36T] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36U] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36V] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36W] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36X]FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36Y] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36Z] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36AA] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36AB] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36AC] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36AD] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36AE] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36AF] FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention. [Figure 36AG]FIG. 1 shows the variable heavy and light chains, and vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2, and vlCDR3 sequences of the anti-PVRIG antibodies of the invention.

[0137] [Figure 37A] FIG. 1 shows the sequences of other PVRIG antibodies of the invention. [Figure 37B] FIG. 1 shows the sequences of other PVRIG antibodies of the invention. [Figure 37C] FIG. 1 shows the sequences of other PVRIG antibodies of the invention. [Figure 37D] FIG. 1 shows the sequences of other PVRIG antibodies of the invention.

[0138] [Figure 38A] FIG. 1 provides additional anti-PVRIG antibodies for use in the present invention. [Figure 38B] FIG. 1 provides additional anti-PVRIG antibodies for use in the present invention. [Figure 38C] FIG. 1 provides additional anti-PVRIG antibodies for use in the present invention. [Figure 38D] FIG. 1 provides additional anti-PVRIG antibodies for use in the present invention. [Figure 38E] FIG. 1 provides additional anti-PVRIG antibodies for use in the present invention. [Figure 38F] FIG. 1 provides additional anti-PVRIG antibodies for use in the present invention. [Figure 38G] FIG. 1 provides additional anti-PVRIG antibodies for use in the present invention. [Figure 38H] FIG. 1 provides additional anti-PVRIG antibodies for use in the present invention. [Figure 38I] FIG. 1 provides additional anti-PVRIG antibodies for use in the present invention. [Figure 38J] FIG. 1 provides additional anti-PVRIG antibodies for use in the present invention. [Figure 38K]FIG. 1 provides additional anti-PVRIG antibodies for use in the present invention. [Figure 38L] FIG. 1 provides additional anti-PVRIG antibodies for use in the present invention. [Figure 38M] FIG. 1 provides additional anti-PVRIG antibodies for use in the present invention. [Figure 38N] FIG. 1 provides additional anti-PVRIG antibodies for use in the present invention. [Figure 38O] FIG. 1 provides additional anti-PVRIG antibodies for use in the present invention. [Figure 38P] FIG. 1 provides additional anti-PVRIG antibodies for use in the present invention. [Figure 38Q] FIG. 1 provides additional anti-PVRIG antibodies for use in the present invention. [Figure 38R] FIG. 1 provides additional anti-PVRIG antibodies for use in the present invention. [Figure 38S] FIG. 1 provides additional anti-PVRIG antibodies for use in the present invention. [Figure 38T] FIG. 1 provides additional anti-PVRIG antibodies for use in the present invention. [Figure 38U] FIG. 1 provides additional anti-PVRIG antibodies for use in the present invention. [Figure 38V] FIG. 1 provides additional anti-PVRIG antibodies for use in the present invention. [Figure 38W] FIG. 1 provides additional anti-PVRIG antibodies for use in the present invention. [Figure 38X] FIG. 1 provides additional anti-PVRIG antibodies for use in the present invention.

[0139] [Figure 39A] FIG. 1 shows the sequences of exemplary anti-PD-1 antibodies. [Figure 39B] FIG. 1 shows the sequences of exemplary anti-PD-1 antibodies.

[0140] [Figure 40A] Figure 1 shows the sequences of exemplary anti-PD-L1 antibodies. [Figure 40B] Figure 1 shows the sequences of exemplary anti-PD-L1 antibodies. [Figure 40C] Figure 1 shows the sequences of exemplary anti-PD-L1 antibodies. [Figure 40D] Figure 1 shows the sequences of exemplary anti-PD-L1 antibodies. [Figure 40E] Figure 1 shows the sequences of exemplary anti-PD-L1 antibodies. [Figure 40F] Figure 1 shows the sequences of exemplary anti-PD-L1 antibodies. [Figure 40G] Figure 1 shows the sequences of exemplary anti-PD-L1 antibodies. [Figure 40H] Figure 1 shows the sequences of exemplary anti-PD-L1 antibodies. [Figure 40I] Figure 1 shows the sequences of exemplary anti-PD-L1 antibodies.

[0141] [Figure 41A-1] Figures 41A-D show Biacore KD measurements performed using biotinylated human / cynomolgus IL18BP-Fc protein coated on a CM5 chip. (Figure 41A-1) Biacore image of anti-IL18BP Fab-human IL18BP interaction; 10 resolution. [Figure 41A-2] A, Biacore image of anti-IL18BP Fab-human IL18BP interaction; 10 resolution. [Figure 41A-3] A, Biacore image of anti-IL18BP Fab-human IL18BP interaction; 10 resolution. [Figure 41B-1] A, Biacore image of anti-IL18BP Fab-human IL18BP interaction; 85 resolution. [Figure 41B-2] A, Biacore image of anti-IL18BP Fab-human IL18BP interaction; 85 resolution. [Figure 41B-3] A, Biacore image of anti-IL18BP Fab-human IL18BP interaction; 85 resolution. [Figure 41C-1] Biacore image of anti-IL18BP Fab-cyno IL18BP interaction, 10 resolution. [Figure 41C-2] Biacore image of anti-IL18BP Fab-cyno IL18BP interaction, 10 resolution. [Figure 41C-3] Biacore image of anti-IL18BP Fab-cyno IL18BP interaction, 10 resolution. [Figure 41D-1] Biacore image of anti-IL18BP Fab-cyno IL18BP interaction, 85 resolution. [Figure 41D-2] Biacore image of anti-IL18BP Fab-cyno IL18BP interaction, 85 resolution. [Figure 41D-3] Biacore image of anti-IL18BP Fab-cyno IL18BP interaction, 85 resolution.

[0142] [Figure 42] FIG. 1 presents a table showing KD values ​​for human / cyno anti-IL18BP Fab-IL18BP interactions measured by Biacore.

[0143] [Figure 43A-1] Figure 1 presents the affinity of the optimized IL18BP antibody accessed using MSD. Figure 2 shows an overlay of the Fab-IL18BP MSD image (black) and the human IL-18-IL18BP MSD image (green). [Figure 43A-2] Figure 1 presents the affinity of the optimized IL18BP antibody accessed using MSD. Figure 2 shows an overlay of the Fab-IL18BP MSD image (black) and the human IL-18-IL18BP MSD image (green). [Figure 43A-3] Figure 1 presents the affinity of the optimized IL18BP antibody accessed using MSD. Figure 2 shows an overlay of the Fab-IL18BP MSD image (black) and the human IL-18-IL18BP MSD image (green). [Figure 43B-1] Figure 1 presents the affinity of the optimized IL18BP antibody accessed using MSD. Figure 2 shows an overlay of the Fab-IL18BP MSD image (black) and the Cyno IL-18-IL18BP MSD image (green). [Figure 43B-2] Figure 1 presents the affinity of the optimized IL18BP antibody accessed using MSD. Figure 2 shows an overlay of the Fab-IL18BP MSD image (black) and the Cyno IL-18-IL18BP MSD image (green). [Figure 43B-3] Figure 1 presents the affinity of the optimized IL18BP antibody accessed using MSD. Figure 2 shows an overlay of the Fab-IL18BP MSD image (black) and the Cyno IL-18-IL18BP MSD image (green).

[0144] [Figure 44] FIG. 1 presents a table showing KD values ​​for human / cyno anti-IL18BP Fab-IL18BP interactions measured by MSD.

[0145] [Figure 45] FIG. 1 presents a table showing KD values ​​for human / cyno IL18-IL18BP interaction measured by MSD.

[0146] [Figure 46] FIG. 1 presents exemplary antibody characteristics for αIL-18BP antibodies of interest (αIL-18BP Abs).

[0147] [Figure 47] Figure 1 shows that IL-18BP levels are elevated in human cancers. Expression of IL18BP transcripts in normal (green) or cancer (red) tissues from the TCGA database. GBM, glioblastoma multiforme; HSNC, head and neck squamous cell carcinoma; KIRC, renal clear cell carcinoma; PAAD, pancreatic adenocarcinoma; SKCM, cutaneous melanoma; STAD, gastric adenocarcinoma (*P<0.01).

[0148] [Figure 48] (a) and (b) show that IL-18BP is expressed in suppressive myeloid populations in the tumor microenvironment (TME), suggesting a mechanism of resistance. Single-cell RNA analysis of tumor-infiltrating myeloid cells, including tumor-associated macrophages (TAMs) and dendritic cells (DCs), in human colorectal cancer (CRC) shows that IL-18BP is expressed in suppressive myeloid populations in tumors, suggesting a mechanism of resistance to immune activation in the tumor microenvironment (TME). Left panel: Peripheral (PBMC), normal tumor (NAT), and intratumoral myeloid cell populations. Right panel: IL18BP is primarily expressed in cDC2-CD1C and TAM-C1QC, suppressive myeloid populations, suggesting that IL18BP may be a mechanism of resistance to immune cell activation in tumors.

[0149] [Figure 49A] Figure 1 shows that αIL-18BP Ab (ADI-71739) enhances the stimulatory activity of human T cells. Schematic diagram of the assay setup. Thawed tumor-infiltrating lymphocytes (TILs) co-cultured with MEL624 cells at a 1:1 ratio were treated with rhIL-18 (R&D systems, 30 ng / mL) and rhIL-18BP (R&D systems, 1 μg / mL) for 30 minutes to allow the formation of IL-18-IL-18BP complexes. After 30 minutes of incubation, cells were treated with ADI-71739 or isotype control (10 μg / mL). [Figure 49B] Figure 1 shows that αIL-18BP Ab (ADI-71739) enhances the stimulatory activity of human T cells. Figure 2 shows that anti-IL-18BP antibodies were able to increase IFNγ secretion from TILs compared to the isotype control.

[0150] [Figure 50A]Figure 1 shows that anti-IL-18BP antibodies fully restored IL-18 activity in a MEL624:TIL assay. Schematic diagram of the assay setup. Thawed tumor-infiltrating lymphocytes (TILs) cocultured with MEL624 cells at a 1:1 ratio were treated with rhIL-18 (30 ng / mL) and rhIL-18BP (1 μg / mL) for 30 minutes to allow for the formation of IL-18-IL-18BP complexes. After 30 minutes of incubation, cells were treated with increasing doses of anti-IL-18BP antibody (ADI-71722) (30 μg / mL to 0.01 μg / mL; 1:3 dilution factor) or isotype control (30 μg / mL). [Figure 50B] Figure 50A shows that anti-IL-18BP antibodies fully restored IL-18 activity in a MEL624:TIL assay. Figure 50B shows that anti-IL-18BP antibodies were able to fully restore IFNγ secretion in a dose-dependent manner. The isotype control was unable to restore IL-18 activity. Figure 50C shows the dose-response curve and calculated EC50 for % rescue by anti-IL-18BP antibodies. Representative data are from one donor. Rescue by anti-IL-18BP Ab is calculated as [(IL-18 + IL-18BP + anti-IL-18BP Ab) - (IL-18 + IL-18BP + isotype)] / [(IL-18) - (IL-18 + IL-18BP + isotype)].

[0151] [Figure 51A]Figure 1 shows that anti-hIL-18BP antibody enhances the activity of PD-1 and DNAM-1 axis blocking in an in vitro CMV recall assay. Anti-IL-18BP antibody alone and in combination with aPVRIG / aTIGIT / pembrolizumab increased IFNγ secretion by CMV-reactive T cells. Schematic diagram of the assay setup. PD-L1-overexpressing MEL-624 cells were loaded with CMV pp65 peptide and plated. Cells were cultured with rhIL-18 (30 ng / mL) and rhIL-18BP (2 μg / mL) for 30 minutes to allow the formation of IL-18-IL-18BP complexes. Cells were then treated with anti-IL-18BP antibody (ADI-71722), anti-PVRIG, anti-TIGIT, pembrolizumab, or an isotype control (all antibodies administered at the same final concentration of 10 μg / mL). After 30 minutes of incubation with antibody, CMV-reactive T cells were added to the cultures. [Figure 51B] Figure 1 shows that anti-hIL-18BP antibody enhances the activity of PD-1 and DNAM-1 axis blocking in an in vitro CMV recall assay. Anti-IL-18BP antibody, alone and in combination with aPVRIG / aTIGIT / pembrolizumab, increased IFNγ secretion by CMV-reactive T cells. ADI-71722, alone and in combination with anti-PVRIG / anti-TIGIT / pembrolizumab, increased IFNγ secretion by CMV-reactive T cells. Left panel: Anti-IL-18BP antibody alone could fully restore IFNγ secretion, and the restoration was even stronger when combined with pembrolizumab / anti-PVRIG. Right panel: ADI-71722 alone could fully restore IFNγ secretion, and the restoration was even stronger when combined with pembrolizumab / anti-TIGIT.

[0152] [Figure 52]

[0023] Figure 1 provides data showing that ADI-71739 binds with high affinity to human and cyno IL-18BP and with low affinity to mouse IL-18BP. ADI-71739 binds with high affinity to human and cyno IL-18BP and with low affinity to mouse IL-18BP: Top panel (left to right): Human IL18-IL18BP interaction, cyno IL18-IL18BP interaction, and mouse IL18-IL18BP interaction measurements in KinExA, with final KDs of 441 fM, 345 fM, and 3.7 pM, respectively. Bottom panel (left to right): ADI-71739-human IL18BP interaction, ADI-71739-cyno IL18BP interaction measurements in KinExA, and ADI-71739-mouse IL18BP interaction measurements by Biacore. The final KDs are 291 fM, 209 fM, and 4 nM, respectively. For each run on KinExA, two or three curves with different column binding protein (CBP) concentrations were run and analyzed using n-curve analysis to determine the Kd.

[0153] [Figure 53] 1 shows the blocking effect of anti-IL18BP Ab on the binding of human IL18BP to human IL-18. The blocking effect of anti-IL18BP Ab was tested by ELISA using 1 ng / mL human IL-18 protein.

[0154] [Figure 54] Figure 1 shows a competitive ELISA using the complex of soluble IL18-IL18BP and anti-IL18BP Ab. Blocking of IL18-IL18BP complex formation was tested by ELISA. MAB1191 shows reduced blocking activity compared to 66716 Ab.

[0155] [Figure 55]Figure 55A shows the expression of IL18Rα on CD8+ and CD4+ T and NK TILs from dissociated human tumors of various cancer types. Each dot represents a distinct tumor from an individual patient. Fold expression values ​​were calculated by dividing the MFI of the target by the MFI of the relevant isotype control (FOI). Means and SEM are shown in tick charts. Figure 55B shows the expression of IL18Rα on CD4+ and CD8+ T and NK cells from donor-matched PBMCs and TME. Statistical analysis was performed using a paired t-test (two-tailed), p<0.05; **p=0.0064.

[0156] [Figure 56] Figures 56A and 56B show that IL18 levels are increased in the serum of cancer patients compared to levels in HD serum. Figure 56A shows the levels of IL18 analytes (IL18 and IL18BP) in the serum of patients across indications. Figure 56B shows dot plots representing IL18 analytes in serum samples from individual patients or HDs. Statistical analysis was performed using a two-tailed t-test, P < 0.0005***.

[0157] [Figure 57]

[0023] Figure 1 shows IL18 analyte (IL18 and IL18BP) levels in tumor-derived supernatants (TDS) across indications. The dot plot represents IL18 analyte in TDS samples. Each dot represents an individual patient sample.

[0158] [Figure 58] Figure 58A shows levels of IL18 in tumor-derived supernatants (TDS) of patients across indications. Mean levels are shown as black lines. Figure 58B shows levels of IL18BP in tumor-derived supernatants (TDS) of patients across indications. Mean levels are shown as black lines.

[0159] [Figure 59] Figure 59A shows that IL-18BP is expressed in suppressive myeloid populations in the TME, suggesting a mechanism of resistance. IL-18BP is expressed in suppressive myeloid populations and correlates with PD-L1 in the TME, suggesting a mechanism of resistance. Figure 59A shows that IL-18BP correlates with PD-L1 at the RNA level (TCGA) in colon and breast cancer, suggesting a mechanism of resistance to immune activation in the tumor microenvironment (TME). Figure 59B shows that single-cell RNA analysis of tumor-infiltrating myeloid cells, including tumor-associated macrophages (TAMs) and dendritic cells (DCs), in colon cancer patients shows that IL-18BP is upregulated in myeloid populations in the TME compared to the periphery (PBMCs), suggesting a mechanism of resistance to immune activation in the TME. Figure 59C shows that single-cell RNA analysis of tumor-infiltrating myeloid cells, including tumor-associated macrophages (TAMs) and dendritic cells (DCs), across indications shows that IL-18BP is upregulated in myeloid populations in the TME compared to the periphery (PBMCs), suggesting a mechanism of resistance to immune activation in the TME.

[0160] [Figure 60]Figures 60A-60D show that IL-18BP is upregulated after immune checkpoint blocking (ICB) treatment. Figure 60A shows that IL-18BP is upregulated after ICB treatment (RNA level). IL-18BP levels are upregulated in the tumor microenvironment (RNA) after treatment with anti-PD-1 (breast cancer), suggesting a potential resistance mechanism. Figure 60B shows that IL-18BP is upregulated after ICB treatment (RNA level). IL-18BP levels are upregulated in the tumor microenvironment (RNA) after treatment with anti-PD-1 plus anti-CTLA-4 (melanoma), suggesting a potential resistance mechanism. Figure 60C shows that IL-18BP is upregulated after ICB treatment (RNA level). IL-18BP levels are upregulated in the tumor microenvironment (RNA) after treatment with anti-PD-1 (basal cell carcinoma), suggesting a potential resistance mechanism. Figure 60D shows that IL-18BP is elevated in NSCLC patient serum after aPD-(L)1 treatment. Quantification of plasma IL-18BP protein levels by ELISA in healthy donors (n=22) and NSCLC patients (n=52) at baseline before treatment and after treatment with anti-PD-(L)1 (n=52).

[0161] [Figure 61] Figure 61A shows that baseline serum levels of IL-18BP may be associated with an inadequate response to anti-PD-1. Supporting data for the role of IL-18BP as a soluble ICP and a potential mechanism of resistance to PD1 blockade in patients with renal cell carcinoma receiving pembrolizumab + lenvatinib. Figure 61B shows that high IL-18BP in the serum of patients pretreated with pembrolizumab + lenvatinib is associated with shorter progression-free survival (PFS). Figure 61C shows that high IL-18BP in the serum of patients pretreated with pembrolizumab + lenvatinib is associated with stable or progressive disease (SD / PD).

[0162] [Figure 62]Figure 1 shows that baseline serum levels of IL-18BP can be associated with an inadequate response to anti-PD-1. Supporting data for the role of IL-18BP as a soluble ICP in melanoma cancer patients undergoing anti-PD-1 therapy and a potential mechanism of resistance to PD1 blocking. High IL-18BP in the serum of melanoma cancer patients pretreated with anti-PD-1 is associated with an inadequate response. Raw Olink data (NPX format). Student's T-test was performed for IL18BP protein after intensity normalization for the target product.

[0163] [Figure 63] Figure 63A shows a principal component analysis (PCA) of IL-18 levels in serum from head and neck cancer patients. Figure 63B shows a principal component analysis (PCA) of IL-18BP levels in serum from head and neck cancer patients. The PCA shows that the tumor location primarily separates between samples with high versus low levels of IL-18. A-B. Tumor location in the tongue correlates with higher levels of IL-18 and lower levels of IL18BP compared to other locations.

[0164] [Figure 64] Figure 1 shows IL-18 and IL-18BP levels in the serum of head and neck patients at different tumor sites (dot plots). Higher levels of IL-18 in the serum of head and neck patients are shown in the tongue.

[0165] [Figure 65A] Figure 1 shows that IL18 and IL18BP plasma levels in NSCLC patients increase after anti-PD-1 monotherapy or anti-PD-1 plus chemotherapy combination. Mean plasma levels of IL18 and IL18BP are higher in responder patients at baseline and increase in NR patients treated with anti-PD1. Figure 2 shows IL18 and IL18BP levels in plasma of R / NR NSCLC patients at baseline. [Figure 65B]Figure 1 shows that IL18 and IL18BP plasma levels in NSCLC patients increase after anti-PD-1 monotherapy or anti-PD-1 plus chemotherapy combination. Mean plasma levels of IL18 and IL18BP are higher in responder patients at baseline and increase in NR patients treated with anti-PD1. Figure 2 shows IL18 and IL18BP levels in the plasma of individual NSCLC patients (R / NR) at baseline and after a single anti-PD-1 treatment. [Figure 65C] Figure 1 shows that IL18 and IL18BP plasma levels in NSCLC patients increase after anti-PD-1 monotherapy or anti-PD-1 plus chemotherapy combination. Mean plasma levels of IL18 and IL18BP are higher in responder patients at baseline and increase in NR patients treated with anti-PD1. Figure 2 shows IL18 and IL18BP levels in plasma of individual NSCLC patients (R / NR) at baseline and after a single anti-PD1 treatment or chemotherapy plus anti-PD-1 combination treatment. [Figure 65D] Figure 1 shows the percentage change from baseline in IL18 and IL18BP in R / NR NSCLC patients after single anti-PD-1 treatment or chemotherapy in combination with anti-PD-1. P values ​​in graphs A-C were obtained according to paired T-test.

[0166] [Figure 66]Figure 1 shows whole blood assay data. The anti-IL-18BP antibody Ab-71709, alone or in combination with nivolumab, showed no signs of systemic immune activation in ID.flow, an ex vivo system mimicking human blood circulation. Fresh whole blood was drawn from six healthy volunteers and immediately transferred to a whole blood loop system. The test item was administered, and the blood was circulated at 37°C to prevent clotting. Blood samples drawn at 24 hours were analyzed for hematology and flow cytometry parameters and then processed to plasma for cytokine analysis. The anti-CD52 antibody alemtuzumab was included as a reference antibody with manageable cytokine release in clinical settings. In contrast to alemtuzumab, the anti-IL-18BP antibody, alone or in combination with the anti-PD1 antibody nivolumab, did not induce any signs of systemic immune activation according to the various readouts used.

[0167] [Figure 67] Figure 67A shows an in vitro experiment testing the effect of ADI-71739 on melanoma cell killing by human TILs. The anti-IL18-BP antibody ADI-71739 increased melanoma cell killing by tumor-infiltrating lymphocytes. Figure 67A shows a schematic diagram of the assay setup. MEL624 cells were co-cultured with human TILs pre-enriched for MART1 or gp100 peptide-specific clones. rhIL-18 (R&D systems, 50 ng / mL) and rhIL-18BP (R&D systems, 1 μg / mL) were added to the co-culture for 30 minutes to allow IL-18:IL-18BP complexes to form before treatment with 10 μg / mL ADI-71739 or isotype control. The co-cultures were monitored for 72 hours using an IncuCyte live-cell imaging instrument. Figure 67B shows that addition of IL-18 (gray) enhanced tumor cell killing, as indicated by lower confluence (left) and increased apoptosis (right) of MEL624 cells over time. In the presence of an isotype control antibody (black), IL-18BP abolished the effects of IL-18, whereas an anti-IL-18BP antibody (turquoise) was able to fully restore these effects.

[0168] [Figure 68] Figure 68A shows an in vitro experiment testing the effect of combining ADI-71739 with other checkpoint blocking antibodies. The anti-IL18-BP antibody ADI-71739, alone and in combination with aPVRIG / aTIGIT / pembrolizumab, increased IFNγ secretion by CMV-specific T cells. Figure 68B shows a schematic of the assay setup. MEL624 cells overexpressing PD-L1 were loaded with the CMV peptide pp65. Cells were cultured with rhIL-18 (R&D systems, 30 ng / mL) and rhIL-18BP (R&D systems, 2 μg / mL) for 30 minutes to allow the formation of IL-18:IL-18BP complexes. Cells were then treated with 10 μg / mL of ADI-71739, aPVRIG (anti-PVRIG), aTIGIT (anti-TIGIT), pembrolizumab (anti-PD-L1), or an isotype control, either alone or in various combinations. CMV-specific T cells were then added to the cultures, and IFNγ secretion was measured after overnight incubation. Figure 68B shows that anti-IL-18BP alone can increase IFNγ secretion by T cells, and this effect was enhanced when combined with pembrolizumab / aPVRIG / aTIGIT.

[0169] [Figure 69]Figure 69A shows an in vitro experiment examining the effect of ADI-71739 on human TIL function in the presence of endogenous IL-18BP levels. The anti-IL18BP antibody ADI-71739 increased IFNγ release by tumor-infiltrating lymphocytes. Figure 69A is a schematic diagram of the assay setup. MEL624 cells were co-cultured with human TILs pre-enriched for MART1 or gp100 peptide-specific clones. IL-18 (3.7 ng / mL) was added to the co-cultures along with 5 μg / mL of ADI-71739 or an isotype control. Co-cultures were performed for 18 hours, after which IFNγ levels were measured in the supernatants. Figure 69B shows that IFNγ levels were increased in co-cultures treated with ADI-71739 (turquoise) compared to isotype-treated samples (black). Representative examples from two TIL donors are shown.

[0170] [Figure 70] Figure 70A shows that bound IL-18 levels in the TME exceed the amount required for in vitro T cell activation. Figure 70A is a schematic diagram of the assay setup. Thawed tumor-infiltrating lymphocytes (TILs) cocultured with MEL624 cells at a 1:1 ratio were treated with rhIL-18 (R&D Systems, 1.23-300 ng / mL) for 24 hours. Figure 70B shows that rhIL-18 increased IFNγ secretion in a dose-dependent manner. rhIL-18 activated TILs at concentrations above approximately 1 ng / mL, reaching saturation at approximately 100 ng / mL. Figure 70C shows that bound IL-18 levels in TDS across indications mostly exceed the levels required for in vitro T cell activation. Bound IL-18 levels were calculated by subtracting free IL-18 from total IL-18 measured for each sample by two separate ELISA kits. The red dashed line represents the level required for functional activity (1.5 ng / mL). The black line represents the median level of bound IL-18 for each tumor type.

[0171] [Figure 71]Figure 71 shows that inflammasome-inducing cytokines such as IL-18 and IL-1β are enriched in the TME, unlike other cytokines. Figure 71A shows that IL-18 and IL-1β are inflammasome-derived cytokines with opposing effects in the TME. IL-18 promotes T cell and NK cell activation, resulting in anti-tumorigenic activity, while IL1β has a dual role, resulting in pro-tumorigenic activity when combined. Figure 71B shows dot plots of cytokine levels in tumor-derived supernatants measured across various indications. Each dot represents one sample. The average is indicated by the short black line. The dashed red lines represent the limit of detection for each cytokine.

[0172] [Figure 72] Figure 1 shows a combination study of anti-IL-18BP antibody and anti-PD-L1 antibody in a mouse tumor model. Anti-IL-18BP Ab in combination with anti-PD-L1 Ab increases tumor growth inhibition and survival in a mouse tumor model. Groups of 10 6-week-old female C57BL / 6 mice were injected subcutaneously with E0771 and administered mIgG1 Synagis isotype control, anti-mouse IL-18BP Ab, anti-PDL1 Ab, or a combination of anti-mouse IL-18BP Ab and anti-PD-L1 Ab (IP), followed by six booster doses. Tumor volumes are expressed as mean volume + SEM. Tumor volumes were measured twice weekly.

[0173] [Figure 73A] Figure 1 shows that administration of anti-IL18BP is expected to have a better therapeutic window than engineered IL-18. C57BL / 6 mice were injected subcutaneously with MC38ova cells and treated twice weekly with the indicated mAb Synagis mIgG1 (IP), anti-IL18bp mIgG1 (IP), PBS (SC), or engineered IL-18 (SC). Figure 2 shows the body weights of mice weighed weekly. [Figure 73B](Figure 1) Administration of anti-IL18BP is expected to have a better therapeutic window than engineered IL-18. C57BL / 6 mice were injected subcutaneously with MC38ova cells and treated twice weekly with the indicated mAb Synagis mIgG1 (IP), anti-IL18bp mIgG1 (IP), PBS (SC), or engineered IL-18 (SC). Mice were bled before the fourth treatment, 4 hours after the fourth treatment, and 24 hours after the fourth treatment. Serum was analyzed for the presence of the indicated molecules - IFNγ, TNF, MCP1, IL6. (Figure 1) [Figure 73C] (Figure 1) Administration of anti-IL18BP is expected to have a better therapeutic window than engineered IL-18. C57BL / 6 mice were injected subcutaneously with MC38ova cells and treated twice weekly with the indicated mAb Synagis mIgG1 (IP), anti-IL18bp mIgG1 (IP), PBS (SC), or engineered IL-18 (SC). Serum was analyzed for IL-18 levels. (Figure 1) [Figure 73D] FIG. 1 shows spleens harvested from mice 24 hours after the fourth treatment and their weights. [Figure 73E] FIG. 1 shows the weight of spleens harvested from mice treated with IL15 or IL15+ILRα.

[0174] [Figure 74] Figure 74 shows evaluation of anti-IL18-BP monotherapy in a mouse syngeneic MC38ova tumor model. C57BL / 6 mice were injected subcutaneously with 1.2M MC38ova cells and treated twice weekly with the indicated mAb Synagis mIgG1 (IP), anti-IL18bp mIgG1 (IP). Figure 74A shows tumor growth measurements for each group. Figure 74B shows a summary of tumor growth measurements for individual mice in each group.

[0175] [Figure 75]Figure 75 shows that anti-IL18bp antibodies modulate the tumor microenvironment without peripheral effects in the MC38ova tumor model. C57BL / 6 mice were injected subcutaneously with MC38ovadim and treated with anti-mouse IL-18BP Ab (IP). Tumors, spleens, and serum were collected to determine immune composition and cytokine concentrations. Figures 75A-G show the tumor microenvironment. Figure 75H shows the spleen. Figure 75I shows the serum.

[0176] [Figure 76] Binding of MAB1191 Ab to human IL18BP, affinity measurement using Biacore.

[0177] [Figure 77] Figure 77A shows the effect of anti-IL18BP in combination with oxaliplatin in the MC38ovadim tumor model. Groups of 10 C57BL / 6 mice were inoculated with MC38OVAdim. At a tumor volume (TV) of 110 mm, mice were treated with 5 mg / kg oxaliplatin or control DDW. At a TV of 140 mm, mice were treated with 15 mg / kg anti-IL18BP mIgG1 Ab or isotype control, followed by five additional doses. Figure 77A shows TV expressed as mean volume ± SEM. Figure 77B shows individual tumor measurements for each mouse (n=10 / group). CR - complete responder, PR - partial responder (TV≦500 mm), CR - complete responder, PR - partial responder.

[0178] [Figure 78]Figure 78A shows anti-IL-18BP Ab inhibits tumor growth in the MC38ova mouse tumor model. Figure 78B shows anti-IL-18BP Ab inhibits tumor growth in the B16F10-hmgp100 mouse tumor model. Tumor volumes are expressed as mean volume ± SEM. Groups of 10 C57BL / 6 mice were inoculated with MC38ovadim or B16F10-hmgp100 cells. Mice were treated with the indicated mAb: anti-IL-18BP Ab or isotype control. Figure 78A shows anti-IL-18BP Ab inhibits tumor growth in the MC38ova mouse tumor model. Figure 78B shows anti-IL-18BP Ab inhibits tumor growth in the B16F10-hmgp100 mouse tumor model. Tumor volumes are expressed as mean volume ± SEM. DETAILED DESCRIPTION OF THE INVENTION

[0179] I. Introduction A. Interleukin-18 binding protein The present invention provides antibodies that specifically bind to interleukin-18 binding protein (IL18-BP). "Protein" in this context is used interchangeably with "polypeptide" and includes peptides as well. The present invention provides antibodies that specifically bind to IL18-BP.

[0180] The IL18-BP gene is located on human chromosome 11, and no exons encoding the transmembrane domain were found in the 8.3-kb genomic sequence containing the IL18-BP gene. Four isoforms of IL18-BP, generated by alternative mRNA splicing, have been identified in humans. They are designated IL18-BP a, b, c, and d, and all share the same N-terminus but differ in their C-terminus (Novick, D. et al., Immunity, 10:127-136, (1999)). These isoforms differ in their ability to bind IL18 (Kim, S.-H. et al., PNAS, 97(3):1190-1195 (2000)). Of the four human IL18-BP (hIL18-BP) isoforms, isoforms a and c are known to have neutralizing activity against IL18. The most abundant IL18-BP isoform, isoform a, exhibits high affinity for IL18, with a fast on-rate, a slow off-rate, and a dissociation constant (K) of approximately 0.4 nM. d) (Kim, S.-H. et al., PNAS, 97(3):1190-1195(2000)). Other researchers have reported that the affinity of IL18-BP for IL-18 is approximately 1 pM or approximately 25 pM (Zhou T. et al., Nature, 583(7817):609-614,(2020), Girard C. et al., Rheumatology 2016; 55:2237-2247(2016)). The IL18-BPb and IL18-BPd isoforms lack the complete Ig domain and lack the ability to bind to or neutralize IL18. Two mouse isoforms of IL18-BP arise from mRNA splicing and have been found in various cDNA libraries, expressed, purified, and evaluated for binding and neutralizing IL18 biological activity (Kim, S.-H. et al., PNAS, 97(3):1190-1195(2000)). Human and mouse IL18-BP share 60.8% amino acid similarity. The mouse IL18-BPc and IL18-BPd isoforms, which share the same Ig domain, also neutralize more than 95% of mouse IL18 at a 2 molar excess. However, mouse IL18-BPd, which shares a common C-terminal motif with human IL18-BPa, also neutralizes human IL18. Molecular modeling has identified a large mixed electrostatic and hydrophobic binding site in the Ig domain of IL18-BP, which may explain its high-affinity binding to ligands (Kim, S.-H. et al., PNAS, 97(3):1190-1195(2000)).

[0181] IL18-BP is a 194 amino acid long secreted protein with a signal peptide (spanning amino acids 1-30) and a secretory chain (spanning amino acids 41-171) and four potential N-glycosylation sites, but no transmembrane domain. The full-length human IL18-BP isoform a protein is shown in FIG. 28 (SEQ ID NO: 254). The present disclosure provides formulations comprising antibodies that specifically bind to the IL18-BP protein. In this context, "protein" is used interchangeably with "polypeptide" and includes peptides as well. The present disclosure provides antibodies that specifically bind to the IL18-BP protein. IL18-BP is a 194 amino acid long secreted protein with a signal peptide (spanning amino acids 1-30) and a secretory chain (spanning amino acids 41-171).

[0182] Thus, as used herein, the terms "IL18BP," "IL-18BP," "IL18BP," "IL18-BP," "IL18-binding protein," or "interleukin-18 binding protein" may optionally include any such protein, including, but not limited to, a known IL18-BP or wild-type IL18-BP as described herein, or a variant, complex, or fragment thereof, as well as any naturally occurring splice variant, amino acid variant, or isoform. The term IL18-BP is used interchangeably with "IL18 binding protein," "interleukin-18 binding protein," "IL18BPa," "interleukin-18 binding protein isoform a," and "interleukin-18 binding protein isoform a precursor." The term "soluble" form of IL18-BP is also used interchangeably with the terms "IL18BP soluble" or "fragment of an IL18-BP polypeptide," and may broadly refer to one or more of the following optional polypeptides: The term "soluble" with respect to forms of IL18-BP is also used interchangeably with the terms "secreted" and "fragment of an IL18-BP polypeptide," which may broadly refer to one or more of the IL18-BP polypeptides disclosed herein.

[0183] IL18-BP is constitutively expressed in the spleen and belongs to the immunoglobulin superfamily. The residues involved in the interaction between IL18 and IL18-BP have been described through computer modeling (Kim, S.-H. et al., PNAS, 97(3):1190-1195(2000)), based on the interaction between the similar proteins IL-1β and IL-1R type I (Vigers, GPA et al., Nature, 386:190-194(1997)). IL18-BP functions as an inhibitor of the inflammatory cytokine IL18. IL-18 regulates immune system functions, including induction of IFN-γ production, Th1 differentiation, NK cell activation, and cytotoxic T lymphocyte (CTL) responses (Tominaga, K., et al., International Immunology, 12(2):151-160(2000) and Senju, H., et al., Int J Biol Sci., 14(3):331-340(2018)). IL18-BP binds to IL18 and prevents IL18 from binding to its receptor, thereby inhibiting IL18-induced activation and proliferation of T cells and NK cells and pro-inflammatory cytokine production, resulting in reduced T cell and NK cell activity and T helper type 1 immune responses. IL18-BP abolishes IL18 induction of IFN-γ and IL18 activation of NF-κB in vitro. Furthermore, IL18-BP inhibits the induction of IFN-γ in mice terminally injected with LPS (Novick, D. et al., Immunity, 10:127-136, (1999)).

[0184] IL18 is constitutively present in many cells (Purenet al., PNAS, 96:2256-2261 (1999)) and circulates in healthy humans (Urushihara et al. 2000), representing a unique phenomenon in cytokine biology. Due to the high affinity of IL18 for IL18-BP (Kd ∼1 pM) and the high concentration of IL18-BP found in the circulation (20-fold molar excess over IL18), it is hypothesized that most, if not all, circulating IL18 molecules are bound to IL18-BP. Thus, circulating IL18-BP, which competes with cell surface receptors for IL18, may act as a natural anti-inflammatory and immunosuppressive molecule.

[0185] According to at least some embodiments of the present invention, anti-IL18-BP antibodies (including antigen-binding fragments) that bind to IL18-BP and block the interaction between IL18 and IL18-BP, thereby releasing increased levels of free IL18, can be used to enhance the activation, proliferation, cytokine and / or chemokine secretion of T cells, NK cells, NKT cells, myeloid cells, dendritic cells, MAIT T cells, γδ T cells, and / or innate lymphoid cells (ILCs), and can be used to treat diseases such as cancer and pathogen infection. These anti-IL18-BP antibodies can be used to treat diseases such as cancer.

[0186] Thus, the present invention provides anti-IL18-BP antibodies provided in Figure 1, Figure 2, and / or Figure 3 (e.g., including anti-IL18-BP antibodies having the same CDRs as those shown in Figure 1, Figure 2, and / or Figure 3). IL18-BP, also known as interleukin-18 binding protein, UniProtKB / Swiss-Prot (O95998) or HGNC (5987) NCBI Entrez Gene (10068), RefSeq accession identifiers: NG_029021.1, NM_001039659.1, NP_001034748.1, NC_000011.10, Chromosome11ReferenceGRCh38.p13Primary Assembly accession identifiers: NM_001039660.2 and NP_001034749.1 and NC_000011.9, Chromosome11ReferenceGRCh38.p13Primary Assembly Accession Identifiers: NP_001034748.1, NM_001039659.2, NP_005690.2, NM_005699.3, NP_001034748.1, NM_001039659.2, NP_005690.2, NM_005699.3, NP_001138529.1, NM_001145057.1, NP_001138527.1, NM_001145055.1. In some embodiments, the antibodies of the invention are specific for IL18-BP.

[0187] II. Anti-IL18-BP antibody Thus, the present invention provides anti-IL18-BP antibodies provided in Figures 1, 2 and 3 (including, for example, anti-IL18-BP antibodies including those having the same CDRs as those shown in Figures 1, 2 and / or 3), as well as antibodies that compete for binding with the antibodies listed in Figures 1, 2 and / or 3.

[0188] As discussed below, the term "antibody" is used generically. Antibodies that find use in the present invention can take many forms, as described herein, including traditional antibodies as well as antibody derivatives, fragments, and mimetics, as described below. In general, the term "antibody" includes any polypeptide that contains at least one antigen-binding domain, as described more fully below. Antibodies may be polyclonal, monoclonal, xenogeneic, allogeneic, syngeneic, or modified forms thereof, as described herein, with monoclonal antibodies finding particular use in many embodiments. In some embodiments, antibodies of the present invention bind specifically or substantially specifically to an IL18-BP molecule. As used herein, the terms "monoclonal antibody" and "monoclonal antibody composition" refer to a population of antibody molecules that contain only one species of antigen-binding site capable of immunoreacting with a particular epitope of an antigen, while the terms "polyclonal antibody" and "polyclonal antibody composition" refer to a population of antibody molecules that contain multiple species of antigen-binding sites capable of interacting with a particular antigen. A monoclonal antibody composition typically exhibits a single binding affinity for a particular antigen with which it immunoreacts.

[0189] Conventional full-length antibody structural units typically comprise a tetramer. Each tetramer typically consists of two identical pairs of polypeptide chains, each pair having one "light" chain (typically having a molecular weight of about 25 kDa) and one "heavy" chain (typically having a molecular weight of about 50-70 kDa). Human light chains are classified as κ and λ light chains. The present invention relates to the IgG class, which has several subclasses, including, but not limited to, IgG1, IgG2, IgG3, and IgG4. Therefore, as used herein, "isotype" refers to any of the immunoglobulin subclasses defined by the chemical and antigenic characteristics of their constant regions. While an exemplary antibody referred to herein as "CPA" is based on an IgG1 heavy chain constant region, as shown in Figure 4, anti-IL18-BP antibodies of the present invention include antibodies using IgG2, IgG3, and IgG4 sequences, or combinations thereof. For example, as is known in the art, different IgG isotypes have different effector functions, which may or may not be desirable. Thus, the CPA antibodies of the invention can also have the IgG1 constant domains swapped for those of IgG2, IgG3, or IgG4 (as shown in FIG. 1E), with IgG2 and IgG4 finding particular use in many situations, for example, for ease of manufacturing or when reduced effector function is desired, the latter being desirable in some situations.

[0190] The amino-terminal portion of each chain contains a variable region of approximately 100-110 or more amino acids primarily responsible for antigen recognition, commonly referred to in the art and herein as the "Fv domain" or "Fv region." In the variable region, three loops from each of the heavy and light chain V domains assemble to form the antigen-binding site. Each loop is called a complementarity-determining region (hereinafter referred to as "CDR"), where amino acid sequence variability is most pronounced. "Variability" refers to the fact that certain segments of the variable region vary significantly in sequence among antibodies. The variability within the variable region is not uniformly distributed. Instead, the V region consists of relatively invariant stretches of 15-30 amino acids called framework regions (FRs) separated by shorter regions of extreme variability called "hypervariable regions."

[0191] Each VH and VL is composed of three hypervariable regions ("complementarity-determining regions", "CDRs") and four FRs, arranged from the amino terminus to the carboxy terminus in the following order: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4.

[0192] The hypervariable regions generally encompass approximately amino acid residues 24-34 (LCDR1; "L" indicates light chain), 50-56 (LCDR2), and 89-97 (LCDR3) in the light chain variable region, and approximately 31-35B (HCDR1; "H" indicates heavy chain), 50-65 (HCDR2), and 95-102 (HCDR3) in the heavy chain variable region, although slight variations in numbering may occur as will be appreciated by those skilled in the art (Kabat et al., SEQUENCES OF PROTEINS OF IMMUNOLOGICAL INTEREST, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)) and / or residues forming hypervariable loops (e.g., residues 26-32 (LCDR1), 50-52 (LCDR2), and 91-96 (LCDR3) in the light chain variable region and 26-32 (HCDR1), 53-55 (HCDR2), and 96-101 (HCDR3) in the heavy chain variable region) (Chothia and Lesk (1987) J. Mol. Biol. 196:901-917). Specific CDRs of the invention are described below and shown in Figures 6A-6D.

[0193] The carboxy-terminal portion of each chain defines a constant region primarily responsible for effector function. Kabat et al. collected a large number of primary sequences of heavy and light chain variable regions. Based on the degree of sequence conservation, they classified each primary sequence into CDRs and frameworks and compiled a list of them (see SEQUENCES OF IMMUNOLOGICAL INTEREST, 5th edition, NIH publication, No. 91-3242, E.A. Kabat et al., which is incorporated by reference in its entirety).

[0194] In the IgG subclass of immunoglobulins, several immunoglobulin domains are present in the heavy chain. As used herein, "immunoglobulin (Ig) domain" refers to a region of an immunoglobulin that has a defined tertiary structure. Of interest in the present invention are heavy chain domains that contain the constant heavy (CH) domain and the hinge domain. In the context of IgG antibodies, each IgG isotype has three CH regions. Thus, the "CH" domains in the context of IgG are as follows: "CH1" refers to positions 118-220 according to the EU index as in Kabat; "CH2" refers to positions 237-340 according to the EU index as in Kabat; and "CH3" refers to positions 341-447 according to the EU index as in Kabat.

[0195] Thus, the present invention provides variable heavy chain domains, variable light chain domains, heavy chain constant domains, light chain constant domains, and Fc domains used as outlined herein. As used herein, "variable region" refers to a region of an immunoglobulin that includes one or more Ig domains substantially encoded by either the Vκ or Vλ and / or VH genes that constitute the κ, λ, and heavy chain immunoglobulin loci, respectively. Thus, the variable heavy chain domain comprises vhFR1-vhCDR1-vhFR2-vhCDR2-vhFR3-vhCDR3-vhFR4, and the variable light chain domain comprises vlFR1-vlCDR1-vlFR2-vlCDR2-vlFR3-vlCDR3-vlFR4. As used herein, "heavy chain constant region" refers to the CH1-hinge-CH2-CH3 portion of an antibody. As used herein, "Fc" or "Fc region" or "Fc domain" refers to a polypeptide comprising the constant region of an antibody excluding the first constant region immunoglobulin domain and, optionally, a portion of the hinge. Thus, Fc refers to the last two constant region immunoglobulin domains of IgA, IgD, and IgG, and the last three constant region immunoglobulin domains of IgE and IgM, as well as the flexible hinge N-terminal to these domains. For IgA and IgM, Fc may include the J chain. For IgG, the Fc domain includes immunoglobulin domains Cγ2 and Cγ3 (Cγ2 and Cγ3) and the lower hinge region between Cγ1 (Cγ1) and Cγ2 (Cγ2). Although the boundaries of the Fc region can vary, the human IgG heavy chain Fc region is usually defined to include residues C226 or P230 at its carboxyl terminus, numbering according to the EU index as in Kabat. In some embodiments, amino acid modifications are made to the Fc region to, for example, alter binding to one or more FcγR or FcRn receptors, as described more fully below.

[0196] Thus, as used herein, "Fc variant" or "variant Fc" refers to a protein comprising an amino acid modification in the Fc domain. Fc variants of the present invention are defined according to the amino acid modifications that comprise them. Thus, for example, N434S or 434S is an Fc variant with a substituted serine at position 434 relative to the parent Fc polypeptide, numbering according to the EU index. Similarly, M428L / N434S defines an Fc variant with the substitutions M428L and N434S relative to the parent Fc polypeptide. The identity of the WT amino acids may not be specified, in which case the variant is referred to as 428L / 434S. Note that the order in which the substitutions are provided is arbitrary, i.e., for example, 428L / 434S is the same Fc variant as M428L / N434S. For all positions discussed in the present invention related to antibodies, unless otherwise specified, the numbering of amino acid positions is according to the EU index.

[0197] As used herein, "Fab" or "Fab region" refers to a polypeptide comprising the VH, CH1, VL, and CL immunoglobulin domains. Fab can refer to this region in isolation or this region in the context of a full-length antibody, antibody fragment, or Fab fusion protein. As used herein, "Fv" or "Fv fragment" or "Fv region" refers to a polypeptide comprising the VL and VH domains of a single antibody. As will be appreciated by those of skill in the art, these are generally composed of two chains.

[0198] Throughout this specification, when referring to residues in the variable domains (approximately residues 1-107 for the light chain variable region and residues 1-113 for the heavy chain variable region), either the IMTG numbering system or the Kabat numbering system is generally used (e.g., Kabat et al., supra (1991)). EU numbering, as in Kabat, is generally used for constant domains and / or Fc domains.

[0199] CDRs contribute to the formation of the antigen-binding site of an antibody, or more specifically, the epitope-binding site. "Epitope" refers to a determinant that interacts with a specific antigen-binding site in the variable region of an antibody molecule, known as a paratope. An epitope is a group of molecules, such as amino acids or sugar side chains, that usually have specific structural and charge characteristics. One antigen can have multiple epitopes.

[0200] An epitope may include amino acid residues that are directly involved in binding (also called the immunodominant component of the epitope) and other amino acid residues that are not directly involved in binding, e.g., amino acid residues that are effectively blocked by the specific antigen-binding peptide, in other words, amino acid residues that lie within the footprint of the specific antigen-binding peptide.

[0201] Epitopes can be either conformational or linear. Conformational epitopes are produced by spatially juxtaposed amino acids from different segments of a linear polypeptide chain. Linear epitopes are produced by adjacent amino acid residues in a polypeptide chain. Conformational and nonconformational epitopes can be distinguished in that the binding to the former is lost in the presence of denaturing solvents, but the binding to the latter is not lost.

[0202] An epitope typically includes at least 3, more usually at least 5, or 8-10 amino acids in a unique spatial conformation. Antibodies that recognize the same epitope can be identified, e.g., "binned," in a simple immunoassay demonstrating the ability of one antibody to block the binding of another antibody to a target antigen. Specific bins are described below.

[0203] Included within the definition of "antibody" is an "antigen-binding portion" of an antibody (also used interchangeably with "antigen-binding fragment," "antibody fragment," and "antibody derivative"). That is, for purposes of the present invention, an antibody of the present invention has the minimal functional requirement of binding to the IL18-BP antigen. As will be appreciated by those skilled in the art, there are numerous antigen-binding fragments and derivatives that retain the ability to bind to the antigen and yet have alternative structures, including, but not limited to: (i) a Fab fragment consisting of the VL, VH, CL, and CH1 domains; (ii) an Fd fragment consisting of the VH and CH1 domains; (iii) an F(ab')2 fragment, a bivalent fragment comprising two linked Fab fragments; (vii) a single-chain Fv molecule (scFv), in which the VH and VL domains are linked by a peptide linker that allows the two domains to associate and form an antigen-binding site (Bird et al., 1988, Science 242:423-426; Huston et al., 1988, Proc. Natl. Acad. Sci. USA 85:5879-5883, entirely incorporated by reference); and (iv) "diabodies" or "triabodies," multivalent or multispecific fragments constructed by gene fusion (Tomlinson et al., 2000, Methods Enzymol. 326:461-479; WO 94 / 13804; Holliger et al., 1993, Proc. Natl. Acad. Sci. USA 90:6444-6448, all of which are incorporated by reference in their entireties), (v) "domain antibodies" or "dAbs" (sometimes called "immunoglobulin single variable domains" and include single antibody variable domains from other species such as rodents (e.g., as disclosed in WO 00 / 29004), nurse shark and camelid V-HH dAbs), (vi) SMIPs (small molecular weight immunopharmaceuticals), camelid antibodies, nanobodies and IgNARs.

[0204] Furthermore, an antibody or antigen-binding portion thereof (antigen-binding fragment, antibody fragment, antibody portion) may be part of a larger immunoadhesion molecule (sometimes called a "fusion protein") formed by covalent or noncovalent attachment of the antibody or antibody portion to one or more other proteins or peptides. Examples of immunoadhesion molecules include the use of a streptavidin core region to generate tetrameric scFv molecules, and the use of cysteine ​​residues, marker peptides, and C-terminal polyhistidine tags to generate bivalent and biotinylated scFv molecules. Antibody portions, such as Fab fragments and F(ab')2 fragments, can be prepared from whole antibodies using conventional techniques, such as papain or pepsin digestion, respectively, of whole antibodies. Furthermore, antibodies, antibody portions, and immunoadhesion molecules can be obtained using standard recombinant DNA techniques, as described herein.

[0205] Generally, the anti-IL18-BP antibodies of the present invention are recombinant. As used herein, "recombinant" refers broadly to a product, e.g., a cell, or a nucleic acid, protein, or vector, and indicates that the cell, nucleic acid, protein, or vector has been modified by the introduction of a heterologous nucleic acid or protein or the modification of a native nucleic acid or protein, or that the cell is derived from a cell so modified. Thus, for example, a recombinant cell expresses genes that are not found within the native (non-recombinant) form of the cell, or expresses native genes that are otherwise aberrantly expressed, under-expressed, or not expressed at all.

[0206] As used herein, the term "recombinant antibody" includes all antibodies prepared, expressed, generated, or isolated by recombinant means, including, for example, (a) antibodies isolated from animals (e.g., mice) transgenic or transchromosomal for human immunoglobulin genes or hybridomas prepared therefrom (described further below); (b) antibodies isolated from host cells transformed to express human antibodies, e.g., from transfectomas; (c) antibodies isolated from recombinant combinatorial human antibody libraries; and (d) antibodies prepared, expressed, generated, or isolated by any other means, including splicing of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies have variable regions in which the framework and CDR regions are derived from human germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies may be subjected to in vitro mutagenesis (or, if animals transgenic for human Ig sequences are used, in vivo somatic mutagenesis), thus allowing for the development of recombinant antibody Vs. H Area and V L The amino acid sequence of the region is human germline V H Sequence and V L These are sequences that are derived from and related to sequences, but which may not naturally occur in vivo within the human antibody germline repertoire.

[0207] A. Anti-IL18-BP binding antibody The present invention provides anti-IL18-BP antibodies (for convenience, the terms "anti-IL18-BP antibody" and "IL18-BP antibody" are used interchangeably). The anti-IL18-BP antibodies of the present invention specifically bind to human IL18-BP, preferably the secretory chain of human IL18-BP, as shown in Figure 28, and include anti-IL18-BP antibodies comprising those having the same CDRs as those shown in Figures 1, 2, and 3, for example.

[0208] As described herein and more fully below, anti-IL18-BP antibodies (including antigen-binding fragments) that bind to IL18-BP and block the interaction of IL18-BP with IL18, thereby releasing increased levels of free IL18, can be used to enhance activation, proliferation, cytokine and / or chemokine secretion of T cells, NK cells, NKT cells, myeloid cells, dendritic cells, MAIT T cells, γδ T cells, and / or innate lymphoid cells (ILCs), and can be used to treat diseases such as cancer and pathogen infections.

[0209] Specific binding to IL18-BP or an IL18-BP epitope is, for example, at least about 10 -5 M, at least about 10 -6 M, at least about 10 -7 M, at least about 10 -8 M, at least about 10 -9 M, or at least about 10 -10 M, at least about 10 -11 M, at least about 10 -12 M, at least about 10 -13 M, at least about 10 -14 A specific binding activity can be exhibited by an antibody having a KD of M or greater, where KD refers to the off-rate of a particular antibody-antigen interaction. Typically, an antibody that specifically binds to an antigen has a KD that is 20-fold, 50-fold, 100-fold, 500-fold, 1000-fold, 5000-fold, 10,000-fold, 100,000-fold, or greater for a control molecule compared to the IL18-BP antigen or epitope.

[0210] However, as evidenced by the Examples, for optimal binding to IL18-BP, antibodies preferably have a KD (also referred to as binding affinity) of less than 0.01 nM, less than 10 nM, and most preferably less than 0.1 pM, less than 1 pM, less than 0.1 pM, and less than 0.01 pM, and find use in the methods of the present invention. In some embodiments, anti-IL-18BP antibodies exhibit a KD of less than 900 pM, less than 850 pM, less than 800 pM, less than 750 pM, less than 700 pM, less than 650 pM, less than 600 pM, less than 550 pM, less than 500 pM, less than 450 pM, less than 400 pM, less than 350 pM, less than 300 pM, less than 250 pM, less than 200 pM, less than 150 pM, less than 100 pM, less than 50 pM, or less than 10 pM. In some embodiments, anti-IL-18BP antibodies exhibit a KD of less than 750 pM. In some embodiments, anti-IL18-BP antibodies of the invention exhibit a KD of 50 nM or less, 10 nM or less, or 1 nM or less (i.e., higher binding affinity), 100 pM or less, 10 pM or less, 1 pM or less, 0.1 pM or less, or 0.01 pM or less. D binds to human IL18-BP at K D is determined by known methods, such as surface plasmon resonance (SPR, e.g., on a Biacore instrument), ELISA, KinExA, most typically by SPR at 25° or 37° C. In some embodiments, an anti-IL18-BP antibody of the invention binds to human IL18-BP with a KD of less than 900 pM, less than 850 pM, less than 800 pM, less than 750 pM, less than 700 pM, less than 650 pM, less than 600 pM, less than 550 pM, less than 500 pM, less than 450 pM, less than 400 pM, less than 350 pM, less than 300 pM, less than 250 pM, less than 200 pM, less than 150 pM, less than 100 pM, less than 50 pM, or less than 10 pM. Dis determined by known methods, such as surface plasmon resonance (SPR, e.g., a Biacore instrument), ELISA, KinExA, most typically SPR at 25°C or 37°C. In some embodiments, the antibody preferably has a KD or binding affinity of less than 0.005 pM, less than 0.01 pM, less than 0.02 pM, less than 0.03 pM, less than 0.04 pM, less than 0.05 pM, less than 0.06 pM, less than 0.07 pM, less than 0.08 pM, less than 0.09 pM, less than 0.10 pM, less than 0.15 pM, less than 0.20 pM, less than 0.25 pM, less than 0.30 pM, less than 0.35 pM, less than 0.40 pM, less than 0.45 pM, less than 0.50 pM, less than 0.55 pM, less than 0.60 pM, less than 0.65 pM, less than 0.70 pM, less than 0.75 pM, less than 0.80 pM, less than 0.85 pM, less than 0.90 pM, less than 0.95 pM, or less than 1 pM.

[0211] Specific binding to a particular antigen or epitope may also be exhibited, for example, by an antibody having a K A or K A for an IL18-BP antigen or epitope that is at least 20-fold, 50-fold, 100-fold, 500-fold, 1000-fold, 5,000-fold, 10,000-fold, 100,000-fold, or more greater than that for the epitope compared to a control, where K A or K A refers to the association rate of a particular antibody-antigen interaction.

[0212] The present invention provides antigen-binding domains, including full-length antibodies, containing any particular enumerated set of six CDRs, as provided in Figure 1, Figure 2, and / or Figure 3. The present invention provides antigen-binding domains, including full-length antibodies, containing any particular enumerated set of six CDRs, as provided in Figure 3.

[0213] The invention further provides variable heavy and light chain domains, as well as full-length heavy and light chains.

[0214] As discussed herein, the present invention further provides variants of the above components, including variants in the CDRs as outlined above. Furthermore, the variable heavy chain may be at least 80%, at least 90%, at least 95%, at least 98%, or at least 99% identical to the "VH" sequences herein, and / or may contain 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid changes if an Fc variant is used. When an Fc variant is used, the variable light chain is provided which may be at least 80%, at least 90%, at least 95%, at least 98%, or at least 99% identical to the "VL" sequences herein, and / or may contain 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid changes. Similarly, heavy and light chains are provided that are at least 80%, at least 90%, at least 95%, at least 98%, or at least 99% identical to the "HC" and "LC" sequences herein, and / or contain 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid changes when Fc variants are used.

[0215] Thus, the present invention provides antibodies (typically full-length domains or scFv domains) comprising the following CHA sets of CDRs, the sequences of which are shown in Figures 1 to 3.

[0216] The 66650 lineage (VH1-03; VL-κ-1-5) consensus sequence of the CDRs (Figure 1A) was generated using the ADI-71701, ADI-71709, ADI-71710, ADI-71707, and ADI-71717 antibodies. The respective sequence alignments are shown in Figure 3B.

[0217] The 66650 lineage (VH1-03; VL-κ-1-5) consensus sequence is a CDR-H1 having the sequence YTFX-X2-YA-X3-H, where X is N, R, D, G or K, X2 is S, H, I or Q, and X3 is M or V; a CDR-H2 having the sequence WIHAGTGXT-X2-YSQKFQG (wherein X is N, A or V and X2 is K or L), a CDR-H3 having the sequence ARGLGXVGPTGTSWFDP (wherein X is S or E), a CDR-L1 having the sequence RASQGISSWLA, a CDR-L2 having the sequence EASSLES; and a CDR-L3 having the sequence QQYRX-X2-PFT (wherein X is S, V, Y, L, or Q and X2 is F, S, or G).

[0218] In some embodiments, the anti-IL18-BP antibody comprises the CDRs, i.e., a CDR-H1 having the sequence YTFX-X2-YA-X3-H, where X is N, R, D, G or K, X2 is S, H, I or Q, and X3 is M or V; a CDR-H2 having the sequence WIHAGTGXT-X2-YSQKFQG (wherein X is N, A or V and X2 is K or L), a CDR-H3 having the sequence ARGLGXVGPTGTSWFDP (wherein X is S or E), a CDR-L1 having the sequence RASQGISSWLA, a CDR-L2 having the sequence EASSLES; and a CDR-L3 having the sequence QQYRX-X2-PFT (wherein X is S, V, Y, L, or Q and X2 is F, S, or G).

[0219] The 66670-family (VH1-69; VL-κ-1-12) consensus sequence of the CDRs (Figure 1B) was generated using the ADI-71719, ADI-71720, ADI-71722, and ADI-71728 antibodies, and the respective sequence alignments are shown in Figure 3C.

[0220] The 66670 lineage (VH1-69; VL-κ-1-12) consensus sequence is a CDR-H1 having the sequence GTFX-X2-Y-X3-IS, where X is S or N, X2 is E or S, and X3 is V or P; a CDR-H2 having the sequence GIIPG-X2-GTA-X3-YAQKFQG (wherein X is G or Y, X2 is A or S, and X3 is N, I, or V); a CDR-H3 having the sequence ARGRHXHET (wherein X is S, G, or F), a CDR-L1 having the sequence RASQGISSWLA, a CDR-L2 having the sequence AASSLQS; and a CDR-L3 having the sequence QQVYX-X2-PWT (wherein X is S or R and X2 is L, I or F).

[0221] In some embodiments, the anti-IL18-BP antibody comprises the CDRs, i.e., a CDR-H1 having the sequence GTFX-X2-Y-X3-IS, where X is S or N, X2 is E or S, and X3 is V or P; a CDR-H2 having the sequence GIIPG-X2-GTA-X3-YAQKFQG (wherein X is G or Y, X2 is A or S, and X3 is N, I, or V); a CDR-H3 having the sequence ARGRHXHET (wherein X is S, G, or F), a CDR-L1 having the sequence RASQGISSWLA, a CDR-L2 having the sequence AASSLQS; and a CDR-L3 having the sequence QQVYX-X2-PWT (wherein X is S or R and X2 is L, I or F).

[0222] The 66692 family of CDRs (VH3-23, VL-κ-1-12) consensus sequence (Figure 1C) was generated using the ADI-71662, ADI-71663, and ADI-66692 antibodies, and the respective sequence alignments are shown in Figure 3A.

[0223] The consensus sequence for strain 66692 (VH3-23, VL-κ-1-12) is a CDR-H1 having the sequence FTFXN-X2-AMS (wherein X is G or D or S and X2 is T or V or Y); a CDR-H2 having the sequence AISX-X1-X2-GSTYYADSVKG (wherein X is G or A, X2 is N or S, and X3 is A or G); a CDR-H3 having the sequence AKGPDRQVFDY; a CDR-L1 having the sequence RASQGIXSWLA (wherein X is S or D), a CDR-L2 having the sequence AASSLQS; and a CDR-L3 having the sequence QHAX-X1-FPYT (wherein X is Y or L and X2 is S or F).

[0224] In some embodiments, the anti-IL18-BP antibody comprises the CDRs, i.e., a CDR-H1 having the sequence FTFXN-X2-AMS (wherein X is G or D or S and X2 is T or V or Y); a CDR-H2 having the sequence AISX-X1-X2-GSTYYADSVKG (wherein X is G or A, X2 is N or S, and X3 is A or G); a CDR-H3 having the sequence AKGPDRQVFDY; a CDR-L1 having the sequence RASQGIXSWLA (wherein X is S or D), a CDR-L2 having the sequence AASSLQS; and a CDR-L3 having the sequence QHAX-X1-FPYT (wherein X is Y or L and X2 is S or F).

[0225] The 66716 lineage (VH1-39; VL-κ-1-12) consensus sequence of the CDRs (Figure 1D) was generated using the ADI-71736, ADI-71739, and ADI-66716 antibodies. The respective sequence alignments are shown in Figure 3D.

[0226] The 66716 lineage (VH1-39; VL-κ-1-12) consensus sequence is a CDR-H1 having the sequence GSISSX-X2-Y-X3-WG, where X is S or P, X2 is E or D, and X3 is G, P, or Y; a CDR-H2 having the sequence SIX-X2-X3-G-X4-TYYNPSLKS (wherein X is Y or V, X2 is Y or N, X3 is Q or S, and X4 is S or A); and a CDR-H3 having the sequence ARGPXRQ-X2-FDY (wherein X is Y or H and X2 is V or L), a CDR-L1 having the sequence RASQGISSWLA, a CDR-L2 having the sequence AASSLQS; and a CDR-L3 having the sequence QQGX-X2-FPYT (wherein X is S or F and X2 is S or V).

[0227] In some embodiments, the anti-IL18-BP antibody comprises the CDRs, i.e., a CDR-H1 having the sequence GSISSX-X2-Y-X3-WG, where X is S or P, X2 is E or D, and X3 is G, P, or Y; a CDR-H2 having the sequence SIX-X2-X3-G-X4-TYYNPSLKS (wherein X is Y or V, X2 is Y or N, X3 is Q or S, and X4 is S or A); and a CDR-H3 having the sequence ARGPXRQ-X2-FDY (wherein X is Y or H and X2 is V or L), a CDR-L1 having the sequence RASQGISSWLA, a CDR-L2 having the sequence AASSLQS; and a CDR-L3 having the sequence QQGX-X2-FPYT (wherein X is S or F and X2 is S or V).

[0228] In some embodiments, the anti-IL18-BP antibody comprises the CDRs, i.e., a CDR-H1 having the sequence YTFX-X2-YA-X3-H, where X is any amino acid, X2 is any amino acid, and X3 is any amino acid; a CDR-H2 having the sequence WIHAGTGXT-X2-YSQKFQG (wherein X is any amino acid and X2 is any amino acid); a CDR-H3 having the sequence ARGLGXVGPTGTSWFDP (wherein X is any amino acid); a CDR-L1 having the sequence RASQGISSWLA, a CDR-L2 having the sequence EASSLES; and a CDR-L3 having the sequence QQYRX-X2-PFT (wherein X is any amino acid and X2 is any amino acid).

[0229] In some embodiments, the anti-IL18-BP antibody comprises the CDRs, i.e., a CDR-H1 having the sequence GTFX-X2-Y-X3-IS, where X is any amino acid, X2 is any amino acid, and X3 is any amino acid; a CDR-H2 having the sequence GIIPG-X2-GTA-X3-YAQKFQG (wherein X is any amino acid and X2 is any amino acid); a CDR-H3 having the sequence ARGRHXHET (where X is any amino acid); a CDR-L1 having the sequence RASQGISSWLA, a CDR-L2 having the sequence AASSLQS; and a CDR-L3 having the sequence QQVYX-X2-PWT (wherein X is any amino acid and X2 is any amino acid).

[0230] In some embodiments, the anti-IL18-BP antibody comprises the CDRs, i.e., a CDR-H1 having the sequence GTFX-X2-Y-X3-IS, where X is any amino acid, X2 is any amino acid, and X3 is any amino acid; a CDR-H2 having the sequence GIIPG-X2-GTA-X3-YAQKFQG (wherein X is any amino acid, X2 is any amino acid, and X3 is any amino acid); a CDR-H3 having the sequence ARGRHXHET (where X is any amino acid); a CDR-L1 having the sequence RASQGISSWLA, a CDR-L2 having the sequence AASSLQS; and a CDR-L3 having the sequence QQVYX-X2-PWT (wherein X is any amino acid and X2 is any amino acid).

[0231] In some embodiments, the anti-IL18-BP antibody comprises the CDRs, i.e., a CDR-H1 having the sequence FTFXN-X2-AMS, where X is any amino acid and X2 is any amino acid; a CDR-H2 having the sequence AISX-X1-X2-GSTYYADSVKG (wherein X is any amino acid, X2 is any amino acid, and X3 is any amino acid); a CDR-H3 having the sequence AKGPDRQVFDY; a CDR-L1 having the sequence RASQGIXSWLA (wherein X is any amino acid); a CDR-L2 having the sequence AASSLQS; and a CDR-L3 having the sequence QHAX-X1-FPYT (wherein X is any amino acid and X2 is any amino acid).

[0232] In some embodiments, the anti-IL18-BP antibody comprises the CDRs, i.e., a CDR-H1 having the sequence GSISSX-X2-Y-X3-WG, where X is any amino acid, X2 is any amino acid, and X3 is any amino acid; a CDR-H2 having the sequence SIX-X2-X3-G-X4-TYYNPSLKS (wherein X is any amino acid, X2 is any amino acid, X3 is any amino acid, and X4 is any amino acid); a CDR-H3 having the sequence ARGPXRQ-X2-FDY (wherein X is any amino acid and X2 is any amino acid); a CDR-L1 having the sequence RASQGISSWLA, a CDR-L2 having the sequence AASSLQS; and a CDR-L3 having the sequence QQGX-X2-FPYT (wherein X is any amino acid and X2 is any amino acid).

[0233] In some embodiments, the antibody i. a heavy chain variable domain, a) CDR-H1 having the sequence YTFX-X2-YA-X3-H, where X is N, R, D, G, T, Q, S, A, or K, X2 is S, H, I, N, L, Y, or Q, and X3 is M or V; and b) a CDR-H2 having the sequence XI-X2-AG-X3-X4-X5-T-X6-YSQKFQG, where X is W or Y, X2 is H or N, X3 is S, T or A, X4 is G or A, X5 is N, A, T or V, and X6 is E, K or L; and c) a heavy chain variable domain comprising a CDR-H3 having the sequence ARGLGXVGPTGTSWFDP (wherein X is S, L, A, K, or E); ii. a light chain variable domain, d) CDR-L1 having the sequence RASQGISSWLA; e) a CDR-L2 having the sequence EASSES (wherein X is L or S); f) a light chain variable domain comprising a CDR-L3 having the sequence QQYRX-X2-PFT (wherein X is S, V, Y, L, T, or Q and X2 is F, S, Y, or G).

[0234] In some embodiments, the antibody i. a heavy chain variable domain, a) a CDR-H1 having the sequence GTFX-X2-Y-X3-IS, where X is S or N, X2 is E or S, and X3 is V or P; b) a CDR-H2 having the sequence GIIPX-X2-GTA-X3-YAQKFQG, where X is G, S, I or Y, X2 is A, V or S, and X3 is N, I or V; and c) a heavy chain variable domain comprising a CDR-H3 having the sequence ARGRHXHET (wherein X is S, G, or F); ii. a light chain variable domain, d) CDR-L1 having the sequence RASQGISSWLA; e) a CDR-L2 having the sequence AASSLQS; f) a light chain variable domain comprising a CDR-L3 having the sequence QQXY-X2-X3-PWT (wherein X is V or L, X2 is S or R, and X3 is L, I, or F).

[0235] In some embodiments, the antibody i. a heavy chain variable domain, a) CDR-H1 having the sequence FTFX-X2-X3-X4-MS, where X is G, S, P or D or S, X2 is N, S or P, X3 is T, V or Y, and X4 is A, H or I; b) a CDR-H2 having the sequence AISX-X2-X3-X4-X5-T-X6-YADSVKG, where X is G or A, X2 is N, T, E or S, X3 is A or G, X4 is A or G, X5 is S or G, and X6 is Y or F; and c) a heavy chain variable domain comprising a CDR-H3 having the sequence AKGPDRQVFDY; ii. a light chain variable domain, d) CDR-L1 having the sequence RASQGIXSWLA (wherein X is S or D); e) a CDR-L2 having the sequence AASSLQS; f) a light chain variable domain comprising a CDR-L3 having the sequence QHX-X2-X3-FPYT (wherein X is A or G, X2 is Y, R or L, and X3 is S, R, L or F).

[0236] In some embodiments, the antibody i. a heavy chain variable domain, a) a CDR-H1 having the sequence GSIXS-X2-X3-Y-X4-W-X5, where X is S or F, X2 is S or P, X3 is E or D, X4 is G, P or Y, and X5 is G or S; and b) a CDR-H2 having the sequence XI-X2-X3-X4-G-X5-TYYNPSLKS, where X is S or V, X2 is Y, V, F or A, X3 is Y, F or N, X4 is Q, A or S, and X5 is S, A or N; and c) a heavy chain variable domain comprising a CDR-H3 having the sequence ARGPXRQ-X2-FDY (wherein X is Y, H, or F, and X2 is V or L); ii. a light chain variable domain, d) CDR-L1 having the sequence RASQGISSWLA; d) a CDR-L2 having the sequence AASSLQS; e) a light chain variable domain comprising a CDR-L3 having the sequence QQGX-X2-FPYT (wherein X is S, N, W, or F and X2 is S or V).

[0237] Anti-IL18-BP antibodies also comprise framework regions. The variable heavy and variable light chain framework regions can be humanized as known in the art (optionally generating variants in the CDRs), thus generating humanized variants of the VH and VL chains of Figure 1, Figure 2, and / or Figure 3. Furthermore, the humanized variable heavy and light chain domains can then be fused to human constant regions, such as those from IgG1, IgG2, IgG3, and IgG4.

[0238] Also included are sequences that may have identical CDRs but may have variations in the variable domain (or entire heavy or light chain). For example, IL18-BP antibodies include those with identical CDRs to those shown in Figures 1-3, but may be less identical along the variable region, e.g., 85%, 88%, 90%, 92%, 95, or 98% identical. For example, IL18-BP antibodies include those with identical CDRs to those shown in Figure 3, but may be less identical along the variable region, e.g., 95 or 98% identical, and in some embodiments, may be at least 95% or at least 98% identical.

[0239] The percent identity between two amino acid sequences can be determined using the algorithm of E. Meyers and W. Miller (Comput. Appl. Biosci., 4:11-17 (1988)), which is incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4. Additionally, the percent identity between two amino acid sequences can be determined using the algorithm of Needleman and Wunsch (J. Mol. Biol. 48:444-453 (1970)), which is incorporated into the GAP program in the commercially available GCG software package, using either a Blossum62 matrix or a PAM250 matrix, and gap weights of 16, 14, 12, 10, 8, 6, or 4 and length weights of 1, 2, 3, 4, 5, or 6.

[0240] Additionally or alternatively, the protein sequences of the present invention can be further used as a "query sequence" to conduct searches against public databases, for example, to identify related sequences. Such searches can be performed using the XBLAST program (version 2.0) of Altschul et al. (1990) J. Mol. Biol. 215:403-10. BLAST protein searches can be performed using the XBLAST program, score=50, wordlength=3, to obtain amino acid sequences homologous to antibody molecules according to at least some embodiments of the present invention. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al. (1997) Nucleic Acids Res. 25(17):3389-3402. When utilizing BLAST and Gapped BLAST programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used.

[0241] Generally, percent identities for comparison between IL18-BP antibodies are at least 75%, at least 80%, at least 90%, with percent identities of at least about 95, 96, 97, 98, or 99% being preferred. The percent identity may be along the entire amino acid sequence, e.g., the entire heavy or light chain, or along a portion of the chain. For example, included within the definition of an anti-IL18-BP antibody of the invention are those that share identity along the entire variable region (e.g., 95 or 98% identical along the variable region, and in some embodiments at least 95% or at least 98% identical), along the entire constant region, or along just the Fc domain.

[0242] B. Specific anti-IL18-BP antibody The present invention provides antigen-binding domains, including full-length antibodies, containing several specific enumerated sets of six CDRs, as well as consensus CDRs (see, e.g., those listed in Figures 1A-1D).

[0243] The antibodies described herein are labeled as follows: An antibody has a reference number, for example, "66650 lineage (VH1-03; VL-κ-1-5)" or "VH1-03" or "ADI-71663 hIgG4 S228Pκ", which represents the CDRs and / or variable heavy and variable light chain combinations as shown in Figures 1, 2, and / or 3. "ADI-71663.VH" refers to the variable heavy chain portion of ADI-71663 hIgG4 S228Pκ, and "ADI-71663.VL" is the variable light chain. "ADI-71663.vhCDR1," "ADI-71663.vhCDR2," "ADI-71663.vhCDR3," "ADI-71663.vlCDR1," "ADI-71663.vlCDR2," and "ADI-71663.vlCDR3" refer to the indicated CDRs. "ADI-71663.HC" refers to the entire heavy chain (e.g., variable and constant domains) of this molecule, and "ADI-71663.LC" refers to the entire light chain (e.g., variable and constant domains) of the same molecule.

[0244] The invention further provides variable heavy and light chain domains, as well as full-length heavy and light chains.

[0245] In many embodiments, the antibodies of the invention are human (phage-derived) and block IL18-BP. As shown in Figures 1A-1D and 2A-2P, anti-IL18-BP antibodies are similarly outlined with their components:

[0246] CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 from lineage 66650 (VH1-03; VL-κ-1-5);

[0247] CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 from lineage 66670 (VH1-69; VL-κ-1-12);

[0248] CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 derived from the 66692 system (VH3-23, VL-κ-1-12);

[0249] CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 derived from the 66716 system (VH1-39; VL-κ-1-12);

[0250] ADI-71663, ADI-71663.VH, ADI-71663.VL, ADI-71663.HC, ADI-71663.LC and ADI-71663.H1, ADI-71663.H2, ADI-71663.H3, ADI-71663.H4; ADI-71663.vhCDR1, ADI-71663.vhCDR2, ADI-71663.vhCDR3, ADI-71663.vlCDR1, ADI-71663.vlCDR2, and ADI-71663.vlCDR3;

[0251] ADI-71662, ADI-71662.VH, ADI-71662.VL, ADI-71662.HC, ADI-71662.LC and ADI-71662.H1, ADI-71662.H2, ADI-71662.H3, ADI-71662.H4; ADI-71662.vhCDR1, ADI-71664.71662, ADI-71662.vhCDR3, ADI-71662.vlCDR1, ADI-71662.vlCDR2, and ADI-71662.vlCDR3;

[0252] ADI-71701, ADI-71701.VH, ADI-71701.VL, ADI-71701.HC, ADI-71701.LC and ADI-71701.H1, ADI-71701.H2, ADI-71701.H3, ADI-71701.H4; ADI-71701.vhCDR1, ADI-71701.vhCDR2, ADI-71701.vhCDR3, ADI-71701.vlCDR1, ADI-71701.vlCDR2, and ADI-71701.vlCDR3;

[0253] ADI-71709, ADI-71709.VH, ADI-71709.VL, ADI-71709.HC, ADI-71709.LC, and ADI-71709.H1, ADI-71709.H2, ADI-71709.H3, ADI-71709.H4; ADI-71709.vhCDR1, ADI-71709.vhCDR2, ADI-71709.vhCDR3, ADI-71709.vlCDR1, ADI-71709.vlCDR2, and ADI-71709.vlCDR3;

[0254] ADI-71710, ADI-71710.VH, ADI-71710.VL, ADI-71710.HC, ADI-71710.LC, and ADI-71710.H1, ADI-71710.H2, ADI-71710.H3, ADI-71710.H4; ADI-71710.vhCDR1, ADI-71710.vhCDR2, ADI-71710.vhCDR3, ADI-71710.vlCDR1, ADI-71710.vlCDR2, and ADI-71710.vlCDR3;

[0255] ADI-71719, ADI-71719.VH, ADI-71719.VL, ADI-71719.HC, ADI-71719.LC, and ADI-71719.H1, ADI-71719.H2, ADI-71719.H3, ADI-71719.H4; ADI-71719.vhCDR1, ADI-71719.vhCDR2, ADI-71719.vhCDR3, ADI-71719.vlCDR1, ADI-71719.vlCDR2, and ADI-71719.vlCDR3;

[0256] ADI-71720, ADI-71720.VH, ADI-71720.VL, ADI-71720.HC, ADI-71720.LC, and ADI-71720.H1, ADI-71720.H2, ADI-71720.H3, ADI-71720.H4; ADI-71720.vhCDR1, ADI-71720.vhCDR2, ADI-71720.vhCDR3, ADI-71720.vlCDR1, ADI-71720.vlCDR2, and ADI-71720.vlCDR3;

[0257] ADI-71722, ADI-71722.VH, ADI-71722.VL, ADI-71722.HC, ADI-71722.LC, and ADI-71722.H1, ADI-71722.H2, ADI-71722.H3, ADI-71722.H4; ADI-71722.vhCDR1, ADI-71722.vhCDR2, ADI-71722.vhCDR3, ADI-71722.vlCDR1, ADI-71722.vlCDR2, and ADI-71722.vlCDR3;

[0258] ADI-71717, ADI-71717.VH, ADI-71717.VL, ADI-71717.HC, ADI-71717.LC, and ADI-71717.H1, ADI-71717.H2, ADI-71717.H3, ADI-71717.H4; ADI-71717.vhCDR1, ADI-71717.vhCDR2, ADI-71717.vhCDR3, ADI-71717.vlCDR1, ADI-71717.vlCDR2, and ADI-71717.vlCDR3;

[0259] ADI-71739, ADI-71739.VH, ADI-71739.VL, ADI-71739.HC, ADI-71739.LC, and ADI-71739.H1, ADI-71739.H2, ADI-71739.H3, ADI-71739.H4; ADI-71739.vhCDR1, ADI-71739.vhCDR&2, ADI-71739.vhCDR3, ADI-71739.vlCDR1, ADI-71739.vlCDR2, and ADI-71739.vlCDR3;

[0260] ADI-71736, ADI-71736.VH, ADI-71736.VL, ADI-71736.HC, ADI-71736.LC, and ADI-71736.H1, ADI-71736.H2, ADI-71736.H3, ADI-71736.H4; ADI-71736.vhCDR1, ADI-71736.vhCDR2, ADI-71736.vhCDR3, ADI-71736.vlCDR1, ADI-71736.vlCDR2, and ADI-71736.vlCDR3;

[0261] ADI-71707, ADI-71707.VH, ADI-71707.VL, ADI-71707.HC, ADI-71707.LC, and ADI-71707.H1, ADI-71707.H2, ADI-71707.H3, ADI-71707.H4; ADI-71707.vhCDR1, ADI-71707.vhCDR2, ADI-71707.vhCDR3, ADI-71707.vlCDR1, ADI-71707.vlCDR2, and ADI-71707.vlCDR3;

[0262] AB-837, AB-837.VH, AB-837.VL, AB-837.HC, AB-837.LC, and AB-837.H1, AB-837.H2, AB-837.H3, AB-837.H4; AB-837.vhCDR1, AB-837.vhCDR2, AB-837.vhCDR3, AB-837.vlCDR1, AB-837.vlCDR2, and AB-837.vlCDR3;

[0263] ADI-66692, ADI-66692.VH, ADI-66692.VL, ADI-66692.HC, ADI-66692.LC, and ADI-66692.H1, ADI-66692.H2, ADI-66692.H3, ADI-66692.H4; ADI-66692.vhCDR1, ADI-66692.vhCDR2, ADI-66692.vhCDR3, ADI-66692.vlCDR1, ADI-66692.vlCDR2, and ADI-66692.vlCDR3;

[0264] ADI-66716, ADI-66716.VH, ADI-66716.VL, ADI-66716.HC, ADI-66716.LC, and ADI-66716.H1, ADI-66716.H2, ADI-66716.H3, ADI-66716.H4; ADI-66716.vhCDR1, ADI-66716.vhCDR2, ADI-66716.vhCDR3, ADI-66716.vlCDR1, ADI-66716.vlCDR2, and ADI-66716.vlCDR3;

[0265] ADI-71728, ADI-71728.VH, ADI-71728.VL, ADI-71728.HC, ADI-71728.LC, and ADI-71728.H1, ADI-71728.H2, ADI-71728.H3, ADI-71728.H4; ADI-71728.vhCDR1, ADI-71728.vhCDR2, ADI-71728.vhCDR3, ADI-71728.vlCDR1, ADI-71728.vlCDR2, and ADI-71728.vlCDR3; or

[0266] ADI-71741, ADI-71741.VH, ADI-71741.VL, ADI-71741.HC, ADI-71741.LC, and ADI-71741.H1, ADI-71741.H2, ADI-71741.H3, ADI-71741.H4; ADI-71741.vhCDR1, ADI-71741.vhCDR2, ADI-71741.vhCDR3, ADI-71741.vlCDR1, ADI-71741.vlCDR2, and ADI-71741.vlCDR3;

[0267] ADI-71742, ADI-71742.VH, ADI-71742.VL, ADI-71742.HC, ADI-71742.LC, and ADI-71742.H1, ADI-71742.H2, ADI-71742.H3, ADI-71742.H4; ADI-71742.vhCDR1, ADI-71742.vhCDR2, ADI-71742.vhCDR3, ADI-71742.vlCDR1, ADI-71742.vlCDR2, and ADI-71742.vlCDR3;

[0268] ADI-71744, ADI-71744.VH, ADI-71744.VL, ADI-71744.HC, ADI-71744.LC, and ADI-71744.H1, ADI-71744.H2, ADI-71744.H3, ADI-71744.H4; ADI-71744.vhCDR1, ADI-71744.vhCDR2, ADI-71744.vhCDR3, ADI-71744.vlCDR1, ADI-71744.vlCDR2, and ADI-71744.vlCDR3;

[0269] ADI-71753, ADI-71753.VH, ADI-71753.VL, ADI-71753.HC, ADI-71753.LC, and ADI-71753.H1, ADI-7l753.H2, ADI-71753.H3, ADI-71753.H4; ADI-71753.vhCDR1, ADI-71753.vhCDR2, ADI-71753.vhCDR3, ADI-71753.vlCDR1, ADI-71753.vlCDR2, and ADI-71753.vlCDR3; or

[0270] ADI-71755, ADI-71755.VH, ADI-71755.VL, ADI-71755.HC, ADI-71755.LC, and ADI-71755.H1, ADI-71755.H2, ADI-71755.H3, ADI-71755.H4; ADI-71755.vhCDR1, ADI-71755.vhCDR2, ADI-71755.vhCDR3, ADI-71755.vlCDR1, ADI-71755.vlCDR2, and ADI-71755.vlCDR3.

[0271] A. IL18-BP antibodies that compete for binding with the listed antibodies The present invention provides not only the recited antibodies, but also additional antibodies that compete with the recited antibodies that specifically bind to the IL18-BP molecule (the VH numbers and ADI numbers recited herein that specifically bind to IL18-BP). The IL18-BP antibodies of the present invention include VH1-03.66650, VH1-69.66670, VH3-23.66692, VH1-39.66716, VL-κ-1-5-66650, VL-κ-1-12, 66670, VL-κ-1-12, 66692, VL-κ-1-12, ADI-71663, ADI-71662, ADI-66692, ADI-71701, ADI-71709, ADI-71710, ADI-71711, ADI-71712, ADI-71713, ADI-71714, ADI-71715, ADI-71716, ADI-71717, ADI-71718, ADI-71719, ADI-71720, ADI-71721, ADI-71722, ADI-71723, ADI-71724, ADI-71725, ADI-71726, ADI-71727, ADI-71728, ADI-71729, ADI-71730, ADI-71731, ADI-71732, ADI-71733, ADI-71734, ADI-71735, ADI-71736, ADI-71737, ADI-71740, ADI-71741, ADI-71742, ADI- 710, ADI-71707, ADI-71717, ADI-71719, ADI-71220, ADI-71722, ADI-71736, ADI-71739, ADI-71728, ADI-66716, ADI-71741, ADI-71742, ADI-71744, ADI-71753 or ADI-71755.

[0272] B. Generation of Further Antibodies Further antibodies against human IL18-BP can be made as is known in the art using well-known methods such as those outlined in the Examples. Thus, further anti-IL18-BP antibodies can be produced by conventional methods, for example, by immunizing mice (possibly using DNA immunization such as that used by Aldevron), followed by screening and hybridoma generation (with antibody purification and collection) against IL18-BP (including human IL18-BP) protein.

[0273] C. Optional Antibody Manipulations Anti-IL18-BP antibodies of the present invention (e.g., anti-IL18-BP antibodies including those having the same CDRs as those shown in Figures 1, 2, and / or 3) may be modified or engineered to alter their amino acid sequence by amino acid substitution.

[0274] As used herein, "amino acid substitution" or "substitution" refers to the replacement of an amino acid at a particular position in a parent polypeptide sequence with a different amino acid. In particular, in some embodiments, the substitution is with an amino acid that does not naturally occur at the particular position, that does not naturally occur in that organism or in any organism. For example, the substitution E272Y refers to a variant polypeptide, in this case an Fc variant, in which glutamic acid at position 272 has been replaced with tyrosine. For clarity, a protein engineered to alter a nucleic acid coding sequence but not change the starting amino acid (e.g., exchanging CGG (which encodes arginine) for CGA (which still encodes arginine) to increase expression levels in a host organism) is not an "amino acid substitution." That is, if a protein has the same amino acid at the particular position where it started, despite the creation of a new gene encoding the same protein, it is not an amino acid substitution.

[0275] As discussed herein, amino acid substitutions can be made to alter the affinity of the CDRs for the IL18-BP protein (including both increasing and decreasing binding, as more fully outlined below) and to alter additional functional properties of the antibody. For example, antibodies can be engineered to contain modifications within the Fc region, typically to alter one or more functional properties of the antibody, such as serum half-life, complement fixation, Fc receptor binding, and / or antigen-dependent cellular cytotoxicity. Furthermore, antibodies according to at least some embodiments of the invention may be chemically modified (e.g., one or more chemical moieties may be attached to the antibody) or modified to alter its glycosylation, again altering one or more functional properties of the antibody. Such embodiments are further described below. The numbering of residues in the Fc region is that of the EU index of Kabat.

[0276] In some embodiments, C H1The hinge region of CH1 is modified such that the number of cysteine ​​residues in the hinge region is altered, e.g., increased or decreased. This approach is further described in U.S. Patent No. 5,677,425 by Bodmer et al. The number of cysteine ​​residues in the hinge region of CH1 is altered, for example, to facilitate assembly of the light and heavy chains or to increase or decrease the stability of the antibody.

[0277] In another embodiment, the Fc-hinge region of the antibody is mutated to decrease the biological half-life of the antibody. More specifically, one or more amino acid mutations are introduced into the CH2-CH3 domain interface region of the Fc-hinge fragment such that the antibody has impaired Staphylococcus protein A (SpA) binding compared to native Fc-hinge domain SpA binding. This approach is described in further detail in U.S. Patent No. 6,165,745 by Ward et al.

[0278] In some embodiments, amino acid substitutions can be made in the Fc region, generally to alter binding to FcγR receptors. As used herein, "Fc gamma receptor," "FcγR," or "Fc gamma R" refers to any member of a family of proteins that bind to the IgG antibody Fc region and are encoded by FcγR genes. In humans, this family includes, but is not limited to, FcγRI (CD64), which includes the isoforms FcγRIa, FcγRIb, and FcγRIC; FcγRII (CD32), which includes the isoforms FcγRIIa (including allotypes H131 and R131), FcγRIIb (including FcγRIIb-1 and FcγRIIb-2), and FcγRIIc; and FcγRIII (CD16), which includes the isoforms FcγRIIIa (including allotypes V158 and F158) and FcγRIIIb (including allotypes FcγRIIIb-NA1 and FcγRIIIb-NA2) (Jefferis et al., 2002, Immunol Lett 82:57-65, incorporated by reference in its entirety), and any undiscovered human FcγR or FcγR isoform or allotype. FcγRs can be derived from any organism, including, but not limited to, humans, mice, rats, rabbits, and monkeys. Mouse FcγRs include, but are not limited to, FcγRI (CD64), FcγRII (CD32), FcγRIII-1 (CD16), and FcγRIII-2 (CD16-2), as well as any unidentified mouse FcγR or FcγR isoform or allotype.

[0279] There are many useful Fc substitutions that can be made to alter binding to one or more FcγR receptors. Substitutions that result in increased binding as well as decreased binding can be useful. For example, increased binding to FcγRIIIa is known to generally increase ADCC (antibody-dependent cell-mediated cytotoxicity; a cell-mediated reaction in which nonspecific cytotoxic cells expressing FcγR recognize bound antibodies on target cells and subsequently cause lysis of the target cells). Similarly, decreased binding to FcγRIIb (an inhibitory receptor) can also be beneficial in some situations. Amino acid substitutions that find use in the present invention include those listed in U.S. Patent Application No. 11 / 124,620 (particularly Figure 41) and U.S. Patent No. 6,737,056, both of which are expressly incorporated by reference in their entirety, particularly with respect to the variants disclosed therein. Particular variants that find use include, but are not limited to, 236A, 239D, 239E, 332E, 332D, 239D / 332E, 267D, 267E, 328F, 267E / 328F, 236A / 332E, 239D / 332E / 330Y, 239D, 332E / 330L, 299T and 297N.

[0280] Additionally, antibodies of the invention are modified to increase their biological half-life. Various approaches are possible. For example, one or more of the following mutations can be introduced: T252L, T254S, T256F, as described in U.S. Patent No. 6,277,375 to Ward. Alternatively, to increase biological half-life, antibodies can be modified to contain salvage receptor binding epitopes taken from two loops of the CH2 domain of the Fc region of IgG, as described in U.S. Patent Nos. 5,869,046 and 6,121,022 to Presta et al. H1 Area or C L Additional mutations to increase serum half-life are disclosed in U.S. Patent Nos. 8,883,973, 6,737,056, and 7,371,826, and include 428L, 434A, 434S, and 428L / 434S.

[0281] In yet other embodiments, the Fc region is altered by substituting at least one amino acid residue with a different amino acid residue to alter the effector function of the antibody. For example, one or more amino acids selected from amino acid residues 234, 235, 236, 237, 297, 318, 320, and 322 can be substituted with a different amino acid residue such that the antibody has altered affinity for an effector ligand but retains the antigen-binding ability of the parent antibody. The effector ligand for which affinity is altered can be, for example, an Fc receptor or the C1 component of complement. This approach is described in further detail in U.S. Patent Nos. 5,624,821 and 5,648,260, both by Winter et al.

[0282] In another example, one or more amino acids selected from amino acid residues 329, 331, and 322 can be substituted with a different amino acid residue such that the antibody has altered C1q binding and / or reduced or abolished complement dependent cytotoxicity (CDC). This approach is described in further detail in U.S. Patent No. 6,194,551 by Idusogie et al.

[0283] In another example, one or more amino acid residues within amino acid positions 231-239 are altered to thereby alter the ability of the antibody to fix complement. This approach is further described in PCT application WO 94 / 29351 by Bodmer et al.

[0284] In yet another example, the Fc region may comprise a nucleotide sequence selected from the group consisting of nucleotides at the following positions: 238, 239, 248, 249, 252, 254, 255, 256, 258, 265, 267, 268, 269, 270, 272, 276, 278, 280, 283, 285, 286, 289, 290, 292, 293, 294, 295, 296, 298, 301, 303, 305, 307, 309, 312, 315, 320, 322, 324, 326, 327, 329, 330 , 331, 333, 334, 335, 337, 338, 340, 360, 373, 376, 378, 382, ​​388, 389, 398, 414, 416, 419, 430, 434, 435, 437, 438, or 439 to increase the antibody's ability to mediate antibody-dependent cell-mediated cytotoxicity (ADCC) and / or increase the antibody's affinity for Fcγ receptors. This approach is further described in PCT application WO 00 / 42072 by Presta. Additionally, the binding sites on human IgG1 for FcγRI, FcγRII, FcγRIII, and FcRn have been mapped, and variants with improved binding have been described (see Shields, R., et al. (2001) J. Biol. Chem. 276:6591-6604). Specific mutations at positions 256, 290, 298, 333, 334, and 339 have been shown to improve binding to FcγRIII. Additionally, the following combination mutations have been shown to improve FcγRIII binding: T256A / S298A, S298A / E333A, S298A / K224A, and S298A / E333A / K334A. Furthermore, mutations such as M252Y / S254T / T256E or M428L / N434S improve binding to FcRn and increase antibody circulating half-life (see Chan CA and Carter PJ (2010) Nature Rev Immunol 10:301-316).

[0285] In yet another embodiment, antibodies can be engineered to abolish in vivo Fab arm exchange. Specifically, this process involves the exchange of IgG4 half molecules (one heavy chain plus one light chain) with other IgG4 antibodies, effectively resulting in a functionally monovalent bispecific antibody. Mutations to the hinge region and constant domain of the heavy chain can abolish this exchange (see Aalberse, RC, Schuurman J., 2002, Immunology 105:9-19).

[0286] In yet another embodiment, the glycosylation of the antibody is modified. For example, an aglycosylated antibody can be generated (i.e., the antibody lacks glycosylation). Glycosylation can be modified, for example, to increase the affinity of the antibody for the antigen or to reduce effector function, such as ADCC. Such carbohydrate modifications can be achieved, for example, by altering one or more of the glycosylation sites within the antibody sequence, e.g., N297. For example, one or more amino acid substitutions can be made that result in the elimination of one or more variable region framework glycosylation sites, thereby eliminating glycosylation at that site.

[0287] Additionally or alternatively, antibodies can be generated with altered glycosylation types, such as hypofucosylated antibodies with reduced amounts of fucosyl residues or antibodies with increased bisecting GlcNAc structures. Such altered glycosylation patterns have been demonstrated to increase the ADCC ability of antibodies. Such carbohydrate modifications can be achieved, for example, by expressing the antibody in a host cell with altered glycosylation machinery. Cells with altered glycosylation machinery have been described in the art and can be used as host cells to express recombinant antibodies according to at least some embodiments of the present invention, thereby producing antibodies with altered glycosylation. For example, cell lines Ms704, Ms705, and Ms709 lack the fucosyltransferase gene FUT8 (α(1,6) fucosyltransferase), and as a result, antibodies expressed in the Ms704, Ms705, and Ms709 cell lines lack fucose on their carbohydrates. The Ms704, Ms705, and Ms709 FUT8 cell lines were generated by targeted disruption of the FUT8 gene in CHO / DG44 cells using two replacement vectors (see U.S. Patent Application Publication No. 20040110704 by Yamane et al. and Yamane-Ohnuki et al. (2004) Biotechnol Bioeng 87:614-22). As another example, European Patent No. 1,176,195 by Hanai et al. describes cell lines with a functionally disrupted FUT8 gene encoding a fucosyltransferase, and antibodies expressed in such cell lines exhibit hypofucosylation by reducing or eliminating the enzyme associated with α1,6 linkages. Hanai et al. also describe cell lines, such as the rat myeloma cell line YB2 / 0 (ATCC CRL1662), that have low or no enzymatic activity for adding fucose to N-acetylglucosamine attached to the Fc region of an antibody.PCT application WO 03 / 035835 by Presta describes a variant CHO cell line, Lec13 cells, that has a reduced ability to attach fucose to Asn(297)-linked carbohydrates, resulting in hypofucosylation of antibodies expressed in the host cells (see also Shields, R. Let al. (2002) J. Biol. Chem. 277:26733-26740). PCT application WO 99 / 54342 by Umana et al. describes cell lines engineered to express glycoprotein-modifying glycosyltransferases (e.g., β(1,4)-N-acetylglucosaminyltransferase III (GnTIII)), and antibodies expressed in the engineered cell lines exhibit increased bisecting GlcNAc structures, resulting in increased ADCC activity of the antibodies (see also Umana et al. (1999) Nat. Biotech. 17:176-180). Alternatively, the fucose residues of the antibody can be cleaved off using a fucosidase enzyme, for example, the fucosidase α-L-fucosidase, which removes fucosyl residues from antibodies (Tarentino, AL et al. (1975) Biochem. 14:5516-23).

[0288] Another modification of the antibodies herein contemplated by the invention is pegylation or the addition of other water-soluble moieties, typically polymers, for example to enhance half-life. Antibodies can be pegylated, for example, to increase the biological (e.g., serum) half-life of the antibody. To pegylate an antibody, the antibody or fragment thereof is typically reacted with polyethylene glycol (PEG), such as a reactive ester or aldehyde derivative of PEG, under conditions such that one or more PEG groups become attached to the antibody or antibody fragment. Preferably, pegylation is carried out via an acylation reaction or an alkylation reaction with a reactive PEG molecule (or an analogous reactive water-soluble polymer). As used herein, the term "polyethylene glycol" refers to a mono(C1-C 10The term "PEG" is intended to encompass any of the forms of PEG that have been used to derivatize other proteins, such as alkoxy- or aryloxy-polyethylene glycol or polyethylene glycol-maleimide. In certain embodiments, the antibody to be PEGylated is an aglycosylated antibody. Methods for PEGylating proteins are known in the art and can be applied to antibodies according to at least some embodiments of the invention. See, for example, European Patent No. 0 154 316 to Nishimura et al. and European Patent No. 0 401 384 to Ishikawa et al.

[0289] In addition to substitutions made to alter binding affinity to FcγR and / or FcRn and / or increase in vivo serum half-life, further antibody modifications can be made, as described in more detail below.

[0290] In some cases, affinity maturation is performed. Amino acid modifications in the CDRs are sometimes referred to as "affinity maturation." An "affinity matured" antibody is an antibody with one or more alterations(s) in one or more CDRs that result in improved affinity of the antibody for the antigen compared to a parent antibody that does not have those alterations(s). In some cases, although rare, it may be desirable to decrease the affinity of an antibody for its antigen, although this is generally not preferred.

[0291] In some embodiments, one or more amino acid modifications are made in one or more of the CDRs of an IL18-BP antibody of the invention. Generally, only one, two, or three amino acids are substituted in any single CDR, and generally no more than one, two, three, four, five, six, seven, eight, nine, or ten changes are made within a set of CDRs. However, it will be understood that any combination of none, one, two, or three substitutions in any CDR can be independently and optionally combined with any other substitution.

[0292] Affinity maturation can be used to increase the binding affinity of an antibody to the IL18-BP antigen by at least about 100% or more, or by at least about 10% or more, compared to the "parent" antibody. 4 That's it, 10 5 That's it, 10 6 That's it, 10 7 Affinity maturation can be performed to increase affinity to or above the nanomolar range. Preferred affinity-matured antibodies have nanomolar or picomolar affinity for the IL18-BP antigen. Affinity-matured antibodies are produced by known procedures. See, for example, Marks et al., 1992, Biotechnology 10:779-783, which describes affinity maturation by variable heavy (VH) and variable light (VL) domain shuffling. Random mutagenesis of CDR and / or framework residues is described, for example, in Barbas, et al., PNAS, USA 91:3809-3813 (1994); Shier et al., Gene, 169:147-155 (1995); Yelton et al., J. Immunol., 155:1994-2004 (1995); Jackson et al., J. Immunol. 154(7):3310-9 (1995); and Hawkins et al., J. Mol. Biol., 226:889-896 (1992).

[0293] Alternatively, amino acid modifications can be made in one or more of the CDRs of an antibody of the invention that are "silent," e.g., that do not significantly alter the affinity of the antibody for antigen. These can be made for several reasons, including to optimize expression (which can be made to nucleic acids encoding an antibody of the invention).

[0294] Thus, included within the definition of the CDRs and antibodies of the invention are variant CDRs and antibodies. That is, the antibodies of the invention may contain amino acid modifications in one or more of the listed CDRs of the antibodies of the invention. Furthermore, as outlined below, amino acid modifications may also be made independently and optionally in any region outside of the CDRs, including framework and constant regions.

[0295] III. Nucleic acid composition Also provided are nucleic acid compositions encoding the anti-IL18-BP antibodies of the invention, as well as expression vectors containing the nucleic acids and host cells transformed with the nucleic acid and / or expression vector compositions. As will be appreciated by those of skill in the art, the protein sequences set forth herein can be encoded by any number of possible nucleic acid sequences due to the degeneracy of the genetic code.

[0296] The nucleic acid composition encoding an IL18-BP antibody will depend on the antibody format. Traditionally, a tetrameric antibody containing two heavy chains and two light chains is encoded by two different nucleic acids, one encoding the heavy chain and the other encoding the light chain. These can be placed into a single expression vector or two expression vectors, as known in the art, and transformed into a host cell, where they are expressed to form the antibody of the present invention. In some embodiments, for example, when an scFv construct is used, a single nucleic acid encoding the variable heavy chain-linker-variable light chain is generally used, which can be inserted into an expression vector for transformation into a host cell. The nucleic acid can be placed into an expression vector containing appropriate transcriptional and translational control sequences (including, but not limited to, signal and secretory sequences, regulatory sequences, promoters, origins of replication, selection genes, etc.).

[0297] Preferred mammalian host cells for expressing recombinant antibodies according to at least some embodiments of the present invention include Chinese hamster ovary (CHO cells), PER.C6, HEK293, and others known in the art.

[0298] Nucleic acids may be present in whole cells, in a cell lysate, or in a partially purified or substantially pure form. A nucleic acid is "isolated" or "substantially purified" when it is purified from other cellular components or other contaminants (e.g., other cellular nucleic acids or proteins) by standard techniques, including alkaline / SDS treatment, CsCl banding, column chromatography, agarose gel electrophoresis, and other techniques well known in the art.

[0299] To generate the scFv gene, H and V L The DNA fragment encoding V H Sequence and V L The sequences are V linked by a flexible linker L Area and V H The fragment is operably linked to another fragment encoding a flexible linker, e.g., encoding the amino acid sequence (Gly4-Ser)3 (SEQ ID NO: 150), so that it can be expressed as a continuous single-chain protein having the region (see, e.g., Bird et al., Science 242:423-426 (1988); Huston et al. PNAS, 85:5879-5883 (1988); McCafferty et al., Nature 348:552-554 (1990)).

[0300] IV. Administration of anti-IL18-BP antibody preparations Administration of pharmaceutical compositions comprising anti-IL18-BP antibodies of the present invention (e.g., anti-IL18-BP antibodies comprising those having the same CDRs as those shown in Figures 1, 2, and / or 3), preferably in the form of a sterile aqueous solution, can be carried out in a variety of ways. As is known in the art, protein therapeutics are often delivered by IV infusion. Antibodies of the present invention can also be delivered using such methods. For example, administration can be intravenous or by intravenous infusion using 0.9% sodium chloride as the infusion vehicle. Such techniques are disclosed in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed., 1980.

[0301] The dosage and frequency of administration are, in some embodiments, selected to be therapeutically or prophylactically effective. As is known in the art, adjustments may be necessary for rates of proteolysis, systemic versus local delivery, and de novo protease synthesis, as well as age, weight, general health, sex, diet, time of administration, drug interactions, and severity of the condition, and can be ascertained by routine experimentation by one of ordinary skill in the art. A therapeutically effective dose of the Fc variants of the invention can be administered to treat a patient. As used herein, a "therapeutically effective dose" refers to a dose that produces an effect and is administered to obtain that effect.

[0302] V. Administration of Anti-IL18-BP Antibody Formulations Administration of pharmaceutical compositions comprising anti-IL18-BP antibodies of the present invention (e.g., anti-IL18-BP antibodies, including those shown in Figures 1, 2, and / or 3), preferably in the form of a sterile aqueous solution, can be accomplished in a variety of ways. As is known in the art, protein therapeutics are often delivered by IV infusion. Antibodies of the present invention can also be delivered using such methods. For example, administration can be intravenous or by intravenous infusion using 0.9% sodium chloride as the infusion vehicle. Such techniques are disclosed in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed., 1980.

[0303] The dosage and frequency of administration are, in some embodiments, selected to be therapeutically or prophylactically effective. As is known in the art, adjustments may be necessary for rates of proteolysis, systemic versus local delivery, and de novo protease synthesis, as well as age, weight, general health, sex, diet, time of administration, drug interactions, and severity of the condition, and can be ascertained by routine experimentation by one of ordinary skill in the art. A therapeutically effective dose of the Fc variants of the invention can be administered to treat a patient. As used herein, a "therapeutically effective dose" refers to a dose that produces an effect and is administered to obtain that effect.

[0304] VI. How to Use Anti-IL18-BP Antibodies A. Therapeutic use Anti-IL18-BP antibodies (e.g., anti-IL18-BP antibodies, including those described in Figures 1, 2, and / or 3) find use in treating patients, such as human subjects, having conditions generally associated with IL18-BP levels or free IL18 levels. As used herein, the term "treatment" refers to both therapeutic treatment and prophylactic or preventative measures, and in this example relates to the treatment of cancer. Those in need of treatment include those already with cancer and those in whom cancer is to be prevented. Thus, the mammal treated herein may have been diagnosed with cancer or may be predisposed or susceptible to cancer. As used herein, the term "treating" refers to preventing, delaying the onset of, curing, reversing, attenuating, mitigating, minimizing, suppressing, arresting, or stabilizing discernible symptoms of the above-mentioned cancerous disease, disorder, or condition. It also includes managing cancer as described above. By "managing" it is meant reducing the severity of the disease, reducing the frequency of disease episodes, shortening the duration of such episodes, reducing the severity of such episodes, slowing / reducing the growth or proliferation of cancer cells, slowing the progression of at least one symptom, improving at least one measurable physical parameter, etc. For example, immunostimulatory anti-IL18-BP immune molecules are said to treat cancer or infectious diseases by promoting cytokine immunity against T cells, NK cells, NKT cells, myeloid cells, dendritic cells, MAIT T cells, γδT cells, and / or innate lymphoid cells (ILCs), or target cells, e.g., cancer, infected cells, or pathogen cells, thereby depleting cells involved in the disease state.

[0305] The IL18-BP antibodies of the present invention are provided in a therapeutically effective dose. A "therapeutically effective dose" of an anti-IL18-BP immune molecule according to at least some embodiments of the present invention preferably results in a reduction in the severity of disease symptoms, an increase in the frequency and duration of disease symptom-free periods, an increase in lifespan, disease remission, or prevention or reduction of functional impairment or disability due to disease affliction. For example, for the treatment of IL18-BP-positive tumors, a "therapeutically effective dose" preferably inhibits cell proliferation or tumor growth by at least about 20%, more preferably at least about 40%, even more preferably at least about 60%, and even more preferably at least about 80%, compared to untreated subjects. The ability of a compound to inhibit tumor growth can be assessed in an animal model system predictive of efficacy in human tumors. Alternatively, this property of a composition can be assessed by testing the compound's ability to inhibit, such as in vitro inhibition, using assays known to those skilled in the art. A therapeutically effective amount of a therapeutic compound may reduce tumor size or otherwise ameliorate symptoms in a subject.

[0306] One of ordinary skill in the art would be able to determine the therapeutically effective amount based on such factors as the subject's size, the severity of the subject's symptoms, and the particular composition or route of administration selected.

[0307] 1. Cancer treatment The IL18-BP antibodies of the present invention, alone or in combination with other therapeutic agents, have particular utility in the treatment of cancer. Generally, the antibodies of the present invention are immunomodulatory in that, rather than directly attacking cancerous cells, the anti-IL18-BP antibodies of the present invention stimulate the immune system, generally by inhibiting the action of IL18-BP. Thus, unlike targeted tumor therapy, which aims to inhibit molecular pathways important for tumor growth and development and / or deplete tumor cells, cancer immunotherapy aims to stimulate the patient's own immune system to eliminate cancer cells and provide long-term tumor destruction. Various approaches can be used in cancer immunotherapy, including therapeutic cancer vaccines to induce tumor-specific T-cell responses and immunostimulatory antibodies (i.e., antagonists of inhibitory receptors = immune checkpoints) to ablate immunosuppressive pathways.

[0308] Clinical responses from targeted therapy or conventional anticancer therapy tend to be transient, as cancer cells develop resistance and tumor recurrence occurs. However, the clinical use of cancer immunotherapy in the past few years has shown that this type of therapy can have durable clinical responses, which have a dramatic impact on long-term survival. However, although the response is long-lasting, only a small number of patients respond (in contrast to conventional therapy or targeted therapy, in which a large number of patients respond, but the response is transient).

[0309] By the time tumors are clinically detected, they have already evaded the immune defense system by acquiring immune tolerance and immunosuppressive properties and creating an immunosuppressive tumor microenvironment through various mechanisms and various immune cells.

[0310] Therefore, the anti-IL18-BP antibodies of the present invention are useful for treating cancer. Due to the nature of tumor immune mechanisms, IL18-BP does not necessarily need to be overexpressed in or correlate with a particular cancer type. That is, the goal is for the anti-IL18-BP antibody to de-inhibit T cell, NK cell, NKT cell, myeloid cell, dendritic cell, MAIT T cell, γδ T cell, and / or innate lymphoid cell (ILC) activation, allowing the immune system to attack cancer.

[0311] As used herein, "cancer" broadly refers to any neoplastic disease (whether invasive or metastatic) characterized by abnormal and uncontrolled cell division (e.g., uncontrolled cell proliferation) that leads to malignant growth or tumors. As used herein, the term "cancer" or "cancerous" should be understood to encompass any neoplastic disease (whether invasive, non-invasive, or metastatic) that is characterized by abnormal and uncontrolled cell division that leads to malignant growth or tumors, non-limiting examples of which are described herein. This typically includes any physiological condition in a mammal that is characterized by uncontrolled cell proliferation.

[0312] "Cancer therapy" herein refers to any method of preventing or treating cancer or ameliorating one or more of the symptoms of cancer. Typically, such therapy will involve administering an immunostimulatory anti-IL18-BP antibody (including an antigen-binding fragment) alone or in combination with chemotherapy or radiotherapy or other biologics to enhance its activity, i.e., in individuals in whom IL18-BP expression suppresses anti-tumor responses and the efficacy or biological effectiveness of chemotherapy or radiotherapy.

[0313] Anti-IL18-BP antibodies of the invention may be used as monotherapy or as part of a combination therapy as described herein in the treatment of solid tumors (including, e.g., cancers of the lung, liver, breast, brain, and GI tract) and hematological cancers (including, e.g., leukemias and preleukemia disorders, lymphomas, and plasma cell disorders), carcinomas, lymphomas, blastomas, sarcomas, and leukemia or lymphoid malignancies. In some embodiments, the cancer is early stage. In some embodiments, the cancer is advanced (including metastatic). In some embodiments, cancers suitable for treatment with the invention include cancers that express IL18-BP, and further include non-metastatic or non-invasive, as well as invasive or metastatic cancers, including cancers in which IL18-BP expression by immune cells, stromal cells, or diseased cells suppresses anti-tumor and anti-invasive immune responses. In some embodiments, anti-IL18-BP antibodies can be used for the treatment of angiogenic tumors. In some embodiments, cancers for treatment using anti-IL18-BP antibodies of the invention include carcinoma, lymphoma, sarcoma, and / or leukemia. In some embodiments, cancers for treatment using anti-IL18-BP antibodies of the invention include angiogenic tumors, melanoma, non-melanoma skin cancer (squamous cell carcinoma and basal cell carcinoma), mesothelioma, squamous cell carcinoma, lung cancer, small cell lung cancer, non-small cell lung cancer, neuroendocrine lung cancer (including pleural mesothelioma and neuroendocrine lung carcinoma), NSCL (large cell), NSCLC large cell adenocarcinoma, non-small cell lung cancer (NSCLC), NSCLC squamous cell, soft tissue sarcoma, Kaposi's sarcoma, adenocarcinoma of the lung, squamous cell carcinoma of the lung, NSCLC with a TPS of PDL1 > 50%, neuroendocrine lung carcinoma, atypical carcinoid lung cancer, cancer of the peritoneum, esophageal cancer, hepatocellular carcinoma, liver cancer (including HCC), gastric cancer, cancer), stomach cancer (including gastrointestinal cancer), pancreatic cancer, glioblastoma, cervical cancer , ovarian cancer, urothelial carcinoma, bladder cancer, liver cancer, glioma, brain cancer (and edema such as that associated with brain tumors), breast cancer (including, for example, triple-negative breast cancer), testicular cancer, testicular germ cell tumor, colon cancer, colorectal cancer (CRC), colorectal cancer with MSS (MSS-CRC); refractory MSS colorectal; MSS (microsatellite stable), primary peritoneal cancer, primary peritoneal ovarian cancer, microsatellite stable primary peritoneal cancer, platinum-resistant microsatellite stable primary peritoneal cancer, CRC (MSS unknown), rectal cancer, endometrial cancer (including endometrioma), uterine cancer, salivary gland cancer, kidney cancer, renal cell carcinoma (RCC), gastroesophageal junction cancer, prostate cancer, vulvar cancer, thyroid cancer, liver cancer, carcinoid cancer, head and neck cancer, B-cell lymphoma (including non-Hodgkin's lymphoma, as well as low-grade / follicular non-Hodgkin's lymphoma (NHL), small lymphocytic (SL) NHL, intermediate-grade / follicular NHL, intermediate-grade diffuse NHL, diffuse large B-cell lymphoma, high-grade immunoblastic NHL, high-grade lymphoblastic NHL, high-grade small non-cleaved cell NHL, giant cell lymphoma, Mass Lesion (including NHL, mantle cell lymphoma, AIDS-related lymphoma, and Waldenstrom's macroglobulinemia), Hodgkin's lymphoma (HD), chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), T-cell acute lymphoblastic leukemia (T-ALL), acute myeloid leukemia (AML), hairy cell leukemia, chronic myeloblastic leukemia, multiple myeloma, post-transplant lymphoproliferative disorder (PTLD), abnormal blood vessel growth associated with phacomatosis, Meigs syndrome, Merkel cell carcinoma, MSI-high cancer, KRAS Includes mutant tumors, adult T-cell leukemia / lymphoma, adenoid cystic carcinoma (including adenoid cystic cell carcinoma), melanoma, malignant melanoma, metastatic melanoma, pancreatic cancer, pancreatic adenocarcinoma, ovarian cancer (including ovarian carcinoma), pleural mesothelioma, cervical squamous cell carcinoma (cervical SCC), anal squamous cell carcinoma (anal SCC), carcinoma of unknown primary, gallbladder carcinoma, pleural mesothelioma, chordoma, endometrial sarcoma, chondrosarcoma, uterine sarcoma, uveal melanoma, amyloidosis, AL-amyloidosis, astrocytoma, and / or myelodysplastic syndrome (MDS).

[0314] In some embodiments, cancers for treatment using the anti-IL18-BP antibodies of the invention include cancers selected from the group consisting of renal clear cell carcinoma (RCC), lung cancer, NSCLC, lung adenocarcinoma, lung squamous cell carcinoma, gastric adenocarcinoma, ovarian cancer, endometrial cancer, breast cancer, triple-negative breast cancer (TNBC), head and neck tumors, colorectal adenocarcinoma, melanoma, and metastatic melanoma.

[0315] 2. Anti-IL-18BP antibody monotherapy The IL-18BP antibodies of the present invention have particular utility in the treatment of cancer as monotherapy. Due to the nature of tumor immune mechanisms, IL18BP does not necessarily need to be overexpressed in or correlate with a particular cancer type. That is, the goal is for the anti-IL18BP antibody to de-inhibit the activation of T cells and NK cells, allowing the immune system to attack the cancer.

[0316] Any of the anti-IL-18 antibodies of Figures 1-3 have use as monotherapy.

[0317] 3. Anti-IL18BP antibody combination therapy As is known in the art, combination therapies involving therapeutic antibodies directed against immunotherapeutic targets and additional therapeutic agents specific to a disease state are very promising. For example, in the field of immunotherapy, there are several promising combination therapies using chemotherapeutic agents (either small molecule drugs or anti-tumor antibodies) or tumor-immune antibodies.

[0318] The terms "in combination" and "co-administration" are not limited to administration of the prophylactic or therapeutic agents at exactly the same time. Instead, it is meant that the antibody and other agent(s) are administered in a sequence and within a time interval such that they act together to provide an increased benefit over treatment with either the antibody of the invention or the other agent(s) alone. It is preferred that the antibody and other agent(s) act additively, and it is particularly preferred that they act synergistically.

[0319] Thus, an antibody of the present invention can be administered simultaneously with one or more other therapeutic regimens or agents. In some embodiments, an antibody of the present invention is administered in the same formulation as one or more other therapeutic regimens or agents. In some embodiments, an antibody of the present invention is administered in a separate and / or different formulation from one or more other therapeutic regimens or agents. The additional therapeutic regimen or agent may be used to improve the efficacy or safety of the antibody. Alternatively, the additional therapeutic regimen or agent may be used to treat the same disease or comorbidity rather than altering the action of the antibody. For example, an antibody of the present invention may be administered to a patient together with chemotherapy, radiation therapy, or both chemotherapy and radiation therapy.

[0320] In some embodiments, the anti-IL18BP antibodies of the present invention can be combined with one of several checkpoint receptor antibodies. In some embodiments, the patient's tumor can be evaluated for receptor expression, and the results can then be used to inform the clinician as to which antibody to administer. Any of the anti-IL-18 antibodies of Figures 1-3 find use as part of a combination therapy.

[0321] a. Immune checkpoint inhibitor combination therapy In some embodiments, the combination or composition further comprises an additional active agent, e.g., a second antigen binding protein. Optionally, the second antigen binding protein binds to a negative regulator of the immune system, an immunosuppressant, or an immune checkpoint protein, including, but not limited to, PD-1, PD-L1, CTLA-4, PD-L2, B7-H3, B7-H4, CEACAM-1, TIGIT, PVR, LAG3, CD112, PVRIG, CD96, TIM3, and / or BTLA, or the costimulatory receptors ICOS, OX40, 41BB, CD27, and / or GITR. All patent documents listed in the following section are incorporated by reference in their entirety for all purposes.

[0322] In some embodiments, the anti-IL18-BP antibody is used in combination with an antibody against an immune checkpoint inhibitory protein, in some embodiments, the immune checkpoint inhibitory protein is selected from the group consisting of an anti-PVRIG antibody, an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-TIGIT antibody, an anti-CTLA-4 antibody, an anti-PD-L2 antibody, an anti-B7-H3 antibody, an anti-B7-H4 antibody, an anti-CEACAM-1 antibody, an anti-PVR antibody, an anti-LAG3 antibody, an anti-CD112 antibody, an anti-CD96 antibody, an anti-TIM3 antibody, an anti-BTLA antibody, an anti-ICOS antibody, an anti-OX40 antibody, an anti-41BB antibody, an anti-CD27 antibody, or an anti-GITR antibody.

[0323] In some embodiments, an anti-IL18-BP antibody is used in combination with one or more anti-PD-1 (e.g., anti-PD-1 targeting antibodies), such as, but not limited to, nivolumab (Opdivo®; BMS; CheckMate078), pembrolizumab (KEYTRUDA®; Merck), TSR-042 (Tesaro), cemiplimab (REGN2810; Regeneron Pharmaceuticals, see U.S. Patent Application Publication No. 20170174779), BMS-936559, spartalizumab (PDR001, Novartis), pidilizumab (CT-011; Pfizer Inc), tislelizumab (BGB-A317, BeiGene), camrelizumab (SHR-1210, Incyte and Jiangsu HengRui), SHR-1210 (CTR20170299 and CTR20170322), SHR-1210 (CTR20160175 and CTR20170090), sintilimab (Tyvyt®; Eli Lily and Innovent Biologics), toripalimab (JS001, Shanghai Junshi Bioscience), JS-001 (CTR20160274), IBI308 (CTR20160735), BGB-A317 (CTR20160872), penprimimab (AK105, Akeso Biopharma), dimvelelimab (Arcus), BAT1306 (Bio-Thera Solutions Ltd), sasanlimab (PF-06801591, Pfizer), dostallimab-gxly (GlaxoSmithKline LLC), prorugolimab (Biocad), kadonilimab (Akeso Inc), geptanolimab (Genor BioPharma Co Ltd), serplulimab (Shanghai Henlius Biotech Inc), balstilimab (Agenus Inc), retifanlimab (Incyte Corp), cetrelimab (Johnson & Johnson), CS-1003 (EQRx Inc), IBI-318 (Innovent Biologics Inc), ivonesimab (Akeso Inc), pucotenlimab (Lepu Biopharma CoLtd), QL-1604 (Qilu Pharmaceutical Co Ltd), SCTI-10A (SinoCelltech Group Ltd), tebotelimab (MacroGenics Inc), AZD-7789 (AstraZeneca Plc), budigalimab (AbbVie Inc), EMB-02 (EpimAb Biotherapeutics Inc), ezabenlimab (Boehringer Ingelheim International GmbH), F-520 (Shandong New Time Pharmaceutical Co Ltd), HX-009 (Waterstone Hanxbio Pty Ltd), zelvalimab (Amgen), peresolimab (Eli Lilly and Co), rosnilimab (AnaptysBio Inc), budalimab (Xencor), izuralimab (Xencor), lorigellimab (MacroGenics Inc), YBL-006 (Y-Biologics Inc), and ONO-4685 (Ono Pharmaceutical Exemplary anti-PD-1 antibody sequences include those described in U.S. Patent Application Publication No. 2017 / 0081409, such as those described in U.S. Pat. No. 6,239,141, and others under development, which can be used in combination with the anti-IL18BP antibodies of the invention. Additional exemplary anti-PD-1 antibody sequences are shown in FIG. 39.

[0324] In some embodiments, pembrolizumab is administered at a dose of about 2 mg / kg to 10 mg / kg. In some embodiments, pembrolizumab is administered at a dose of about 2 mg / kg. In some embodiments, pembrolizumab is administered at a dose of about 2 mg / kg. In some embodiments, pembrolizumab is administered at a dose of about 3 mg / kg. In some embodiments, pembrolizumab is administered at a dose of about 4 mg / kg. In some embodiments, pembrolizumab is administered at a dose of about 5 mg / kg. In some embodiments, pembrolizumab is administered at a dose of about 6 mg / kg. In some embodiments, pembrolizumab is administered at a dose of about 7 mg / kg. In some embodiments, pembrolizumab is administered at a dose of about 8 mg / kg. In some embodiments, pembrolizumab is administered at a dose of about 9 mg / kg. In some embodiments, pembrolizumab is administered at a dose of about 10 mg / kg.

[0325] In some embodiments, pembrolizumab is administered at a dose of about 2 mg / kg or less. In some embodiments, pembrolizumab is administered at a dose of about 1 mg / kg to 2 mg / kg. In some embodiments, pembrolizumab is administered at a dose of about 0.1 mg / kg to 1 mg / kg. In some embodiments, pembrolizumab is administered at a dose of about 0.01 mg / kg to 0.1 mg / kg.

[0326] In some embodiments, pembrolizumab is administered at a dose of at least about 10 mg / kg. In some embodiments, pembrolizumab is administered at a dose of about 10 mg / kg to 20 mg / kg. In some embodiments, pembrolizumab is administered at a dose of about 20 mg / kg to 30 mg / kg. In some embodiments, pembrolizumab is administered at a dose of about 30 mg / kg to 40 mg / kg. In some embodiments, pembrolizumab is administered at a dose of about 40 mg / kg to 50 mg / kg.

[0327] In some embodiments, pembrolizumab is administered about every week to every six weeks. In some embodiments, pembrolizumab is administered about every week. In some embodiments, pembrolizumab is administered about every two weeks. In some embodiments, pembrolizumab is administered about every three weeks. In some embodiments, pembrolizumab is administered about every four weeks. In some embodiments, pembrolizumab is administered about every five weeks. In some embodiments, pembrolizumab is administered about every six weeks.

[0328] In some embodiments, pembrolizumab is administered at a dose of about 2 mg / kg every 3 weeks. In some embodiments, pembrolizumab is administered at a dose of about 10 mg / kg every 3 weeks. In some embodiments, pembrolizumab is administered at a dose of about 200 mg every 3 weeks. In some embodiments, pembrolizumab is administered at a dose of about 400 mg every 6 weeks.

[0329] In some embodiments, pembrolizumab is administered over about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, about 30 minutes, about 35 minutes, or about 40 minutes, hi some embodiments, pembrolizumab is administered over about 30 minutes + / - 10 minutes.

[0330] Further disclosure of pembrolizumab is provided at https: / / www.accessdata.fda.gov / spl / data / 157262d6-15e0-4b0a-968f-b99bab4aef50 / 157262d6-15e0-4b0a-968f-b99bab4aef50.xml, which is incorporated herein by reference in its entirety.

[0331] In some embodiments, an anti-IL18-BP antibody is used in combination with one or more anti-PD-L1 antibodies (e.g., anti-PD-L1 targeting antibodies). There are three approved anti-PD-L1 antibodies: atezolizumab (TECENTRIQ®; MPDL3280A; IMpower110; Roche / Genentech), avelumab (BAVENCIO®, MSB001071 8C, EMD Serono & Pfizer), and durvalumab (MEDI4736, IMFINZI®, AstraZeneca). Additionally, other antibodies are in development, such as lodapolimab (LY3300054, Eli Lily), pimivalimab (Jounce Therapeutics Inc), SHR-1316 (Jiangsu Hengrui Medicine Co Ltd), emvafolimab (Jiangsu Simcere Pharmaceutical Co Ltd), sugemalimab (CStone Pharmaceuticals Co Ltd), cosibelimab (Checkpoint Therapeutics Inc), and pakmilimab (CytomX Therapeutics Inc)、IBI-318、IBI-322、IBI-323(Innovent Biologics Inc)、INBRX-105(Inhibrx Inc)、KN-046(Alphamab Oncology)、6MW-3211(Mabwell Shanghai Bioscience Co Ltd)、BNT-311(BioNTech SE)、FS-118(F-star Therapeutics Inc)、GNC-038(Systimmune Inc)、GR-1405(Genrix(Shanghai)Biopharmaceutical Co Ltd)、HS-636(Zhejiang Hisun Pharmaceutical Co Ltd)、LP-002(Lepu Biopharma Co Ltd)、PM-1003(Biotheus Inc)、PM-8001(Biotheus Inc)、STIA-1015(ImmuneOncia Therapeutics LLC)、ATG-101(Antengene Corp Ltd)、BJ-005(BJ Bioscience Inc)、CDX-527(Celldex Therapeutics Inc)、GNC-035(Systimmune Inc)、GNC-039(Systimmune Inc)、HLX-20(Shanghai Henlius Biotech Inc)、JS-003(Shanghai Junshi Bioscience Co Ltd), LY-3434172 (Eli Lilly and Co), MCLA-145 (Merus NV), MSB-2311 (Transcenta Holding Ltd), PF-07257876 (Pfizer Inc), Q-1802 (QureBio Ltd), QL-301 (QLSF Biotherapeutics Inc), QLF-31907 (Qilu Pharmaceutical Co Ltd), RC-98 (RemeGen Co Ltd), TST-005 (Transcenta Holding Ltd), atezolizumab (IMpower133), BMS-936559 / MDX-1105, and / or RG-7446 / MPDL3280A, and YW243.55.S70. In some embodiments, the PD-L1 antibody is one described in U.S. Patent Application Publication No. 2017 / 0281764, and WO 2013 / 079174 (avelumab) and WO 2010 / 077634 (or U.S. Patent Application Publication No. 2016 / 0222117 or U.S. Patent No. 8,217,149; atezolizumab). In some embodiments, the PD-L1 antibody comprises the heavy chain sequence of SEQ ID NO: 34 and the light chain sequence of SEQ ID NO: 36 (from U.S. Patent Application Publication No. 2017 / 281764), as well as others in development that can be used in combination with the anti-IL18BP antibodies of the invention. Further exemplary anti-PD-L1 antibody sequences are shown in Figure 40.

[0332] In some embodiments, an anti-IL18-BP antibody is used in combination with one or more anti-PD-L2 antibodies (e.g., anti-PD-L2 targeting antibodies). Examples of anti-PD-L2 antibodies include, but are not limited to, the anti-PD-L2 antibodies described in WO 2010 / 036959, WO 20140 / 22758, and other antibodies in development that can be used in combination with the anti-IL18-BP antibodies of the invention.

[0333] In some embodiments, an anti-IL18-BP antibody is used in combination with one or more anti-CTLA-4 antibodies (e.g., anti-CTLA-4 targeting antibodies), such as, but not limited to, the FDA-approved antibodies ipilimumab and tremelimumab. In some embodiments, the anti-CTLA-4 antibody may be, for example, Yervoy® (ipilimumab or antibody 10D1, described in PCT Application WO 01 / 14424), tremelimumab (formerly ticilimumab, CP-675206), a monoclonal antibody, or any of the antibodies described in the following publications: WO 98 / 42752; WO 00 / 37504; U.S. Pat. No. 6,207,156; Hurwitz et al. (1998) Pro. Natl. Acad. Sci. USA 95(17):10067-10071; Camacho et al. (2004) J. Clin. Oncology 22(145):antibodiestract No. 2505 (antibody CP-675206); and Mokyr et al. (1998) Cancer Any of the anti-CTLA-4 antibodies disclosed in WO 2013 / 173223 can be used, including, but not limited to, the anti-CTLA-4 antibodies described in Res. 58:5301-5304, as well as other antibodies in development that can be used in combination with the anti-IL18-BP antibodies of the invention.

[0334] In some embodiments, an anti-IL18-BP antibody is used in combination with one or more anti-B7H3 antibodies (e.g., anti-B7H3 targeting antibodies). Examples of anti-B7H3 antibodies include antibodies in clinical trials, such as enoblituzumab (MGA271; MacroGenics), and the anti-B7H3 antibodies described in WO 2016 / 033225, the anti-B7H3 antibodies outlined in U.S. Pat. No. 9,441,049, and other antibodies in development that can be used in combination with the anti-IL18BP antibodies of the invention.

[0335] In some embodiments, an anti-IL18-BP antibody is used in combination with one or more anti-B7H4 antibodies (e.g., anti-B7H4 targeting antibodies). Examples of anti-B7H4 antibodies include, but are not limited to, the anti-B7H4 monoclonal antibody from FivePrime, FPA150, which is currently in Phase I clinical trials, the antibodies described in WO 2022 / 002012, and other antibodies in development that can be used in combination with the anti-IL18BP antibodies of the invention.

[0336] In some embodiments, an anti-IL18-BP antibody is used in combination with one or more anti-carcinoembryonic antigen-related cell adhesion molecule-1 antibodies (also known as anti-CEACAM1 antibodies or anti-CD66a antibodies). Examples of anti-CEACAM-1 antibodies include, but are not limited to, antibodies in clinical trials, such as besilesomab (TheraPharm), AMG211 (Amgen), and CM-24 (MK-6018, KitovPharma). Examples of anti-CEACAM-1 antibodies also include the antibodies outlined in U.S. Patent Application Publication No. 20200277398 (A1) (CM-24, under development by Famewave Ltd), the antibodies outlined in U.S. Patent No. 9,072,797 (B2) (CD66-binding moiety and radionuclide yttrium-90 (90Y)), and other antibodies under development that can be used in combination with the anti-IL18-BP antibodies of the present invention.

[0337] In some embodiments, an anti-IL18-BP antibody is used in combination with one or more anti-PVR antibodies (e.g., anti-PVR targeting antibodies). Examples of anti-PVR antibodies include, but are not limited to, those described in WO 2017 / 149538, and examples of anti-PVR antibodies include those described in WO 2021 / 070181. In some embodiments, the second agent is an antagonist of PVRL1, PVRL2, PVRL3, PVRL4, and CD155, such as ASG-22CE (Astellas), which can be used in combination with the anti-IL18-BP antibodies of the invention. Pharm / a Inc), enfortumab (Astellas Pharma), as well as others in development.

[0338] In some embodiments, an anti-IL18-BP antibody is used in combination with one or more anti-LAG3 antibodies (e.g., anti-LAG3 targeting antibodies). Examples of anti-LAG3 antibodies include, for example, antibodies in clinical trials, such as LAG525 (Novartis), TSR-033 (Tesaro), fianlimab (REGN3767, Regeneron), BI-754111 (Boehringer Ingelheim), and BI-754111 (Boehringer Ingelheim). Ingelheim), Sym-022 (Symphogen), RO7247669 (Roch), BMS-986016 (see WO 2010 / 019570), GSK2831781 (see U.S. Patent Application Publication No. 2016 / 0017037), and Merck clones 22D2, 11C9, 4A10, and / or 19E8 (see WO 2016 / 028672), as well as antibodies comprising the CDRs or variable regions of antibodies 25F7, 26H10, 25E3, 8B7, 11F2, or 17E5 described in U.S. Patent Application Publication Nos. 2011 / 0150892, WO 2010 / 19570, and 2014 / 008218. Other art-recognized anti-LAG-3 antibodies that can be used include IMP731 and IMP-321, which are described in U.S. Patent Application Publication Nos. 2011 / 007023, WO 2008 / 132601, and WO 2009 / 44273. Anti-LAG-3 antibodies that compete with and / or bind to the same epitope as any of these antibodies can also be used in combination therapy. Additionally, other antibodies in development can be used in combination with the anti-IL18-BP antibodies of the invention.

[0339] In some embodiments, an anti-IL18-BP antibody is used in combination with one or more anti-CD112 (also referred to as PVRL2, including, for example, anti-CD112 targeting antibodies) antibodies. Examples of anti-CD112 antibodies include, but are not limited to, those outlined in, for example, U.S. Patent Application Publication No. 2020 / 0040081, U.S. Patent Application Publication No. 2019 / 0040154, or WO 2017 / 021526, as well as other antibodies under development that can be used in combination with the anti-IL18-BP antibodies of the invention.

[0340] In some embodiments, an anti-IL18-BP antibody is used in combination with one or more anti-CD96 antibodies (e.g., anti-CD96 targeting antibodies). Examples of anti-CD96 antibodies include, but are not limited to, those outlined in WO 2019 / 091449, WO 2021042019, and other antibodies under development that can be used in combination with the anti-IL18-BP antibodies of the invention.

[0341] In some embodiments, an anti-IL18-BP antibody is used in combination with one or more anti-TIM3 antibodies (e.g., anti-TIM3 targeting antibodies). Examples of anti-TIM3 antibodies include antibodies in clinical trials, such as sabatolimab (Novartis), TSR-022 (Tesaro), INCAGN02385 (Incyte Corporation), INCAGN02390 (Incyte Corporation), BGB-A425 (BeiGene), LY3321367 (Eli Lilly), and BMS986258, as well as other antibodies in development that can be used in combination with the anti-IL18-BP antibodies of the invention.

[0342] In some embodiments, an anti-IL18-BP antibody is used in combination with one or more anti-BTLA antibodies (e.g., anti-BTLA targeting antibodies). Examples of anti-BTLA antibodies include, but are not limited to, JS004 (Shanghai Junshi Bioscience), the anti-BTLA antibodies disclosed in WO 2011 / 014438, as well as other antibodies under development that can be used in combination with the anti-IL18-BP antibodies of the invention.

[0343] In some embodiments, an anti-IL18-BP antibody is used in combination with one or more anti-ICOS antibodies (e.g., anti-ICOS targeting antibodies). Examples of anti-ICOS antibodies include, but are not limited to, anti-ICOS antibodies in clinical trials, such as MEDI-570 (MedImmune), vopratelimab (Jounce Therapeutics), KY1044 (Kymab Limited), and ferazilimab (GlaxoSmithKline). Examples of anti-ICOS antibodies also include the anti-ICOS antibodies outlined in U.S. Pat. No. 9,957,323, the anti-ICOS antibodies outlined in WO 2016 / 120789, and the anti-ICOS antibodies outlined in WO 2016 / 154177, as well as other antibodies under development that can be used in combination with the anti-IL18-BP antibodies of the present invention.

[0344] In some embodiments, anti-IL18-BP antibodies are used in combination with one or more anti-OX40 antibodies (e.g., anti-OX40 targeting antibodies). Examples of anti-OX40 antibodies include, but are not limited to, anti-OX40 antibodies in clinical trials, such as PF-04518600 (Pfizer), BAT6026 (Bio-Thera Solutions), MEDI6469, MEDI-0562, MEDI6962 (MedImmune), BMS986178, GSK3174998, ABBV-368 (AbbVie), and ATOR-1015 (Alligator Bioscience). Examples of anti-OX40 antibodies also include the anti-OX40 antibodies outlined in U.S. Patent No. 10,730,951, the anti-OX40 antibodies outlined in U.S. Patent No. 10,851,173, as well as other antibodies under development that can be used in combination with the anti-IL18-BP antibodies of the present invention.

[0345] In some embodiments, an anti-IL18-BP antibody is used in combination with one or more anti-41BB antibodies (e.g., anti-41BB targeting antibodies). Examples of anti-41BB antibodies include, but are not limited to, utomilumab (Pfizer, PF-05082566), LVGN6051 (Lyvgen Biopharma), ATOR-1017 (Alligator Bioscience), BMS-663513, the anti-41BB antibodies reviewed in U.S. Patent No. 10,501,551, as well as other antibodies under development that can be used in combination with the anti-IL18-BP antibodies of the present invention.

[0346] In some embodiments, the anti-IL18-BP antibody is used in combination with one or more anti-CD27 antibodies (e.g., anti-CD27 targeting antibodies). Examples of anti-CD27 antibodies include, but are not limited to, varlilumab (CDX-1127, Leap Therapeutics), the anti-CD27 antibodies outlined in U.S. Patent Application Publication No. 2020 / 0277393, the anti-CD27 antibodies outlined in WO 2019 / 195452, as well as other antibodies under development that can be used in combination with the anti-IL18-BP antibodies of the present invention.

[0347] In some embodiments, an anti-IL18-BP antibody is used in combination with one or more anti-GITR antibodies (e.g., anti-GITR targeting antibodies). Examples of anti-GITR antibodies in clinical trials include, but are not limited to, MK-4166, MK-1248 (Merck Sharp & Dohme), BMS-986156, INCAGN01876 (Incyte Corporation), OMP-336B11 (OncoMed Pharmaceuticals), and MEDI1873 (MedImmune). Examples of anti-GITR antibodies also include, but are not limited to, the anti-GITR antibodies described in WO 2016 / 196792 and WO 2015 / 187835 (the contents of which are incorporated herein by reference), such as antibodies having the heavy and light chain variable region CDRs, heavy and light chain variable regions, or heavy and light chains of antibodies 28F3, 19D3, 18E10, 3C3-1, 3C3-2, 2G6, 9G7-1, 9G7-2, 14E3, 19H8-1, 19H8-2, and / or 6G10, and variants thereof. The sequences of the antibodies described in WO 2015 / 187835 are provided in Table 2 (see SEQ ID NOS: 5-14 and 27-228). Patients may also be treated with any other anti-GITR antibody, such as TRX518 (Leap Therapeutics), MK4166 (Merck), LKZ-145 (Novartis), GWN-323 (Novartis Pharmaceuticals Corp.), Medi 1873 (MedImmune), INBRX-110 (Inhibrx), GITR-Fc protein (OncoMed), and antibodies described in WO 2006 / 105021, WO 2009 / 009116, WO 2011 / 028683, U.S. Patent Application Publication Nos. 2014 / 0072565, 2014 / 0072566, 2014 / 0065152, WO 2015 / 031667, WO 2015 / 184099, WO 2015 / 184099, or WO 2016 / 054638.

[0348] In some embodiments, an anti-IL18-BP antibody is used in combination with one or more anti-TIGIT antibodies, such as CPA.9.083.H4(S241P), CPA.9.086.H4(S241P), CHA.9.547.7.H4(S241P), CHA.9.547.13.H4(S241P), CPA.9.018, CPA.9.027, CPA.9.049, and CPA.9.057, as described in WO 2018 / 220446. , CPA.9.059, CPA.9.083, CPA.9.086, CPA.9.089, CPA.9.093, CPA.9.101, CPA.9.103, CHA.9.536.3 .1, CHA.9.536.3, CHA.9.536.4, CHA.9.536.5, CHA.9.536.7, CHA.9.536.8, CHA.9.560.1, CHA.9.56 0.3, CHA.9.560.4, CHA.9.560.5, CHA.9.560.6, CHA.9.560.7, CHA.9.560.8, CHA.9.546.1, CHA.9. 547.1, CHA.9.547.2, CHA.9.547.3, CHA.9.547.4, CHA.9.547.6, CHA.9.547.7, CHA.9.547.8, CHA.9 CHA.9.547.9, CHA.9.547.13, CHA.9.541.1, CHA.9.541.3, CHA.9.541.4, CHA.9.541.5, CHA.9.541.6, CHA.9.541.7 and CHA.9.541.8, CHA.9.547.18, and other antibodies in clinical trials, e.g., EOS-448 (GlaxoSmithKline, iTeos) Therapeutics), BMS-986207, domvanalimab (AB154, Arcus Biosciences, Inc.), AB308 (Arcus Bioscience), osipelimab (aBGB-A1217, BeiGene), tiragolumab (MTIG7192A, Roche), BAT6021 (Bio-Thera Solutions), BAT6005 (Bio-Thera Solutions), IBI939 (Innovent Biologics, U.S. Patent Application Publication No. 2021 / 00040201), JS006 (Junshi Bioscience / COHERUS),Anti-TIGIT antibodies include, but are not limited to, ASP8374 (Astellas Pharma Inc), vibostolimab (MK-7684, Merck Sharp & Dohme), M6332 (Merck KGAA), etigilimab (OMP-313M32, Mereo BioPharma), SEA-TGT (Seagen)y, HB0030 (Huabo Biopharma), AK127 (AKESO), or Genentech antibody (MTIG7192A). In some embodiments, the anti-TIGIT antibody is as described in U.S. Pat. No. 9,713,364 (including MAB1, MAB2, MAB3, MAB4, MAB5, MAB6, MAB7, MAB8, MAB9, MAB10, MAB11, MAB12, MAB13, MAB14, MAB15, MAB16, MAB17, MAB18, MAB19, MAB20, and / or MAB21), and the anti-TIGIT antibody is as described in U.S. Pat. The anti-TIGIT antibody is as described in WO 2016 / 191643, the anti-TIGIT antibody is as described in WO 2017 / 053748, the anti-TIGIT antibody is as described in WO 2016 / 191643, the anti-TIGIT antibody is as described in WO 2016 / 028656, the anti-TIGIT antibody is as described in WO 2016 / 028656, the anti-TIGIT antibody is as described in WO 2016 / 191643 ...191643, the anti-TIGIT antibody is as described in WO 2016 / 028656, the anti-TIGIT antibody is as described in WO 2016 / 191643, the anti-TIGIT antibody is as described in WO 2016 / 028656, the anti-TIGIT antibody is as described in WO 2016 / 191643, the anti-TIGIT antibody is as described in WO 201 The antibody is as described in U.S. Patent Application Publication No. 2017 / 030823, the anti-TIGIT antibody is as described in U.S. Patent Application Publication No. 2016 / 0176963, the anti-TIGIT antibody is as described in WO 2017 / 037707, the anti-TIGIT antibody is as described in WO 2017 / 059095, the anti-TIGIT antibody is as described in U.S. Patent Application Publication No. 2017281764 The anti-TIGIT antibody is as described in WO 2015 / 009856; the anti-TIGIT antibody is any of the antibodies described in U.S. Patent Application Publication No. 2017 / 0037133; the anti-TIGIT antibody is any of the antibodies described in WO 2017 / 048824 (including 10A7, 1F4, 14A6, 28H5, 31C6, 15A6, 22G2, 11G11, and / or 10D7); the anti-TIGIT antibody isand one of the antibodies described in the same application Ser. No. 2016 / 028656. In some embodiments, the anti-TIGIT antibody, usually a full-length or scFv domain, comprises the following CHA set of CDRs, the sequences of which are shown in Figure 30A: CPA.9.083.H4(S241P)vhCDR1, CPA.9.083.H4(S241P)vhCDR2, CPA.9.083.H4(S241P)vhCDR3, CPA.9.083.H4(S241P)vlCDR1, CPA.9.083.H4(S241P)vlCDR2, and CPA.9.083.H4(S241P)vlCDR3. In some embodiments, anti-TIGIT antibodies, typically full-length or scFv domain, comprise the following CHA set of CDRs, the sequences of which are shown in Figure 30B: CPA.9.086.H4(S241P)vhCDR1, CPA.9.086.H4(S241P)vhCDR2, CPA.9.086.H4(S241P)vhCDR3, CPA.9.086.H4(S241P)vlCDR1, CPA.9.086.H4(S241P)vlCDR2, and CPA.9.086.H4(S241P)vlCDR3. Such anti-TIGIT antibodies can be used in combination with anti-IL18-BP antibodies of the invention. Additional exemplary anti-TIGIT antibody sequences are shown in Figure 34.

[0349] In some embodiments, an anti-IL18-BP antibody is used in combination with one or more anti-PVRIG antibodies, such as CHA.7.518.1.H4(S241P), CHA.7.538.1.2.H4(S241P), and CHA.7.502, CHA.7.503, CHA.7.506, CHA.7.508, CHA.7.510, CHA.7.512, CHA.7.514, CHA.7.516, CHA.7.518.1.H4(S241P), CHA. 7.518, CHA.7.518.4, CHA.7.520.1, CHA.7.520.2, CHA.7.522, CHA.7.524, CHA.7.526, CHA.7.527, CHA.7.528, CHA.7.5 30, CHA.7.534, CHA.7.535, CHA.7.537, CHA.7.538.1.2.H4(S241P), CHA.7.538.1, CHA.7.538.2, CHA.7.543, CHA.7.54 4, CHA.7.545, CHA.7.546, CHA.7.547, CHA.7.548, CHA.7.549, CHA.7.550, CPA.7.001, CPA.7.003, CPA.7.004, CPA.7.0 06, CPA.7.008, CPA.7.009, CPA.7.010, CPA.7.011, CPA.7.012, CPA.7.013, CPA.7.014, CPA.7.015, CPA.7.017, CPA.7. In some embodiments, the antibody sequences are derived from WO 201 / 6134333, including, but not limited to, CPA.018, CPA.7.019, CPA.7.021, CPA.7.022, CPA.7.023, CPA.7.024, CPA.7.033, CPA.7.034, CPA.7.036, CPA.7.040, CPA.7.046, CPA.7.047, CPA.7.049, and CPA.7.050, as well as other antibodies in clinical trials, such as GSK4381562 / SRF816 (GSK / Surface), NTX2R13 (Nectin Therapeutics).In some embodiments, the anti-PVRIG antibody is typically a full-length or scFv domain and comprises the following CHA set of CDRs, the sequences of which are shown in Figure 29A: CHA.7.518.1.H4(S241P)vhCDR1, CHA.7.518.1.H4(S241P)vhCDR2, CHA.7.518.1.H4(S241P)vhCDR3, CHA.7.518.1.H4(S241P)vlCDR1, CHA.7.518.1.H4(S241P)vlCDR2, and CHA.7.518.1.H4(S241P)vlCDR3. In some embodiments, the anti-PVRIG antibody, typically a full-length or scFv domain, comprises the following CHA set of CDRs, the sequences of which are shown in Figure 30B: CHA.7.538.1.2.H4(S241P)vhCDR1, CHA.7.538.1.2.H4(S241P)vhCDR2, CHA.7.538.1.2.H4(S241P)vhCDR3, CHA.7.538.1.2.H4(S241P)vlCDR1, CHA.7.538.1.2.H4(S241P)vlCDR2, and CHA.7.538.1.2.H4(S241P)vlCDR3. Such anti-PVRIG antibodies can be used in combination with the anti-IL18-BP antibodies of the invention. Further exemplary anti-PVRIG antibody sequences are shown in Figures 36, 37, and 38.

[0350] b. Other cancer combination therapy The anti-IL-18BP antibodies of the present invention may be administered in combination with one or more other prophylactic or therapeutic agents, including, but not limited to, cytotoxic agents, chemotherapeutic agents, cytokines, growth inhibitors, antihormones, kinase inhibitors, anti-angiogenic agents, cardioprotective agents, immunostimulatory agents, immunosuppressants, agents that promote the proliferation of blood cells, angiogenesis inhibitors, protein tyrosine kinase (PTK) inhibitors, or other therapeutic agents.

[0351] In this context, a "chemotherapeutic agent" is a chemical compound useful in the treatment of cancer. Examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclosphosphamide, alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carboquone, metredopa, and uredopa; ethylenimines and methylameramines, including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolmelamine; acetogenins, especially Bullatacin and bullatacinone; delta-9-tetrahydrocannabinol (dronabinol, MARINOL'); beta-lapachone; lapachol; colchicine; betulinic acid; camptothecins (including synthetic analogs topotecan (HYCAMTN®), CPT-11 (irinotecan, CAMPTOSAR®), acetylcamptothecin, scopolectin, and 9-aminocamptothecin); bryostatin; kallistatin; CC-1065 (adozelesin, calstatin) Zelesin, and bizelesin synthetic analogs; podophyllotoxin; podophyllic acid; teniposide; cryptophycins (especially cryptophycin 1 and cryptophycin 8); dolastatins; duocarmycins (including synthetic analogs, KW-2189 and CB1-TM1); eleutherobin; pancratistatin; sarcodictyin; spongistatin; nitrogen mustards, e.g., chlorambucil, chlornaphazine, chlorophosphamide, estramusti benzodiazepine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, nobembine, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosoureas, e.g., carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimustine; antibiotics, e.g., enediyne antibiotics (e.g., calicheamicin, particularly calicheamicin γ1I and calicheamicin ω1I (see, e.g., Agnew, Chem Intl. Ed. Engl., 33:183-186 (1994)); dynemicin, including dynemicin A; esperamicin;and neocarzinostatin chromophore and related chromoprotein enediyne antibiotic chromophores), aclacinomycin, actinomycin, autramycin, azaserine, bleomycin, cactinomycin, carabicin, carminomycin, carzinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin (including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin, and deoxydoxorubicin), epinephrine, erythromycin ... Mitomycins such as rubicin, esorubicin, idarubicin, marcelomycin, mitomycin C, mycophenolic acid, nogalamycin, olivomycin, peplomycin, porfiromycin, puromycin, chelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs Antibodies, such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogues, such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens, such as calsterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone; antiadrenergics, such as aminoglutethimide, mitotane, trilostane; folic acid supplements, such as furoic acid, aceglatone; aldophosphamide Doglycosides; Aminolevulinic acid; Eniluracil; Amsacrine; Bestravcil; Bisantrene; Edatraxate; Defofamine; Demecolcine; Diazicon; Elforornithine; Elliptinium acetate; Epothilone; Etoglucide; Gallium nitrate; Hydroxyurea; Lentinan; Lonidynin; Maytansinoids, such as maytansine and ansamitocin; Mitoguazone; Mitoxantrone; Mopidanmol; Nitraerin; Pentostatin; Fenamet; Pirarubicin; Rosoxantrone; 2-Ethylhydrazide; Procarbazine;PSK® polysaccharide complex (JHS Natural Products, Eugene, Oreg.); razoxane; rhizoxin; schizofiran; spirogermanium; tenuazonic acid; triazicone; 2,2',2''-trichlorotriethylamine; trichothecenes (especially T-2 toxin, veracrine A, roridin A, and anguidine); urethane; vindesine (ELDISINE®, FILDESIN®); dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); thiotepa; taxoids, such as paclitaxel (TAXOL®; Bristol-Myers Squibb Oncology, Princeton, NJ), ABRAXANE®, a cremophor-free albumin-modified nanoparticle formulation of paclitaxel (American Pharmaceutical Partners, Schaumberg, Ill.), and docetaxel (TAXOTERE®; Rhone-Poulenc Rorer, Antony, France); chlorambucil; gemcitabine (GEMZARM®); 6-thioguanine; mercaptopurine; methotrexate; platinum analogs, such as cisplatin and carboplatin; vinblastine (VELBAN®); platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine (ONCOVIN®); oxaliplatin; leucovorin; vinorelbine (NAVELBINE®); novantrone; edatrexate; daunomycin; aminopterin; ibandronate; the topoisomerase inhibitor RFS2 000; difluoromethylornithine (DMFO); retinoids, such as retinoic acid; capecitabine (XELODA®); pharmaceutically acceptable salts, acids, or derivatives of any of the above; and combinations of two or more of the above, such as CHOP (an abbreviation for combination therapy of cyclophosphamide, doxorubicin, vincristine, and prednisolone); CVP (an abbreviation for combination therapy of cyclophosphamide, vincristine, and prednisolone); and FOLFOX (an abbreviation for a treatment regimen using oxaliplatin (ELOXATIN®) in combination with 5-FU and leucovorin).

[0352] In some embodiments, the chemotherapeutic agent is selected from the group consisting of platinum, oxaliplatin, cisplatin, paclitaxel (taxol), sorafenib, doxorubicin, sorafenib, 5-FU, and gemcitabine, irinotecan (CPT-11).

[0353] In some embodiments, the other therapeutic agent is an agent used in radiation therapy for the treatment of cancer. Thus, in some embodiments, the active agents described herein are administered in combination with one or more of a platinum coordination compound, a topoisomerase inhibitor, an antibiotic, an antimitotic alkaloid, and a difluoronucleoside.

[0354] In some embodiments, the anti-IL18BP antibody is combined with one or more inflammasome activators. In some embodiments, the inflammasome activator is a CD39 inhibitor. In some embodiments, the CD39 inhibitor is an anti-CD39 antibody.

[0355] According to at least some embodiments, anti-IL18BP antibodies can be used in combination with any standard of care cancer treatment known in the art (e.g., as can be found on the world wide web at cancer.gov / cancertopics). [Example]

[0356] Example 1: Expression of IL18 and IL18BP in the tumor microenvironment IL18-BP is a blocker for IL18, resulting in the inhibition of IL18 activity (Dinarello, et al., Front. Immunol., 1:1-10 (2013)). Therefore, for IL18-BP blocking to be effective, both the antibody targets, IL18-BP and IL18, must be present in the tumor microenvironment (TME). Figure 4 shows the expression of both IL18 (Figure 4A) and IL18-BP (Figure 4B), demonstrating that both proteins are expressed across all TCGA tumors, with only pheochromocytomas and paragangliomas (see Table 1 for TCGA tumor type abbreviations; showing somewhat limited expression for IL18 in a subset of these tumor types (reference line at 1 RPKM indicates background expression level below). The ultimate target cells of free IL18 are leukocytes / lymphocytes (Tominaga, K., et al., International Immunology, 12(2):151-160 (2000) and Senju, H., et al., Int J Biol Sci., 14(3):331-340 (2018)), these experiments target tumors with a higher immune presence, as evidenced by an IFNγ inflammatory signature (see, e.g., U.S. Patent Application Publication No. 2016 / 0312295 (A1)). As seen in Figure 5, both IL18 (Figure 5A) and IL18-BP (Figure 5B) were predominant across all tumor subsets with inflammation (high IFNγ signature values). Significant expression of both proteins was detected even in the lowest IFNγ subset. Because IL18 depends on the inflammasome for its secretion, a signature of core inflammasome genes was generated. These genes are common to multiple types of inflammasome-activating signals (Chauhan, D., et al., Immunological Reviews, 297:123-138 (2020)). Signature values ​​are calculated as the log10 of the genes listed in Table 2. Calculated as the mean of RPKM expression values.As shown in Figure 6A, the core inflammasome signature was highly expressed in all TMEs, generally with higher expression in the inflammatory (high IFNγ signature) subset. This pattern is very similar to that presented for IL18 (Figure 5A), indicating that the IL18 mechanism was present in all inflammatory (high IFNγ signature) tumors and most low IFNγ signature tumors, except for LGG and PCPG. To verify that IL18 and the inflammasome core signature genes were indeed expressed in the same cells in the TME, we used single-cell data to calculate cosine similarities between IL18, IL18-BP, and core inflammasome genes, as well as additional upstream genes and IL18 receptors. Figure 6B presents the cosine similarity matrix for macrophages in NSCLC, demonstrating that IL18 and the core inflammasome signature were indeed present in the same cells. Similar results were obtained in additional tumor types (data not shown). Specifically, triple-negative breast cancers have been reported to have more inflammatory features in approximately 30% of cases compared with approximately 5-10% in hormone receptor-positive breast cancers (Thomas, F., et al., Frontiers in Oncology, 10:1-17 (2021)). Therefore, we specifically examined the expression of IL18 and IL18-BP in single-cell breast cancers (raw data adopted from Bassez, A., et al., Nature Medicine, 27:820-832 (2021)). Figure 7A shows that both IL18 and IL18-BP are more abundant in TNBC, both by the percentage of expressing cells and the average level of expression, and are shed by the HER2+ subset, with minimal expression in the hormone receptor-positive subset, particularly in pretreatment samples. In this particular dataset, patients were sampled pre-treatment and then administered aPD1 neoadjuvant treatment or aPD1 with chemotherapy, followed by tumor resection (at the time of treatment biopsy). The authors measured T cell clonal expansion after treatment.T cell clonal expansion could be considered a surrogate measure of response to aPD1 treatment (Bassez et al., Nature Medicine, 27:820-832 (2021)). Figure 7B shows that baseline levels of IL18 were lower in non-expanding patients, but IL18-BP was higher in these patients. This may indicate a potential role for IL18-BP in attenuating IL18 activity and preventing the activity of immune checkpoint blocking (ICB) therapy. Both genes were upregulated after aPD1 treatment. These observations reinforce the selection of more inflammatory indications, both in general and specifically in TNBC.

[0357] Materials and Methods Preprocessing, filtering, and normalization UMIs were quantified using Cellranger 3.0.2 (10x Genomics) with the reference transcriptome GRCh38. Subsequent analyses were performed using "Seurat" (https: / / satijalab.org / seurat / ) unless otherwise noted.

[0358] Clustering and cell type annotation The top 15 principal components were used to construct the SNN graph and UMAP embedding. Cell annotation in Thomas et al. 2021 was based on metadata submitted by the authors.

[0359] Cells are clustered by nested PCA, and clusters are annotated by their gene signature expression.

[0360] The cosine similarity matrix for the desired gene set is calculated by calculating the value of equation (i) for each pair of genes and their expression vectors.

number

[0361] The IFNγ inflammation signature (as described in US Patent Application Publication No. 2016 / 0312295(A1), which is incorporated by reference in its entirety) is calculated in three steps. 1. The IFNγ_up signature is calculated as the average of the log10 RPKM expression values ​​of the following genes: CCR5, HLA-DRA, CXCL13, CCL5, STAT1, KLRK1, NKG7, CXCL9, LAIR1, LAG3, CXCR6, KLRD1, GZMA, PRF1, SIGLEC14, PTPN22, CD86, SLA, SIRPG, CD72, HAVCR2, PSTPIP2, SLAMF6, CD84, CD300LF , CD3D, IFNG, CXCL11, CD2, CTSZ, GZMB, IL2RG, CXCL10, LILRB4, PDCD1, CCL8, CIITA, CCL4, IGSF6, PTPRC, CLEC9A, CST7, MYLIP, ITGAL, CDH1, PSTPIP1, GZMK, HLA-E, CD3E, TAGAP, TNFRSF9 2. The IFNγ_down signature is calculated as the mean of the log10 RPKM expression values ​​of the following genes: CLEC3B, NR4A2, EEF1G, PIK3CA, TYRO3, CX3CL1, ING1, BST1, ACKR3, UBB, PPARG, PTEN, THY1, CLCA1, EFEMP1, GAS6, ITM2A, CD55, NFATC1, BCL6, RETNLB, PDCD4, TIMP3, CDO1, POL R1B, DDR1, F2R, CTSG, LILRA5, CX3CR1, TBP, CLEC1B, RGS16, PTPN13, IRF1, MON1B, CPD, PHACTR2, OAZ1, CASP3, IFI16, ITGA1, RPL19, CCR6, LTK, C10orf54, SLAMF1, and TNFAIP8L2 3. Finally, the IFNγ inflammatory signature is calculated as the difference between the two signatures: IFNγ = IFNγ_up - IFNγ_down.

[0362] Clustering and cell type annotation The top 15 principal components were used to construct the SNN graph and UMAP embedding. [Table 1] [Table 2]

[0363] Example 2: IL-18BP is a soluble immune checkpoint - RNA expression data Upregulation of IL-18BP in TME-TCGA versus GTEX Figure 47: Expression of IL18BP transcripts in normal (green) or cancer (red) tissues from the TCGA and GTEX databases. GBM, glioblastoma multiforme; HSNC, head and neck squamous cell carcinoma; KIRC, renal clear cell carcinoma; PAAD, pancreatic adenocarcinoma; SKCM, cutaneous melanoma; STAD, gastric adenocarcinoma ( * P<0.01).

[0364] IL-18BP is expressed in suppressive myeloid populations and correlates with PD-L1 in the TME, suggesting a mechanism of resistance As shown in Figure 59A, IL-18BP correlates with PD-L1 at the RNA level (TCGA) in colon and breast cancer, suggesting a mechanism of resistance to immune activation in the tumor microenvironment (TME).

[0365] As shown in Figure 59B and Figure 48, single-cell RNA analysis of tumor-infiltrating myeloid cells, including tumor-associated macrophages (TAMs) and dendritic cells (DCs), in colon cancer patients showed that IL-18BP was upregulated in myeloid populations in the TME compared to the periphery (PBMCs), suggesting a mechanism of resistance to immune activation in the TME.

[0366] As shown in Figure 59C, single-cell RNA analysis of tumor-infiltrating myeloid cells, including tumor-associated macrophages (TAMs) and dendritic cells (DCs) across indications, showed that IL-18BP was upregulated in myeloid populations in the TME compared to the periphery (PBMCs), suggesting a mechanism of resistance to immune activation in the TME.

[0367] Upregulation of IL-18BP in response to ICB treatment - scRNA / bulk RNA data Figures 60A-60C: IL-18BP is upregulated (RNA level) after immune checkpoint blocking (ICB) treatment. IL-18BP levels are upregulated in the tumor microenvironment (RNA) after treatment with anti-PD-1 (breast cancer and basal cell carcinoma) or anti-PD-1 + anti-CTLA-4 (melanoma), suggesting a potential resistance mechanism.

[0368] Figure 60D: IL-18BP is elevated in NSCLC patient serum after aPD-(L)1 treatment. Quantification of plasma IL-18BP protein levels by ELISA for healthy donors (n=22) and NSCLC patients (n=52) at baseline before treatment and at subsequent CT scans after receiving treatment with anti-PD-(L)1 (n=52). Association of IL-18BP levels with inadequate response to aPD-(L)1 blockade: 1) IL18BP in RCC (pembrolizumab + lenvatinib combination) and 2) melanoma responders / NR (Olink)

[0369] Supportive data for the role of IL-18BP as a soluble ICP and a potential mechanism of resistance to PD1 blockade in patients with renal cell carcinoma receiving pembrolizumab + lenvatinib. As seen in Figure 61A, high IL-18BP in the serum of patients pretreated with pembrolizumab + lenvatinib is associated with shorter progression-free survival (PFS). As seen in Figure 61B, high IL-18BP in the serum of patients pretreated with pembrolizumab + lenvatinib is associated with stable or progressive disease (SD / PD).

[0370] Supporting data for the role of IL-18BP as a soluble ICP in melanoma cancer patients undergoing anti-PD-1 therapy and a potential mechanism of resistance to PD1 blocking. As seen in Figure 62, high IL-18BP in the serum of melanoma cancer patients pretreated with anti-PD-1 is associated with poor response. Raw Olink data (NPX format). Student's T-test was performed for IL18BP protein after intensity normalization for target product.

[0371] Example 3: Inflammasome-inducing cytokines such as IL-18 and IL-1β are abundant in the TME. Unlike other cytokines, inflammasome-inducing cytokines such as IL-18 and IL-1β are abundant in the TME.

[0372] method: Tumors were cut into small pieces with a scalpel and transferred to GentleMACs™ C tubes (Miltenyi Biotec) containing enzyme mix using a human tumor dissociation kit (Miltenyi Biotec) according to the manufacturer's protocol. After dissociation, samples were centrifuged at 300 g for 5 minutes, the supernatant collected, and recentrifuged at 3130 g for 10 minutes. After centrifugation, the supernatant was recollected and aliquoted for storage at -80°C. On the day of the assay, samples were thawed at room temperature, followed by centrifugation at 14,000 RPM for 10 minutes, and the supernatant collected for immediate use in ELISA or CBA using the following kits: Human IL18 ELISA kit (MBL, 7620) Human Th1 / Th2 / Th17 Cytokine Cytometric Bead Array (CBA) (BD560484)

[0373] Human Inflammatory Cytokine Cytometric Bead Array (CBA) (BD551811)

[0374] result: Figure 71A: IL-18 and IL-1β are inflammasome-derived cytokines with opposing effects in the TME. IL-18 promotes T cell and NK cell activation, resulting in anti-tumorigenic activity, whereas IL1β has a dual role, with combined effects resulting in pro-tumorigenic activity.

[0375] Figure 71B: Dot plot graph shows cytokine levels measured in tumor-derived supernatants across various indications. Each dot represents one sample. The mean is indicated by the short black line. All other cytokines except IL-1β and IL-18 were below the lower limit of detection.

[0376] Example 4: IL18 and IL18BP protein levels in serum of patients across indications compared to healthy donors method: Serum samples from healthy donors and cancer patients were thawed and the levels of IL18 analytes (IL18 total, IL18BP) were measured by the following ELISA kits according to the manufacturer's protocol. Human IL18 ELISA kit (MBL, 7620) Human IL18BP ELISA Kit (R&D DBP180)

[0377] result: Figure 56A. IL18 analyte levels in patient serum across indications. Figure 56B. Dot plots represent IL18 analyte levels in serum samples from individual patients or healthy donors. Statistical analysis was performed using a two-tailed t-test, P<0.001. *** IL-18 expression was significantly increased in the serum of cancer patients compared with healthy donors, confirming that IL-18 levels are enhanced in the periphery during malignancy.

[0378] Example 5: High levels of IL-18 protein in the serum of head and neck cancer patients with tumor sites in the tongue method: Serum samples from head and neck cancer patients were thawed and the levels of IL18 analyte were measured by the following ELISA kits according to the manufacturer's protocols: Human IL18 ELISA kit (MBL, 7620) and Human IL18BP ELISA kit (R&D DBP180).

[0379] result: Figures 63A-63B: Principal component analysis (PCA) shows that primarily tumor location separates samples with high and low levels of IL-18. Tumor location in the tongue correlates with high levels of IL-18 and low levels of IL18BP compared to other sites. Figure 63C. Individual patient serum levels of IL-18 and IL18BP are shown in dot plots at different tumor sites.

[0380] Example 6: IL18BP and IL18 protein levels in plasma of NSCLC patients treated with anti-PD1 / anti-PD1+ chemotherapy method: Plasma samples from NSCLC patients were thawed, and the levels of IL18 and IL18BP were measured by the following ELISA kits according to the manufacturer's protocols: human IL18 ELISA kit (MBL, 7620) and human IL18BP ELISA kit (R&D DBP180).

[0381] result: Figure 65: Plasma from NSCLC patients was collected at baseline and after a single dose of anti-PD-1 (Keytruda) (n=8) or chemotherapy plus a single dose of anti-PD-1 (n=14). Clinical assessment of patient response (response / non-response, R / NR) was performed according to PET-CT scans after several treatment cycles.

[0382] The mean plasma levels of IL18BP and IL18 measured at baseline were higher in patients responding to therapy (anti-PD-1 monotherapy or anti-PD-1 plus chemotherapy combination) compared to non-responding patients (Figure 65A).

[0383] As shown in Figures 65B and 65D, patients who did not respond clinically to anti-PD-1 monotherapy exhibited higher plasma levels of IL18 and IL18BP compared to baseline levels, whereas IL18 and IL18BP levels did not change significantly from baseline in patients who responded to anti-PD-1 monotherapy. In contrast, only patients who responded clinically to the anti-PD-1 + chemotherapy combination exhibited higher levels of IL18 and IL18BP compared to baseline (Figures 65C and 65D).

[0384] Consideration: NSCLC patients treated with anti-PD-1 (Keytruda) were most likely to develop a PDL1-CPS of greater than 50%, potentially indicating an increase in immune infiltrate and subsequent increase in IFNγ expression in the TME. Considering that IL18BP is an IFNγ-inducible gene, this may suggest a potential immune resistance mechanism in anti-PD-1-treated patients and provide a rationale for combining anti-IL18BP and anti-PD-1 blockade to further increase patients' potential anti-tumor responses. Patients receiving chemotherapy plus anti-PD-1 combination therapy tend to have a PDL1-CPS of less than 50% and larger tumor masses. Patients who clinically respond to the anti-PD-1 plus chemotherapy combination may have a potential increase in the infiltration of immune cells capable of secreting IL18 and a subsequent induction of IFNγ levels, potentially leading to increased IL18BP secretion. Clinical anti-tumor responses in these patients could be enhanced with anti-IL18BP antibodies.

[0385] Example 7: IL18 and IL18BP protein levels in tumor-derived supernatants (TDS) method: Tumors were cut into small pieces with a scalpel and transferred to GentleMACs™ C tubes (Miltenyi Biotec) containing enzyme mix using a human tumor dissociation kit (Miltenyi Biotec) according to the manufacturer's protocol. After dissociation, samples were centrifuged at 300 g for 5 minutes, the supernatant was collected, and recentrifuged at 3130 g for 10 minutes. After centrifugation, the supernatant was recollected and aliquoted for storage at -80°C. On the day of the assay, samples were thawed at room temperature, followed by centrifugation at 14,000 RPM for 10 minutes, and the supernatant was collected for immediate use in ELISA using the following kit: Human IL18 ELISA kit (MBL, 7620) Human IL18BP ELISA Kit (R&D DBP180)

[0386] result: IL-18 and IL-18BP were detected in TDS across a variety of indications.

[0387] Figure 57. Dot plots represent IL18 and IL18BP in TDS samples from individual patients. Figure 58. IL18 and IL18BP levels in TDS of patients across indications.

[0388] Example 8: IL18Rα is expressed on TILs in the TME and its expression is induced on CD4 TILs compared to the periphery method: Tumor samples were cut into small pieces with a scalpel and transferred to GentleMACs™ C tubes (Miltenyi Biotec) containing an enzyme mix. After dissociation, cells were filtered through a 70 μm filter. Single-cell suspensions were seeded into 96-well V-bottom plates, and Fc receptors were blocked using a cocktail of antibodies (Abs) against CD16 (BioLegend), CD32 (Thermo Fisher), and CD64 (BioLegend). Immune populations were stained with anti-human IL18Rα or its isotype control (BioLegend). After washing (1% BSA, 0.1% sodium azide in PBS), cells were acquired on a FACS Fortessa cytometer (BD Bioscience). Analysis was performed using FlowJo.

[0389] result: IL-18Rα expression was induced on tumor-infiltrating T cells compared to matched PBMCs, with statistical significance on CD4+ T cells and a trend towards CD8+ T cells.

[0390] Figure 55A. CD8 from dissociated human tumors of various cancer types + and CD4 + Figure 55B shows the expression of IL18Rα on CD4 TILs and NK TILs. Each dot represents a separate tumor from an individual patient. Fold expression values ​​were calculated by dividing the MFI of the target by the MFI of the relevant isotype control (FOI). Means and SEM are shown in the tick chart. Figure 55B shows CD4 TILs from donor-matched PBMCs and TME. + and CD8 + Expression of IL18Rα on T and NK cells. Statistical analysis was performed using paired t test (two-tailed), P < 0.05; ** p=0.0064

[0391] Example 9: Co-expression of TIGIT and IL18Rα within the TME Tumor samples were mechanically dissociated and enzymatically digested using a Milteny human tumor dissociation kit (according to the manufacturer's instructions). Single-cell suspensions were stained with Zombie-Nir to exclude dead cells, and with antibodies against CD45, CD3, CD4, CD8, CD56, TIGIT, or IL18Rα. Cells were acquired using a FACS Fortessa cytometer (BD Bioscience) and analyzed using FlowJo software (V10). Cell surface markers were used to detect the following immune populations: CD8 (CD3+CD8+), CD4 (CD3+CD4+), NK (CD3-CD56+), and NKT (CD3+CD56+).

[0392] The results are shown in Figure 33, which displays flow cytometry dot plots showing co-expression of IL18Rα and TIGIT on CD8 T cells, CD4 T cells, NK, and NKT cells in the endometrial and colonic TME. Co-expression of TIGIT and IL18Rα on the same cells indicates that targeting both pathways by combined administration of inhibitory anti-IL18BP and anti-TIGIT antibodies can have beneficial effects.

[0393] Example 10: Generation and characterization of custom AB against human IL18-BP protein from Adimab Ltd Generation of anti-IL18-BP hIgG1-N297A Ab against human IL18-BP protein antigen preparation Antigens were biotinylated using the EZ-Link Sulfo-NHS-Biotinylation Kit (Thermo Scientific, Cat. No. 21425).

[0394] The antigen was concentrated to approximately 1 mg / mL and buffer exchanged into PBS, followed by the addition of a biotinylation reagent at a molar ratio of 1:7.5. The mixture was kept overnight at 4°C before another buffer exchange to remove free biotin in solution. Biotinylation was confirmed by streptavidin sensor binding of the labeled protein on a ForteBio.

[0395] Naive Library Selection About 10 each 9 Eight naive human synthetic yeast libraries of diversity were grown as previously described (see, e.g., Y. Xu et al., PEDS 26(10), 663-70 (2013); WO 2009036379; WO 2010105256; and WO 2012009568).

[0396] For the first two rounds of selection, a magnetic bead sorting technique utilizing the Miltenyi MACS system was performed as previously described (see, e.g., Siegel et al., J Immunol Methods 286(1-2), 141-153 (2004)). Briefly, yeast cells (approximately 10 10 Yeast cells (1000 cells / library) were incubated with 10 nM biotinylated human IL18-BP-Fc fusion in wash buffer (phosphate-buffered saline (PBS) / 0.1% bovine serum albumin (BSA)) for 30 min at 30°C. After washing once with 40 mL of ice-cold wash buffer, the cell pellet was resuspended in 20 mL of wash buffer, and streptavidin microbeads (500 μL) were added to the yeast and incubated for 15 min at 4°C. The yeast were then pelleted, resuspended in 5 mL of wash buffer, and loaded onto a Miltenyi LS column. After loading 5 mL, the column was washed three times with 3 mL of wash buffer. The column was then removed from the magnetic field, and the yeast were eluted with 5 mL of growth medium and then grown overnight.

[0397] Subsequent rounds of selection were performed using flow cytometry (FACS). Yeast were pelleted, washed three times with wash buffer, and incubated at 30°C with 10 nM biotinylated human IL18-BP-Fc fusion, 10 nM biotinylated cyno IL18-BP-Fc fusion, 100 nM human IL18-BP-Fc monomer, 100 nM biotinylated cyno IL18-BP monomer, or polyspecificity reagent (PSR) to remove nonspecific antibodies. Some selections were also performed to enrich for IL18-competing antibodies by incubation with biotinylated human IL18-BP-Fc fusion pre-complexed to human IL18. For PSR depletion, the library was incubated with a 1:10 dilution of biotinylated PSR reagent as previously described (see, e.g., Y. Xu. Xu et al., PEDS 26(10), 663-70 (2013)). Yeast were then washed twice with wash buffer and incubated with goat (Fab')2 anti-human kappa-FITC (LC-FITC) (Southern Biotech, Cat. No. 2062-02) diluted 1:100 and streptavidin-AF633 (SA-633) (Life Technologies, Inc.) diluted 1:500. The cells were stained with either secondary reagent (Sigma-Aldrich, Cat. No. S21375) or Extravidin-Phycoerythrin (EA-PE) (Sigma-Aldrich, Cat. No. E4011) diluted 1:50 for 15 minutes at 4°C. After two washes with ice-cold wash buffer, the cell pellet was resuspended in 0.3 mL of wash buffer and transferred to a strainer-capped sort tube. Sorting was performed using a FACS ARIA sorter (BD Biosciences), and sort gates were determined to select for antibodies with the desired characteristics. Selection rounds were repeated until a population possessing all of the desired characteristics was obtained. After the final round of sorting, the yeast was plated, and individual colonies were picked for characterization.

[0398] Antibody Optimization Antibody optimization was performed via light chain batch shuffling and then by introducing diversity into the heavy and light chain variable regions as described below. Some combination of these approaches was used for each antibody.

[0399] Light chain batch shuffling: Heavy chains from the naive output were used to prepare light chain diversified libraries. Selections were performed on these libraries as described above, i.e., using one round of MACS and four rounds of FACS. In different FACS selection rounds, the libraries were evaluated for PSR binding and affinity pressure, e.g., by antigen titration. Sorting was performed to obtain populations with the desired characteristics. Individual colonies from each final FACS selection round were picked for sequencing and characterization.

[0400] CDRH1 and CDRH2 selection: CDRH3 of a single antibody was selected from approximately 10 8 The CDRH1 and CDRH2 variants were recombined into a pre-generated library with a diversity of 1000 and selected using one round of MACS and four rounds of FACS as described for naive selection. For each FACS round, the library was tested for PSR binding and affinity pressure and sorted to obtain populations with the desired characteristics.

[0401] Selection of CDRH3 and CDRL3: Oligos containing CDRH3 and CDRL3 and the flanking regions on either side of CDR3 were ordered from IDT. Each oligo altered one or two amino acids in CDR3 via NNK diversity. Approximately 10 8For combined library diversity, the CDRH3 oligo was recombined with the heavy chain FR1-FR3 variable region containing variants selected from the CDRH1 and CDRH2 selections, and the CDRL3 oligo was recombined with the light chain FR1-FR3 variable region from the parent antibody. Selections were performed using one round of MACS and four rounds of FACS, as described for the naive selection. For each FACS round, the library was examined for PSR binding and affinity pressure, and sorted to obtain populations with the desired characteristics. For these selections, affinity pressure was applied by preincubating the antigen with the parent IgG for 30 minutes, then applying the precomplexed mixture to the yeast library for a period of time allowing the selection to reach equilibrium. Antibodies with higher affinities could then be selected.

[0402] Antibody production and purification Yeast clones were grown to saturation and then induced for 48 hours at 30°C with shaking. After induction, yeast cells were pelleted and the supernatant was collected for purification. IgG was purified using a protein A column and eluted with acetic acid (pH 3.5).

[0403] Size Exclusion Chromatography A TSKgel SuperSW mAb HTP column (22855) was used for high-speed SEC analysis of mammalian-produced mAbs at 0.4 mL / min with a cycle time of 6 min / run. 200 mM sodium phosphate and 250 mM sodium chloride were used as the mobile phase.

[0404] Dynamic scanning fluorimetry Add 10 μL of 20x Sypro Orange to 20 μL of 0.2–1 mg / mL mAb or Fab solution. Using an RT-PCR instrument (BioRad CFX96 RT PCR), increase the sample plate temperature from 40°C to 95°C in 0.5°C increments, allowing 2 minutes to equilibrate at each temperature. Extract the Tm by taking the negative of the first derivative of the raw data. [Table 3]

[0405] Anti-IL18-BP hIgG1 Ab analysis included the following steps. Affinity measurement of anti-human Abs to human IL18-BP-Fc protein and cynomolgus monkey IL18-BP-Fc protein using ForteBio Octet-Naive output Octet affinity measurements were generally performed on an Octet HTX as previously described (see, e.g., Estep et al., Mabs 5(2), 270-278 (2013)). Briefly, ForteBio affinity measurements were performed by loading IgG online onto the AHC sensor. The sensor was equilibrated offline in assay buffer for 30 minutes and then monitored online for 60 seconds to establish a baseline. The IgG-loaded sensor was exposed to 100 nM antigen for 3 minutes, then transferred to assay buffer for 3 minutes for off-rate measurements. All kinetics were analyzed using a 1:1 binding model.

[0406] SPR measurement Surface plasmon resonance K D measurement Kinetic analysis was performed using a Biacore 8K optical biosensor (Global Life Sciences Solutions USA, Marlborough, MA) in an HBS-EP+ running buffer system (10 mM HEPES (pH 7.4), 150 mM NaCl, 3 mM EDTA, 0.05% surfactant P20) at 25°C. The sample compartment was maintained at 10°C during each experiment.

[0407] For antibody capture experiments, goat anti-human Fc antibodies (Jackson ImmunoResearch) were covalently coupled to flow cells 1 and 2 of a CM5 sensor chip surface via standard amine coupling (1:1 EDC:NHS) and then blocked with ethanolamine (1.0 M, pH 8.5). The antibody (10.0 nM in running buffer) was injected into flow cell 2 (40 s at 10 μL / min). IL18-BP-Fc monomers at concentrations ranging from 27.0 to 0.111 nM were injected into flow cells 1 and 2 (300 s at 30 μL / min). Dissociation of the IL18-BP-Fc monomer was monitored for 600 s or 5130 s. Several blank buffer samples were injected into flow cells 1 and 2 (300 s at 30 μL / min) and used for reference surface subtraction. All surfaces (flow cells 1 and 2) were regenerated with two injections (20 seconds at 30 μL / min) of 10 mM glycine (pH 1.5).

[0408] Biotinylated antigen capture in solution experiments—For Fab or full Ab, each experimental cycle consisted of the application of biotin capture reagent (Global Biotin) in running buffer onto flow cells 1 and 2. The experiment started with the injection of a 1:20 solution of IL18-BP (Life Sciences Solutions USA) (2 μL / min for 150 s). This was followed by the injection of biotinylated IL18-BP-Fc fusion (10.0 nM) onto flow cell 2 (1.0 μL / min for 120 s). After capture of the biotinylated IL18-BP-Fc fusion onto the sensor surface, a series of Fab concentrations (24.3–0.1 nM, 3-fold dilutions) and total Ab concentrations (12.5–0.8 nM, 2-fold dilutions) were injected onto flow cells 1 and 2 (300 s at 30 μL / min). Dissociation of the Fab or Ab was monitored for 600 s or 5130 s. Several blank buffer samples were injected onto flow cells 1 and 2 (300 s at 30 μL / min) and used for reference surface subtraction. Finally, the sensor surface was prepared for another cycle by injection of regeneration solution (6 M guanidine-HCl in 0.25 M NaOH) over flow cells 1 and 2 (10 μL / min for 120 seconds).

[0409] For data processing and fitting, sensorgrams were trimmed to include only the association and dissociation steps. The trimmed data were then aligned, double-referenced, and fitted to a 1:1 binding model using Biacore Insight Evaluation software (version 3.0.11.15423).

[0410] The results are shown in Figures 41 and 42.

[0411] Figure 41A: Biacore image of anti-IL18BP Fab-human IL18BP interaction; 10 resolution separation.

[0412] Figure 41B: Biacore image of anti-IL18BP Fab-human IL18BP interaction; 85 resolution.

[0413] Figure 41C: Biacore image of anti-IL18BP Fab-cyno IL18BP interaction, 10 resolution.

[0414] Figure 41D: Biacore image of anti-IL18BP Fab-cyno IL18BP interaction, 85 resolution.

[0415] FIG. 42 shows a table showing KD values ​​for human / cyno anti-IL18BP Fab-IL18BP interactions measured by Biacore.

[0416] MSD-SET K D measurement Equilibrium affinity measurements were performed as previously described (Estep et al., 2013). Solution equilibrium titrations (SET) were performed in PBS + 0.1% IgG-free BSA (PBSF) containing antigen (biotinylated IL18-BP-Fc fusion) held constant at 50 pM and incubated with 1.5- to 3-fold serial dilutions of Fab starting at 10 nM and reaching 500 pM (experimental conditions depended on the sample). Antibody (20 nM in PBS) was coated onto standard-binding MSD-ECL plates overnight at 4°C or at room temperature for 30 minutes. The plates were ...

Claims

[Claim 1] The invention described in the specification.

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