US2020283500A1PendingUtilityA1
Variant icos ligand immunomodulatory proteins and related compositions and methods
Assignee: ALPINE IMMUNE SCIENCES INCPriority: Oct 18, 2017Filed: Oct 17, 2018Published: Sep 10, 2020
Est. expiryOct 18, 2037(~11.3 yrs left)· nominal 20-yr term from priority
C07K 2319/43C07K 2319/21A61P 35/00A61K 38/00C12N 15/85C07K 2319/30C07K 14/7051A61K 47/50A61K 38/1774C12P 21/02C07K 2319/41C07K 2319/03A61P 17/00A61K 35/54C12N 5/0681C07K 2319/31C07K 2319/02A61P 11/06A61K 40/31A61K 40/11A61K 40/4211A61K 40/421A61K 40/4205A61K 40/418A61K 40/22A61K 2239/31C07K 14/70532
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Claims
Abstract
Provided herein are immunomodulatory proteins comprising ICOSL variants and nucleic acids encoding such proteins. The immunomodulatory proteins provide therapeutic utility for a variety of immunological and oncological conditions. Compositions and methods for making and using such proteins are provided.
Claims
exact text as granted — not AI-modifiedWhat is claimed:
1 . A variant ICOS Ligand (ICOSL) polypeptide, comprising one or more amino acid modifications in an immunoglobulin superfamily (IgSF) domain of an ICOSL reference polypeptide, wherein the ICOSL reference polypeptide is a truncated extracellular domain comprising a contiguous sequence of amino acids comprising amino acids 1-112 and a C-terminal truncation of at least 25 amino acids with reference to the ICOSL extracellular domain sequence set forth in SEQ ID NO:32.
2 . The variant ICOSL polypeptide of claim 1 , wherein the variant ICOSL polypeptide exhibits increased binding to the ectodomain(s) of ICOS or CD28 compared to the binding of the ICOSL reference polypeptide for the same ectodomain(s).
3 . The variant ICOSL polypeptide of claim 1 or claim 2 , wherein the C-terminal truncation is of at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, at least 100, at least 125 amino acid residues.
4 . The variant ICOSL polypeptide of any of claims 1 - 3 , wherein the ICOSL reference polypeptide is altered in or lacks a protease cleavage site set forth as amino acids 204-209 of SEQ ID NO:32.
5 . The variant ICOSL polypeptide of any of claims 1 - 4 , wherein the ICOSL reference polypeptide comprises the sequence of amino acids set forth in SEQ ID NO:545.
6 . The variant ICOSL polypeptide of any of claims 1 - 4 , wherein the ICOSL reference polypeptide consists of the sequence of amino acids set forth in SEQ ID NO:545.
7 . A variant ICOSL Ligand (ICOSL) polypeptide, comprising one or more amino acid modifications in an ICOSL reference polypeptide, wherein the ICOSL reference polypeptide consists of the sequence of amino acids set forth in SEQ ID NO:545.
8 . A variant ICOS Ligand (ICOSL) polypeptide, comprising one or more amino acid modifications in an immunoglobulin superfamily (IgSF) domain of an ICOSL reference polypeptide, wherein the ICOSL reference polypeptide is altered in one or more amino acids corresponding to amino acids 204-209 with reference to SEQ ID NO:32.
9 . The variant ICOSL polypeptide of any of claims 1 - 8 , wherein the one or more amino acid modifications are in a position corresponding to position(s) 52, 57 or 100, with reference to numbering of SEQ ID NO:32.
10 . The variant ICOSL polypeptide of any of claims 1 - 9 , wherein the one or more amino acid modifications are selected from N52A, N52C, N52D, N52G, N52H, N52K, N52L, N52M, N52Q, N52R, N52S, N52T, N52V, N52Y, N52K, N57A, N57D, N57E, N57F, N57H, N57K, N57L, N57M, N57P, N57Q, N57S, N57T, N57V, N57Y, N57W, Q100A, Q100D, Q100G, Q100K, Q100L, Q100M, Q100N, Q100P, Q100R, Q100S, Q100T or Q100V, with reference to numbering of SEQ ID NO:32.
11 . The variant ICOSL polypeptide of any of claims 1 - 10 , wherein the one or more amino acid modifications are selected from among N52Y/N57Y/F138L/L203P, N52H/N57Y/Q100P, N52S/Y146C/Y152C, N52H/C198R, N52H/C140D/T225A, N52H/C198R/T225A, N52H/K92R, N52H/S99G, N57Y/Q100P, N52S/S130G/Y152C, N52S/Y152C, N52S/C198R, N52Y/N57Y/Y152C, N52Y/N57Y/H129P/C198R, N52H/L161P/C198R, N52S/T113E, N52D/S54P, N52K/L208P, N52S/Y152H, N52D/V151A, N52H/1143T, N52S/L80P, N52S/R75Q/L203P, N52S/D158G, N52D/Q133H, N52S/N57Y/H94D/L96F/L98F/Q100R, N52S/N57Y/H94D/L96F/L98F/Q100R/G103E/F120S, N52S/G103E, N52H/F78L/Q100R, N52H/N57Y/Q100R/V110D, N52H/N57Y/R75Q/Q100R/V110D, N52H/N57Y/Q100R, N52H/N57Y/L74Q/Q100R/V10D, N52H/Q100R, N52H/S121G, A20V/N52H/N57Y/Q100R/S109G, N52H/N57Y/R61S/Q100R/V110D/L173S, N52H/N57Y/Q100R/V122A, N52H/N57Y/Q100R/F172S, N52H/N57Y, N52S/F120S, N52S/V97A, N52S/G72R, N52S/A71T/A117T, N52S/E220G, Y47H/N52S/V107A/F120S, N52H/N57Y/Q100R/V110D/S132F/M175T, E16V/N52H/N57Y/Q100R/V110D/H115R/Y152C/K156M/C198R, Q37R/N52H/N57Y/Q100R/V110N/S142F/C198R/D217V/R221G, N52H/N57Y/Q100R/V110D/C198R, N52H/N57Y/Q100R/V1D/V16A/L161M/F172S/S192G/C198R, F27S/N52H/N57Y/V11N, N52S/H94E/L96I/S109N/L166Q, S18R/N52S/F93L/I143V/R221G, A20T/N52D/Y146C/Q164L, V11E/N30D/N52H/N57Y/H94E/L96I/L98F/N194D/V210A/I218T, N52S/H94E/L96I/V122M, N52H/N57Y/H94E/L96I/F120I/S126T/W153R/I218N, M10V/S18R/N30D/N52S/S126R/T139S/L203F, S25G/N30D/N52S/F120S/N227K, N30D/N52S/L67P/Q100K/D217G/R221K/T225S, N52H/N57Y/Q100R/V110D/A117T/T190S/C198R, N52H/N57Y/Q100R/V110D/F172S/C198R, S25G/F27C/N52H/N57Y/Q100R/V110D/E135K/L173S/C198R, N52H/N57Y/V110A/C198R/R221I, M10I/S13G/N52H/N57Y/D77G/V110A/H129P/I143V/F172S/V193M, C198R, N52H/N57Y/R61C/Y62F/Q100R/V110N/F120S/C198R, N52H/N57Y/Q100R/V110D/H115R/C198R, N52H/N57Y/Q100R/V110D/N144D/F172S/C198R, N52S/H94E/L98F/Q100R, N52S/E90A, N30D/K42E/N52S, N52S/F120S/I143V/I224V, N52H/N57Y/Q100R/V110D/C198R/S212G, N52H/N57Y/Q100R/C198R, N52S/N194D, N52H/N57Y/Q100R/L102R/V110D/H115R/C198R, N52H/N57Y/Q100R/V110D/C198R/S212G, N52H/N57Y/Q100R/C198R, N52S/N194D, N52H/N57Y/Q100R/L102R/V110D/H115R/C198R, N52S/S54P, T38P/N52S/N57D, N52H/C140del/T225A, N52H/F78L/Q100R/C198R, N52H/N57Y/R75Q/Q100P/V110D, N52H/N57Y/L74Q/V110D/S192G, N52H/S121G/C198R, N52S/F120S/N227K, N52S/A71T/A117T/T190A/C198R, T43A/N52H/N57Y/L74Q/D89G/V110D/F172S, N52H/N57Y/Q100R/V110D/S132F/M175T, N52D, N52H/N57Y/Q100R/V107I/V110D/I154F/C198R/R221G, N52Q/N207Q, N168Q/N207Q, N52Q/N168Q, N52Q/N84Q, N52Q/N119Q, N52Q/N84Q/N168Q, N52Q/N84Q/N207Q, N52Q/N119Q/N155Q, N52H/N84Q/N119Q, N52H/N84Q, N52H/N84Q/N168Q/N207Q, N52Q/N84Q/N155Q/N168Q, N52Q/N84Q/N119Q/N168Q, N52Q/N84Q/N119Q/N207Q, N52Q/N84Q/N119Q/N155Q, N52Q/N84Q/N119Q/N155Q/N207Q, N52Y/F138L/L203P, N57Y/Q100R/C198R, N57Y/F138L/L203P, Q100R/F138L, N52H/N57Y/Q100R/H115R/C198R, N52H/N57Y/Q100R/F172S/C198R, N52H/N57Y/Q100R/H115R/F172S/C198R, N52H/N57Y/Q100R/H115R/I143V/F172S/C198R, N52H/N57Y/Q100R/L102R/H115R/F172S/C198R, N52H/V122A/F172S/C198R, N52H/N57Y/Q100R/H115R/F172S/N194D, N52H/N57Y/H115R/F172S/C198R, N52H/N57Y/Q100R/H115R/C198R, N52H/N57Y/H115R, N52H/N57Y/Q100R/H115R, N52H/N57Y/Q100R/H115R/F172S/I224V, N52H/N57Y/Q100R/H115R/F172S, N52H/N57Y/Q100R/F172S, N52H/Q100R/H115R/I143T/F172S, N52H/N57Y/Q100P/H115R/F172S, N52Y/N57Y/Q100P/F172S, E16V/N52H/N57Y/Q100R/V110D/H115R/C198R, E16V/N52H/N57Y/Q100R/V110D/H115R/Y152C/K156M/F172S/C198R, N52S/E90A/H115R, N30D/K42E N52S/H115R, N30D/K42E/N52S/H115R/C198R/R221I, N30D/K42E/N52S/H115R/C198R, N30D/K42E/N52S/H115R/F172S/N194D, N52S/H115R/F120S/I143V/C198R, N52S/H115R/F172S/C198R, N52H/N57Y/Q100P/C198R, N52H/N57Y/Q100P H115R/F172S/C198R, N52H/N57Y/Q100P/F172S/C198R, N52H/N57Y/Q100P/H115R, N52H/N57Y/Q100P/H115R/C198R, N52H/Q100R/C198R, N52H/Q100R/H115R/F172S, N52H/Q100R/F172S/C198R, N52H/Q100R/H115R/F172S/C198R, N52H/N57Y/Q100R/F172S/C198R, N52A/N57F/Q100S, N52A/N57H/Q100S, N52A/N57Y/Q100A, N52D/N57A/Q100A, N52D/Q100S, N52G/Q100A, N52H/Q100A, N52M/N57H/Q100S, N52M/N57W/Q100P, N52Q/N57F, N52Q/N57S/Q100A, N52R/N57L/Q100A, N52R/N57Y/Q100P, N52R/N57Y/Q100S, N52S/N57A/Q100A, N52S/N57H/Q100E, N52S/N57L/Q100S, N52S/N57M/Q100S, N52S/N57Y/Q100S, N52S/N57Y/Q100M, N52S/N57Y/Q100V, N52T/N57H/Q100S, N52T/N57H/Q100A, N52T/N57Y/Q100A, N52V/N57L/Q100A, N52H/N57Y/Q100K, N52K/N57Y/Q100R, N52L/N57H/Q100R, N52R/N57F/Q100N, N52R/N57F/Q100P, N52R/N57F/Q100R, N52R/N57F/Q100T, N52R/N57H/Q100K, N52R/N57L/Q100S, N52R/N57W/Q100K, N52R/N57W, N52R/N57Y/Q100R, N52C/N57E/Q100S, N52G/N57P/Q100D, N52G/N57V/Q100G, N52G/N57V, N52L/N57V, N52P/N57P, N52P/N57S/Q100G, N52S/N57L/Q100G, N52T/N57K/Q100P, N52V/N57T/Q100L, N57Q/Q100P, or R26S/N52H/N57Y/V110D/T137A/C198R, with reference to numbering of SEQ ID NO:32.
12 . The variant ICOSL polypeptide of any of claims 1 - 11 , wherein the one or more amino acid modifications are selected from among N52A/N57Y/Q100A, N52D/Q100S, N52G/Q100A, N52M/N57H/Q100S, N52M/N57W/Q100P, N52Q/N57S/Q100A, N52R/N57L/Q100A, N52S/N57H/Q100E, N52S/N57L/Q100S, N52S/N57M/Q100S, N52S/N57Y/Q100M, N52T/N57H/Q100S, N52R/N57F/Q100P, N52R/N57F/Q100T, N52R/N57W/Q100K, N52R/N57W, N52G/N57V, N52L/N57V, N52S/N57L/Q100G, N52T/N57K/Q100P, N52S, N52H, N52D, N52Y/N57Y/F138L/L203P, N52H/N57Y/Q100P, N52S/Y146C/Y152C, N52H/C198R, N52H/C198R/T225A, N52H/K92R, N57Y, N52S/C198R, N52S/T113E, S54A, N52D/S54P, N52K/L208P, N52H/I143T, N52S/D158G, N52D/Q133H, N52H/N57Y/Q100R/V110D/C198R/S212G, N52H/N57Y/Q100R/V122A, N52H/N57Y/Q100R/F172S, N52H/N57Y/Q100R, N52S/N194D, N52H/N57Y/Q100R/L102R/V110D/H115R/C198R, N52S/E90A, N52S/F120S/I143V/I224V, N52H/N57Y/Q100R/F172S/C198R, N52H/N57Y/Q100R/H115R/F172S/C198R, N52Y/N57Y/Q100P/F172S, E16V/N52H/N57Y/Q100R/V110D/H115R/Y152C/K156M/F172S/C198R, N52S/H115R/F120S/I143V/C198R, N52H/N57Y/Q100P/C198R, N52H/N57Y/Q100P/H115R/F172S/C198R, N52H/N57Y/Q100P/F172S/C198R, N52H/N57Y/Q100P/H115R, N52H/N57Y/Q100P/H115R/C198R, N52H/Q100R/C198R, N52H/Q100R/H115R/F172S, N52H/Q100R/H115X/F172S/C198R, N52H/Q100R/H115R/F172S/C198R, N52H/N57Y/Q100R/H115R/F172S/C198R, N52H/N57Y/Q100R/H115R/F172S, N52H/N57Y/Q100R/H115R/F172S/C198R, Q100R, N52Y/F138L/L203P, N57Y/Q100R/C198R, N57Y/F138L/L203P, N52H, N57Y, N57Y/Q100P, Q100R/F138L, N52H/N57Y/Q100R/H115R, N52H/N57Y/Q100R/F172S, N52H/N57Y/Q100R/H115R/F172S/I224V, N52H/N57Y/Q100R/H115R/F172S, N52H/N57Y/Q100R/H115R/C198R, N52H/N57Y/Q100R/F172S/C198R, N52H/N57Y/Q100R/H115R/F172S/C198R, N52H/N57Y/Q100R/H115R/I143V/F172S/C198R, N52H/N57Y/Q100R/L102R H115R/F172S/C198R, N52H/N57Y/Q100R/H115R F172S/N194D, N52H/N57Y/H115R/F172S/C198R, N52H/N57Y/Q100R/H115R/C198R, N52H/N57Y/H115R, N52H/Q100R/H115R/I143T/F172S, N52H/N57Y/Q100P/H115R/F172S, E16V/N52H/N57Y/Q100R/V110D/H115R/C198R, N52S/E90A/H115R, N52S/E90A/H115R, or N30D/K42E/N52S/H115R, with reference to numbering of SEQ ID NO:32.
13 . The variant ICOSL polypeptide of any of claims 1 - 12 , comprising one or more amino acid modifications N52H/Q100R.
14 . The variant ICOSL polypeptide of claim 13 , wherein the variant ICOSL polypeptide has the sequence set forth in SEQ ID NO:567.
15 . The variant ICOSL polypeptide of any of claims 1 - 13 , comprising one or more amino acid modifications are N52H/N57Y/Q100R.
16 . The variant ICOSL polypeptide of claim 15 , wherein the polypeptide has the sequence set forth in SEQ ID NO:565.
17 . The variant ICOSL polypeptide of any of claims 1 - 12 , comprising one or more amino acid modifications are N52L/N57H/Q100R.
18 . The variant ICOSL polypeptide of claim 17 , wherein the polypeptide has the sequence set forth in SEQ ID NO:761.
19 . The variant ICOSL polypeptide of any of claims 1 - 12 , comprising the amino acid modification is N52D.
20 . The variant ICOSL polypeptide of claim 19 , wherein the polypeptide has the sequence set forth in SEQ ID NO:548.
21 . The variant ICOSL polypeptide of any of claims 8 - 12 , wherein:
the alteration comprises a deletion of one or more contiguous amino acids corresponding to amino acids 204-209 with reference to SEQ ID NO:32; or the alteration comprises at least one amino acid substitution at one or both of position 207 and 208 corresponding to positions set forth in SEQ ID NO:32.
22 . The variant ICOSL polypeptide of claim 21 , wherein the at least one amino acid substitution is N207A, N207G or L208G with reference to numbering of SEQ ID NO:32, or a conservative amino acid substitution thereof.
23 . The variant ICOSL polypeptide of any of claims 1 - 22 , wherein the variant ICOSL polypeptide exhibits reduced proteolytic cleavage when expressed from a cell compared to a full-length extracellular domain of the variant ICOSL polypeptide when expressed from the same cell.
24 . The variant ICOSL polypeptide of any of claims 1 - 8 and 21 - 23 , wherein the one or more amino acid modifications are in a position corresponding to position(s) selected from 10, 11, 13, 16, 18, 20, 25, 26, 27, 30, 33, 37, 38, 42, 43, 47, 52, 54, 57, 61, 62, 67, 71, 72, 74, 75, 77, 78, 80, 84, 89, 90, 92, 93, 94, 96, 97, 98, 99, 100, 102, 103, 107, 109, 110, 111, 113, 115, 116, 117, 119, 120, 121, 122, 126, 129, 130, 132, 133, 135, 137, 138, 139, 140, 142, 143, 144, 146, 151, 152, 153, 154, 155, 156, 158, 161, 164, 166, 168, 172, 173, 175, 190, 192, 193, 194, 198, 201, 203, 207, 208, 210, 212, 217, 218, 220, 221, 224, 225, or 227 with reference to numbering of SEQ ID NO:32.
25 . The variant ICOSL polypeptide of any of claims 1 - 8 and 21 - 24 , wherein the one or more amino acid modifications are selected from M10V, M10I, V11E, S13G, E16V, S18R, A20T, A20V, S25G, R26S, F27C, F27S, N30D, Y33del, Q37R, T38P, K42E, T43A, Y47H, N52A, N52C, N52D, N52G, N52H, N52K, N52L, N52M, N52P, N52Q, N52R, N52S, N52T, N52V, N52Y, S54A, S54F, S54P, N57A, N57D, N57E, N57F, N57H, N57K, N57L, N57M, N57P, N57Q, N57S, N57T, N57V, N57W, N57Y, R61C, R61S, Y62F, L67P, A71T, G72R, L74Q, R75Q, D77G, F78L, L80P, N84Q, D89G, E90A, K92R, F93L, H94D, H94E, L96F, L96I, V97A, L98F, S99G, Q100A, Q100D, Q100E, Q100G, Q100K, Q100L, Q100M, Q100N, Q100P, Q100R, Q100S, Q100T, Q100V, L102R, G103E, V107A, V107I, S109G, S109N, V110A, V110D, V110N, E111del, T113E, H115Q, H115R, V116A, A117T, N119Q, F120I, F120S, S121G, V122A, V122M, S126R, S126T, H129P, S130G, S132F, Q133H, E135K, T137A, F138L, T139S, C140del, C140D, S142F, I143T, I143V, N144D, Y146C, V151A, Y152C, Y152H, W153R, I154F, N155H, N155Q, K156M, D158G, L161M, L161P, Q164L, L166Q, N168Q, F172S, L173S, M175T, T190A, T190S, S192G, V193A, V193M, N194D, C198R, N201S, L203F, L203P, N207Q, L208P, V210A, S212G, D217G, D217V, I218N, I218T, E220G, R221G, R221I, R221K, I224V, T225A, T225S, N227K with reference to numbering of SEQ ID NO:32, or a conservative amino acid substitution thereof.
26 . The variant ICOSL polypeptide of any of claims 1 - 8 and 21 - 25 , wherein the one or more amino acid modifications are selected from among F120S/Y152H/N201S, E111del, Y33del, N168Q/N207Q, N84Q/N207Q, N155Q/N207Q, N119Q/N168Q, N119Q/N207Q, N119Q/N155Q, N84Q/N119Q, N84Q/N155Q/N168Q, N84Q/N168Q/N207Q, N84Q/N155H/N207Q, N155Q/N168Q/N207Q, N119Q N155Q/N168Q, N119Q/N168Q/N207Q, N84Q/N119Q/N207Q, N119Q/N155H/N207Q, N84Q/N119Q/N155Q, N84Q/N119Q/N155Q/N168Q, N84Q/N155Q/N168Q/N207Q, N84Q/N119Q/N155Q/N207Q, N84Q/N119Q/N155Q/N168Q/N207Q or F138L/L203P, with reference to numbering of SEQ ID NO:32.
27 . The variant ICOSL polypeptide of any of claims 1 - 8 and 21 - 25 , wherein the one or more amino acid modifications are selected from amino acid modifications N52Y/N57Y/F138L/L203P, N52H/N57Y/Q100P, N52S/Y146C/Y152C, N52H/C198R, N52H/C140D/T225A, N52H/C198R/T225A, N52H/K92R, N52H/S99G, N57Y/Q100P, N52S/S130G/Y152C, N52S/Y152C, N52S/C198R, N52Y/N57Y/Y152C, N52Y/N57Y/H129P/C198R, N52H/L161P/C198R, N52S/T113E, N52D/S54P, N52K/L208P, N52S/Y152H, N52D/V151A, N52H/I143T, N52S/L80P, F120S/Y152H/N201S, N52S/R75Q/L203P, N52S/D158G, N52D/Q133H, N52S/N57Y/H94D/L96F/L98F/Q100R, N52S/N57Y/H94D/L96F/L98F/Q100R/G103E/F120S, N52S/G103E, N52H/F78L/Q100R, N52H/N57Y/Q100R/V110D, N52H/N57Y/R75Q/Q100R/V110D, N52H/N57Y/Q100R, N52H/N57Y/L74Q/Q100R/V110D, N52H/Q100R, N52H/S121G, A20V/N52H/N57Y/Q100R/S109G, N52H/N57Y/R61S/Q100R/V110D/L173S, N52H/N57Y/Q100R/V122A, N52H/N57Y/Q100R/F172S, N52H/N57Y, N52S/F120S, N52S/V97A, N52S/G72R, N52S/A71T/A117T, N52S/E220G, Y47H/N52S/V107A/F120S, N52H/N57Y/Q100R/V110D/S132F/M175T, E16V/N52H/N57Y/Q100R/V110D/H115R/Y152C/K156M/C198R, Q37R/N52H/N57Y/Q100R/V110N/S142F/C198R/D217V/R221G, N52H/N57Y/Q100R/V110D/C198R, N52H/N57Y/Q100R/V110D/V116A/L161M/F172S/S192G/C198R, F27S/N52H/N57Y/V110N, N52S/H94E/L96I/S109N/L166Q, S18R/N52S/F93L/I143V/R221G, A20T/N52D/Y146C/Q164L, V11E/N30D/N52H/N57Y/H94E/L96I/L98F/N194D/V210A/I218T, N52S/H94E/L96I/V122M, N52H/N57Y/H94E/L96I/F120I/S126T/W153R/I218N, M10V/S18R/N30D/N52S/S126R/T139S/L203F, S25G/N30D/N52S/F120S/N227K, N30D/N52S/L67P/Q100K/D217G/R221K/T225S, N52H/N57Y/Q100R/V110D/A117T/T190S/C198R, N52H/N57Y/Q100R/V110D/F172S/C198R, S25G/F27C/N52H/N57Y/Q100R/V110D/E135K/L173S/C198R, N52H/N57Y/V110A/C198R/R221I, M1I/S13G/N52H/N57Y/D77G/V110A/H129P/I143V/F172S/V193M, C198R, N52H/N57Y/R61C/Y62F/Q100R/V110N/F120S/C198R, N52H/N57Y/Q100R/V110D/H115R/C198R, N52H/N57Y/Q100R/V110D/N144D/F172S/C198R, N52S/H94E/L98F/Q100R, N52S/E90A, N30D/K42E/N52S, N52S/F120S/I143V/I224V, N52H/N57Y/Q100R/V110D/C198R/S212G, N52H/N57Y/Q100R/C198R, N52S/N194D, N52H/N57Y/Q100R/L102R/V110D/H115R/C198R, N52H/N57Y/Q100R/V110D/C198R/S212G, N52H/N57Y/Q100R/C198R, N52S/N194D, N52H/N57Y/Q100R/L102R/V110D/H115R/C198R, N52S/S54P, T38P/N52S/N57D, N52H/C140del/T225A, N52H/F78L/Q100R/C198R, N52H/N57Y/R75Q/Q100P/V110D, N52H/N57Y/L74Q/V110D/S192G, N52H/S121G/C198R, N52S/F120S/N227K, N52S/A71T/A117T/T190A/C198R, T43A/N52H/N57Y/L74Q/D89G/V110D/F172S, N52H/N57Y/Q100R/V110D/S132F/M175T, N52H/N57Y/Q100R/V107I/V110D/I154F/C198R/R221G, N52Q/N207Q, N52Q/N168Q, N52Q/N84Q, N52Q/N119Q, N52Q/N84Q/N168Q, N52Q/N84Q/N207Q, N52Q/N119Q/N155Q, N52H/N84Q/N119Q, N52H/N84Q, N52H/N84Q/N168Q/N207Q, N52Q/N84Q/N155Q/N168Q, N52Q/N84Q/N119Q/N168Q, N52Q/N84Q/N119Q/N207Q, N52Q/N84Q/N119Q/N155Q, N52Q/N84Q/N119Q/N155Q/N207Q, N52Y/F138L/L203P, N57Y/Q100R/C198R, N57Y/F138L/L203P, Q100R/F138L, N52H/N57Y/Q100R/H115R/C198R, N52H/N57Y/Q100R/F172S/C198R, N52H/N57Y/Q100R/H115R/F172S/C198R, N52H/N57Y/Q100R/H115R/I143V/F172S/C198R, N52H/N57Y/Q100R/L102R/H115R/F172S/C198R, N52H/V122A/F172S/C198R, N52H/N57Y/Q100R/H115R/F172S/N194D, N52H/N57Y/H115R/F172S/C198R, N52H/N57Y/H115R, N52H/N57Y/Q100R/H115R, N52H/N57Y/Q100R/H115R/F172S/I224V, N52H/N57Y/Q100R/H115R/F172S, N52H/N57Y/Q100R/F172S, N52H/Q100R/H115R/I143T/F172S, N52H/N57Y/Q100P/H115R/F172S, N52Y/N57Y/Q100P/F172S, E16V/N52H/N57Y/Q100R/V110D/H115R/C198R, E16V/N52H/N57Y/Q100R/V110D/H115R/Y152C/K156M/F172S/C198R, N52S/E90A/H115R, N30D/K42E N52S/H115R, N30D/K42E/N52S/H115R/C198R/R221I, N30D/K42E/N52S/H115R/C198R, N30D/K42E/N52S/H115R/F172S/N194D, N52S/H115R/F120S/I143V/C198R, N52S/H115R/F172S/C198R, N52H/N57Y/Q100P/C198R, N52H/N57Y/Q100P/H115R/F172S/C198R, N52H/N57Y/Q100P/F172S/C198R, N52H/N57Y/Q100P/H115R, N52H/N57Y/Q100P/H115R/C198R, N52H/Q100R/C198R, N52H/Q100R/H115R/F172S, N52H/Q100R/F172S/C198R, N52H/Q100R/H115R/F172S/C198R, N52H/N57Y/Q100R/F172S/C198R, N52A/N57F/Q100S, N52A/N57H/Q100S, N52A/N57Y/Q100A, N52D/N57A/Q100A, N52D/Q100S, N52G/Q100A, N52H/Q100A, N52M/N57H/Q100S, N52M/N57W/Q100P, N52Q/N57F, N52Q/N57S/Q100A, N52R/N57L/Q100A, N52R/N57Y/Q100P, N52R/N57Y/Q100S, N52S/N57A/Q100A, N52S/N57H/Q100E, N52S/N57L/Q100S, N52S/N57M/Q100S, N52S/N57Y/Q100S, N52S/N57Y/Q100M, N52S/N57Y/Q100V, N52T/N57H/Q100S, N52T/N57H/Q100A, N52T/N57Y/Q100A, N52V/N57L/Q100A, N52H/N57Y/Q100K, N52K/N57Y/Q100R, N52L/N57H/Q100R, N52R/N57F/Q100N, N52R/N57F/Q100P, N52R/N57F/Q100R, N52R/N57F/Q100T, N52R/N57H/Q100K, N52R/N57L/Q100S, N52R/N57W/Q100K, N52R/N57W, N52R/N57Y/Q100R, N52C/N57E/Q100S, N52G/N57P/Q100D, N52G/N57V/Q100G, N52G/N57V, N52L/N57V, N52P/N57P, N52P/N57S/Q100G, N52S/N57L/Q100G, N52T/N57K/Q100P, N52V/N57T/Q100L, N57Q/Q100P, or R26S/N52H/N57Y/V110D/T137A/C198R, with reference to numbering of SEQ ID NO:32.
28 . A variant ICOS Ligand (ICOSL) polypeptide, comprising an IgV domain or specific binding fragment thereof, an IgC domain or a specific binding fragment thereof, or both, wherein the variant ICOSL polypeptide comprises one or more amino acid modifications in an ICOSL reference polypeptide or a specific binding fragment thereof corresponding to amino acid modifications are selected from N52A, N52C, N52G, N52K, N52L, N52M, N52R, N52T, N52V, N57A, N57E, N57F, N57H, N57K, N57L, N57M, N57P, N57Q, N57S, N57T, N57V, N57W, Q100A, Q100D, Q100G, Q100L, Q100M, Q100N, Q100S, Q100T or Q100V with reference to SEQ ID NO:32.
29 . The variant ICOSL polypeptide of claim 28 , wherein the one or more amino acid modifications are selected from among N52A/N57F/Q100S, N52A, /N57H/Q100S, N52A/N57Y/Q100A, N52D/N57A/Q100A, N52D/Q100S, N52G/Q100A, N52H/Q100A, N52M/N57H/Q100S, N52M/N57W/Q100P, N52Q/N57F, N52Q/N57S/Q100A, N52R/N57L/Q100A, N52R/N57Y/Q100P, N52R/N57Y/Q100S, N52S/N57A/Q100A, N52S/N57H/Q100E, N52S/N57L/Q100S, N52S/N57M/Q100S, N52S/N57Y/Q100S, N52S/N57Y/Q100M, N52S/N57Y/Q100V, N52T/N57H/Q100S, N52T/N57H/Q100A, N52T/N57Y/Q100A, N52V/N57L/Q100A, N52H/N57Y/Q100K, N52K/N57Y/Q100R, N52L/N57H/Q100R, N52R/N57F/Q100N, N52R/N57F/Q100P, N52R/N57F/Q100R, N52R/N57F/Q100T, N52R/N57H/Q100K, N52R/N57L/Q100S, N52R/N57W/Q100K, N52R/N57W, N52R/N57Y/Q100R, N52C/N57E/Q100S, N52G/N57P/Q100D, N52G/N57V/Q100G, N52G/N57V, N52L/N57V, N52P/N57P, N52P/N57S/Q100G, N52S/N57L/Q100G, N52T/N57K/Q100P, N52V/N57T/Q100L or N57Q/Q100P.
30 . The variant ICOSL polypeptide of claim 28 or claim 29 , wherein the ICOSL reference polypeptide comprises (i) the sequence of amino acids set forth in SEQ ID NO:32, (ii) a sequence of amino acids that has at least 95% sequence identity to SEQ ID NO:32; or (iii) a portion of (i) or (ii) comprising an IgV domain or IgC domain or specific binding fragments thereof or both.
31 . The variant ICOSL polypeptide of any of claims 28 - 30 , wherein the variant ICOSL polypeptide comprises the IgV domain or a specific binding fragment thereof.
32 . The variant ICOSL polypeptide of any of claims 28 - 31 , wherein the IgV domain or specific binding fragment thereof is the only ICOSL portion of the variant ICOSL polypeptide.
33 . The variant ICOSL polypeptide of any of claims 28 - 32 , wherein the ICOSL reference polypeptide comprises the sequence of amino acids set forth in SEQ ID NO:545.
34 . The variant ICOSL polypeptide of any of claims 28 - 32 , wherein the ICOSL reference polypeptide consists of the sequence of amino acids set forth in SEQ ID NO:545.
35 . The variant ICOSL polypeptide of any of claims 1 - 34 , wherein the variant ICOSL polypeptide exhibits increased binding to the ectodomain(s) of ICOS or CD28 compared to the binding of the ICOSL reference polypeptide for the same ectodomain(s).
36 . The variant ICOSL polypeptide of any of claims 1 - 34 , wherein the variant ICOSL polypeptide exhibits increased binding to the ectodomain(s) of ICOS and CD28 compared to the binding of the ICOSL reference polypeptide for the same ectodomain(s).
37 . The variant ICOSL polypeptide of any of claims 2 - 6 , 9 - 27 , 35 and 36 , wherein the binding is increased more than 1.2-fold, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold or 60-fold.
38 . The variant ICOSL polypeptide of any of claims 2 - 6 , 9 - 27 , 35 , 36 and 37 , wherein the ICOS is a human ICOS.
39 . The variant ICOSL polypeptide of any of claims 2 - 6 , 9 - 27 , 35 , 36 , 37 and 38 , wherein the CD28 is a human CD28.
40 . The variant ICOSL polypeptide of any of claims 1 - 39 , wherein the variant ICOSL polypeptide comprises up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 amino acid modifications, optionally amino acid substitutions, insertions and/or deletions.
41 . The variant ICOSL polypeptide of any of claims 1 - 40 that is a soluble protein.
42 . The variant ICOSL polypeptide of any of claims 1 - 41 , wherein:
the variant ICOSL polypeptide lacks a transmembrane domain and intracellular signaling domain; and/or when expressed from a cell, the variant ICOSL polypeptide is not expressed on the surface of the cell.
43 . The variant ICOSL polypeptide of any of claims 1 - 40 , wherein the variant ICOSL polypeptide further comprises a transmembrane domain.
44 . The variant ICOSL polypeptide of claim 43 , further comprising a cytoplasmic signaling domain linked to the transmembrane domain.
45 . An immunomodulatory protein, comprising the variant ICOSL polypeptide of any of claims 1 - 44 and a half-life extending moiety.
46 . The immunomodulatory protein of claim 45 , wherein the half-life extending moiety comprises a multimerization domain, albumin, an albumin-binding polypeptide, Pro/Ala/Ser (PAS), a C-terminal peptide (CTP) of the beta subunit of human chorionic gonadotropin, polyethylene glycol (PEG), long unstructured hydrophilic sequences of amino acids (XTEN), hydroxyethyl starch (HES), an albumin-binding small molecule, or a combination thereof.
47 . The immunomodulatory protein of claim 45 or claim 46 , wherein the half-life extending moiety is or comprises a multimerization domain.
48 . The immunomodulatory protein of claim 47 , wherein the multimerization domain is or comprises an Fc region of an immunoglobulin.
49 . The immunomodulatory protein of claim 47 or claim 48 , wherein the variant ICOSL polypeptide is linked, directly or indirectly via a linker, to the multimerization domain.
50 . The immunomodulatory protein of any of claims 47 - 49 , wherein the immunomodulatory protein is a multimer comprising a first variant ICOSL polypeptide linked to a first multimerization domain and a second variant ICOSL polypeptide linked to a second multimerization domain, wherein the first and second multimerization domains interact to form a multimer comprising the first and second variant ICOSL polypeptide.
51 . The immunomodulatory protein of claim 50 , wherein the multimer is a dimer.
52 . The immunomodulatory protein of claim 50 or claim 51 , wherein the first variant ICOSL polypeptide and the second variant ICOSL polypeptide are the same.
53 . The immunomodulatory protein of claim 51 or claim 52 , wherein the dimer is a homodimer.
54 . The immunomodulatory protein of claim 51 , wherein the dimer is a heterodimer.
55 . The immunomodulatory protein of any of claims 48 - 54 , wherein the Fc region is a human IgG or is a variant Fc region comprising one or more amino acid substitutions compared to the wildtype human IgG1.
56 . The immunomodulatory protein of any of claims 48 - 55 , wherein the Fc region comprises the sequence of amino acids set forth in SEQ ID NO: 226 or a variant thereof that exhibits at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO:226.
57 . The immunomodulatory protein of any of claims 48 - 56 , wherein the Fc region exhibits one or more effector functions.
58 . The immunomodulatory protein of any of claims 48 - 56 , wherein the Fc region is a variant Fc region that exhibits one or more reduced effector function compared to an Fc of a wildtype human IgG1.
59 . The immunomodulatory protein of claim 58 , wherein the variant Fc region comprises one or more amino acid substitutions selected from N297G, E233P/L234V/L235A/G236del/S267K or L234A/L235E/G237A, wherein the residue is numbered according to the EU index of Kabat.
60 . The immunomodulatory protein of claim 59 , wherein the variant Fc region further comprises the amino acid substitution C220S, wherein the residues are numbered according to the EU index of Kabat.
61 . The immunomodulatory protein of claim 59 or claim 60 , wherein the Fc region comprises K447del, wherein the residue is numbered according to the EU index of Kabat.
62 . The immunomodulatory protein of any of claims 59 - 61 , wherein the Fc region comprises the sequence of amino acids set forth in SEQ ID NO:476 or SEQ ID NO:632.
63 . The immunomodulatory protein of any of claims 59 - 61 , wherein the Fc region comprises the sequence of amino acids set forth in SEQ ID NO:478 or SEQ ID NO:634.
64 . The immunomodulatory protein of any of claims 59 - 61 , wherein the Fc region comprises the sequence of amino acids set forth in SEQ ID NO:477.
65 . The immunomodulatory protein of any of claims 59 - 61 , wherein the Fc region comprises the sequence of amino acids set forth in SEQ ID NO:633.
66 . The immunomodulatory protein of any of claims 59 - 61 , wherein the Fc region comprises the sequence of amino acids set forth in SEQ ID NO:474.
67 . The immunomodulatory protein of any of claims 59 - 61 , wherein the Fc region comprises the sequence of amino acids set forth in SEQ ID NO:637.
68 . The immunomodulatory protein of any of claims 49 - 67 , wherein the variant ICOSL polypeptide is linked via a linker to the Fc region.
69 . The immunomodulatory protein of any of claims 49 - 68 , wherein the linker comprises 1 to 10 amino acids.
70 . The immunomodulatory protein of any of claims 49 - 69 , wherein the linker is AAA.
71 . The immunomodulatory protein of any of claims 49 - 69 , wherein the linker is G4S (SEQ ID NO:636).
72 . The immunomodulatory protein of any of claims 49 - 69 , wherein the linker is (G 4 S) 2 (SEQ ID NO:229).
73 . The immunomodulatory protein of any of claims 49 - 69 , wherein the linker is GSGGGGS linker (SEQ ID NO: 635).
74 . An immunomodulatory protein, comprising the variant ICOSL polypeptide of any of claims 1 - 44 linked to a second polypeptide comprising an immunoglobulin superfamily (IgSF) domain.
75 . The immunomodulatory polypeptide of claim 74 , wherein the IgSF domain of the second polypeptide exhibits increased binding to one or more of its cognate binding partner(s) compared to the unmodified or wild-type IgSF domain.
76 . The immunomodulatory protein of claim 74 or claim 75 , wherein the variant ICOSL polypeptide is capable of specifically binding to CD28 or ICOS and the IgSF domain of the second polypeptide is capable of binding to a binding partner other than one specifically bound by the ICOSL variant polypeptide.
77 . The immunomodulatory polypeptide of any of claims 74 - 76 , wherein the IgSF domain of the second polypeptide is a tumor-localizing moiety that binds to a ligand expressed on a tumor.
78 . The immunomodulatory polypeptide of claim 77 , wherein the ligand is B7H6.
79 . The immunomodulatory polypeptide of claim 77 or claim 78 , wherein the IgSF domain is from NKp30.
80 . The immunomodulatory polypeptide of any of claims 74 - 79 , wherein the IgSF domain of the second polypeptide is or comprises an IgV domain.
81 . The immunomodulatory polypeptide of any of claims 74 - 80 , wherein the IgSF domain of the second polypeptide has the sequence set forth in SEQ ID NO:504.
82 . The immunomodulatory polypeptide of any of claims 45 - 81 , wherein the variant ICOSL polypeptide is or comprise an IgV domain.
83 . The immunomodulatory polypeptide of any of claims 45 - 82 , wherein the variant ICOSL polypeptide comprises amino acid modifications N52H/Q100R.
84 . The immunomodulatory polypeptide of claim 83 , wherein the variant ICOSL polypeptide has the sequence set forth in SEQ ID NO:567.
85 . The immunomodulatory polypeptide of any of claims 45 - 83 , wherein the variant ICOSL polypeptide comprises amino acid modifications N52H/N57Y/Q100R.
86 . The immunomodulatory polypeptide of claim 85 , wherein the variant ICOSL polypeptide has the sequence set forth in SEQ ID NO:565.
87 . The immunomodulatory polypeptide of any of claims 45 - 82 , wherein the variant ICOSL polypeptide comprises amino acid modifications are N52L/N57H/Q100R.
88 . The immunomodulatory polypeptide of claim 87 , wherein the variant ICOSL polypeptide has the sequence set forth in SEQ ID NO:761.
89 . The immunomodulatory polypeptide of any of claims 45 - 82 , wherein the variant ICOSL polypeptide comprises the amino acid modification is N52D.
90 . The immunomodulatory polypeptide of claim 89 , wherein the variant ICOSL polypeptide has the sequence set forth in SEQ ID NO:548.
91 . The immunomodulatory protein of any of claims 74 - 90 , wherein the immunomodulatory protein comprises a multimerization domain linked to one or both of the variant ICOSL polypeptide or the second polypeptide comprising the IgSF domain.
92 . The immunomodulatory protein of claim 91 , wherein the multimerization domain is an Fc region.
93 . The immunomodulatory protein of any of claims 74 - 92 that is dimeric.
94 . The immunomodulatory protein of claim 93 that is homodimeric.
95 . The immunomodulatory protein of claim 93 that is heterodimeric.
96 . The immunomodulatory protein of any of claims 92 - 95 , wherein the Fc region is a human IgG or is a variant Fc region comprising one or more amino acid substitutions compared to the wildtype human IgG1.
97 . The immunomodulatory protein of any of claims 92 - 96 , wherein the Fc region comprises the sequence of amino acids set forth in SEQ ID NO: 226 or a variant thereof that exhibits at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO:226.
98 . The immunomodulatory protein of any of claims 92 - 97 wherein the Fc region exhibits one or more effector functions.
99 . The immunomodulatory protein of any of claims 92 - 97 , wherein the Fc region is a variant Fc region that exhibits one or more reduced effector function compared to an Fc of a wildtype human IgG1.
100 . The immunomodulatory protein of claim 99 , wherein the variant Fc region comprises one or more amino acid substitutions selected from N297G, E233P/L234V/L235A/G236del/S267K or L234A/L235E/G237A, wherein the residue is numbered according to the EU index of Kabat.
101 . The immunomodulatory protein of claim 100 , wherein the variant Fc region further comprises the amino acid substitution C220S, wherein the residues are numbered according to the EU index of Kabat.
102 . The immunomodulatory protein of claim 100 or claim 101 , wherein the Fc region comprises K447del, wherein the residue is numbered according to the EU index of Kabat.
103 . The immunomodulatory protein of any of claims 100 - 102 , wherein the Fc region comprises the sequence of amino acids set forth in SEQ ID NO:476 or SEQ ID NO:632.
104 . The immunomodulatory protein of any of claims 100 - 102 , wherein the Fc region comprises the sequence of amino acids set forth in SEQ ID NO:478 or SEQ ID NO:634.
105 . The immunomodulatory protein of any of claims 100 - 102 , wherein the Fc region comprises the sequence of amino acids set forth in SEQ ID NO:477.
106 . The immunomodulatory protein of any of claims 100 - 102 , wherein the Fc region comprises the sequence of amino acids set forth in SEQ ID NO:633.
107 . The immunomodulatory protein of any of claims 100 - 102 , wherein the Fc region comprises the sequence of amino acids set forth in SEQ ID NO:474.
108 . The immunomodulatory protein of any of claims 100 - 102 , wherein the Fc region comprises the sequence of amino acids set forth in SEQ ID NO:637.
109 . The immunomodulatory protein of any of claims 74 - 108 , wherein the variant ICOSL polypeptide and the IgSF domain of the second polypeptide are linked by a linker.
110 . The immunomodulatory protein of claim 109 , wherein the linker is 3×GGGGS (SEQ ID NO: 228).
111 . The immunomodulatory protein of any of claims 91 - 110 , wherein the multimerization domain is linked via a linker to one or both of the variant ICOSL polypeptide or the second polypeptide comprising the IgSF domain.
112 . The immunomodulatory protein of claim 111 , wherein the linker is GSGGGGS (SEQ ID NO: 635).
113 . A conjugate comprising the variant ICOSL polypeptide of any of claims 1 - 44 or immunomodulatory protein of any of claims 45 - 112 and a heterologous moiety.
114 . The conjugate of claim 113 , wherein the conjugate is a fusion protein.
115 . The conjugate of claim 113 or claim 114 , wherein the moiety is a targeting moiety that specifically binds to a molecule on the surface of a cell.
116 . The conjugate of claim 115 , wherein the targeting moiety specifically binds to a molecule on the surface of an immune cell.
117 . The conjugate of claim 116 , wherein the immune cell is an antigen presenting cell or a lymphocyte.
118 . The conjugate of claim 115 , wherein the targeting moiety is a tumor-localizing moiety that binds to a molecule on the surface of a tumor.
119 . The conjugate of any of claims 115 - 118 , wherein the targeting moiety is an antibody or antigen-binding fragment.
120 . The conjugate of claim 119 , wherein the antibody is selected from cetuximab, panitumumab, zalutumumab, nimotuzumab, trastuzumab, Ado-trastuzumab emtansine, Tositumomab (Bexxar®), Rituximab (Rituxan, Mabthera), Ibritumomab tiuxetan (Zevalin), Daclizumab (Zenapax), Gemtuzumab (Mylotarg), Alemtuzumab, CEA-scan Fab fragment, OC125 monoclonal antibody, ab75705, B72.3, Bevacizumab (Avastin®), Afatinib, Axitinib, Bosutinib, Cabozantinib, Ceritinib, Crizotinib, Dabrafenib, Dasatinib, Dinutuximab, Erlotinib, Everolimus, Ibrutinib, Imatinib, Lapatinib, Lenvatinib, Nilotinib, Olaparib, Olaratumab, Palbociclib, Pazopanib, Pertuzumab, Ramucirumab, Regorafenib, Ruxolitinib, Sorafenib, Sunitinib, Temsirolimus, Trametinib, Vandetanib, Vemurafenib, Vismodegib, Basiliximab, Ipilimumab, Nivolumab, pembrolizumab, MPDL3280A, Pidilizumab (CT-011), AMP-224, MSB001078C, or MEDI4736, BMS-935559, LY3300054, atezolizumab, avelumab or durvalumab or is an antigen-binding fragment thereof.
121 . A monovalent fusion protein comprising:
(a) a variant ICOSL polypeptide of any of claims 1 - 44 ; and (b) a label for detection or purification of the variant ICOSL polypeptide.
122 . The fusion protein of claim 121 , wherein the label for detection or purification is selected from a poly-histidine (His) tag, a FLAG-tag, a Myc-tag, or a fluorescent protein-tag.
123 . A nucleic acid molecule(s), encoding a variant ICOSL polypeptide of any of claims 1 - 44 , an immunomodulatory protein of any of claims 45 - 112 or a fusion protein of any of claims 114 - 122 .
124 . A vector, comprising the nucleic acid molecule(s) of claim 123 .
125 . A cell, comprising the nucleic acid molecule(s) of claim 123 or the vector of claim 124 .
126 . A method of producing an immunomodulatory protein comprising a variant ICOSL polypeptide, comprising introducing the nucleic acid molecule of claim 123 or vector of claim 124 into a host cell under conditions to express the protein in the cell.
127 . The method of claim 126 that is a mammalian cell.
128 . The cell of claim 126 or claim 127 that is a Chinese Hamster Ovary (CHO) cell or a derivative thereof.
129 . The cell of any of claims 126 - 128 that is CHO DG44.
130 . The method of any of claims 126 - 129 , further comprising isolating or purifying the protein from the cell.
131 . A protein produced by the method of any of claims 126 - 130 .
132 . A composition comprising a protein comprising a variant ICOSL polypeptide of any of claims 1 - 44 or an immunomodulatory protein of any of claims 45 - 112 , wherein at least 95%, 96%, 97%, 98%, 99% of the individual sequences of the protein or the immunomodulatory protein in the composition have an identical sequence length.
133 . The composition of claim 132 , wherein the protein or immunomodulatory protein is purified from Chinese Hamster Ovary Cells or a derivative thereof.
134 . A polynucleotide comprising a nucleic acid encoding a variant ICOSL polypeptide comprising a transmembrane domain of claim 43 or claim 44 and one or more nucleic acid encoding one or more chain of a recombinant antigen receptor.
135 . The polynucleotide of claim 134 , wherein the recombinant antigen receptor is a chimeric antigen receptor (CAR) or an engineered T cell receptor (TCR).
136 . The polynucleotide of claim 134 or claim 135 , wherein each of the nucleic acid encoding the variant ICOSL polypeptide and the one or more nucleic acid encoding one or more chain of the recombinant receptor is separated by a nucleic acid encoding a self-cleaving peptide or a peptide that causes ribosome skipping.
137 . An engineered cell comprising the variant ICOSL polypeptide of any of claims 1 - 44 , the immunomodulatory protein of any of claims 45 - 112 , or the fusion protein of any of claims 114 - 122 .
138 . The engineered cell of claim 137 , wherein:
the nucleic acid encoding the variant ICOSL polypeptide, immunomodulatory protein or fusion protein encodes a signal peptide; the variant ICOSL polypeptide, immunomodulatory protein or fusion protein does not comprise a transmembrane domain and/or is not expressed on the surface of the cell; and/or the variant ICOSL polypeptide, immunomodulatory protein or fusion protein is secreted from the engineered cell.
139 . The engineered cell of claim 137 , wherein the engineered cell comprises a variant ICOSL polypeptide comprising a transmembrane domain of claim 43 or claim 44 .
140 . The engineered cell of any of claims 137 - 139 , wherein the cell is an immune cell.
141 . The engineered cell of claim 140 , wherein the immune cell is an antigen presenting cell (APC) or a lymphocyte.
142 . The engineered cell of any of claims 137 - 141 that is a primary human cell.
143 . The engineered cell of any of claims 137 - 142 , further comprising a chimeric antigen receptor (CAR) or an engineered T-cell receptor.
144 . An infectious agent, comprising a nucleic acid molecule encoding a variant ICOSL polypeptide of any of claims 1 - 44 or an immunomodulatory protein of any of claims the immunomodulatory protein of any of claims 45 - 112 , or the fusion protein of any of claims 114 - 122 .
145 . The infectious agent of claim 144 , wherein the infectious agent is a bacteria or a virus.
146 . A pharmaceutical composition, comprising the variant ICOSL polypeptide of any of claims 1 - 44 , the immunomodulatory protein of any of claims 45 - 112 , a conjugate or fusion protein of any of claims 113 - 122 or an engineered cell of any of claims 137 - 143 or an infectious agent of claim 144 or claim 145 .
147 . The pharmaceutical composition of claim 146 , comprising a pharmaceutically acceptable excipient.
148 . An article of manufacture comprising the pharmaceutical composition of claim 146 or claim 147 in a vial.
149 . A kit comprising the composition of claim 146 or claim 147 or the article of manufacture of claim 148 , and instructions for use.
150 . A method of modulating an immune response in a subject, comprising administering the pharmaceutical composition of claim 146 or claim 147 to the subject.
151 . A method of modulating an immune response in a subject, comprising administering the engineered cells of any of claims 137 - 143 to the subject.
152 . The method of claim 151 , wherein the engineered cells are autologous to the subject.
153 . The method of claim 151 , wherein the engineered cells are allogenic to the subject.
154 . The method of any of claims 150 - 153 , wherein modulating the immune response treats a disease or condition in the subject.
155 . A method of treating a disease or condition in a subject, the method comprising administering the pharmaceutical composition of claim 146 or claim 147 to the subject.
156 . The method of any of claims 150 - 155 , wherein the immune response is increased in the subject.
157 . The method of any of claims 150 and 154 - 156 , wherein the pharmaceutical composition comprises an immunomodulatory protein or conjugate comprising a variant ICOSL polypeptide linked to a tumor-localizing moiety.
158 . The method of claim 157 , wherein the tumor-localizing moiety is or comprises a binding molecule that recognizes a tumor antigen.
159 . The method of any of claims 150 and 154 - 158 , wherein the pharmaceutical composition comprises the immunomodulatory protein of any of claims 77 - 112 or the conjugate or fusion protein of any of claims 113 - 120 is administered to the subject.
160 . The method of any of claims 150 - 156 , wherein the pharmaceutical composition comprises an engineered cell comprising a variant ICOSL polypeptide that is a transmembrane immunomodulatory protein of claim 43 or claim 44 .
161 . The method of any of claims 154 - 160 , wherein the disease or condition is a tumor or cancer.
162 . The method of any one of claims 154 - 161 , wherein the disease or condition is selected from melanoma, lung cancer, bladder cancer, a hematological malignancy, liver cancer, brain cancer, renal cancer, breast cancer, pancreatic cancer, colorectal cancer, spleen cancer, prostate cancer, testicular cancer, ovarian cancer, uterine cancer, gastric carcinoma, a musculoskeletal cancer, a head and neck cancer, a gastrointestinal cancer, a germ cell cancer, or an endocrine and neuroendocrine cancer.
163 . The method of any of claims 150 - 155 , wherein the immune response is decreased.
164 . The method of any of claims 150 - 155 and 163 , wherein the pharmaceutical composition comprises a variant ICOSL polypeptide immunomodulatory protein that is an immunomodulatory Fc fusion protein.
165 . The method of any of claims 150 - 155 , 163 and 164 , wherein the pharmaceutical composition comprises a variant ICOSL polypeptide of any of claims 1 - 42 or an immunomodulatory protein of any of claims 45 - 76 .
166 . The method of any of claims 150 - 155 and 163 , wherein the pharmaceutical composition comprises an engineered cell comprising a secretable variant ICOSL polypeptide is administered to the subject.
167 . The method of any of claims 150 - 155 , 163 and 166 , wherein the pharmaceutical composition comprises an engineered cell of claim 137 or claim 138 .
168 . The method of any of claims 150 - 155 and 163 , wherein the pharmaceutical composition comprises an infectious agent encoding a variant ICOSL polypeptide that is a secretable immunomodulatory protein is administered to the subject, optionally under conditions in which the infectious agent infects a tumor cell or immune cell and the secretable immunomodulatory protein is secreted from the infected cell.
169 . The method of any of claims 154 - 155 and 163 - 168 , wherein the disease or condition is an inflammatory or autoimmune disease or condition.
170 . The method of any of claims 154 - 155 and 163 - 169 , wherein the disease or condition is an Antineutrophil cytoplasmic antibodies (ANCA)-associated vasculitis, a vasculitis, an autoimmune skin disease, transplantation, a Rheumatic disease, an inflammatory gastrointestinal disease, an inflammatory eye disease, an inflammatory neurological disease, an inflammatory pulmonary disease, an inflammatory endocrine disease, or an autoimmune hematological disease.
171 . The method of claim 169 or claim 170 , wherein the disease or condition is selected from inflammatory bowel disease, transplant, Crohn's disease, ulcerative colitis, multiple sclerosis, asthma, rheumatoid arthritis, or psoriasis.Cited by (0)
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