ANTI-PVRIG ANTIBODIES AND METHODS OF USE

1 . An antibody that binds to human PVRIG, or antigen-binding fragment thereof, comprising:

(i)

three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:44,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:45,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:46, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:47,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:48,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:49;

(ii)

three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:34,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:35,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:36, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:37,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:38,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:39;

(iii)

three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:14,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:15,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:16, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:17,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:18,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:19;

(iv)

three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:4,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:5,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:6, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:7,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:8,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:9;

(v)

three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:24,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:25,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:26, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:27,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:28,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:29;

(vi)

three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:54,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:55,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:56, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:57,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:58,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:59;

(vii)

three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:64,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:65,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:66, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:67,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:68,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:69;

(viii)

three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:74,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:75,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:76, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:77,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:78,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:79;

(ix)

three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:84,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:85,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:86, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:87,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:88,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:89;

(x)

three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:94,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:95,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:96, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:97,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:98,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:99;

(xi)

three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:104,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO: 105,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO: 106, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:107,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:108,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:109;

(xii)

three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:114,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:115,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:116, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:117,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:118,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:119;

(xiii)

three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:124,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:125,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO: 126, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:127,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:128,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:129;

(xiv)

three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO: 134,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:135,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:136, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:137,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO: 138,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:139;

(xv)

three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:144,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:145,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO: 146, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:147,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:148,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:149; or

(xvi)

three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:187,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO: 188,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:46, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:47,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:48,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:49.

2 . The antibody or antigen-binding fragment thereof of claim 1 , comprising:

(i) a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:50, and a light chain variable region (VL) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:51;

(ii) a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:40, and a light chain variable region (VL) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:41;

(iii) a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:20, and a light chain variable region (VL) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:21;

(iv) a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:10, and a light chain variable region (VL) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:11;

(v) a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:30, and a light chain variable region (VL) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:31;

(vi) a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:60, and a light chain variable region (VL) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:61;

(vii) a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:70, and a light chain variable region (VL) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:71;

(viii) a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:80, and a light chain variable region (VL) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:81;

(ix) a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:90, and a light chain variable region (VL) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:91;

(x) a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:100, and a light chain variable region (VL) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:101;

(xi) a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:110, and a light chain variable region (VL) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:111;

(xii) a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:120, and a light chain variable region (VL) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:121;

(xiii) a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:130, and a light chain variable region (VL) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:131;

(xiv) a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:140, and a light chain variable region (VL) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:141; or

(xv) a heavy chain variable region (VH) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:150, and a light chain variable region (VL) comprising an amino acid sequence at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to SEQ ID NO:151.

3 . The antibody or antigen-binding fragment thereof of claim 2 , wherein one, two, three, four, five, six, seven, eight, nine, or ten amino acids within SEQ ID Nos: 50 and 51, SEQ ID Nos: 40 and 41, SEQ ID Nos: 20 and 21, SEQ ID Nos: 10 and 11, SEQ ID Nos: 30 and 31, SEQ ID Nos: 60 and 61, SEQ ID Nos: 70 and 71, SEQ ID Nos: 80 and 81, SEQ ID Nos: 90 and 91, SEQ ID Nos: 100 and 101, SEQ ID Nos: 110 and 111, SEQ ID Nos: 120 and 121, SEQ ID Nos: 130 and 131, SEQ ID Nos: 140 and 141, or SEQ ID Nos: 150 and 151, have been inserted, deleted or substituted.

4 . The antibody or antigen-binding fragment thereof of claim 1 , that comprises:

(i) a heavy chain variable region (VH) comprising an amino acid sequence as set forth in SEQ ID NO: 50, and a light chain variable region (VL) comprising an amino acid sequence as set forth in SEQ ID NO:51;

(ii) a heavy chain variable region (VH) comprising an amino acid sequence as set forth in SEQ ID NO: 40, and a light chain variable region (VL) comprising an amino acid sequence as set forth in SEQ ID NO:41;

(iii) a heavy chain variable region (VH) comprising an amino acid sequence as set forth in SEQ ID NO: 20, and a light chain variable region (VL) comprising an amino acid sequence as set forth in SEQ ID NO:21;

(iv) a heavy chain variable region (VH) comprising an amino acid sequence as set forth in SEQ ID NO: 10, and a light chain variable region (VL) comprising an amino acid sequence as set forth in SEQ ID NO:11;

(v) a heavy chain variable region (VH) comprising an amino acid sequence as set forth in SEQ ID NO: 30, and a light chain variable region (VL) comprising an amino acid sequence as set forth in SEQ ID NO:31;

(vi) a heavy chain variable region (VH) comprising an amino acid sequence as set forth in SEQ ID NO: 60, and a light chain variable region (VL) comprising an amino acid sequence as set forth in SEQ ID NO:61;

(vii) a heavy chain variable region (VH) comprising an amino acid sequence as set forth in SEQ ID NO: 50, and a light chain variable region (VL) comprising an amino acid sequence as set forth in SEQ ID NO:51;

(viii) a heavy chain variable region (VH) comprising an amino acid sequence as set forth in SEQ ID NO: 70, and a light chain variable region (VL) comprising an amino acid sequence as set forth in SEQ ID NO:71;

(ix) a heavy chain variable region (VH) comprising an amino acid sequence as set forth in SEQ ID NO: 80, and a light chain variable region (VL) comprising an amino acid sequence as set forth in SEQ ID NO:81;

(x) a heavy chain variable region (VH) comprising an amino acid sequence as set forth in SEQ ID NO: 90, and a light chain variable region (VL) comprising an amino acid sequence as set forth in SEQ ID NO:91;

(xi) a heavy chain variable region (VH) comprising an amino acid sequence as set forth in SEQ ID NO: 100, and a light chain variable region (VL) comprising an amino acid sequence as set forth in SEQ ID NO:101;

(xii) a heavy chain variable region (VH) comprising an amino acid sequence as set forth in SEQ ID NO: 110, and a light chain variable region (VL) comprising an amino acid sequence as set forth in SEQ ID NO:111;

(xiii) a heavy chain variable region (VH) comprising an amino acid sequence as set forth in SEQ ID NO: 120, and a light chain variable region (VL) comprising an amino acid sequence as set forth in SEQ ID NO:121;

(xiv) a heavy chain variable region (VH) comprising an amino acid sequence as set forth in SEQ ID NO: 140, and a light chain variable region (VL) comprising an amino acid sequence as set forth in SEQ ID NO:141;

(xv) a heavy chain variable region (VH) comprising an amino acid sequence as set forth in SEQ ID NO: 150, and a light chain variable region (VL) comprising an amino acid sequence as set forth in SEQ ID NO:151.

5 . The antibody or antigen-binding fragment thereof of claim 1 , further comprising an antigen binding domain that specifically binds to human PVRIG at an epitope comprising S27, S31, L32, H52, F99, K95, P100, E101, S103 and E105 of SEQ ID NO: 154.

6 . An isolated human monoclonal antibody, or an antigen-binding portion thereof, wherein the antibody or antigen-binding portion thereof cross-competes for binding to PVRIG with a reference antibody, or a reference antigen-binding portion thereof, comprising the antibody or antigen-binding fragment thereof of claim 1 .

7 . (canceled)

8 . The antibody or antigen-binding portion thereof of claim 6 , wherein the antibody binds to a discontinuous epitope on PVRIG, wherein the discontinuous epitope comprises amino acid residues S27, S31 and L32 of human PVRIG comprising SEQ ID NO: 154.

9 . The antibody or antigen-binding portion thereof of claim 8 , wherein the discontinuous epitope further comprises amino acid residues H52, K95, F99, P100, E101, S103 and E105 of human PVRIG comprising SEQ ID NO:154.

10 . The antibody or antigen-binding fragment thereof of claim 1 , which is a monoclonal antibody, a chimeric antibody, a humanized antibody, a human engineered antibody, a single chain antibody (scFv), a Fab fragment, a Fab′ fragment, or a F(ab′) 2 fragment.

11 . The antibody or antigen-binding fragment thereof of claim 1 , wherein the antibody or antigen-binding fragment thereof: (a) has reduced glycosylation or no glycosylation or is hypofucosylated, (b) comprises increased bisecting GlcNac structures, or (c) comprises an IgG1 Fc domain.

12 . (canceled)

13 . (canceled)

14 . A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof of claim 1 , and further comprising a pharmaceutically acceptable carrier.

15 . An isolated nucleic acid that encodes the antibody or antigen-binding fragment thereof of claim 1 .

16 . A vector comprising the nucleic acid of claim 15 .

17 . A host cell comprising the nucleic acid of claim 15 .

18 . A process for producing an antibody or antigen-binding fragment thereof comprising cultivating the host cell of claim 17 in culture media and recovering the antibody or antigen-binding fragment thereof from the culture media.

19 . A purified composition comprising an anti-human PVRIG antibody or antigen binding fragment thereof produced by the process of claim 18 .

20 . (canceled)

21 . (canceled)

22 . A method of treating cancer comprising administering to a patient in need thereof an effective amount of the pharmaceutical composition of claim 14 .

23 .- 32 . (canceled)

33 . A method of stimulating an immune response in a subject comprising administering to the subject the pharmaceutical composition of claim 14 .

34 . (canceled)

35 . A method of inducing trogocytosis of PVRIG from a donor cell to an acceptor cell, comprising contacting the donor cell with an antibody or antigen-binding fragment of claim 1 for a time sufficient to induce trogocytosis from the donor cell to the acceptor cell.

36 . A method of activating NK-cells comprising administering to a patient in need thereof an effective amount of the pharmaceutical composition of claim 14 .

37 .- 39 . (canceled)