Ex vivo gamma delta t cell populations

Abstract

The invention relates to ex vivo methods of modulating gamma variable 4 (Vγ4) T cells using antibodies or fragments thereof, which specifically bind to a Vγ4 chain of a γδ T cell receptor (TCR) and not to a gamma variable 2 (Vγ2) chain of a γδ TCR. Methods of treatment and other uses of expanded populations of Vγ4 T cells produced by said methods are also provided.

1 . An ex vivo method of modulating gamma variable 4 (Vγ4) T cells comprising administering an antibody or fragment thereof, which specifically binds to a Vγ4 chain of a γδ T cell receptor (TCR) and not to a gamma variable 2 (Vγ2) chain of a γδ TCR, to a cell population comprising Vγ4 T cells. 
     
     
         2 . The method as defined in  claim 1 , wherein the Vγ4 chain of the γδ TCR is human Vγ4 and the Vγ2 chain of the γδ TCR is human Vγ2. 
     
     
         3 . The method as defined in  claim 1  or  2 , wherein the antibody or fragment thereof binds to an epitope of the Vγ4 chain of the γδ TCR comprising one or more amino acid residues within amino acid region 67-82 of SEQ ID NO: 1. 
     
     
         4 . The method as defined in  claim 3 , wherein the epitope comprises at least one of amino acid residues 71, 73, 75, 76, 79 of SEQ ID NO: 1. 
     
     
         5 . The method as defined in any one of  claims 1 to 4 , wherein the epitope comprises or consists of K76 and/or M80 of SEQ ID NO: 1. 
     
     
         6 . The method as defined in any one of  claims 1 to 5 , wherein the epitope is an activating epitope of a γδ T cell. 
     
     
         7 . The method as defined in  claim 6 , wherein binding of the activating epitope: (i) downregulates the γδ TCR; (ii) activates degranulation of the γδ T cell; (iii) activates γδ T cell-mediated killing; and/or (iv) activates or increases Vγ4 chain-mediated cell signalling. 
     
     
         8 . An ex vivo method of modulating gamma variable 4 (Vγ4) T cells comprising administering an anti-Vγ4 antibody or fragment thereof, which comprises one or more of:
 a CDR3 comprising a sequence having at least 80% sequence identity with any one of SEQ ID NOs: 2-47, preferably with SEQ ID NO: 10 and/or 33; 
 a CDR2 comprising a sequence having at least 80% sequence identity with any one of SEQ ID NOs: 48-70 and SEQUENCES: A1-A23 (of Figure 1), preferably with SEQ ID NO: 56 and/or SEQUENCE A9; and/or 
 a CDR1 comprising a sequence having at least 80% sequence identity with any one of SEQ ID NOs: 71-116, preferably with SEQ ID NO: 79 and/or 102, 
to a cell population comprising Vγ4 T cells. 
     
     
         9 . The method as defined in  claim 8 , wherein the antibody of fragment thereof comprises a VH region comprising a CDR3 comprising a sequence having at least 80% sequence identity with any one of SEQ ID NOs: 2-24, such as SEQ ID NOs: 10, 4, 14, 15, 17, 19 or 23. 
     
     
         10 . The method as defined in  claim 8  or  claim 9 , wherein the antibody of fragment thereof comprises a VL region comprising a CDR3 comprising a sequence having at least 80% sequence identity with any one of SEQ ID NOs: 25-47, such as SEQ ID NOs: 33, 27, 37, 38, 40, 42 or 46. 
     
     
         11 . An ex vivo method of modulating gamma variable 4 (Vγ4) T cells comprising administering an anti-Vγ4 antibody or fragment thereof, which comprises an amino acid sequence having at least 80% sequence identity with any one of SEQ ID NOs: 117-162 or 261-283, to a cell population comprising Vγ4 T cells. 
     
     
         12 . An ex vivo method of modulating gamma variable 4 (Vγ4) T cells comprising administering an anti-Vγ4 antibody or fragment thereof comprises one or more of:
 (a) a VH comprising a HCDR1 having SEQ ID NO: 79, a HCDR2 having SEQ ID NO: 56 and a HCDR3 having SEQ ID NO: 10, optionally wherein the VH comprises SEQ ID NO: 125; and 
 a VL comprising a LCDR1 having SEQ ID NO: 102, a LCDR2 having SEQUENCE A9 (of  FIG.  1   ) and a LCDR3 having SEQ ID NO: 33, optionally wherein the VL comprises SEQ ID NO: 148 or 269; 
 
 (b) a VH comprising a HCDR1 having SEQ ID NO: 86, a HCDR2 having SEQ ID NO: 63 and a HCDR3 having SEQ ID NO: 17, optionally wherein the VH comprises SEQ ID NO: 132; and 
 a VL comprising a LCDR1 having SEQ ID NO: 109, a LCDR2 having SEQUENCE A16 (of  FIG.  1   ) and a LCDR3 having SEQ ID NO: 40, optionally wherein the VL comprises SEQ ID NO: 155 or 276; 
 
 (c) a VH comprising a HCDR1 having SEQ ID NO: 73, a HCDR2 having SEQ ID NO: 50 and a HCDR3 having SEQ ID NO: 4, optionally wherein the VH comprises SEQ ID NO: 119; and 
 a VL comprising a LCDR1 having SEQ ID NO: 96, a LCDR2 having SEQUENCE A3 (of  FIG.  1   ) and a LCDR3 having SEQ ID NO: 27, optionally wherein the VL comprises SEQ ID NO: 142 or 263; 
 
 (d) a VH comprising a HCDR1 having SEQ ID NO: 83, a HCDR2 having SEQ ID NO: 60 and a HCDR3 having SEQ ID NO: 14, optionally wherein the VH comprises SEQ ID NO: 129; and
 a VL comprising a LCDR1 having SEQ ID NO: 106, a LCDR2 having SEQUENCE A13 (of  FIG.  1   ) and a LCDR3 having SEQ ID NO: 37, optionally wherein the VL comprises SEQ ID NO: 152 or 273; 
 
 (e) a VH comprising a HCDR1 having SEQ ID NO: 84, a HCDR2 having SEQ ID NO: 61 and a HCDR3 having SEQ ID NO: 15, optionally wherein the VH comprises SEQ ID NO: 130; and
 a VL comprising a LCDR1 having SEQ ID NO: 107, a LCDR2 having SEQUENCE A14 (of  FIG.  1   ) and a LCDR3 having SEQ ID NO: 38, optionally wherein the VL comprises SEQ ID NO: 153 or 274; 
 
 (f) a VH comprising a HCDR1 having SEQ ID NO: 88, a HCDR2 having SEQ ID NO: 65 and a HCDR3 having SEQ ID NO: 19, optionally wherein the VH comprises SEQ ID NO: 134; and
 a VL comprising a LCDR1 having SEQ ID NO: 111, a LCDR2 having SEQUENCE A18 (of  FIG.  1   ) and a LCDR3 having SEQ ID NO: 42, optionally wherein the VL comprises SEQ ID NO: 157 or 278; 
 
 (g) a VH comprising a HCDR1 having SEQ ID NO: 92, a HCDR2 having SEQ ID NO: 69 and a HCDR3 having SEQ ID NO: 23, optionally wherein the VH comprises SEQ ID NO: 138; and
 a VL comprising a LCDR1 having SEQ ID NO: 115, a LCDR2 having SEQUENCE A22 (of  FIG.  1   ) and a LCDR3 having SEQ ID NO: 46, optionally wherein the VL comprises SEQ ID NO: 161 or 282; 
 
 (h) a VH comprising a HCDR1 having SEQ ID NO: 71, a HCDR2 having SEQ ID NO: 48 and a HCDR3 having SEQ ID NO: 2, optionally wherein the VH comprises SEQ ID NO: 117; and
 a VL comprising a LCDR1 having SEQ ID NO: 94, a LCDR2 having SEQUENCE A1 (of  FIG.  1   ) and a LCDR3 having SEQ ID NO: 25, optionally wherein the VL comprises SEQ ID NO: 140 or 261; 
 
 (i) a VH comprising a HCDR1 having SEQ ID NO: 72, a HCDR2 having SEQ ID NO: 49 and a HCDR3 having SEQ ID NO: 3, optionally wherein the VH comprises SEQ ID NO: 118; and
 a VL comprising a LCDR1 having SEQ ID NO: 95, a LCDR2 having SEQUENCE A2 (of  FIG.  1   ) and a LCDR3 having SEQ ID NO: 26, optionally wherein the VL comprises SEQ ID NO: 141 or 262; 
 
 (j) a VH comprising a HCDR1 having SEQ ID NO: 74, a HCDR2 having SEQ ID NO: 51 and a HCDR3 having SEQ ID NO: 5, optionally wherein the VH comprises SEQ ID NO: 120; and
 a VL comprising a LCDR1 having SEQ ID NO: 97, a LCDR2 having SEQUENCE A4 (of  FIG.  1   ) and a LCDR3 having SEQ ID NO: 28, optionally wherein the VL comprises SEQ ID NO: 143 or 264; 
 
 (k) a VH comprising a HCDR1 having SEQ ID NO: 75, a HCDR2 having SEQ ID NO: 52 and a HCDR3 having SEQ ID NO: 6, optionally wherein the VH comprises SEQ ID NO: 121; and
 a VL comprising a LCDR1 having SEQ ID NO: 98, a LCDR2 having SEQUENCE A5 (of  FIG.  1   ) and a LCDR3 having SEQ ID NO: 29, optionally wherein the VL comprises SEQ ID NO: 144 or 265; 
 
 (I) a VH comprising a HCDR1 having SEQ ID NO: 76, a HCDR2 having SEQ ID NO: 53 and a HCDR3 having SEQ ID NO: 7, optionally wherein the VH comprises SEQ ID NO: 122; and
 a VL comprising a LCDR1 having SEQ ID NO: 99, a LCDR2 having SEQUENCE A6 (of  FIG.  1   ) and a LCDR3 having SEQ ID NO: 30, optionally wherein the VL comprises SEQ ID NO: 145 or 266; 
 
 (m) a VH comprising a HCDR1 having SEQ ID NO: 77, a HCDR2 having SEQ ID NO: 54 and a HCDR3 having SEQ ID NO: 8, optionally wherein the VH comprises SEQ ID NO: 123; and
 a VL comprising a LCDR1 having SEQ ID NO: 100, a LCDR2 having SEQUENCE A7 (of  FIG.  1   ) and a LCDR3 having SEQ ID NO: 31, optionally wherein the VL comprises SEQ ID NO: 146 or 267; 
 
 (n) a VH comprising a HCDR1 having SEQ ID NO: 78, a HCDR2 having SEQ ID NO: 55 and a HCDR3 having SEQ ID NO: 9, optionally wherein the VH comprises SEQ ID NO: 124; and
 a VL comprising a LCDR1 having SEQ ID NO: 101, a LCDR2 having SEQUENCE A8 (of  FIG.  1   ) and a LCDR3 having SEQ ID NO: 32, optionally wherein the VL comprises SEQ ID NO: 147 or 268; 
 
 (o) a VH comprising a HCDR1 having SEQ ID NO: 80, a HCDR2 having SEQ ID NO: 57 and a HCDR3 having SEQ ID NO: 11, optionally wherein the VH comprises SEQ ID NO: 126; and
 a VL comprising a LCDR1 having SEQ ID NO: 103, a LCDR2 having SEQUENCE A10 (of  FIG.  1   ) and a LCDR3 having SEQ ID NO: 34, optionally wherein the VL comprises SEQ ID NO: 149 or 270; 
 
 (p) a VH comprising a HCDR1 having SEQ ID NO: 81, a HCDR2 having SEQ ID NO: 58 and a HCDR3 having SEQ ID NO: 12, optionally wherein the VH comprises SEQ ID NO: 127; and
 a VL comprising a LCDR1 having SEQ ID NO: 104, a LCDR2 having SEQUENCE A11 (of  FIG.  1   ) and a LCDR3 having SEQ ID NO: 35, optionally wherein the VL comprises SEQ ID NO: 150 or 271; 
 
 (q) a VH comprising a HCDR1 having SEQ ID NO: 82, a HCDR2 having SEQ ID NO: 59 and a HCDR3 having SEQ ID NO: 13, optionally wherein the VH comprises SEQ ID NO: 128; and
 a VL comprising a LCDR1 having SEQ ID NO: 105, a LCDR2 having SEQUENCE A12 (of  FIG.  1   ) and a LCDR3 having SEQ ID NO: 36, optionally wherein the VL comprises SEQ ID NO: 151 or 272; 
 
 (r) a VH comprising a HCDR1 having SEQ ID NO: 85, a HCDR2 having SEQ ID NO: 62 and a HCDR3 having SEQ ID NO: 16, optionally wherein the VH comprises SEQ ID NO: 131; and
 a VL comprising a LCDR1 having SEQ ID NO: 108, a LCDR2 having SEQUENCE A15 (of  FIG.  1   ) and a LCDR3 having SEQ ID NO: 39, optionally wherein the VL comprises SEQ ID NO: 154 or 275; 
 
 (s) a VH comprising a HCDR1 having SEQ ID NO: 87, a HCDR2 having SEQ ID NO: 64 and a HCDR3 having SEQ ID NO: 18, optionally wherein the VH comprises SEQ ID NO: 133; and
 a VL comprising a LCDR1 having SEQ ID NO: 110, a LCDR2 having SEQUENCE A17 (of  FIG.  1   ) and a LCDR3 having SEQ ID NO: 41, optionally wherein the VL comprises SEQ ID NO: 156 or 277; 
 
 (t) a VH comprising a HCDR1 having SEQ ID NO: 89, a HCDR2 having SEQ ID NO: 66 and a HCDR3 having SEQ ID NO: 20, optionally wherein the VH comprises SEQ ID NO: 135; and
 a VL comprising a LCDR1 having SEQ ID NO: 112, a LCDR2 having SEQUENCE A19 (of  FIG.  1   ) and a LCDR3 having SEQ ID NO: 43, optionally wherein the VL comprises SEQ ID NO: 158 or 279; 
 
 (u) a VH comprising a HCDR1 having SEQ ID NO: 90, a HCDR2 having SEQ ID NO: 67 and a HCDR3 having SEQ ID NO: 21, optionally wherein the VH comprises SEQ ID NO: 136; and
 a VL comprising a LCDR1 having SEQ ID NO: 113, a LCDR2 having SEQUENCE A20 (of  FIG.  1   ) and a LCDR3 having SEQ ID NO: 44, optionally wherein the VL comprises SEQ ID NO: 159 or 280; 
 
 (v) a VH comprising a HCDR1 having SEQ ID NO: 91, a HCDR2 having SEQ ID NO: 68 and a HCDR3 having SEQ ID NO: 22, optionally wherein the VH comprises SEQ ID NO: 137; and 
 a VL comprising a LCDR1 having SEQ ID NO: 114, a LCDR2 having SEQUENCE A21 (of  FIG.  1   ) and a LCDR3 having SEQ ID NO: 45, optionally wherein the VL comprises SEQ ID NO: 160 or 281; and/or 
 
 (w) a VH comprising a HCDR1 having SEQ ID NO: 93, a HCDR2 having SEQ ID NO: 70 and a HCDR3 having SEQ ID NO: 24, optionally wherein the VH comprises SEQ ID NO: 139; and
 a VL comprising a LCDR1 having SEQ ID NO: 116, a LCDR2 having SEQUENCE A23 (of  FIG.  1   ) and a LCDR3 having SEQ ID NO: 47, optionally wherein the VL comprises SEQ ID NO: 162 or 283, 
 
 to a cell population comprising Vγ4 T cells. 
     
     
         13 . An ex vivo method of modulating gamma variable 4 (Vγ4) T cells comprising administering an anti-Vγ4 antibody or fragment thereof which comprises an amino acid sequence having at least 80% sequence identity with any one of SEQ ID NOs: 163-185, to a cell population comprising Vγ4 T cells. 
     
     
         14 . An ex vivo method of modulating gamma variable 4 (Vγ4) T cells comprising administering an anti-Vγ4 antibody which comprises an amino acid sequence having at least 80% sequence identity with any one of SEQ ID NOs: 233-255, to a cell population comprising Vγ4 T cells. 
     
     
         15 . An ex vivo method of modulating gamma variable 4 (Vγ4) T cells comprising administering an anti-Vγ4 antibody or fragment thereof comprising a heavy chain amino acid sequence having at least 80% sequence identity with any one of SEQ ID NOs: 284-306 and/or a light chain amino acid sequence having at least 80% sequence identity with any one of SEQ ID NOs: 307-329, to a cell population comprising Vγ4 T cells. 
     
     
         16 . The method as defined in any one of  claims 1 to 15 , wherein the anti-Vγ4 antibody or fragment thereof is:
 (i) an scFv or a full length antibody; and/or 
 (ii) a human antibody or fragment thereof. 
 
     
     
         17 . The method as defined in any one of  claims 1 to 16 , wherein the modulation comprises expansion of Vγ4 T cells. 
     
     
         18 . The method as defined in  claim 17 , wherein the method provides an expanded population of Vγ4 T cells which contains greater than about 60% Vγ4 T cells, such as greater than about 70% Vγ4 T cells. 
     
     
         19 . The method as defined in any one of  claims 1 to 18 , wherein the method comprises culturing the cell population for at least 5 days. 
     
     
         20 . The method as defined in any one of  claims 1 to 19 , wherein the method comprises culturing the cell population in the presence of IL-2 and/or IL-15. 
     
     
         21 . The method as defined in any one of  claims 1 to 20 , wherein the cell population is obtained from a haematopoietic sample or a fraction thereof. 
     
     
         22 . The method as defined in  claim 21 , wherein the haematopoietic sample consists of peripheral blood mononuclear cells (PBMCs) or low density mononuclear cells (LDMCs). 
     
     
         23 . The method as defined in any one of  claims 1 to 20 , wherein the cell population is obtained from a non-haematopoietic tissue sample, such as a skin, colon, gut, mammary gland, lung, prostate, liver, spleen, pancreas, uterus, vagina or other cutaneous, mucosal or serous membrane sample. 
     
     
         24 . The method as defined in any one of  claims 1 to 23 , wherein the cell population is obtained from human or non-human animal tissue. 
     
     
         25 . A Vγ4 T cell population obtained by the ex vivo method as defined in any one of  claims 1 to 24 . 
     
     
         26 . A composition comprising the Vγ4 T cell population as defined in  claim 25 . 
     
     
         27 . A pharmaceutical composition comprising the Vγ4 T cell population as defined in  claim 25 , together with a pharmaceutically acceptable diluent or carrier. 
     
     
         28 . The pharmaceutical composition as defined in  claim 27 , for use as a medicament. 
     
     
         29 . The pharmaceutical composition as defined in  claim 27  for use in the treatment of cancer, an infectious disease or an inflammatory disease. 
     
     
         30 . A method of treating a cancer, an infectious disease or an inflammatory disease in a subject in need thereof, comprising administering a therapeutically effective amount of the Vγ4 T cell population as defined in  claim 25  or the pharmaceutical composition as defined in  claim 27 .