Il-18 polypeptides and uses thereof

Abstract

Provided herein are IL- 18 polypeptide variants that have binding specificity for the IL- 18 receptor. Also provided herein are fusion proteins, and conjugates comprising the IL- 18 polypeptide variants.

Description

[0001] Attorney Docket No.: 761146.272320 IL-18 POLYPEPTIDES AND USES THEREOF

[0001] The present application claims the benefit of U.S. Provisional Application No.: 63 / 618,829, filed on January 8, 2024, and U.S. Provisional Application No.: 63 / 566,650, filed on March 18, 2024, and U.S. Provisional Application No.: 63 / 575,078, filed on April 5, 2024, each of which are incorporated by reference in their entirety. 1. BACKGROUND

[0002] Interleukin-18 (IL-18) is a pro-inflammatory cytokine that plays a crucial role in the immune system. It is known for its ability to induce cell-mediated immunity following infections by various pathogens. IL-18 is involved in numerous immune responses, including the production of interferon- gamma (IFN-γ) in T-cells and natural killer cells. See, Ihim et al., (2022), Front Immunol., 13:919973.

[0003] IL-18 has potential as an anticancer drug, but its interactions with the inhibitory decoy receptor IL-18 binding protein (IL-18BP), which is frequently upregulated in tumors, has limited its clinical utility as an antitumor agent. See, Deckers et al., (2023), Nature, 1:286-3033.

[0004] As such, there remains a need for IL-18 variants that demonstrate improved properties for clinical and research use. 2. SUMMARY

[0005] The disclosure relates to variants of IL-18 that have binding specificity for the IL-18 receptor, methods of making, and uses thereof. Such variants of IL-18 can be used, for example, as a therapeutic agents. Also provided herein are IL-18-scFc (single chain fragment crystallizable) fusion proteins that include an IL-18 polypeptide as disclosed herein operably linked to a single chain Fc domain.

[0006] Human IL-18 polypeptides disclosed herein can comprise SEQ ID NO: 1 or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 1, in which the amino acid at position 1 is tryptophan.

[0007] Human IL-18 polypeptides disclosed herein can comprise SEQ ID NO: 1 or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 1, in which the amino acid at position 3 is not glycine, the amino acid at position 6 is not glutamic acid, the amino acid at position 10 is not serine, the amino acid at position 49 is not isolucine, the amino acid at position 54 is not aspartic acid, the amino acid at position 62 is not valine, and / or the amino acid at position 91 is not asparagine.

[0008] The amino acid position 3 can be selected from the group consisting of proline, aspartic acid, arginine, glutamine, and valine. The amino acid at position 6 can be selected from the group consisting of 1 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 arginine, glutamine, asparagine, tryptophan, and lysine. The amino acid at position 10 can be selected from the group consisting of S10L, S10N, S10A, S10T, S10Q, S10H, S10L, S10I, S10H, S10A, S10K, S10R, S10G, S10F, and S10V. The amino acid substitution at position 49 can be selected from the group consisting of I49L, I49T, I49Q, I49K, I49R, and I49S. The amino acid substitution at position 54 can be selected from the group consisting of D54F, D54P, and D54H. The amino acid substitution at position 56 can be Q56P. The amino acid substitution at position 57 can be P57H. The amino acid substitution at position 62 can be selected from the group consisting of V62S, V62A, and V62L. The amino acid at position 91 can be selected from the group consisting of N91R, N91L, N91M, N91Y, N91L, N91T, N91Y, N91K, N91G, N91W, N91A, N91I, N91V, N91S, and N91H.

[0009] Human IL-18 polypeptides disclosed herein can comprise SEQ ID NO: 1 or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 1, in which the amino acid at position 40 is asparagine or glutamic acid.

[0010] Human IL-18 polypeptides disclosed herein can comprise SEQ ID NO: 1 or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 1, in which the amino acid at position 56 is proline.

[0011] Human IL-18 polypeptides disclosed herein can comprise SEQ ID NO: 1 or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 1, in which the amino acid at position 57 is histidine.

[0012] IL-18 polypeptides disclosed herein have binding specificity for the human IL-18 receptor.

[0013] The IL-18 polypeptides disclosed herein (e.g., SEQ ID NO: 1) can comprises amino acid substitutions selected from the group consisting of: a) S10X, I49X, and N91X; b) K53X, M51X, and Y1X; c) M60X, Q56X, and P57X; d) S10X and I49X; e) S10X and I49X; f) V62X and N91X; g) I49X and N91X; h) S10X and N91X; i) M60X and N9X; j) D54X and G3X; k) E6X and G3X; l) D40X and E6X; m) K53X and M51; n) S10X; o) V62X; p) N91X; q) I49X; r) D54X; s) E6X; t) G3X; or u) D40X.

[0014] The IL-18 polypeptides disclosed herein (e.g., SEQ ID NO: 1) can comprise amino acid substitutions comprising a) S10L, I49L, and N91R; b) S10L, I49Q, and N91Y; c) S10I, I49K, and N91K; d) Q56P, P57H, and M60K; e) S10A, I49L, and V62Y; f) Y1W, M51R, K53S; and / or g) S10T, I49T, and N91L.

[0015] IL-18 polypeptides disclosed herein (e.g., SEQ ID NO: 1) can comprise amino acid substitutions comprising a) S10A and I49L; b) S10R and I49T; c) S10I, I49K; d) V62A and N91M; e) I49L and N91A; f) I49T and N91R; g) I49I and N91K; h) I49R and N91Y; i) I49S and N91R; j) S10H and N91Y; k) S10H and N91L; l) S10H and N91R; m) S10H and N91T; n) S10T and N91L; o) S10T and N91R; p) S10F and N91M; q) G3P and D45P; r) G3D and D54F; s) G3P and 6EQ; t) G3Q and 6EW; u) E6K and 2 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 D40N; v) K53S and M51R; w) K53S and M51K; x) K53T and M51R; y) M60K and N91L; z) G3P and D54H; or za) G3R and E6N.

[0016] The IL-18 polypeptides disclosed herein (e.g., SEQ ID NO: 1) can comprise amino acid substitutions comprising S10R and I49T, S10H and N91T, or G3R and E6N.

[0017] The IL-18 polypeptides disclosed herein (e.g., SEQ ID NO: 1) can comprise amino acid substitutions comprising

[0018] a) S10N; b) S10H; c) S10G; d) S10Q; e) S10K; f) S10R; g) V62S; h) N91G; i) N91R; j) N91W; k) N91L; l) N91T; m) N91I; n) N91V; o) N91S; p) N91M; q) N91K; r) N91Y; s) N91K; t) N91A; u) I49R; v) D54P; w) E6Q; x) G3R; y) G3V; z) D40N; and za) D40E.

[0019] The IL-18 polypeptides disclosed herein (e.g., SEQ ID NO: 1) can comprise one amino acid substitutions at positions S10K, N91V, or I49R.

[0020] The IL-18 polypeptides disclosed herein can comprise an amino acid sequence that is at least about 95% identical to the amino acid sequence of any one of SEQ ID NO: 2-170, 200-209, and 211-245.

[0021] The IL-18 polypeptide can comprise the amino acid sequence of SEQ ID NO: 2 or an amino acid sequence that is at least about 95% identical to the amino acid sequence of SEQ ID NO: 2.

[0022] The IL-18 polypeptide can comprise the amino acid sequence of SEQ ID NO: 13 or an amino acid sequence that is at least about 95% identical to the amino acid sequence of SEQ ID NO: 13.

[0023] The IL-18 polypeptide can comprise the amino acid sequence of SEQ ID NO: 27 or an amino acid sequence that is at least about 95% identical to the amino acid sequence of SEQ ID NO: 27.

[0024] The IL-18 polypeptide can comprise the amino acid sequence of SEQ ID NO: 40 or an amino acid sequence that is at least about 95% identical to the amino acid sequence of SEQ ID NO: 40.

[0025] The IL-18 polypeptide can comprise the amino acid sequence of SEQ ID NO: 53 or an amino acid sequence that is at least about 95% identical to the amino acid sequence of SEQ ID NO: 53.

[0026] The IL-18 polypeptide can comprise the amino acid sequence of SEQ ID NO: 60 or an amino acid sequence that is at least about 95% identical to the amino acid sequence of SEQ ID NO: 60.

[0027] The IL-18 polypeptide can comprise the amino acid sequence of SEQ ID NO: 69 or an amino acid sequence that is at least about 95% identical to the amino acid sequence of SEQ ID NO: 69.

[0028] The IL-18 polypeptides disclosed herein can have substantially reduced binding to the IL-18 binding protein compared to wildtype human IL-18. Preferably, IL-18 polypeptides has binding specificity for IL-18Rα.

[0029] The disclosure further provides nucleic acids encoding the IL-18 polypeptides disclosed herein, vectors, host cells, and methods of making the IL-18 polypeptides. 3 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0030] The disclosure further relates to fusion polypeptides comprising an IL-18 polypeptide disclosed herein; and an amino acid sequence of interest. The amino acid sequence of interest can be an antibody or antibody binding fragment thereof, such as an a Fc domain.

[0031] The amino acid sequence of interest can be a polypeptide with biological activity. The polypeptide with biological activity can comprise a cytokine, a chemokine, a growth factor, a soluble receptor, a tumor antigen binding domain, a peptide hormone, a hormone receptor agonist, or muteins, variants or combinations of any of the foregoing.

[0032] Also provided herein are pharmaceutical compositions comprising an IL-18 polypeptide as disclosed herein or a fusion protein as disclosed herein and a pharmaceutically acceptable carrier.

[0033] Also provided herein are methods for treating cancer in a subject in need thereof. The method comprises administering to the subject an effective amount of the IL-18 variant polypeptide as disclosed herein, a fusion protein as disclosed herein, or the pharmaceutical composition. 3. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] The drawings are not necessarily to scale or exhaustive. Instead, the emphasis is generally placed upon illustrating the principles of the inventions described herein. The accompanying drawings, which constitute a part of the specification, illustrate several embodiments consistent with the disclosure and, together with the description, serve to explain the principles of the disclosure. In the drawings:

[0035] FIGs.1A and 1B are tables showing sequence alignments of IL-18 polypeptide variants. Alignment of IL-18 polypeptide variants that have reduced binding affinity to IL-18 binding protein relative to wild-type IL-18 is shown in FIG.1A. Alignment of IL-18 polypeptide variants that have improved binding affinity for IL-18R1 / RAP relative to wild-type IL-18 is shown in FIG.1B. FIG.1A discloses SEQ ID NOS 247, 1, 48, 65-67, 69, 21-22, 27-28, 31-33 and 45 and FIG.1B discloses SEQ ID NOS 248, 1 and 58-64, all respectively, in order of appearance.

[0036] FIGs.2A and 2B are graphs depicting results from a HEK-Blue IL-18 reporter assay. The reporter assay compares free cytokine with scFc tagged cytokines. Analysis was performed based on quantification of Secreted Alkaline Phosphatase (SEAP) activity using the reagent QUANTI-Blue (InvivoGen). Control IL-18 proteins are shown in FIG.2A. FIG.2B shows a dose response of wild-type IL-18 (circles, a fixed concentration (approximately EC70) of wild-type IL-18 with a dose response of IL- 18 binding protein (squares), and a fixed concentration (approximately EC70) of control binding protein resistant (BPR) IL-18 with a dose response of IL-18 binding protein (triangles) . 4 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0037] FIGs.3A-3D are graphs depicting results from a HEK-Blue IL-18 reporter assay performed on human IL-18 variants. Analysis was performed based on quantification of Secreted Alkaline Phosphatase (SEAP) activity using the reagent QUANTI-Blue (InvivoGen).

[0038] FIGs.4A and 4B are graphs depicting results from a HEK-Blue IL-18 reporter assay performed on IL-18 binding protein inhibition on human IL-18 variants. Analysis was performed based on quantification of Secreted Alkaline Phosphatase (SEAP) activity using the reagent QUANTI-Blue (InvivoGen).

[0039] FIGs.5A and 5B are graphs showing the activity of IL-18 proteins in an IL-18 PBMC assay. FIG.5A is a graph depicting the activity of BPR IL-18 (WW50991) in comparison to the activity of BPR IL-18 WW50991 in the presence of human IL-18BP. FIG.5B is a graph depicting the activity of BPR IL- 18 WW50993 in comparison to the activity of BPR IL-18 WW50993 in the presence of human IL-18BP. Circles represent recombinant human BPR IL-18, squares represent human BPR IL-18 in the presence of titrated human IL-18BP. EC50 values for each protein are shown. Analysis was performed based on quantification of IFNγ in cell supernatants. Results confirm human BPR IL-18 proteins are active in the absence of human IL-18BP and are resistant to human IL-18BP binding and subsequent loss of activity. This is shown as a straight line near the EC70 for each BPR IL-18 protein.

[0040] FIGs.6A-6E are a graphs showing the activity of IL-18 proteins in a IL-18 HEK Blue reporter assay. FIG.6A depicts the activity of WT IL-18-scFc (CTL-88716) in comparison to the activity of WT IL-18-scFc in the presence of human IL-18BP. FIG.6B depicts the activity of BPR IL-18 WW50991 in comparison to the activity of BPR IL-18 WW50991in the presence of human IL-18BP. FIG.6C depicts the activity of BPR IL-18 WW50993 in comparison to the activity of BPR IL-18 WW50993 in the presence of human IL-18BP. FIG.6D depicts the activity of BPR IL-18 WW50992 in comparison to the activity of BPR IL-18 WW50992 in the presence of human IL-18BP. FIG.6E depicts the activity of BPR IL-18 WW50994 in comparison to the activity of BPR IL-18 WW50994 in the presence of human IL- 18BP. Circles represent recombinant human IL-18 (WT or BPR), squares represent human IL-18 (WT or BPR) in the presence of titrated human IL-18BP. Analysis was performed based on quantification of IFNγ in cell supernatants. EC50 values for each protein are shown. The IC50 value for WT IL-18 in the presence of human IL-18BP is also shown in the figure. IC50 values for the BRP IL-18 proteins were unable to be calculated. Results confirm human WT IL-18 is active in the absence of human IL-18BP, but that activity is lost in the presence of human IL-18BP. All human BPR IL-18 proteins are active without human IL-18BP and are resistant to human IL-18BP binding and subsequent loss of activity.

[0041] FIGs.7A-7C are graphs showing the activity of wild type (WT) or BPR mIL-18 in a murine splenocyte proliferation assay. The graphs show proliferation of cells stimulated with WT mIL-18 (FIG. 5 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 7A), or BPR mIL-18 (FIG.7B and 7C). In order to measure resistance to IL-18BP, cells were treated with titrated amounts of recombinant mIL-18BP in the presence of 0.1nM WT mIL-18 or BPR mIL-18.

[0042] FIGs.8A and 8B are graphs showing the activity of wild-type (FIG.8A) or BPR IL-18 (FIG. 8B) assessed by IFNg production from PBMCs in the presence of 10 ng / mL IL-12 + / - IL-18BP at various concentrations for 48 hr. Cell culture supernatants were harvested, and IFNg production was measured using a hIFNg AlphaLISA. 4. DETAILED DESCRIPTION

[0001] The disclosure relates to variants of IL-18 that have binding specificity for the IL-18 receptor, methods of making, and uses thereof. Such variants of IL-18 can be used, for example, as a therapeutic agents. Also provided herein are IL-18-scFc (single chain fragment crystallizable) fusion proteins that include an IL-18 polypeptide as disclosed herein operably linked to a single chain Fc domain.

[0002] The IL-18 polypeptides of this disclosure differ from naturally occurring human IL-18 in amino acid sequence and have desirable characteristics. For example, and without wishing to be bound by any particular theory, the IL-18 polypeptides disclosed herein have binding specificity for human IL-18 receptor but have reduced binding to human IL-18 binding protein compared to naturally occurring or wild-type human IL-18. The IL-18 polypeptides of this disclosure comprise minimal amino acid modifications, generally limited to about four or fewer amino acid substitutions. This contrasts with known IL-18 variants, which usually have more amino acid modifications. Without wishing to be bound by any particular theory, it is believed that the IL-18 polypeptides of this disclosure have improved safety in comparison to IL-18 polypeptide that have more extensive modification. In particular, immunogenicity is significantly reduced, which may thereby decrease immune response risk. A. IL-18 Polypeptides

[0003] The IL-18 polypeptide variants disclosed herein comprise an amino acid sequence that has at least about 90% identity to the wild-type IL-18 polypeptide. For instance, the IL-18 polypeptides disclosed herein have at least about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity to SEQ ID NO: 1. This disclosure also relates to fragments of such polypeptides that retain IL-18 receptor binding activity.

[0004] The IL-18 polypeptide variants disclosed herein can comprise one or more amino acid modifications. The amino acid modification can include amino acid substitutions, insertions, and / or deletions.

[0005] The IL-18 polypeptide variants typically comprise one or more amino acid substitutions relative to SEQ ID NO: 1. The IL-18 polypeptide variant or fragment thereof can comprise at least 1, at least 2, at 6 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10 or more amino acid substitutions. Certain preferred IL-18 polypeptide variants or fragments thereof comprise no more than four amino acid substitutions (e.g., one, two, three, or four amino acid substitutions) relative to SEQ ID NO: 1. Certain preferred IL-18 polypeptide variants or fragments thereof comprise no more than 8 amino acid substitutions (e.g., one, two, three, four, five, six, seven, or eight amino acid substitutions) relative to SEQ ID NO: 1.

[0006] Numbering of the amino acid positions of IL-18 and amino acid modifications disclosed herein are based on SEQ ID NO: 1. For clarity, amino acid substitutions relative to SEQ ID NO: 1 can be noted as Xaa1(amino acid position in SEQ ID NO: 1) Xaa2, wherein Xaa1 is the amino acid at the indicated position of SEQ ID NO: 1, and Xaa2 is the replacement amino acid. For example, Y1W indicates that the tyrosine at position 1 of SEQ ID NO: 1 is replaced with tryptophan, and Y1X indicates that the tyrosine at position 1 of SEQ ID NO: 1 is replaced with any other amino acid.

[0007] The IL-18 polypeptide variant or fragment thereof can comprise at least one or more amino acid substitutions selected from Y1X, G3X, E6X, S10X, S36X, D37X, D40X, I49X, M51X, K53X, D54X, Q56X, P57X, M60X, V62X, and N91X of SEQ ID NO: 1. Preferably, the IL-18 polypeptide variant comprises no more than three amino acid substitutions, which are selected from the group consisting of Y1X, G3X, E6X, S10X, S36X, D37X, D40X, I49X, M51X, K53X, D54X, Q56X, P57X, M60X, V62X, and N91X of SEQ ID NO: 1.

[0008] The IL-18 polypeptide variants disclosed herein can comprise one or more amino acid modifications that decrease binding affinity for IL-18 binding protein (IL-18BP). Residues that may be modified to reduce binding affinity for IL-18BP include amino acid modifications at positions 3, 6, 10, 49, 51, 53, 54, 56, 57, 60, 6291, 126, or 149. Amino acid residues that may be modified to decrease binding affinity for IL-18BP can be selected from S10L, S10N, S10A, S10T, S10Q, S10H, S10L, S10I, S10K, S10R, S10G, S10F, S10V, I49L, I49S, I49K, I49R, I49K, I49Q, I49T, M51R, M51K, M60K, K53S, K53T, D54F, D54P, D54H, Q57P, Q56P, P57H, P57H, P57S, M60K, V62Y, V62S, V62A, V62L, N91L, N91M, N91Y, N91R, N91T, N91K, N91G, N91W, N91A, N91I, N91V, N91S, N91L, N91H, A126C or I149C.

[0009] The IL-18 polypeptide variants disclosed herein can comprise one or more amino acid modifications that increase binding affinity for IL-18 receptor (IL-18R). Residues that may be modified to increase binding affinity for IL-18R include one or more amino acid modifications at positions 6, 36, 37, or 40. Amino acid residues that may be modified to increase binding affinity for IL-18R include E6K, S36E, S36Q, S36A, D37E, D40N, and D40E. 7 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0010] The IL-18 polypeptide variants disclosed herein can comprise one or more amino acid modifications that reduce binding affinity for IL-18BP and one or more amino acid modifications that increase binding affinity for IL-18R.

[0011] The IL-18 polypeptide variant can comprise SEQ ID NO: 1 in which the amino acid at position 1 is modified. The amino acid modification can be Y1W. The IL-18 polypeptide variant having the amino acid modification Y1W can further comprise one or more additional amino acid modifications. For example, the IL-18 polypeptide variant having amino acid modification Y1W can further comprise one additional amino acid modification. For example, the IL-18 polypeptide variant having amino acid modification Y1W can further comprise two amino acid modifications. Such amino acid modifications can be at any desired position. In particular, the amino acid modifications are preferably at positions 51 or 53. The amino acid modification at position 51 is preferably M51R or M51K. The amino acid modification at position 53 is K53S or K53T.

[0012] The IL-18 polypeptide variant can comprise SEQ ID NO: 1 in which the amino acid at any one of positions 3, 6, 10, 49, 54, 62, and 91 is modified. The amino acid modification can be any one or more of G3X, E6X, S10X, I49X, D54X, V62X, and N91X. The amino acid “X” is not the same as the amino acid at the corresponding position in SEQ ID NO: 1.

[0013] The IL-18 polypeptide variant in which any one of positions G3, E6, S10, I49, D54, V62, and N91 is modified, can further comprise one or more additional amino acid modifications. Typically, the IL-18 polypeptide variant will have no more than three amino acid modifications (e.g., one, two, or three amino acid modifications). For example, the IL-18 polypeptide variant can comprise one amino acid modification. For example, the IL-18 polypeptide variant can comprise two amino acid modifications. For example, the IL-18 polypeptide variant can comprise three amino acid modifications.

[0014] When the IL-18 polypeptide variant comprises an amino acid modification at position G3, the amino acid modification is preferably G3D, G3P, G3R, G3Q, or G3V. The IL-18 polypeptide variant can further comprise one or more additional amino acid modifications. The one or more additional amino acid modifications can be at any desired position and any desired amino acid. Generally, an IL-18 polypeptide variant comprising an amino acid modification at position 3 comprises either no additional amino acid modifications or one additional modifications (e.g., the IL-18 polypeptide variant comprises one or two amino acid modifications). Preferred additional amino acid modifications are at position 6 and position 54. Amino acid modifications at position 54 include, but are not limited to D54F, D54P, and D54H. Amino acid modifications at position 6 include, but are not limited to E6R, E6Q, E6N, E6W, and E6K.

[0015] When the IL-18 polypeptide variant comprises an amino acid modification at position 6, the amino acid modification is preferably E6R, E6Q, E6N, E6W, or E6K. The IL-18 polypeptide variant can 8 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 further comprise one or more additional amino acid modifications. The one or more additional amino acid modifications can be at any desired position and any desired amino acid. Generally, an IL-18 polypeptide variant comprising an amino acid modification at position 6 comprises either no additional amino acid modifications or one additional modifications (e.g., the IL-18 polypeptide variant comprises one or two amino acid modifications). Preferred additional amino acid modifications are at position 3 and position 54. Amino acid modifications at position 54 include, but are not limited to D54F, D54P, and D54H. Amino acid modifications at position 3 include, but are not limited to G3D, G3P, G3R, G3Q, or G3V.

[0016] When the IL-18 polypeptide variant comprises an amino acid modification at position 10, the amino acid modification is preferably S10L, S10N, S10A, S10T, S10Q, S10H, S10I, S10K, S10R, S10G, S10V, or S10F. The IL-18 polypeptide variant can further comprise one or more additional amino acid modifications. The one or more additional amino acid modifications can be at any desired position and any desired amino acid. Generally, an IL-18 polypeptide variant comprising an amino acid modification at position 10 comprises either no additional amino acid modifications, one additional amino acid modifications, or two additional amino acid modifications (e.g., the IL-18 polypeptide variant comprises one, two, or three amino acid modifications). Preferred additional amino acid modifications are at position 49, position 62, and position 91. Amino acid modifications at position 49 include, but are not limited to I49L, I49T, I49Q, I49K, I49R, and I49S. Amino acid modifications at position 62 include, but are not limited to V62Y, V62S, V62A, and V62L. Amino acid modifications at position 91 include, but are not limited to, N91R, N91L, N91M, N91Y, N91T, N91K, N91G, N91W, N91A, N91I, N91V, N91S, and N91H.

[0017] When the IL-18 polypeptide variant comprises an amino acid modification at position 49, the amino acid modification is preferably I49L, I49T, I49Q, I49K, I49R, and I49S. The IL-18 polypeptide variant can further comprise one or more additional amino acid modifications. The one or more additional amino acid modifications can be at any desired position and any desired amino acid. Generally, an IL-18 polypeptide variant comprising an amino acid modification at position 49 comprises either no additional amino acid modifications, one additional amino acid modifications, or two additional amino acid modifications (e.g., the IL-18 polypeptide variant comprises one, two, or three amino acid modifications). Preferred additional amino acid modifications are at position 10 or position 91. Amino acid modifications at position 10 include, but are not limited to, S10L, S10N, S10A, S10T, S10Q, S10H, S10I, S10K, S10R, S10G, S10V, or S10F. Amino acid modifications at position 91 include, but are not limited to, N91R, N91L, N91M, N91Y, N91T, N91K, N91G, N91W, N91A, N91I, N91V, N91S, and N91H. 9 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0018] When the IL-18 polypeptide variant comprises an amino acid modification at position 45, the amino acid modification is preferably D54F, D54P, or D54H. The IL-18 polypeptide variant can further comprise one or more additional amino acid modifications. The one or more additional amino acid modifications can be at any desired position and any desired amino acid. Generally, an IL-18 polypeptide variant comprising an amino acid modification at position 54 comprises either no additional amino acid modifications, or one additional amino acid modifications (e.g., the IL-18 polypeptide variant comprises one or two amino acid modifications). Preferred additional amino acid modifications are at position 3. Amino acid modifications at position 3 include, but are not limited to G3D, G3P, G3R, G3Q, or G3V.

[0019] When the IL-18 polypeptide variant comprises an amino acid modification at position 62, the amino acid modification is V62Y, V62S, V62L, or V62A. The IL-18 polypeptide variant can further comprise one or more additional amino acid modifications. The one or more additional amino acid modifications can be at any desired position and any desired amino acid. Generally, an IL-18 polypeptide variant comprising an amino acid modification at position 62 comprises either no additional amino acid modifications, one additional amino acid modifications, or two additional amino acid modifications (e.g., the IL-18 polypeptide variant comprises one, two, or three amino acid modifications). Preferred additional amino acid modifications are at position 10, position 49, and position 91. Amino acid modifications at position 10 include, but are not limited to, S10L, S10N, S10A, S10T, S10Q, S10H, S10I, S10K, S10R, S10G, S10V, or S10F. Amino acid modifications at position 91 include, but are not limited to, N91R, N91L, N91M, N91Y, N91T, N91K, N91G, N91W, N91A, N91I, N91V, N91S, and N91H. Amino acid modifications at I49 include, but are not limited to I49L, I49T, I49Q, I49K, I49R, and I49S.

[0020] When the IL-18 polypeptide variant comprises an amino acid modification at position 91, the amino acid modification is N91R, N91L, N91M, N91Y, N91T, N91K, N91G, N91W, N91A, N91I, N91V, N91S, or N91H. The IL-18 polypeptide variant can further comprise one or more additional amino acid modifications. The one or more additional amino acid modifications can be at any desired position and any desired amino acid. Generally, an IL-18 polypeptide variant comprising an amino acid modification at position N91 comprises either no additional amino acid modifications, one additional amino acid modifications, or two additional amino acid modifications (e.g., the IL-18 polypeptide variant comprises one, two, or three amino acid modifications). Preferred additional amino acid modifications are at position 10, position 49, position 62, or position 60. Amino acid modifications at position 10 include, but are not limited to, S10L, S10N, S10A, S10T, S10Q, S10H, S10I, S10K, S10R, S10G, S10V, or S10F. Amino acid modifications at position 49 include, but are not limited to I49L, I49T, I49Q, I49K, I49R, and I49S. Amino acid modifications at position 62 include, but are not limited to V62Y, V62S, VS2A, and V62L. Amino acid modifications at position 60 include, but are not limited to M60K. 10 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0021] The IL-18 polypeptide variant can comprise SEQ ID NO: 1 in which the amino acid at position 40 is modified. The amino acid modification can be D40N or D40E. The IL-18 polypeptide variant having the amino acid modification D40N or D40E can further comprise one or more additional amino acid modifications. For example, the IL-18 polypeptide variant having amino acid modification D40N or D40E can further comprise one or two additional amino acid modifications. Such amino acid modifications can be at any desired position. In particular, the amino acid modifications are preferably at position 6. The amino acid modification at position 6 is preferably E6R, E6Q, E6N, E6W, or E6K.

[0022] The IL-18 polypeptide variant can comprise SEQ ID NO: 1 in which the amino acid at position 56 is modified. The amino acid modification can be Q56P. The IL-18 polypeptide variant having the amino acid modification Q10P can further comprise one or more additional amino acid modifications. For example, the IL-18 polypeptide variant having amino acid modification Q56P can further comprise one, two, or three additional amino acid modifications. Such amino acid modifications can be at any desired position. In particular, the amino acid modifications are preferably at position 60 or position 57. Amino acid modifications at position 60 include, but are not limited to, M60K. Amino acid modifications at position 57 include, but are not limited to 57H.

[0023] The IL-18 polypeptide variant can comprise SEQ ID NO: 1 in which the amino acid modification at position 57 is modified. The amino acid modification can be P57H, P57A, P57T, or P57S. The IL-18 polypeptide variant having the amino acid modification P57H, P57A, P57T, or P57S can further comprise one or more additional amino acid modifications. For example, the IL-18 polypeptide variant having amino acid modification P57H , P57A, P57T, or P57S can further comprise one, two, or three additional amino acid modifications. Such amino acid modifications can be at any desired position. In particular, the amino acid modifications are preferably at position 56, position 60, position 62, or position 91. Amino acid modifications at position 60 include, but are not limited to M60K. Amino acid modifications at position 56 include, but are not limited to Q56P. Amino acid modifications at position 62 include, but are not limited to V62S. Amino acid modifications at position 91 include, but are not limited to N91L and N91T.

[0024] The IL-18 polypeptide variant can comprise SEQ ID NO: 1 in which the amino acid at position 36 is modified. The amino acid modification can be S36E, S36A, or S36Q. The IL-18 polypeptide variant having the amino acid modification S36E, S36A, or S36E can further comprise one or more additional amino acid modifications. For example, the IL-18 polypeptide variant having amino acid modification S36E, S36A, or S36E can further comprise one or two additional amino acid modifications. Such amino acid modifications can be at any desired position. In particular, the amino acid modifications are 11 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 preferably at position 37 or 40. The amino acid modification at position 37 is preferably D37E. The amino acid modification at position 40 is preferably D40N or D40E.

[0025] The IL-18 polypeptide variant can comprise SEQ ID NO: 1 in which the amino acid at position 36 is modified. The amino acid modification can be S37E. The IL-18 polypeptide variant having the amino acid modification S37E can further comprise one or more additional amino acid modifications. For example, the IL-18 polypeptide variant having amino acid modification S37E can further comprise one or two additional amino acid modifications. Such amino acid modifications can be at any desired position. In particular, the amino acid modifications are preferably at position 36 or 40. The amino acid modification at position 36 is preferably S36E, S36A, or S36Q. The amino acid modification at position 40 is preferably D40N or D40E.

[0026] The IL-18 polypeptide variant can comprise SEQ ID NO: 1 in which the amino acid at positions 36 and 37 is modified. The amino acid modification can be S36E, S36Q, S36A, S36D, S36R, S36T, S36Q and / or D37E. The IL-18 polypeptide variant comprising amino acid at positions 36 and 37 can further comprise one or more additional amino acid modifications. Such amino acid modifications can be at any desired position. In particular, the amino acid modifications are preferably at position 49, 51, 53, 54, 56, 57, 59, 60, and / or 133. Amino acid modifications at position 49 include, but are not limited to, I49R Amino acid modifications at position 51 include, but are not limited to K51R. Amino acid modifications at position 53 include, but are not limited to K53S. Amino acid modifications at position 54 include, but are not limited to S54P. Amino acid modifications at position 56 include, but are not limited to Q56S. Amino acid modifications at position 57 include, but are not limited to P57G, P57T, P57H, or P57S. Amino acid modifications at position 59 include, but are not limited to G59N, G50A. Amino acid modifications at position 60 include, but are not limited to M60K, M60R, or M60Y. Amino acid modifications at position 133 include, but are not limited to L133A, L133H, L133M, L133R.

[0027] The IL-18 polypeptide variant can comprise amino acid modifications relative to SEQ ID NO: 1 selected from the group consisting of S36E / D37E / M60K, S36E / D37E / Q56R / P57S / M60K, S36E / D37E / Q56H / M60K / L133R, S36E / D37E / Q56H / M60K, S36E / D37E / P57T / M60K, S36E / D37E / P57T / M60K, S36E / D37E / P57S / M60K / L133M, S36E / D37E / P57D / M60Y, S36E / D37E / P57H / M60R, S36Q / D37E / P57A / M60K, S36Q / D37E / M60K, S36Q / D37E / M60K / L133A, S36Q / D37E / P57V / M60K, S36Q / D37E / G59N / M60K, S36Q / D37E / P57M / M60K, S36A / D37E / M60K, S36A / D37E / P57G / G59A / M60K, S36D / D37E / K51R / K53S, S36R / D37E / K51R / K53S, S36T / D37E / I49R / P57S, S36E / D37E / S54P, or S36E / D37E / I49R / P57S.

[0028] The IL-18 polypeptide variant can comprise SEQ ID NO: 1 in which the amino acid at position 36 is modified. The amino acid modification can be S36Q or S36E. The IL-18 polypeptide variant 12 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 comprising amino acid at position 36 can further comprise one or more additional amino acid modifications. Such amino acid modifications can be at any desired position. In particular, the amino acid modifications are preferably at position 49, 51, 53, 57, 93, and / or 134. Amino acid modifications at position 49 include, but not limited to I49R Amino acid modifications at position 51 include, but not limited to M51R. Amino acid modifications at position 53 include, but not limited to K53S. Amino acid modifications at position 57 include, but not limited to P57S. Amino acid modifications at position 93 include, but not limited to K93L. Amino acid modifications at position 134 include, but not limited to F134S. The IL-18 polypeptide variant can comprise amino acid modifications relative to SEQ ID NO: 1 selected from the group consisting of S36Q / M51R / K53S, S36Q / I49R / K53S, or S36Q / I49R / K93L / F134S.

[0029] The IL-18 polypeptide variant can comprise SEQ ID NO: 1 in which the amino acid at position 51 is modified. The amino acid modification can be M51R. The IL-18 polypeptide variant comprising amino acid at position 51 can further comprise one or more additional amino acid modifications. Such amino acid modifications can be at any desired position. In particular, the amino acid modifications are preferably at position 34, 49, 53, 57, 111, 129, 131, 132, 134, 145, 149, or 150. Amino acid modifications at position 34 include, but not limited to T34S. Amino acid modifications at position 49 include, but not limited to I49Q or I49R. Amino acid modifications at position 53 include, but not limited to K53S or K53T. Amino acid modifications at position 90 include, but not limited to D90A. Amino acid modifications at position 93 include, but not limited to K93N. Amino acid modifications at position 129 include, but not limited to K129L or K129R. Amino acid modifications at position 131 include, but not limited to R131A, R131G or R131V. Amino acid modifications at position 132 include, but not limited to D132G. Amino acid modifications at position 134 include, but not limited to F134D. Amino acid modifications at position 145 include, but not limited to G145R. Amino acid modifications at position 150 include, but not limited to M150L. The IL-18 polypeptide variant can comprise amino acid modifications relative to SEQ ID NO: 1 selected from the group consisting of I49Q / M51R / K53S / K129L / R131G / F134D, M51R / K53S / G145R, M51R / K53S / M150L, M51R / K53S / R131A / D132G, M51R / K53T / M150L,I49R / K53S / R131V, T34S / I49R / K53S, I49R / K53S / K129R, or I49R / K53S / K129R / D90A / K93N.

[0030] Typically, the IL-18 polypeptide variant will have no more than four additional amino acid modifications (e.g., one, two, three, or four amino acid modifications). Certain preferred additional amino acid modifications are at position 36, position 38, position 40, position 126, and position 149. The amino acid modification at position 36 can be S36E. The amino acid modification at position 38 can C38S. The amino acid modification at position 40 can be D40N. The amino acid modification at position 126 can be A126C. The amino acid modification at position 149 can be S149C. 13 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0031] The IL-18 polypeptide variant disclosed herein can further comprise one additional amino acid modification selected from the group consisting of S36E, C38S, D40N, A126C, and I149C.

[0032] The IL-18 polypeptide variant disclosed herein can comprise two additional amino acid modifications selected from the group consisting of S36E / D40N, S36E / C38S, S36E / A126C, S36E / I149C, C38S / D40N, C38S / A126C, C38S / I149C, A126C / I149C, D40N / A126A, and D40N / I149C.

[0033] The IL-18 polypeptide variant disclosed herein can comprise three additional amino acid modifications selected from the group consisting of S36E / D40N / C38S, S36E / D40N / A126C, S36E / D40N / IY49C, C38S / D40N / A126C, C38S / D40N / I149C, C38S / A126C / I149C, and D40N / A126C / I149C.

[0034] The IL-18 polypeptide variant disclosed herein can comprise four additional amino acid modifications selected from the group consisting of S36E / C38S / D40N / A126C, S36E / C38S / D40N / I149C, C38S / D40N / A126C / I149C, S36E / D40N / A126C / I149C and S36E / C38S / A126C / I149C.

[0035] The IL-18 polypeptide variants disclosed herein can comprise five additional amino acid modifications S36E / C38S / D40N / A126C / I149C.

[0036] In certain preferred IL-18 polypeptide variants, the IL-18 polypeptide variant comprises at least one amino acid modification to cysteine. Without wishing to be bound by theory, it is believed that when an amino acid modification is a cysteine, it may introduce the potential for new disulfide bonds in the IL- 18.

[0037] The IL-18 polypeptide variant can comprise SEQ ID NO: 1 in which one to three amino acids are modified relative to SEQ ID NO: 1. In examples, the IL-18 polypeptide variant can comprise SEQ ID NO:1 with one or more amino acid substitutions selected from the group consisting of S10X / I49X / N91X, K53X / M51X / Y1X, S10X / I49X / V62X, K53X / M51X / Y1X, M60X / Q56X / P57X, S10X / I49X, , V62X / N91X, I49X / N91X, S10X / N91X, M60X / N91X, D54X / 3GX, E6X / G3X, D40X / E6X, I49X / P57X, K53X / M51X, S10X, V62X, N91X, I49X, D54X, E6X, G3X, and D40X.

[0038] The IL-18 polypeptide variant can comprise three amino acid modifications relative to SEQ ID NO: 1 selected from the group consisting of S10L / I49L / N91R, S10L / I49Q / N91Y, S10I / I49K / N91K, Q56P / P57H / M60K, S10A / I49L / V62Y, Y1W / M51R / K53S, and S10T / I49T / N91L. Certain preferred IL- 18 polypeptide variants comprise three amino acid modifications at positions S10L / I49L / N91R, S10L / I49Q / N91Y, and S10T / I49T / N91L of SEQ ID NO: 1.

[0039] The IL-18 polypeptide variant can comprise two amino acid modifications relative to SEQ ID NO: 1 selected from the group consisting of S10A / I49L, S10R / I49T, S10I / I49K, V62A / N91M, I49L / N91A, I49T / N91R, I49I / N91K, I49R / N91Y, I49S / N91R, S10H / N91Y, S10H / N91L, S10H / N91R, S10H / N91T, S10T / N91L, S10T / N91R, S10F / N91M, G3P / D45P, G3D / D54F, G3P / 6EQ, G3Q / E6W, 14 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 E6K / D40N, I49R / P57S, K53S / M51R, K53S / M51K, K53T / M51R, M60K / N91L, G3P / D54H, and G3R / E6N. Certain preferred IL-18 polypeptide variants comprise two amino acid modifications at positions G3R / E6N, M60K / N91L, S10R / I49T, I49R / P57S, K53S / M51R or S10H / N91T of SEQ ID NO: 1.

[0040] The IL-18 polypeptide variant can comprise one amino acid modification relative to SEQ ID NO: 1 selected from the group consisting of S10N, S10H, S10G, S10Q, S10K, S10R, V62S, N91G, N91R, N91W, N91L, N91T, N91I, N91V, N91S, N91M, N91K, N91Y, N91K, N91A, I49R, D54P, E6Q, G3R, G3V, D40N, AND D40E. Certain preferred IL-18 polypeptide variants comprise one amino acid modification at position S10K, D40N, I49R, N91V, or N91R of SEQ ID NO: 1.

[0041] The IL-18 polypeptide variant can comprise SEQ ID NO: 1 in which two to eight amino acids are modified relative to SEQ ID NO: 1. In examples, the IL-18 polypeptide variant can comprise SEQ ID NO: 1 with one or more amino acid substitutions selected from the group consisting of S10X / I49X / P57X, S10X / I49X / P57X / S36X, S10X / I49X / P57X / S36X / D40X, I49X / K53X / S36X, I49X / K53X / S36X / D40X, S10X / I49X / P57X / C38X, S10X / I49X / P57X / S36X / C38X, S10X / I49X / P57X / S36X / D40X / C38X, M51X / P57X / C38SX, M51X / P57X / S36X / X38S, M51X / P57X / S36X / D40X / C38X, I49X / K53X / C38X, I49X / K53X / S36X / C38X, I49X / K53X / D40X, A126X / 1149X, S10X / I49X / P57X / A126X / I49X, S10X / I49X / P57X / S36X / A126X / I149X, S10X / I49X / P57X / S36X / D40X / A126X / I149X, M51X / P57X / A126X / I189X, M51X / P57X / S36X / A126X / I149X, M51X / P57X / S36X / D40X / A126X / I149X, I49X / K53X / A162X / I149X, I49X / K53X / S36X / A126X / I149X, I49X / K53X / S36X / D40X / A126X / I149X, C38X / A126X / I149X, S10X / I49X / P57X / C38X / A126X / I149X, S10X / I49X / P57X / S36X / C38X / A126X / I149X, S10X / I49X / P57 X / S36 X / D40 X / C38X / A126X / I149X,

[0042] M51X / P57 X / S36X / C38X / A126X / I149X, M51X / P57X / S36X / D40X / C38X / A126X / I49X, I49X / P53X / C38X / A126X / I149X, I49X / P53X / C38X / A162X / I149X, I49X / P53X / S36X / C38X / A126X / I149X and I49X / K53X / S36X / I40N / C38X / A126X / I149X.

[0043] The IL-18 polypeptide variant can comprise two amino acid modifications relative to SEQ ID NO: 1 selected from A126C / I149C.

[0044] The IL-18 polypeptide variant can comprise three amino acid modifications relative to SEQ ID NO: 1 selected from the group consisting of S10K / I49R / P57S, M51R / P57S / C38S, C38S / A126C / I149C, and I49R / K53S / C38S.

[0045] The IL-18 polypeptide variant can comprise four amino acid modifications relative to SEQ ID NO: 1 selected from the group consisting of S10K / I49R / P57S / S36E, S10K / I49R / P57S / S36E, I49R / K53S / S36E / D40N, S10K / I49R / P57S / C38S, M51R / P57S / S36E / C38S, I49R / K53S / S36E / C38S, M51R / K57S / A126C / I149C, S36E / D40N / I49R / P57S, S36E / D40N / M51R / K53S and 15 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 I49R / K53S / A126C / I49C. Certain preferred amino acid modifications are S36E / D40N / I49R / P57S S36E / D40N / M51R / K53S.

[0046] The IL-18 polypeptide variant can comprise five amino acid modifications relative to SEQ ID NO: 1 selected from the group consisting of S10K / I49R / P57S / S36E / D40N, S10K / I49R / P57S / S36E / C38S, M51R / P57S / S36E / D40N / C38S, I49R / K53S / S36E / D40N / C38S, S10K / I49R / P57S / A126C / I149C, M51R / P57S / S36E / A126 / I149C, I49R / K53S / S36E / A126C / I149C, and I49R / K53S / C38S / A126C / I149C.

[0047] The IL-18 polypeptide variant can comprise six amino acid modifications relative to SEQ ID NO: 1 selected from the group consisting of S10K / I49R / P57S / S36E / D40N / C38S, S10K / I49R / P57S / S36E / A126C / I185C, M51R / P57S / S36E / D40N / A126C / I149C, S10K / I49R / P57S / C38S / A162C / I185C, M51R / P57S / S36E / C38S / A126C / I149C, I49R / K53S / S36E / C38S / A126C / I149C, and M51R / P57S / S36E / D40N / A126C / I149C.

[0048] The IL-18 polypeptide variant can comprise seven amino acid modifications relative to SEQ ID NO: 1 selected from the group consisting of I49R / K53S / S36E / D40N / C38S / A126C / I149C, S10K / I49R / P57S / S36E / D40N / A126C / I149C, S10K / I49R / P57S / S36E / C38S / A126C / I149C, M51R / P57S / S36E / D40N / C38S / A126C / I149C, and I49R / K53S / S36E / D40N / C38S / A126C / I149C.

[0049] The IL-18 polypeptide variant can comprise eight amino acid modifications relative to SEQ ID NO: 1 selected from S10K / I49R / P57S / S36E / D40N / C38S / A126C / I185C.

[0050] The IL-18 polypeptide variants disclosed herein can comprise one or more deletions at any desired position. In examples, the IL-18 polypeptide variant can comprise SEQ ID NO:1 with an amino acid deletion at position 1. When the IL-18 polypeptide variant comprises a deletion, it can be referred to as a truncated IL-18 polypeptide variant. The truncated IL-18 polypeptide variant can further comprise one or more amino acid modifications. Typically, the truncated IL-18 polypeptide variant will comprise no more than 4 amino acid modifications. For example, one amino acid modification, two amino acid modifications, three amino acid modifications, or four amino acid modifications.

[0051] In examples, the truncated IL-18 polypeptide variant can comprise SEQ ID NO: 1 with a deletion at position one and one or more amino acid substitutions selected from the group consisting of S36X, S36X / D40X, C38X, S36X / C38X, S36X / D40X / C38X / A126X / I149X / S36X, A126X / I149X, S36S / D40N / A126X / I149X.

[0052] In examples, the truncated IL-18 polypeptide variant can comprise SEQ ID NO: 1 with a deletion at position one and one amino acid substitutions selected from the group consisting of C38S and S36E.

[0053] In examples, the truncated IL-18 polypeptide variant can comprise SEQ ID NO: 1 with a deletion at position one and one amino acid substitution selected from the group consisting of C38S and S36E. 16 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0054] In examples, the truncated IL-18 polypeptide variant can comprise SEQ ID NO: 1 with a deletion at position one and two amino acid substitutions selected from the group consisting of S36E / D40N, S36E / C38S, and A126C / I149C.

[0055] In examples, the truncated IL-18 polypeptide variant can comprise SEQ ID NO: 1 with a deletion at position one and three amino acid substitutions selected from the group consisting of S36E / D40N / C38S, S36E / A126C / I149C.

[0056] In examples, the truncated IL-18 polypeptide variant can comprise SEQ ID NO: 1 with a deletion at position one and four amino acid substitutions selected from the group consisting of S36E / D40N / A126C / I149C.

[0057] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 2, or an amino acid sequence that has at least about 95% identity, about 90%, preferably about 95%, about 96%, about 97%, about 98%, or about 99% to SEQ ID NO: 2. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 2, wherein the amino acid at position 56 is deleted.

[0058] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 3, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 3. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 3 wherein the amino acids at positions 10, 49 and 91 of SEQ ID NO: 3 are not substituted.

[0059] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 4, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 4. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 4 wherein the amino acid at position 10 of SEQ ID NO: 4 is not substituted.

[0060] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 5, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 5. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 5 wherein the amino acids at positions 10, 49 and 62 of SEQ ID NO: 5 are not substituted.

[0061] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 6, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 6. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 6 wherein the amino acids at positions 10, 49 and 91 of SEQ ID NO:6 are not substituted. 17 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0062] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 7, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 7. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 7 wherein the amino acid at position 10 of SEQ ID NO: 7 is not substituted.

[0063] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 8, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 8. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 8 wherein the amino acids at positions 57 and 62 of SEQ ID NO: 8 are not substituted.

[0064] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 9, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 9. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 9 wherein the amino acids at positions 62 and 91 of SEQ ID NO: 9 are not substituted.

[0065] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 10, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 10. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 10 wherein the amino acids at positions 10 and 91 of SEQ ID NO: 10 are not substituted.

[0066] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 11, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 11. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 11 wherein the amino acids at positions 10 and 91 of SEQ ID NO: 11 are not substituted.

[0067] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 12, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 12. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 12 wherein the amino acids at positions 10 and 91 of SEQ ID NO: 12are not substituted.

[0068] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 13, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 13. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 18 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 95% identity to SEQ ID NO: 13 wherein the amino acids at positions 10 and 91 of SEQ ID NO: 13 are not substituted.

[0069] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 14, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 14. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 14 wherein the amino acids at positions 10, 49 and 91 of SEQ ID NO: 14 are not substituted.

[0070] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 15, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 15. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 15 wherein the amino acids at positions 10, 49 and 91 of SEQ ID NO: 15 are not substituted.

[0071] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 16, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 16. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 16 wherein the amino acid at position 10 of SEQ ID NO: 16 is not substituted.

[0072] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 17, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 17. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 17 wherein the amino acids at positions 10, 49, 62 and 91 of SEQ ID NO: 17 are not substituted.

[0073] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 18, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 18. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 18 wherein the amino acid at position 91 of SEQ ID NO: 18 is not substituted.

[0074] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 19, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 19. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 19 wherein the amino acid at position 91 of SEQ ID NO: 19 is not substituted. 19 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0075] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 20, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 20. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 20 wherein the amino acid at position 91 of SEQ ID NO: 20 is not substituted.

[0076] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 21, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 21. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 21 wherein the amino acid at positions 10 and 91 of SEQ ID NO: 21 is not substituted.

[0077] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 22, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 22. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 22 wherein the amino acids at position 10, 51, and 91 of SEQ ID NO: 22 is not substituted.

[0078] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 23, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 23. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 23 wherein the amino acids at positions 49 and 91 of SEQ ID NO: 23 are not substituted.

[0079] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 24, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 24. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 24 wherein the amino acid at position 91 of SEQ ID NO: 24 is not substituted.

[0080] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 25, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 25. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 25 wherein the amino acids at positions 49 and 91 of SEQ ID NO: 25 are not substituted.

[0081] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 26, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 26. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 20 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 95% identity to SEQ ID NO: 26 wherein the amino acid at position 91 of SEQ ID NO: 26 is not substituted.

[0082] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 27, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 27. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 27 wherein the amino acids at positions 49 and 57 of SEQ ID NO: 27 is not substituted.

[0083] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 28, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 28. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 28 wherein the amino acid at position 10 or 57 of SEQ ID NO: 28 is not substituted.

[0084] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 29, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 29. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 29 wherein the amino acid at position 10 of SEQ ID NO: 29 is not substituted.

[0085] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 30, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 30. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 30 wherein the amino acid at position 91 of SEQ ID NO: 30 is not substituted.

[0086] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 31, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 31. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 31 wherein the amino acid at position 49 and 57 of SEQ ID NO: 31 is not substituted.

[0087] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 32, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 32. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 32 wherein the amino acids at positions 60 and 91 of SEQ ID NO: 32 are not substituted. 21 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0088] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 33, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 33. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 33 wherein the amino acids at positions 10, 49, 57, and 91 of SEQ ID NO: 33 are not substituted.

[0089] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 34, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 34. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 34 wherein the amino acids at positions 10, 49 and 91 of SEQ ID NO: 34 are not substituted.

[0090] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 35, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 35. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 35 wherein the amino acid at position 91 of SEQ ID NO: 35 is not substituted.

[0091] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 36, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 36. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 36 wherein the amino acid at position 91 of SEQ ID NO: 36 is not substituted.

[0092] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 37, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 37. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 37 wherein the amino acid at position 91 of SEQ ID NO: 37 is not substituted.

[0093] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 38, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 38. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 38 wherein the amino acids at position 10 of SEQ ID NO: 38 is not substituted.

[0094] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 39, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 39. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 22 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 95% identity to SEQ ID NO: 39 wherein the amino acid at position 91 of SEQ ID NO: 39 is not substituted.

[0095] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 40, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 40. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 40 wherein the amino acids at positions 10 and 91 of SEQ ID NO: 50 are not substituted.

[0096] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 41, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 41. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 41 wherein the amino acids at positions 10 and 91 of SEQ ID NO: 41 are not substituted.

[0097] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 42, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 42. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 42 wherein the amino acids at positions 10 and 91 of SEQ ID NO: 42 are not substituted.

[0098] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 43, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 43. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 43 wherein the amino acids at positions 49 and 91 of SEQ ID NO: 43 are not substituted.

[0099] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 44, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 44. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 44 wherein the amino acids at positions 10 and 49 of SEQ ID NO: 44 are not substituted.

[0100] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 45, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 45. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 45 wherein the amino acid at position 60 and 91 of SEQ ID NO: 45 is not substituted. 23 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0101] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 46, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 46. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 46 wherein the amino acids at positions 10, 49 and 91 of SEQ ID NO: 46 are not substituted.

[0102] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 47, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 47. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 47 wherein the amino acids at positions 3 and 54 of SEQ ID NO: 47 are not substituted.

[0103] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 48, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 48. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 48 wherein the amino acid at position 54 of SEQ ID NO: 48 is not substituted.

[0104] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 49, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 49. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 49 wherein the amino acids at positions 3 and 54 of SEQ ID NO: 49 are not substituted.

[0105] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 50, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 50. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 50 wherein the amino acids at positions 3 and 6 of SEQ ID NO: 50 are not substituted.

[0106] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 51, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 51. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 51 wherein the amino acid at positions 3 and 6 of SEQ ID NO: 51 is not substituted.

[0107] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 52, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 52. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 24 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 95% identity to SEQ ID NO: 52 wherein the amino acid at position 3 of SEQ ID NO: 52 is not substituted.

[0108] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 53, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 53. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 53 wherein the amino acids at positions 3 and 6 of SEQ ID NO: 53 are not substituted.

[0109] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 54, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 54. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 54 wherein the amino acid at position 3 of SEQ ID NO: 54 is not substituted.

[0110] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 55, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 55. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 55 wherein the amino acids at positions 3 and 54 of SEQ ID NO: 55 are not substituted.

[0111] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 56. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 56 wherein the amino acids at positions 3 and 6 of SEQ ID NO: 56 are not substituted.

[0112] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 57, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 57. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 57 wherein the amino acid at position 3 of SEQ ID NO: 57 is not substituted.

[0113] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 58, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 58. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 58 wherein the amino acids at positions 36 and 37 of SEQ ID NO: 58 is not substituted.

[0114] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 59, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 59. For example, 25 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 59 wherein the amino acid at positions 36 and 40 of SEQ ID NO: 59 is not substituted.

[0115] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 60, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 60. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 60 wherein the amino acids at positions 6, 36, and 40 of SEQ ID NO: 60 are not substituted.

[0116] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 61, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 61. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 61 wherein the amino acids at positions 36 and 37 of SEQ ID NO: 61 are not substituted.

[0117] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 62, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 62. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 62 wherein the amino acids at positions 36 and 37 of SEQ ID NO: 62 are not substituted.

[0118] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 63, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 63. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 63 wherein the amino acid at positions 36 of SEQ ID NO: 63 is not substituted.

[0119] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 64, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 64. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 64 wherein the amino acid at position 36 and 40 of SEQ ID NO: 64 is not substituted.

[0120] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 65, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 65. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 65 wherein the amino acids at positions 1, 51 and 53 of SEQ ID NO: 65 are not substituted. 26 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0121] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 66, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 66. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 66 wherein the amino acids at positions 51 and 53 of SEQ ID NO: 66 are not substituted.

[0122] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 67, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 67. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 67 wherein the amino acids at positions 51 and 53 of SEQ ID NO: 67 are not substituted.

[0123] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 68, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 68. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 68 wherein the amino acids at positions 56, 57 and 60 of SEQ ID NO: 68 are not substituted.

[0124] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 69, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 69. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 69 wherein the amino acids at positions 51 and 53 of SEQ ID NO: 6 are not substituted.

[0125] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 70, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 70. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 70 wherein the amino acids at positions 36, 49, and 57 of SEQ ID NO: 70 are not substituted.

[0126] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 71, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 71. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 71 wherein the amino acids at positions 36, 40, 49, and 57 of SEQ ID NO: 71 are not substituted.

[0127] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 72, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 72. For example, the IL-18 27 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 72 wherein the amino acids at positions 36 and 54 of SEQ ID NO: 72 are not substituted.

[0128] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 73, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 73. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 73 wherein the amino acids at positions 36, 40, and 54 of SEQ ID NO: 73 are not substituted.

[0129] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 74, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 74. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 74 wherein the amino acids at positions 36, 51, and 53 of SEQ ID NO: 74 are not substituted.

[0130] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 75, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 75. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 75 wherein the amino acids at positions 36, 40, 51, and 53 of SEQ ID NO: 75 are not substituted.

[0131] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 76, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 76. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 76 wherein the amino acids at positions 36, 51, and 53 of SEQ ID NO: 76 are not substituted.

[0132] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 77, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 77. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 77 wherein the amino acids at positions 36, 40, 51, and 53 of SEQ ID NO: 77 are not substituted.

[0133] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 78, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 78. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 78 wherein the amino acids at positions 10, 49, and 57 of SEQ ID NO: 78 are not substituted. 28 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0134] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 79, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 79. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 79 wherein the amino acids at positions 10, 36, 49, and 57 of SEQ ID NO: 79 are not substituted.

[0135] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 80, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 80. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 80 wherein the amino acids at positions 10, 36, 40, 49, and 57 of SEQ ID NO: 80 are not substituted.

[0136] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 81, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 81. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 80 wherein the amino acids at positions 1 is deleted.

[0137] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 82, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 82. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 82 wherein the amino acids at position 36 of SEQ ID NO: 82 is not substituted.

[0138] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 83, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 83. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 83 wherein the amino acids at positions 36 and 40 of SEQ ID NO: 83 are not substituted.

[0139] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 84, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 84. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 84 wherein the amino acids at positions 51 and 57 of SEQ ID NO: 84 are not substituted.

[0140] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 85, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 85. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 85 wherein the amino acids at positions 36, 51 and 57 of SEQ ID NO: 85 are not substituted.

[0141] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 86, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 86. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to 29 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 SEQ ID NO: 86 wherein the amino acids at positions 36, 40, 51 and 57 of SEQ ID NO: 86 are not substituted.

[0142] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 87, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 87. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 87 wherein the amino acids at positions 49 and 53 of SEQ ID NO: 87 are not substituted.

[0143] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 88, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 88. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 88 wherein the amino acids at positions 36, 49 and 53 of SEQ ID NO: 88 are not substituted.

[0144] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 89, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 89. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 89 wherein the amino acids at positions 36, 40, 49 and 53 of SEQ ID NO: 89 are not substituted.

[0145] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 90, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 90. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 90 wherein the amino acid at position 38 SEQ ID NO: 90 is not substituted.

[0146] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 91, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 91. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 91 wherein the amino acids at positions 38, 49 and 57 of SEQ ID NO: 91 are not substituted.

[0147] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 92, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 92. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 92 wherein the amino acids at positions 36, 38, 49 and 57 of SEQ ID NO: 92 are not substituted.

[0148] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 93, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 93. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 93 wherein the amino acids at positions 36, 38, 40, 49 and 57 of SEQ ID NO: 93 are not substituted. 30 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0149] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 94, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 94. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 94 wherein the amino acids at positions 38 and 54 of SEQ ID NO: 94 are not substituted.

[0150] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 95, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 95. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 95 wherein the amino acids at positions 36, 38 and 54 of SEQ ID NO: 95 are not substituted.

[0151] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 96, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 96. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 96 wherein the amino acids at positions 36, 38, 40 and 54 of SEQ ID NO: 96 are not substituted.

[0152] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 97, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 97. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 97 wherein the amino acids at positions 38, 51 and 53 of SEQ ID NO: 97 are not substituted.

[0153] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 98, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 98. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 98 wherein the amino acids at positions 36, 38, 51 and 53 of SEQ ID NO: 98 are not substituted.

[0154] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 99, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 99. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 99 wherein the amino acids at positions 36, 38, 40, 51 and 53 of SEQ ID NO: 99 are not substituted.

[0155] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 100, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 100. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 100 wherein the amino acids at positions 38, 51 and 53 of SEQ ID NO: 100 are not substituted. 31 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0156] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 101, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 101. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 101 wherein the amino acids at positions 36, 38, 51 and 53 of SEQ ID NO: 101 are not substituted.

[0157] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 102, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 102. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 102 wherein the amino acids at positions 36, 38, 40, 51 and 53 of SEQ ID NO: 102 are not substituted.

[0158] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 103, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 103. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 103 wherein the amino acids at positions 10, 49, 57, and 38 of SEQ ID NO: 103 are not substituted.

[0159] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 104, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 104. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 104 wherein the amino acids at positions 10, 36, 38, 49, and 57 of SEQ ID NO: 104 are not substituted.

[0160] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 105, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 105. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 105 wherein the amino acids at positions 10, 36, 38, 40, 49, and 57 of SEQ ID NO: 105 are not substituted.

[0161] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 106, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 106. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 106 wherein the amino acid at position 38 of SEQ ID NO: 106 is not substituted.

[0162] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 107, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 107. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 107 wherein the amino acids at positions 36 and 38 of SEQ ID NO: 107 are not substituted. 32 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0163] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 108, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 108. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 108 wherein the amino acids at positions 36, 38 and 40 of SEQ ID NO: 108 are not substituted.

[0164] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 109, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 109. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 109 wherein the amino acids at positions 36, 38 and 40 of SEQ ID NO: 109 are not substituted.

[0165] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 110, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 110. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 110 wherein the amino acids at positions 36 and 38 of SEQ ID NO: 110 are not substituted.

[0166] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 111, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 111. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 111 wherein the amino acids at positions 38, 51 and 57 of SEQ ID NO: 111 are not substituted.

[0167] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 112, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 112. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 112 wherein the amino acids at positions 36, 38, 51 and 57 of SEQ ID NO: 112 are not substituted.

[0168] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 113, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 113. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 113 wherein the amino acids at positions 36, 38, 40, 51 and 57 of SEQ ID NO: 113 are not substituted.

[0169] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 114, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 114. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to 33 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 SEQ ID NO: 114 wherein the amino acids at positions 38, 49 and 53 of SEQ ID NO: 114 are not substituted.

[0170] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 115, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 115. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 115 wherein the amino acids at positions 36, 38, 49 and 53 of SEQ ID NO: 115 are not substituted.

[0171] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 116, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 116. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 116 wherein the amino acids at positions 36, 38, 40, 49 and 53 of SEQ ID NO: 116 are not substituted.

[0172] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 117, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 117. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 117 wherein the amino acid at position 185 of SEQ ID NO: 117 is not substituted.

[0173] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 118, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 118. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 118 wherein the amino acids at positions 49, 57, 162 and 185 of SEQ ID NO: 118 are not substituted.

[0174] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 119, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 119. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 119 wherein the amino acids at positions 36, 49, 57, 162 and 185 of SEQ ID NO: 119 are not substituted.

[0175] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 120, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 120. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 120 wherein the amino acids at positions 36, 40, 49, 57, 162 and 185 of SEQ ID NO: 120 are not substituted.

[0176] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 121, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 121. For example, the IL- 34 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 121 wherein the amino acids at positions 54, 162 and 185 of SEQ ID NO: 121 are not substituted.

[0177] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 122, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 122. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 122 wherein the amino acids at positions 36, 54, 162 and 185 of SEQ ID NO: 122 are not substituted.

[0178] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 123, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 123. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 123 wherein the amino acids at positions 36, 40, 54, 162 and 185 of SEQ ID NO: 123 are not substituted.

[0179] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 124, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 124. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 124 wherein the amino acids at positions 51, 53, 162 and 185 of SEQ ID NO: 124 are not substituted.

[0180] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 125, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 125. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 125 wherein the amino acids at positions 36, 51, 53, 162 and 185 of SEQ ID NO: 125 are not substituted.

[0181] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 126, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 126. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 126 wherein the amino acids at positions 36, 40, 51, 53, 162 and 185 of SEQ ID NO: 126 are not substituted.

[0182] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 127, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 127. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 127 wherein the amino acids at positions 51, 53, 162 and 185 of SEQ ID NO: 127 are not substituted. 35 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0183] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 128, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 128. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 128 wherein the amino acids at positions 36, 162 and 185 of SEQ ID NO: 128 are not substituted.

[0184] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 129, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 129. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 129 wherein the amino acids at positions 36, 40, 51, 53, 162 and 185 of SEQ ID NO: 129 are not substituted.

[0185] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 130, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 130. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 130 wherein the amino acids at positions 10, 49, 57, 162, and 185 of SEQ ID NO: 130 are not substituted.

[0186] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 131, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 131. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 131 wherein the amino acids at positions 10, 36, 49, 57, 162, and 185 of SEQ ID NO: 131 are not substituted.

[0187] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 132, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 132. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 132 wherein the amino acids at positions 10, 36, 40, 49, 57162, and 185 of SEQ ID NO: 132 are not substituted.

[0188] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 133, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 133. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 133 wherein the amino acids at positions 162 and 185 of SEQ ID NO: 133 are not substituted.

[0189] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 134, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 134. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to 36 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 SEQ ID NO: 134 wherein the amino acids at positions 36, 162 and 185 of SEQ ID NO: 134 are not substituted.

[0190] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 135, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 135. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 135 wherein the amino acids at positions 36, 40, 162 and 185 of SEQ ID NO: 135 are not substituted.

[0191] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 136, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 136. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 136 wherein the amino acids at positions 36, 40, 162 and 185 of SEQ ID NO: 136 are not substituted.

[0192] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 137, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 137. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 137 wherein the amino acids at positions 36, 162 and 185 of SEQ ID NO: 137 are not substituted.

[0193] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 138, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 138. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 138 wherein the amino acids at positions 51, 162 and 185 of SEQ ID NO: 138 are not substituted.

[0194] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 139, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 139. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 139 wherein the amino acids at positions 36, 51, 57, 162 and 185 of SEQ ID NO: 139 are not substituted.

[0195] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 140, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 140. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 140 wherein the amino acids at positions 36, 51, 57, 40, 162 and 185 of SEQ ID NO: 140 are not substituted. 37 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0196] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 141, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 141. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 141 wherein the amino acids at positions 49, 53, 162 and 185 of SEQ ID NO: 141 are not substituted.

[0197] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 142, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 142. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 142 wherein the amino acids at positions 36, 49, 53, 162 and 185 of SEQ ID NO: 142 are not substituted.

[0198] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 143, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 143. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 143 wherein the amino acids at positions 36, 40, 49, 53, 162 and 185 of SEQ ID NO: 143 are not substituted.

[0199] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 144, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 144. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 144 wherein the amino acids at positions 162 and 185 of SEQ ID NO: 144 are not substituted.

[0200] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 145, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 145. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 145 wherein the amino acids at positions 49, 38, 57, 162 and 185 of SEQ ID NO: 145 are not substituted.

[0201] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 146, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 146. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 146 wherein the amino acids at positions 36, 38, 49, 57, 162 and 185 of SEQ ID NO: 146 are not substituted.

[0202] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 147, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 147. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to 38 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 SEQ ID NO: 147 wherein the amino acids at positions 36, 38, 40, 49, 57, 162 and 185 of SEQ ID NO: 147 are not substituted.

[0203] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 148, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 148. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 148 wherein the amino acids at positions 38, 54, 162 and 185 of SEQ ID NO: 148 are not substituted.

[0204] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 149, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 149. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 149 wherein the amino acids at positions 36, 38, 54, 162 and 185 of SEQ ID NO: 149 are not substituted.

[0205] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 150, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 150. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 150 wherein the amino acids at positions 36, 38, 40, 54, 162 and 185 of SEQ ID NO: 150 are not substituted.

[0206] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 151, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 151. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 151 wherein the amino acids at positions 38, 51, 53, 162 and 185 of SEQ ID NO: 151 are not substituted.

[0207] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 152, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 152. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 152 wherein the amino acids at positions 36, 38, 51, 53, 162 and 185 of SEQ ID NO: 152 are not substituted.

[0208] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 153, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 153. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 153 wherein the amino acids at positions 36, 38, 40, 51, 53, 162 and 185 of SEQ ID NO: 153 are not substituted. 39 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0209] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 154, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 154. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 154 wherein the amino acids at positions 38, 51, 53, 162 and 185 of SEQ ID NO: 154 are not substituted.

[0210] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 155, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 155. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 155 wherein the amino acids at positions 36, 38, 51, 53, 162 and 185 of SEQ ID NO: 155 are not substituted.

[0211] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 156, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 156. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 156 wherein the amino acids at positions 36, 38, 40, 51, 53, 162 and 185 of SEQ ID NO: 156 are not substituted.

[0212] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 157, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 157. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 157 wherein the amino acids at positions 10, 38, 49, 57, 162 and 185 of SEQ ID NO: 157 are not substituted.

[0213] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 158, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 158. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 158 wherein the amino acids at positions 10, 36, 38, 49, 57, 162 and 185 of SEQ ID NO: 158 are not substituted.

[0214] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 159, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 159. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 159 wherein the amino acids at positions 10, 36, 38, 40, 49, 57, 162 and 185 of SEQ ID NO: 159 are not substituted.

[0215] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 160, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 160. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to 40 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 SEQ ID NO: 160 wherein the amino acids at positions 38, 162 and 185 of SEQ ID NO: 160 are not substituted.

[0216] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 161, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 161. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 161 wherein the amino acids at positions 36, 38, 162 and 185 of SEQ ID NO: 161 are not substituted.

[0217] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 162, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 162. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 162 wherein the amino acids at positions 36, 38, 40, 162 and 185 of SEQ ID NO: 162 are not substituted.

[0218] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 163, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 163. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 163 wherein the amino acids at positions 36, 38, 40, 162 and 185 of SEQ ID NO: 163 are not substituted.

[0219] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 164, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 164. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 164 wherein the amino acids at positions 36, 38, 162 and 185 of SEQ ID NO: 164 are not substituted.

[0220] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 165, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 165. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 165 wherein the amino acids at positions 38, 51, 57, 162 and 185 of SEQ ID NO: 165 are not substituted.

[0221] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 166, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 166. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 166 wherein the amino acids at positions 36, 38, 51, 57, 162 and 185 of SEQ ID NO: 166 are not substituted. 41 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0222] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 167, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 167. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 167 wherein the amino acids at positions 36, 38, 40, 51, 57, 162 and 185 of SEQ ID NO: 167 are not substituted.

[0223] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 168, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 168. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 168 wherein the amino acids at positions 38, 49, 53, 162, and 185 of SEQ ID NO: 168 are not substituted.

[0224] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 169, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 169. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 169 wherein the amino acids at positions 36, 38, 49, 53, 162, and 185 of SEQ ID NO: 169 are not substituted.

[0225] The IL-18 polypeptide can comprise or consist of the amino acid sequence of SEQ ID NO: 170, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 170. For example, the IL- 18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 170 wherein the amino acids at positions 36, 38, 40, 49, 53, 162, and 185 of SEQ ID NO: 170 are not substituted.

[0226] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 200, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 200.

[0227] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 201, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 201.

[0228] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 202, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 202.

[0229] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 203, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 203.

[0230] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 204, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 204.

[0231] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 205, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 205. 42 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0232] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 206, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 206.

[0233] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 207, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 207.

[0234] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 208, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 208.

[0235] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 209, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 209.

[0236] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 211, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 211. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 211 wherein the amino acids at positions 51, 53, and 60 of SEQ ID NO: 211 are not substituted.

[0237] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 212, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 212. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 212 wherein the amino acids at positions 36, 37, 56, 57, and 60 of SEQ ID NO: 212 are not substituted.

[0238] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 213, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 213. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 213 wherein the amino acids at positions 36, 37, 56, 60, and 133 of SEQ ID NO: 213 are not substituted.

[0239] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 214, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 214. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 214 wherein the amino acids at positions 36, 37, 56, and 60 of SEQ ID NO: 214 are not substituted.

[0240] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 215, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 215. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 215 wherein the amino acids at positions 36, 37, 56, 60, and 133 of SEQ ID NO: 215 are not substituted. 43 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0241] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 216, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 216. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 216 wherein the amino acids at positions 36, 37, 56, 60, and 133 of SEQ ID NO: 216 are not substituted.

[0242] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 217, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 217. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 217 wherein the amino acids at positions 36, 37, 56, and 60 of SEQ ID NO: 217 are not substituted.

[0243] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 218, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 218. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 218 wherein the amino acids at positions 36, 37, 56, and 60 of SEQ ID NO: 218 are not substituted.

[0244] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 219, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 219. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 219 wherein the amino acids at positions 36, 37, 57, and 60 of SEQ ID NO: 219 are not substituted.

[0245] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 220, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 220. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 220 wherein the amino acids at positions 36, 37, and 60 of SEQ ID NO: 220 are not substituted.

[0246] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 221, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 221. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 221 wherein the amino acids at positions 36, 37, 60, and 133 of SEQ ID NO: 221 are not substituted.

[0247] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 222, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 222. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least 44 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 about 95% identity to SEQ ID NO: 222 wherein the amino acids at positions 36, 37, 60, and 133 of SEQ ID NO: 222 are not substituted.

[0248] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 223, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 223. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 223 wherein the amino acids at positions 36, 37, 57, and 60 of SEQ ID NO: 223 are not substituted.

[0249] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 224, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 224. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 224 wherein the amino acids at positions 36, 37, 59, and 60 of SEQ ID NO: 224 are not substituted.

[0250] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 225, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 225. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 225 wherein the amino acids at positions 36, 37, 57, and 60 of SEQ ID NO: 225 are not substituted.

[0251] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 226, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 226. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 226 wherein the amino acids at positions 36, 37, and 60 of SEQ ID NO: 226 are not substituted.

[0252] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 227, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 227. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 227 wherein the amino acids at positions 36, 37, 57, 59, and 60 of SEQ ID NO: 227 are not substituted.

[0253] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 228, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 228. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 228 wherein the amino acids at positions 36, 37, 57, 59, and 60 of SEQ ID NO: 228 are not substituted. 45 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0254] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 229, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 229. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 229 wherein the amino acids at positions 36, 51, and 53 of SEQ ID NO: 229 are not substituted.

[0255] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 230, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 230. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 230 wherein the amino acids at positions 51, 53, 129, 131, and 134 of SEQ ID NO: 230 are not substituted.

[0256] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 231, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 231. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 231 wherein the amino acids at positions 36, 37, 51, and 53 of SEQ ID NO: 231 are not substituted.

[0257] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 232, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 232. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 232 wherein the amino acids at positions 51, 53, and 145 of SEQ ID NO: 232 are not substituted.

[0258] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 233, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 233. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 233 wherein the amino acids at positions 51, 53, and 150 of SEQ ID NO: 233 are not substituted.

[0259] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 234, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 234. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 234 wherein the amino acids at positions 51, 53, 131, and 132 of SEQ ID NO: 234 are not substituted.

[0260] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 235, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 235. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least 46 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 about 95% identity to SEQ ID NO: 235 wherein the amino acids at positions 51, 53, and 149 of SEQ ID NO: 235 are not substituted.

[0261] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 236, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 236. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 236 wherein the amino acids at positions 36, 37, 49, and 57 of SEQ ID NO: 236 are not substituted.

[0262] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 237, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 237. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 237 wherein the amino acids at positions 36, 49, and 57 of SEQ ID NO: 237 are not substituted.

[0263] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 238, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 238. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 238 wherein the amino acids at positions 37, 49, 57, 93, and 134 of SEQ ID NO: 238 are not substituted.

[0264] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 239, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 239. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 239 wherein the amino acids at positions 24, 27, 57, 49, and 111 of SEQ ID NO: 239 are not substituted.

[0265] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 240, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 240. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 240 wherein the amino acids at positions 49 and 57 of SEQ ID NO: 240 are not substituted.

[0266] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 241, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 241. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 241 wherein the amino acids at positions 34, 49, and 57 of SEQ ID NO: 241 are not substituted. 47 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0267] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 242, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 242. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 242 wherein the amino acids at positions 49, 57, and 129 of SEQ ID NO: 242 are not substituted.

[0268] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 243, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 243. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 243 wherein the amino acids at positions 49, 57, 129, 131, and 134 of SEQ ID NO: 243 are not substituted.

[0269] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 244, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 244. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 244 wherein the amino acids at positions 36, 37, and 54 of SEQ ID NO: 244 are not substituted.

[0270] The IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 245, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 245. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 245 wherein the amino acids at positions 36, 37, 49, and 57 of SEQ ID NO: 245 are not substituted.

[0271] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 8, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 8. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 8 wherein the amino acids at positions 57 and 62 of SEQ ID NO: 8 are not substituted.

[0272] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 14, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 14. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 14 wherein the amino acids at positions 10, 49 and 91 of SEQ ID NO: 14 are not substituted.

[0273] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 18, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 18. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least 48 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 about 95% identity to SEQ ID NO: 18 wherein the amino acid at position 91 of SEQ ID NO: 18 is not substituted.

[0274] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 25, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 25. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 25 wherein the amino acids at positions 49 and 91 of SEQ ID NO: 25 are not substituted.

[0275] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 27, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 27. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 27 wherein the amino acids at positions 49 and 57 of SEQ ID NO: 27 is not substituted.

[0276] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 66, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 66. For example, the IL-18 polypeptide variant can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 66 wherein the amino acids at positions 51 and 53 of SEQ ID NO: 66 are not substituted.

[0277] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 71, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 71. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 71 wherein the amino acids at positions 36, 40, 49, and 57 of SEQ ID NO: 71 are not substituted.

[0278] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 75, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 75. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 75 wherein the amino acids at positions 36, 40, 51, and 53 of SEQ ID NO: 75 are not substituted.

[0279] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 73, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 73. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 73 wherein the amino acids at positions 36, 40, and 54 of SEQ ID NO: 73 are not substituted. 49 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0280] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 83, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 83. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 83 wherein the amino acids at positions 36 and 40 of SEQ ID NO: 83 are not substituted.

[0281] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 96, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 96. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 96 wherein the amino acids at positions 36, 38, 40 and 54 of SEQ ID NO: 96 are not substituted.

[0282] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 103, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 103. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 103 wherein the amino acids at positions 10, 49, 57, and 38 of SEQ ID NO: 103 are not substituted

[0283] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 120, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 120. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 120 wherein the amino acids at positions 36, 40, 49, 57, 162 and 185 of SEQ ID NO: 120 are not substituted.

[0284] Certain preferred IL-18 polypeptide variants can comprise or consist of the amino acid sequence of SEQ ID NO: 150, or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 150. For example, the IL-18 polypeptide can comprise or consist of an amino acid sequence that has at least about 95% identity to SEQ ID NO: 150 wherein the amino acids at positions 36, 38, 40, 54, 162 and 185 of SEQ ID NO: 150 are not substituted.

[0285] In certain preferred aspects, the IL-18 polypeptide variant of this disclosure comprises SEQ ID NO:1 wherein one or more amino acids are modified (e.g., substituted or omitted) as described herein, with the proviso that the IL-18 polypeptide variant does not include amino acid modification at one or more of positions 5, 8, 17, 31, 34, 35, 41, 59, 77, 103, 105, 109, 110, 111, 113, 153 and 155. In other preferred aspects, the IL-18 polypeptide variant of this disclosure comprises SEQ ID NO:1 wherein one or more amino acids are modified (e.g., substituted or omitted) as described herein, with the proviso that the IL-18 polypeptide variant does not include amino acid modification at one or more of positions 1 through 8, 17, 31, 34 through 41, and 51 through 60. In some instances, the IL-18 polypeptide variant 50 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 does not include amino acid modifications at positions 1, 5, 8, 17, 31, 34, 35, 36, 37, 40, 41, 51, 53, 56, 57, 59, 60, 103, 105, 110, 111, and 113.

[0286] Certain preferred IL-18 variant polypeptides of this disclosure comprises SEQ ID NO:1 wherein one or more amino acids are modified (e.g., substituted or omitted) as described herein, with the proviso IL-18 polypeptide variants do not include one or more, including all, of the substitutions Y1R, Y1H, Y1D, Y1L, Y1F, L5H, L5I, L5Y, L5F, L5H, L5F, K8Q, K8R, D17G, D17R, D17A, E31K, E31A, E31T, E31G, E31R, T34E, T34K, T34A, D35A, D35S, D35Y, D35A, S36N, S36K, S36R, D37P, D37R, D37A, D37L, D37H, D37V, D40A, D40S, D40Y, N41R, N41S, N41K, M51T, M51D, M51N, M51E, M51F, M51I, M51L, K53R, K53G, Q56E, Q56A, Q56R, Q56V, Q56G, Q56L, Q56K, Q56H, P57L, P57G, P57A, P57K, G59A, G59T, M60Q, M60K, M60R, M60L, M60I, M60F, E77D, Q103K, Q103E, Q103A, Q103R, Q103I, Q103L, S105D, S105A, S105N, H109A, H109P, H109D, D110K, D110H, D110Q, D110E, D110N, D110S, D110G, N111H, N111Y, N111D, N111R, N111S, N111G, M113V, M113R, M113T, M113F, M113I, M113L, V153I, V153T, V153A, N155K, and N155H.

[0287] The IL-18 polypeptide variants disclosed herein preferably bind to the IL-18 receptor (human IL- 18 receptor) and induced signaling through the IL-18 receptor. Preferred IL-18 polypeptide variants of this disclosure bind the IL-18 receptor with an affinity that is about the same as or stronger than naturally occurring human IL-18 (SEQ ID NO:1). More preferably, the IL-18 polypeptide variants disclosed herein also bind to IL-18 binding protein with affinity that is weaker than naturally occurring human IL-18 (SEQ ID NO:1). The IL-18 polypeptide variants disclosed herein can have a binding affinity for IL-18R of about 10 nM, about 5 nM, about 1 nM, about 0.5 nM or less. The IL-18 polypeptide variants disclosed herein can have a binding affinity for IL-18BP of about 0.5 nM, about 1 nM, about 5 nM, about 100 nM, about 599 nM, about 1000 nM or no binding.

[0288] The IL-18 polypeptide variants disclosed herein can comprise one or more amino acid modifications to improve binding to the IL-18 receptor. For instance, the IL-18 polypeptide variant can have higher potency and increased binding affinity to the IL-18 receptor relative to SEQ ID NO: 1. The IL-18 polypeptide variants disclosed herein can comprise one or more amino acid modifications to reduce binding to the IL-18 binding protein. For instance, the IL-18 polypeptide variant can have decreased binding affinity to the IL-18 binding protein relative to SEQ ID NO: 1.

[0289] The IL-18 polypeptide variants disclosed herein are mammalian IL-18 polypeptide variants. For example, the IL-18 polypeptide variant can be a human IL-18 polypeptide variant or a murine IL-18 polypeptide variant. Preferably, the IL-18 polypeptide variant is a human IL-18 polypeptide variant. 51 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0290] The IL-18 polypeptide variant can be further modified. For example, post-translationally modified, for example by prenylation, acetylation, amidation, carboxylation, glycosylation, or PEGylation (covalent attachment of polyethylene glycol (PEG) polymer chains). B. IL-18 Polypeptide Fusion Proteins and Conjugates

[0291] The IL-18 polypeptide variants disclosed herein can be formatted into a variety of suitable polypeptide structures, for example, proteins, fusion proteins, conjugates or any other suitable format. i. Fusion Proteins and Conjugates

[0292] The IL-18 polypeptide variants disclosed herein are particularly suitable for use in fusion polypeptides and conjugates. The fusion polypeptide can comprise a continuous polypeptide chain that comprises an IL-18 polypeptide variant as disclosed herein and an amino acid sequence of interest. The IL-18 polypeptide variants can be fused to the amino acid sequence of interest directly, or indirectly, for example through an amino acid linker. If desired, the fusion polypeptide can associate with one or more other polypeptides to form a functional polypeptide complex.

[0293] In non-limiting examples, the fusion polypeptide can be represented by Formula V or VI: [A1]-[L1]-[D1] (V) [D1]-[L1]-[A1] (VI), wherein A1 is the IL-18 polypeptide variants disclosed herein, L1 is a linker that connects or links A1 to D1 or is absent, and D1 is an amino acid sequence of interest.

[0294] The fusion polypeptide can comprise an IL-18 polypeptide variant and an amino acid sequence of interest. For example, the fusion polypeptide can comprise an IL-18 polypeptide comprising the amino acid sequence of any one of SEQ ID NO: 2-170, and an amino acid sequence of interest.

[0295] The amino acid sequence of interest can be any desired amino acid sequence, typically it is the amino acid sequence of a therapeutic agent, targeting agent, an agent to enhance its efficacy, or an agent to improve its safety. Exemplary amino acid sequences of interest include, but are not limited to those of, cytokines, receptors (e.g., soluble receptors that bind cytokines, growth factors, hormones, Toll-like receptor agonists, and the like), enzymes, growth factors, hormones (e.g., peptide hormones), chemokine, tumor antigen binding domains, matrix proteins, pathogen antigens, immunoglobulin formats (e.g., antigen-binding portions of antibodies) or fragments thereof, half-life extension elements (e.g., polyethylene glycol or human serum albumin), antibiotics, cytotoxic drugs, or other recombinant polypeptide complexes. 52 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0296] Suitable amino acid sequences of interest (e.g., cytokines, growth factors, hormones, antibodies and other polypeptides) include, but are not limited to, 4-1BBL, ApoE, Apo-SAA, BDNF, cardiotrophin- 1, CD40L, CD70, EGF, EGF receptor, ENA-79, exotoxin, Epo-R, FGF-1, FGF-2, FGF-3, FGF-4, FGF-5, FGF-6, FGF-7, FGF-8a, FGF-8b, FGF-8e, FGF-8f, FGF-9, FGF-10, FGF-11, FGF-12, FGF-13, FGF-14, FGF-16, FGF-17, FGF-18, FGF-19, FGF-20, FGF-21, FGF-22, FGF-23, FLT3 ligand, Fractalkine (CX3C), GDNF, G-CSF, GITRL, GM-CSF, CF-beta1, insulin, IFN-alpha, IFN-beta, IFN-gamma, IGF-I, IGF-II, IL-1, IL-2, IL-4, IL-5, IL-6, IL-10, IL-18, IL-22, IL-23p19, IL-11, IL-12, IL-13, IL-15, IL-12p35, IL-12p40, IL-21, IL-30 (IL27p28), IL-34, IL-35, IL-36, inhibin α, Inhibin β, IP-10, keratinocyte growth factor-2 (KGF-2), KGF, LIF, Lymphotactin, Mullerian inhibitory substance, monocyte colony inhibitory factor, monocyte attractant protein, M-CSF, MDC (67 a.a.), MDC (69 a.a.), MCP-I (MCAF), MCP-2, MCP-3, MCP- 4, MDC (67 a.a.), MDC (69 a.a.), MIG, MIP-Ia, MIP-I β, MIP-3α, MIP-3β, MIP-4, myeloid progenitor inhibitor factor- 1 (MPIF-I), NAP -2, Neurturin, Nerve growth factor, β-NGF, NT-3, NT-4, Oncostatin M, OX40L, PDGF-AA, PDGF-AB, PDGF-BB, PF-4, RANTES, SDFl α, SDFl β, SCF, SCGF, stem cell factor (SCF), TARC, TGF-α, TGF-β, TGF-β2, TGF-β3, tumour necrosis factor (TNF), TNF-α, TNF-β, TNIL-I, TPO, VEGF, GCP-2, GRO / MGSA, GRO-β, GRO-γ, HCCl, 1-309, TNF-alpha, IFN-gamma, matrix metalloproteinase, CEA, PsmAr,. It will be appreciated that this list is by no means exhaustive.

[0297] Suitable hormones and hormone receptor agonists, include, Leptin, GLP-1, GLP-1 receptor agonists, GIP, Ghrelin, asprosin, CGRP, insulin and the like.

[0298] Suitable chemokines can include, for example, CXCL9, CXCL10, CXCL111, CXCL13, CCL19, CCL20, CCL21, XCL1 or functional fragments.

[0299] Suitable tumor antigen binding domains can include, but are not limited to domains that bind EpCAM, EGFR, HER-2, HER-3, c-Met, FolR, PSMA, CD38, BCMA, CEA, 5T4, AFP, B7-H3, Cadherin-6, CAIX, CD117, CD123, CD138, CD166, CD19, CD20, CD205, CD22, CD30, CD33, CD352, CD37, CD44, CD52, CD56, CD70, CD71, CD74, CD79b, DLL3, EphA2, FAP, FGFR1, FGFR2, FGFR3, FGFR4, FOLH1 (PSMA), GPC3, gpA33, FLT-3, gpNMB, HPV-16 E6, HPV-16 E7, ITGA2, ITGA3, SLC39A6, MAGE, mesothelin, Muc1, Muc16, NaPi2b, Nectin-4, P-cadherin, NY-ESO-1, PRLR, PSCA, PTK7, ROR1, SLC44A4, SLTRK5, SLTRK6, STEAP1, TIM1, Trop2, WT1.

[0300] Examples of suitable receptors include, but are not limited to, Type I cytokine receptors, such as GM-CSF receptor, G-CSF receptor, Type I IL receptors, Epo receptor, LIF receptor, CNTF receptor, TPO receptor; Type II Cytokine receptors, such as IFN-alpha receptor (IFNAR1, IFNAR2), IFB-beta receptor, IFN-gamma receptor (IFNGR1, IFNGR2), Type II IL receptors; chemokine receptors, such as CC chemokine receptors, CXC chemokine receptors, CX3C chemokine receptors, XC chemokine receptors; 53 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 tumor necrosis receptor superfamily receptors, such as TNFRSF5 / CD40, TNFRSF8 / CD30, TNFRSF7 / CD27, TNFRSF1A / TNFR1 / CD120a, TNFRSF1B / TNFR2 / CD120b; TGF-beta receptors, such as TGF-beta receptor 1, TGF-beta receptor 2; Ig super family receptors, such as IL-1 receptors, CSF- 1R, PDGFR (PDGFRA, PDGFRB), SCFR, VEGF receptor 1, VEGF receptor 2, VEGF receptor 3, internalizing receptors that are over-expressed on certain cells, such as the epidermal growth factor receptor (EGFR), ErBb2 receptor on tumor cells, an internalizing cellular receptor, LDL receptor, FGF2 receptor, ErbB2 receptor, transferrin receptor, PDGF receptor, human chemokine receptors CXCR4 or CCR5, and HER 1, HER 2, HER 3, HER 4, and CD4.

[0301] The amino acid sequence of interest preferably encodes a cytokine. Suitable cytokines, include for example, IFN-alpha, IFN-beta, IFN-gamma, IL-1, IL-2, IL-4, IL-5, IL-6, IL-10, IL-18, IL-22, IL- 23p19, IL-11, IL-12, IL-13, IL-15, IL-12p35, IL-21, IL-30 (IL27p28), IL-34, IL-35, IL-36 and the like. Preferable cytokines include IL-2, IL-12 (IL-12 p35 and / or IL-12p40), IL-10, IL-18, and IL-21. For example, the preferred cytokine can be IL-2. For example, the preferred cytokine can be IL-12. For example, the preferred cytokine can be IL-2. If desired, the amino acid of interest can encode an inducible cytokine prodrug. For example, the inducible cytokine prodrug can be an inducible IL-2 prodrug, an inducible IL-12 prodrug, a inducible IL-10 prodrug, an inducible IL-21 prodrug, an inducible IL-21 prodrug, or an inducible IFN prodrug. Inducible IL-12, IL-2, and interferon prodrugs have been described in International Publication Nos.: WO2019 / 222294, WO2019 / 222295, WO2019 / 222296, WO2021 / 097376.

[0302] Suitable antibodies or antibody fragments of interest can have binding specificity for any desired antigen, and include, for example, anti-CD20 antibodies, anti-HER2 antibodies, anti-PD-1 / PD-L1 antibodies, anti-CD19 antibodies, and anti-VEGEF antibodies. Other suitable antibodies or antibody fragments of interest can have binding specificity for IL-18 (i.e. an anti-IL-18 antibody or antibody fragment).

[0303] The amino acid sequence of interest can be an immunoglobulin format. Suitable immunoglobulin formats are well-known in the art and include bispecific and multi-specific formats that can be bivalent or multivalent. Suitable immunoglobulin formats include, for example, heavy chain antibody, Fab, Fab’, F(ab’)2, Fv, disulfide stabilized Fv fragment (dsFv), (dsFV) 2, bispecific dsFv (dsFv-dsFv’), a disulfide diabody, a single-chain Fv (scFv), a diabody, triabodies, tetrabodies, tribodies, a multispecific antibody formed from a portion of an antibody comprising one or more CDRs, sdAb, a bivalent domain antibody, or any other antibody fragment that binds to an antigen but does not comprise a complete IgG antibody structure (e.g., an antibody fragment from a monoclonal antibody, a polyclonal antibody, a recombinant antibody, a human antibody, a humanized antibody). 54 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0304] The amino acid of interest can be immunoglobulin formats that are known in the art as bispecific, trispecific or multispecific cell engagers (or an amino acid chain of a such a cell engager). Such cell engagers, including bispecific T-cell engagers (BiTEs), dual-affinity re-targering molecules (DARTs), bispecific killer cell engagers (BiKEs) and trispecific killer cell engagers (TriKEs) are well-known in the art and include a binding region (typically a sdAb or two variable regions that form a VH / VL binding sight) that binds an antigen on an immune cell (e.g, an effector cell such as a T-cell or NK cell) and a binding region that binds a tumor-associated antigen. See, e.g., Allen et al., (2021), Life (Basel), 11(6):465. For example, such bispecific, trispecific or multispecific cell engagers can engage T-cells and tumor cells, by binding to CD3 on the T cell and to a tumor associated antigen on a tumor cell. For example, such bispecific, trispecific or multispecific cell engagers can engage NK cells and tumor cells, by binding to CD16 on the NK cell and to a tumor associated antigen on a tumor cell.

[0305] Of specific interest are fusion polypeptides that are suitable for use with payloads that target or are targeted to tumor microenvironments.

[0306] The IL-18 polypeptide variants disclosed herein can be linked directly or indirectly to the immune cell (e.g., T-cell or NK cell) binding domain and / or a target antigen (e.g., a tumor associated antigen) binding domain. The IL-18 polypeptide variants disclosed herein can be linked to the immune cell (e.g., T cell or NK cell binding domain and / or the target antigen (e.g., tumor associated antigen) binding domain through a linker, such as a protease cleavable linker as described herein, or a non-cleavable linker. The IL-18 polypeptide disclosed herein can be linked to the immune cell binding domain. The IL-18 polypeptide variants disclosed herein can be linked to the target antigen binding domain. The IL-18 polypeptide variants disclosed herein can be linked both the immune cell binding domain and the target antigen binding domain. The bispecific, trispecific or multispecific T-cell engager can comprise a) a IL- 18 polypeptide variant as disclosed herein, b) a binding region (e.g., sdAb, scFv etc.) that has specificity for the TCR complex including CD3, and c) a binding region that has specificity for a target antigen, such as a tumor associated antigen.

[0307] The binding region with specificity for the TCR complex can specifically bind to CD3, preferably human CD3. The binding region can have binding specificity for one or more chain of the CD3 complex, such as CD3 epsilon, CD3 gamma, CD3 delta, or CD3 zeta. The binding region can have binding specificity for CD3 epsilon. The binding region can have binding specificity for CD3 gamma. The binding region can have binding specificity for CD3 delta. The binding region can have binding specificity for CD3 zeta. The binding region having binding specificity for the TCR can further comprise a domain that specifically bind the alpha chain of the TCR and / or the beta chain of the TCR. 55 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0308] The binding region having binding specificity to CD3 can be any suitable immunoglobulin format that binds CD3. Examples include, but are not limited to, an antigen binding fragments, such as single domain antibodies, Fab, Fab’, F(ab)2, single chain Fv fragments, disulfide stabilizd Fv fragments, Fv fragments, heavy chain monomers or dimers, light chain monomers or dimers. Suitable binding fragments that have specificity for CD3 can be obtained, for example, from any anti-CD3 antibodies. Many such antibodies and binding regions are well-known, such as muromonab-CD3 (OKT3), otelixizumab (TRX4), teplizumab (MGA031), visilizumab (Nuvion), SP34, TR-66 or X35-3, VIT3, BMA030 (BW264 / 56), CLB-T3 / 3, CRIS7, YTH12.5M, F111-409, CLB-T3.4.2M TR-66, WT32, SPv-T3b, 11D8, XIII-141, XIII-141, XIII-87, T3 / RW2-8C8, T3 / RW2-8C8, T3 / RW2-4B6, OKT3D, M-T301, SMC2, F101.01, UCHT-1, and WT-31.

[0309] In addition, the bispecific, trispecific or multispecific cell engager can further comprise a region (e.g., sdAb, scFv) that binds to a target antigen. A target antigen is involved in and / or associated with a disease, disorder, or condition. In particular, the target antigen can be associated with or involved with a disease, disorder, or condition. Specifically, the target antigen can be associated with cancer, an inflammatory disease, an immunological disorder, an autoimmune disease, a viral disease, or a bacterial disease. The target antigen binding region can be any suitable immunoglobulin format that binds to the target antigen. Examples include, but are not limited to, an antigen binding fragments, such as single domain antibodies, Fab, Fab’, F(ab)2, single chain Fv fragments, disulfide stabilizd Fv fragments, Fv fragments, heavy chain monomers or dimers, light chain monomers or dimers. This disclosure also relates to conjugates. Like the fusion polypeptides described herein, conjugates comprise an IL-18 polypeptide variant as described herein and an amino acid sequence of interest. In the conjugates of this disclosure, the IL-18 polypeptide variant is covalently bonded directly or indirectly to the amino acid sequence of interest. The IL-18 polypeptide variant can be conjugated to the amino acid sequence of interest at the C-terminus of the amino acid sequence of interest, at the N-terminus of the amino acid sequence of interest, or internally, for example through, an amino acid side chain of an amino acid in the amino acid sequence of interest. Many suitable linkers and functional groups that can be used to prepare conjugates as described herein are well-known in the art. For example, homo- or hetero-bifunctional cross-linkers can be used to link a IL-18 polypeptide variant disclosed herein to an amino acid sequence of interest, for example through an amine group on the IL-18 polypeptide variant and a carboxyl group on the amino acid sequence of interest. In some aspects, the IL-18 polypeptide variant is indirectly conjugated to the amino acid sequence of interest through a linker that includes a peptide or polypeptide amino acid sequence. Such linkers can be non-cleavable or cleavable, for example by protease (e.g., a protease with activity that is associated with diseased cells or tissues as disclosed herein). 56 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0310] The IL-18 polypeptide variant and the amino acid sequence of interest in the fusion polypeptides and conjugates of this disclosure can be linked to each other directly or indirectly. The IL-18 polypeptide variant and the amino acid sequence of interest may be linked through a suitable amino acid or peptide or polypeptide linker. Such linker sequences can be a naturally occurring sequence or a non-naturally occurring sequence. Preferably, the linker sequence is a non-naturally occurring sequence.

[0311] The IL-18 polypeptide variant and the amino acid sequence of interest can be linked by a non- cleavable linker. The IL-18 polypeptide variant and the amino acid sequence of interest is preferably linked by a cleavable linker. The cleavable linker can comprise one or more cleavage sites for one or more desired protease. Preferably, the desired protease activity is enriched or selectively expressed at the desired target site of the amino acid sequence of interest, such as a desired cite of cytokine activity (e.g., the tumor microenvironment). The linker is preferentially or selectively cleaved at the target site.

[0312] Suitable linkers are typically less than about 100 amino acids. Such linkers can be of different lengths, such as from 1 amino acid (e.g., Gly) to 30 amino acids, from 1 amino acid to 40 amino acids, from 1 amino acid to 50 amino acids, from 1 amino acid to 60 amino acids, from 1 to 70 amino acids, from 1 to 80 amino acids, from 1 to 90 amino acids, and from 1 to 100 amino acids. In some embodiments, the linker is at least about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22, about 23, about 24, about 25, about 26, about 27, about 28, about 29, about 30, about 31, about 32, about 33, about 34, about 35, about 36, about 37, about 38, about 39, about 40, about 41, about 42, about 43, about 44, about 45, about 46, about 47, about 48, about 49, about 50, about 51, about 52, about 53, about 54, about 55, about 56, about 57, about 58, about 59, about 60, about 61, about 62, about 63, about 64, about 65, about 66, about 67, about 68, about 69, about 70, about 71, about 72, about 73, about 74, about 75, about 76, about 77, about 78, about 79, about 80, about 81, about 83, about 83, about 84, about 85, about 86, about 87, about 88, about 89, about 90, about 91, about 92, about 93, about 94, about 95, about 96, about 97, about 98, about 99m or about 100 amino acids in length. Preferred linkers are typically from about 5 amino acids to about 30 amino acids.

[0313] Preferably the lengths of linkers vary from 2 to 30 amino acids, optimized for each condition so that the linker does not impose any constraints on the conformation or interactions of the linked amino acid sequence of interest. In a preferred embodiment, the linker is cleavable by a cleaving agent, e.g., an enzyme. Preferably, the linker comprises a protease cleavage site. In some cases, the linker comprises one or more cleavage sites. The linker can comprise a single protease cleavage site. The linker can also comprise 2 or more protease cleavage sites. For example, 2 cleavage sites, 3 cleavage sites, 4, cleavage sites, 5 cleavage sites, or more. In cases where the linker comprises 2 or more protease cleavage sites, the 57 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 cleavage sites can be cleaved by the same protease or different proteases. A linker comprising two or more cleavage sites is referred to as a “tandem linker.” The two or more cleavage sites can be arranged in any desired orientation, including, but not limited tom one cleavage site adjacent to another cleavage site, one cleavage site overlapping another cleavage site, or one cleavage site following by another cleavage site with intervening amino acids between the two cleavage sites.

[0314] The IL-18 polypeptide variant can be linked, either directly or indirectly, to a small molecule. Suitable small molecules of interest include, but are not limited to, methotrexate, tyrosine kinase inhibitors, corticosteroids, immunomodulators, and chemotherapeutics.

[0315] When the IL-18 polypeptide variant is fused to an amino acid sequence of interest it can be a fragment of the IL-18 polypeptide variant. In some cases, the IL-18 polypeptide variant fragment can be about 100 amino acids or more in length up to full-length the IL-18 polypeptide variant. For example, the IL-18 polypeptide variant fragment can be about 100 amino acids, about 110 amino acids, about 115 amino acids, about 120 amino acids, about 125 amino acids, about 130 amino acids, about 135 amino acids, about 140 amino acids, about 145 amino acids, about 150 amino acids, or about 155 amino acids or more. ii. Half-Life Extended IL-18 Polypeptide Variants

[0316] Exemplary fusion polypeptides and conjugates disclosed herein can have increased in vivo half- life. The fusion polypeptide or a conjugate can comprise an IL-18 polypeptide variant as disclosed herein, a half-life extension element (e.g., an Fc-domain), and optionally a linker (e.g., cleavable or non- cleavable linker). The optional linker fuses the IL-18 polypeptide variant and the half-life extension element indirectly or directly.

[0317] The half-life extension element can improve tissue targeting, tissue penetration, diffusion within the tissue, and enhanced efficacy as compared with a protein without a half-life extension element. Without being bound by theory, an exemplary way to improve the pharmacokinetics of a polypeptide is by expression of an element in the polypeptide chain that binds to receptors that are recycled to the plasma membrane of cells rather than degraded in the lysosomes, such as the FcRn receptor on endothelial cells and transferrin receptor. Three types of proteins, e.g., human IgGs, HSA (or fragments), and transferrin, persist for much longer in human serum than would be predicted just by their size, which is a function of their ability to bind to receptors that are recycled rather than degraded in the lysosome. These proteins, or fragments that retain FcRn binding, are routinely linked to other polypeptides to extend their serum half-life. HSA may also be directly bound to the pharmaceutical compositions or bound via a short linker. Fragments of HSA may also be used. HSA and fragments thereof can function as both a 58 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 blocking element and a half-life extension element. Human IgGs and immunoglobulin Fc or fragments thereof can also carry out a similar function.

[0318] The serum half-life extension element can also be an antigen-binding polypeptide that binds to a protein with a long serum half-life such as serum albumin (e.g., HSA), transferrin and the like. Examples of such polypeptides include antibodies and fragments thereof including, a polyclonal antibody, a recombinant antibody, a human antibody, a humanized antibody, a single chain variable fragment (scFv), an antigen binding fragment (Fab), single-domain antibody such as a heavy chain variable domain (VH), a light chain variable domain (VL) and a variable domain of camelid-type nanobody (VHH), a dAb and the like. Other suitable antigen-binding domains include non-immunoglobulin proteins that mimic antibody binding and / or structure such as, anticalins, affilins, affibody molecules, affimers, affitins, alphabodies, avimers, DARPins, fynomers, kunitz domain peptides, monobodies, and binding domains based on other engineered scaffolds such as SpA, GroEL, fibronectin, lipocalin and CTLA4 scaffolds. Further examples of antigen-binding polypeptides include a ligand for a desired receptor, a ligand-binding portion of a receptor, a lectin, and peptides that bind to or associate with one or more target antigens.

[0319] The half-life extension element can be an immunoglobulin Fc domain or a fragment thereof. For example, the immunoglobulin Fc domain can comprise a CH2 and CH3 domain or fragment thereof. For example, the immunoglobulin Fc domain can comprise a constant domain of the heavy chain polypeptide.

[0320] The immunoglobulin Fc domain or a fragment thereof can further comprise one or more amino acid modifications, if desired. A preferred amino acid modification is glycine to serine.

[0321] The half-life extended IL-18 polypeptide variants can further comprise a linker as described herein. The linker can be a cleavable linker as disclosed herein. Preferably, the cleavable linker is a protease cleavable linker. The desired protease activity is enriched or selectively present at the desired target site of the cytokine activity (e.g., the tumor microenvironment). The linker is preferentially or selectively cleaved at the target site.

[0322] The half-life extended IL-18 polypeptide variants can comprise an IL-18 polypeptide that comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 70-170, 172-209, or 210- 245, or an amino acid sequence that is at least 95% identical to the amino acid sequence of any one of SEQ ID NOs: 70-170, 172-209, or 210-245 and a scFc linked to the C-terminus of the IL-18 polypeptide. The scFc can comprise or consist of the amino acid sequence of SEQ ID NO: 210, or an amino acid sequence that has at least 85% identity to SEQ ID NO: 110. The half-life extended IL-18 polypeptide variants can comprise an IL-18 polypeptide that comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 70-170, 172-209, or 210-245, or an amino acid sequence that is at least 95% identical to the amino acid sequence of any one of SEQ ID NOs: 70-170, 172-209, or 210-245 and an 59 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 scFc that comprises or consists of the amino acid sequence of SEQ ID NO: 210, or an amino acid sequence that has at least 85% identity to SEQ ID NO: 110.

[0323] The half-life extended IL-18 polypeptide variants can comprise or consist of the amino acid sequence of any one of SEQ ID NOs: 172-181, or an amino acid sequence that has at least about 85% identity to any one of SEQ ID NOs: 172-181.

[0324] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 172, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 172.

[0325] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 173, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 173.

[0326] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 174, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 174.

[0327] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 175, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 175.

[0328] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 176, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 176.

[0329] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 177, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 177.

[0330] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 178, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 178.

[0331] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 179, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 179.

[0332] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 180, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 180. 60 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0333] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 181, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 181.

[0334] The half-life extended IL-18 polypeptide variants can comprise or consist of the amino acid sequence of any one of SEQ ID NOs: 182-197, or an amino acid sequence that has at least about 85% identity to any one of SEQ ID NOs: 182-197.

[0335] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 182, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 182.

[0336] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 183, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 183.

[0337] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 184, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 184.

[0338] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 185, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 185.

[0339] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 186, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 186.

[0340] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 187, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 187.

[0341] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 188, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 188.

[0342] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 189, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 189.

[0343] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 190, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 190. 61 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0344] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 191, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 191.

[0345] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 192, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 192.

[0346] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 193, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 193.

[0347] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 194, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 194.

[0348] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 195, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 195.

[0349] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 196, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 196.

[0350] The half-life extended IL-18 polypeptide variant can comprise or consist of the amino acid sequence of SEQ ID NO: 197, or an amino acid sequence that has at least about 85% identity to SEQ ID NO: 197. iii. Adoptive Cell Therapy and Immune Cells

[0351] This disclosure further relates to immune cells that can be engineered to express a IL-18 polypeptide variant disclosed herein. For example, a T cell (e.g., TIL, CAR-T, TCR-T) can be engineered to express a IL-18 polypeptide variant as disclosed herein. Similarly, other immune cells, such as NK cells can be engineered to express an IL-18 polypeptide variant disclosed herein, and if desired, an antigen binding protein that directs NK cell activity to cells that express a selected antigen.

[0352] An immune cell may be engineered to express an IL-18 polypeptide variant using any suitable method, such as via one or more viral vectors. An immune cell engineered to express an IL-18 polypeptide variant may be autologous or allogeneic with respect to a subject receiving it as a therapy. In the case of an allogeneic cell, it may be advantageous to further engineer the cell such that it lacks functional endogenous TCRs, MHCs, or both. An immune cell may express an IL-18 polypeptide variant temporarily (such as if the immune cell is transduced with mRNA encoding the IL-18 polypeptide 62 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 variant) or over the course of its lifespan (such as if the immune cell is transduced with DNA encoding the IL-18 polypeptide variant). An immune cell may be engineered to express an IL-18 polypeptide variant and an antigen binding protein (such as a CAR) by transducing it with a single polynucleotide encoding both the IL-18 polypeptide variant and the antigen binding protein. Alternatively, an immune cell may be engineered to express an IL-18 polypeptide variant and an antigen binding protein by transducing it with two or more polynucleotides, for example, one or more polynucleotide encoding the IL-18 polypeptide variant and one or more polynucleotide encoding the antigen binding protein.

[0353] In some embodiments, an immune cell expresses an IL-18 polypeptide variant as described herein. In some embodiments, a cell contains a single polynucleotide encoding both an antigen binding protein and an IL-18 polypeptide variant as described herein. In some embodiments, a cell contains a first polynucleotide encoding an antigen binding protein and a second polynucleotide encoding an IL-18 polypeptide variant as described herein. In some embodiments, a cell contains a first polynucleotide encoding an antigen binding protein and a second polynucleotide encoding a IL-18 polypeptide variant as described herein. Preferably, an antigen binding protein expressed by an immune cell directs the immune cell and its immune effector functions to cells that express a desired antigen. In some embodiments, a cell contains one or more polynucleotides encoding the IL-18 polypeptide variant and one or more polynucleotides encoding the antigen binding protein. The polynucleotides encoding the antigen binding protein and / or the IL-18 polypeptide variant are typically engineered for expression in a desired cell and are introduced into the cell using suitable methods, e.g., transduction, transfection. Accordingly, such polynucleotides are not found in the naturally occurring cell type or species, e.g., human. For example, an immune cell can contain at least three or more polynucleotides that collectively encode the antigen binding protein and the IL-18 polypeptide variant. The immune cell may contain two or more polynucleotides encoding an IL-18 polypeptide variant and at least one polypeptide encoding an antigen binding protein.

[0354] The IL-18 polypeptide variant can be expressed by the cell as a soluble protein, which is secreted by the cell into the extracellular space. The IL-18 polypeptide variant can also be expressed by the cell as a membrane-associated protein, which can optionally be removed from the membrane by proteolytic cleavage. Methods for designing membrane-associated proteins are well-known in the art and include IL- 18 polypeptide variants that include a suitable transmembrane or membrane anchoring sequence, and optionally an intracellular domain that is fused to the IL-18 polypeptide variant, optionally through a suitable spacer sequence. The spacer sequence can include a cleavage site for a protease that has greater activity in the tumor microenvironment than in other locations. Upon cleavage of such a spacer sequence, the IL-18 polypeptide variant can be released from the cell membrane. Suitable spacer sequences include 63 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 the linkers disclosed herein, and amino acid sequences that include cleavage sites disclosed herein. Suitable transmembrane or membrane anchoring sequences are well-known in the art and include the transmembrane segments of a platelet derived growth factor receptor (PDGFR,e.g., PDGFRbeta, PDGFRalpha), CD4, CD8, IL-2R and the like. Preferably, the membrane associate IL-18 polypeptide variant lacks a cytoplasmic portion or has a cytoplasmic portion that does not transduce activation signals to the engineered cell. In one example, a CAR-T cell also expresses an IL-18 polypeptide variant that can be a secreted molecule or membrane-associated. The engineered cell can be a T cell that expresses an IL- 18 polypeptide variant. Preferably the engineered T cell is a TIL or a CAR-T cell. The i IL-18 polypeptide variant can be soluble or membrane tethered. An IL-18 polypeptide variant can be membrane tethered. The IL-18 polypeptide variant can contain a suitable transmembrane domain or membrane anchoring sequence. Typically, the suitable transmembrane domain or membrane anchoring sequence is linked to the IL-18 polypeptide variant, directly or indirectly, through a protease cleavable linker and can be released from the membrane upon cleavage of such protease cleavable linker. C. Pharmaceutical Compositions

[0355] The disclosure further relates to pharmaceutical compositions comprising a IL-18 polypeptide variant disclosed herein or a fusion polypeptide or conjugate as disclosed herein. The pharmaceutical formulations or compositions comprise an IL-18 polypeptide variant as described herein, the fusion polypeptides or conjugates as described herein and typically a pharmaceutically acceptable carrier.

[0356] Compositions comprising the IL-18 polypeptide variants described herein or the fusion polypeptides or conjugates described herein are suitable for administration in vitro or in vivo. The term "pharmaceutically acceptable carrier" includes, but is not limited to, any carrier that does not interfere with the effectiveness of the biological activity of the ingredients and that is not toxic to the subject to whom it is administered. Examples of suitable pharmaceutical carriers are well known in the art and include phosphate buffered saline solutions, water, emulsions, such as oil / water emulsions, various types of wetting agents, sterile solutions etc. Such carriers can be formulated by conventional methods and can be administered to the subject at a suitable dose. Preferably, the compositions are sterile. These compositions may also contain adjuvants such as preservative, emulsifying agents and dispersing agents. Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents.

[0357] Suitable carriers and their formulations are described in Remington: The Science and Practice of Pharmacy, 21st Edition, David B. Troy, ed., Lippicott Williams & Wilkins (2005). Typically, an appropriate amount of a pharmaceutically-acceptable salt is used in the formulation to render the 64 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 formulation isotonic, although the formulate can be hypertonic or hypotonic if desired. Examples of the pharmaceutically-acceptable carriers include, but are not limited to, sterile water, saline, buffered solutions like Ringer's solution, and dextrose solution. The pH of the solution is generally about 5 to about 8 or from about 7 to 7.5. Other carriers include sustained release preparations such as semipermeable matrices of solid hydrophobic polymers containing the immunogenic polypeptides. Matrices are in the form of shaped articles, e.g., films, liposomes, or microparticles. Certain carriers may be more preferable depending upon, for instance, the route of administration and concentration of composition being administered. Carriers are those suitable for administration of the IL-18 polypeptide variants described herein, or the fusion polypeptides or conjugates described herein or nucleic acid sequences encoding the IL-18 polypeptide variants, or the fusion polypeptides or conjugates described herein to humans or other subjects.

[0358] In some embodiments of the pharmaceutical compositions, the IL-18 polypeptide variants described herein, the fusion polypeptides or conjugates described herein as encapsulated in nanoparticles. In some embodiments, the nanoparticles are fullerenes, liquid crystals, liposome, quantum dots, superparamagnetic nanoparticles, dendrimers, or nanorods. In other embodiments of the pharmaceutical compositions, the IL-18 polypeptide variants, the fusion polypeptides or conjugates as described herein. In some instances, the IL-18 polypeptide variants, the fusion polypeptides or conjugates described herein are conjugated to the surface of liposomes. In some instances, the IL-18 polypeptide variants as described herein, or the fusion polypeptides or conjugates described herein are encapsulated within the shell of a liposome. In some instances, the liposome is a cationic liposome.

[0359] The IL-18 polypeptide variants described herein, or the fusion polypeptides or conjugates described herein are suitable for use as a medicament and can be administered by an appropriate route of administration, e.g. by intravenous, intraperitoneal, subcutaneous, intramuscular, topical or intradermal administration. In some embodiments, the route of administration depends on the kind of therapy and the kind of compound contained in the pharmaceutical composition. The dosage regimen will be determined by the attending physician and other clinical factors. Dosages for any one patient depends on many factors, including the patient's size, body surface area, age, sex, the particular compound to be administered, time and route of administration, the kind of therapy, general health and other drugs being administered concurrently. An "effective dose" refers to amounts of the active ingredient that are sufficient to affect the course and the severity of the disease, leading to the reduction or remission of such pathology and may be determined using known methods.

[0360] The pharmaceutical compositions can comprise nucleic acids that encode the IL-18 polypeptide variants disclosed herein. For example, nucleic acids include DNA and RNA molecules. Exemplary RNA 65 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 molecules include, but are not limited to mRNA, siRNA, circular RNA, self-replication RNA, transfer RNA, small RNA, miRNA, short hairpin RNAs, double-stranded RNAs, long noncoding RNAs, small cell nuclear RNAs (Y NRA), short interfering RNAs (siRNA), antisense oligonucleotide (ASO), messenger RNA (mRNA), or guide RNA on a ribonucleoprotein (RNP).

[0361] While parenteral administration is generally preferred, optionally, the IL-18 polypeptide variants described herein, or the fusion polypeptides or conjugates described herein, or nucleic acid sequences encoding the IL-18 polypeptide variants or the fusion polypeptides or conjugates are administered by a vector. There are a number of compositions and methods which can be used to deliver the nucleic acid molecules and / or polypeptides to cells, either in vitro or in vivo via, for example, expression vectors. These methods and compositions can largely be broken down into two classes: viral based delivery systems and non-viral based delivery systems. Such methods are well known in the art and readily adaptable for use with the compositions and methods described herein. Such compositions and methods can be used to transfect or transduce cells in vitro or in vivo, for example, to produce cell lines that express and preferably secrete the encoded chimeric polypeptide or to therapeutically deliver nucleic acids to a subject.

[0362] As used herein, plasmid or viral vectors are agents that transport the disclosed nucleic acids into the cell without degradation and include a promoter yielding expression of the nucleic acid molecule and / or polypeptide in the cells into which it is delivered. Viral vectors are, for example, Adenovirus, Adeno-associated virus, herpes virus, Vaccinia virus, Polio virus, Sindbis, and other RNA viruses, including these viruses with the HIV backbone. Also preferred are any viral families which share the properties of these viruses which make them suitable for use as vectors. Retroviral vectors, in general and methods of making them are described by Coffin et al., Retroviruses, Cold Spring Harbor Laboratory Press (1997). The construction of replication-defective adenoviruses has been described (Berkner et al., J. Virol.61:1213-20 (1987); Massie et al., Mol. Cell. Biol.6:2872-83 (1986); Haj-Ahmad et al., J. Virol. 57:267-74 (1986); Davidson et al., J. Virol.61:1226-39 (1987); Zhang et al., BioTechniques 15:868-72 (1993)). The benefit and the use of these viruses as vectors is that they are limited in the extent to which they can spread to other cell types, since they can replicate within an initial infected cell, but are unable to form new infectious viral particles. Recombinant adenoviruses have been shown to achieve high efficiency after direct, in vivo delivery to airway epithelium, hepatocytes, vascular endothelium, CNS parenchyma, and a number of other tissue sites. Other useful systems include, for example, replicating and host-restricted non-replicating vaccinia virus vectors. Suitable viral vectors be a monocistronic vector, a bicistronic vector, or a multicistronic vector. 66 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0363] The IL-18 polypeptide variants, the fusion polypeptides and / or nucleic acid molecules can be delivered via virus like particles. Virus like particles (VLPs) consist of viral protein(s) derived from the structural proteins of a virus. Methods for making and using virus like particles are described in, for example, Garcea and Gissmann, Current Opinion in Biotechnology 15:513-7 (2004).

[0364] The IL-18 polypeptide variants described herein, or the fusion polypeptides or conjugates described herein can be delivered by subviral dense bodies (DBs). DBs transport proteins into target cells by membrane fusion. Methods for making and using DBs are described in, for example, Pepperl- Klindworth et al., Gene Therapy 10:278-84 (2003). The provided polypeptides can be delivered by tegument aggregates. Methods for making and using tegument aggregates are described in International Publication No. WO 2006 / 110728.

[0365] Non-viral based delivery methods, can include expression vectors comprising nucleic acid molecules and nucleic acid sequences encoding polypeptides, wherein the nucleic acids are operably linked to an expression control sequence. Suitable vector backbones include, for example, those routinely used in the art such as plasmids, artificial chromosomes, BACs, YACs, or PACs. Numerous vectors and expression systems are commercially available from such corporations as Novagen (Madison, Wis.), Clonetech (Pal Alto, Calif.), Stratagene (La Jolla, Calif.), and Invitrogen / Life Technologies (Carlsbad, Calif.). Vectors typically contain one or more regulatory regions. Regulatory regions include, without limitation, promoter sequences, enhancer sequences, response elements, protein recognition sites, inducible elements, protein binding sequences, 5′and 3′ untranslated regions (UTRs), transcriptional start sites, termination sequences, polyadenylation sequences, and introns. Such vectors can also be used to make the IL-18 polypeptide variants disclosed herein, or the fusion polypeptides or conjugates described herein by expression in a suitable host cell, such as CHO cells.

[0366] Preferred promoters controlling transcription from vectors in mammalian host cells may be obtained from various sources, for example, the genomes of viruses such as polyoma, Simian Virus 40 (SV40), adenovirus, retroviruses, hepatitis B virus, and most preferably cytomegalovirus (CMV), or from heterologous mammalian promoters, e.g., β-actin promoter or EF1α promoter, or from hybrid or chimeric promoters (e.g., CMV promoter fused to the β-actin promoter). Of course, promoters from the host cell or related species are also useful herein.

[0367] Enhancer generally refers to a sequence of DNA that functions at no fixed distance from the transcription start site and can be either 5′ or 3′ to the transcription unit. Furthermore, enhancers can be within an intron as well as within the coding sequence itself. They are usually between 10 and 300 base pairs (bp) in length, and they function in cis. Enhancers usually function to increase transcription from nearby promoters. Enhancers can also contain response elements that mediate the regulation of 67 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 transcription. While many enhancer sequences are known from mammalian genes (globin, elastase, albumin, fetoprotein, and insulin), typically one will use an enhancer from a eukaryotic cell virus for general expression. Preferred examples are the SV40 enhancer on the late side of the replication origin, the cytomegalovirus early promoter enhancer, the polyoma enhancer on the late side of the replication origin, and adenovirus enhancers.

[0368] The promoter and / or the enhancer can be inducible (e.g., chemically or physically regulated). A chemically regulated promoter and / or enhancer can, for example, be regulated by the presence of alcohol, tetracycline, a steroid, or a metal. A physically regulated promoter and / or enhancer can, for example, be regulated by environmental factors, such as temperature and light. Optionally, the promoter and / or enhancer region can act as a constitutive promoter and / or enhancer to maximize the expression of the region of the transcription unit to be transcribed. In certain vectors, the promoter and / or enhancer region can be active in a cell type specific manner. Optionally, in certain vectors, the promoter and / or enhancer region can be active in all eukaryotic cells, independent of cell type. Preferred promoters of this type are the CMV promoter, the SV40 promoter, the β-actin promoter, the EF1α promoter, and the retroviral long terminal repeat (LTR).

[0369] The vectors also can include, for example, origins of replication and / or markers. A marker gene can confer a selectable phenotype, e.g., antibiotic resistance, on a cell. The marker product is used to determine if the vector has been delivered to the cell and once delivered is being expressed. Examples of selectable markers for mammalian cells are dihydrofolate reductase (DHFR), thymidine kinase, neomycin, neomycin analog G418, hygromycin, puromycin, and blasticidin. When such selectable markers are successfully transferred into a mammalian host cell, the transformed mammalian host cell can survive if placed under selective pressure. Examples of other markers include, for example, the E. coli lacZ gene, green fluorescent protein (GFP), and luciferase. In addition, an expression vector can include a tag sequence designed to facilitate manipulation or detection (e.g., purification or localization) of the expressed polypeptide. Tag sequences, such as GFP, glutathione S-transferase (GST), polyhistidine, c-myc, hemagglutinin, or FLAG™ tag (Kodak; New Haven, Conn.) sequences typically are expressed as a fusion with the encoded polypeptide. Such tags can be inserted anywhere within the polypeptide including at either the carboxyl or amino terminus.

[0370] The compositions disclosed herein can comprise nucleic acids encoding the IL-18 polypeptide variants described herein, or the fusion polypeptides or conjugates described herein. Nucleic acids include DNA and RNA molecules. Exemplary RNA molecules include, but are not limited to mRNA, siRNA, circular RNA, self-replication RNA, transfer RNA, small RNA, miRNA, short hairpin RNAs, double- stranded RNAs, long noncoding RNAs, small cell nuclear RNAs (Y NRA), short interfering RNAs 68 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 (siRNA), antisense oligonucleotide (ASO), messenger RNA (mRNA), or guide RNA on a ribonucleoprotein (RNP).

[0371] The composition can comprise DNA encoding the IL-18 polypeptide variants described herein, or the fusion polypeptides or conjugates described herein. The composition can comprise RNA encoding the IL-18 polypeptide variants described herein, or the fusion polypeptides or conjugates described herein. In particular, the composition can comprise mRNA encoding the IL-18 polypeptide variants described herein, or the fusion polypeptides or conjugates described herein. The composition can comprise an RNA sequence (e.g., mRNA) transcribed from a DNA sequence encoding the IL-18 polypeptide variants described herein, or the fusion polypeptides or conjugates described herein.

[0372] The RNA can comprise a 5’ untranslated region (UTR). The 5’ UTR can be upstream of an initiation codon. Without wishing to be bound by theory or mechanism, the 5’UTR can regulate translation of RNA, stabilizes the RNA, and / or increases the half-life of the RNA. Suitable 5’ UTRs are well-known in the art. The RNA can comprise a 3’ untranslated region (UTR). The 3’ UTR can follow the translation termination codon. Without wishing to be bound by theory or mechanism, the 3’ UTR can regulate polyadenylation, translation efficiency, localization, or stability of the RNA. Suitable 3’ UTRs are well-known in the art. The composition generally comprises both a 5’ UTR and a 3’ UTR. However, if desired the composition can comprise only a 5’ UTR or only a 3’ UTR.

[0373] The RNA can comprise a poly-A tail. The poly-A tail can be at least about 25 nucleotides, at least about 30 nucleotides, at least about 40 nucleotides, at least about 50 nucleotides, at least about 70 nucleotides, at least about 80 nucleotides, at least about 90 nucleotides, at least about 100 nucleotides, or more. Suitable poly-A tails are well known in the art.

[0374] In certain embodiments, the RNA can comprise a 5’ cap, a 5’UTR, a nucleic acid encoding the IL-18 polypeptide variants disclosed herein, or the fusion polypeptides or conjugates described herein, a 3’ UTR, and a poly-A tail.

[0375] The nucleic acids and compositions disclosed herein can comprise any desired modification. For example, one or more nucleosides can be modified or one or more ribose can be modified. The modifications can improve stability of the nucleic acids and compositions disclosed herein. Suitable modifications are well-known in the art.

[0376] For instance, one or more uridine in the RNA can be replaced by a modified nucleoside. The modified nucleoside can be a modified uridine. The modified uridine replacing uridine is pseudouridine (ψ), N1-methyl-pseudouridine (m1ψ), or 5-methyl-uridine (m5U).

[0377] One or more cytosine, adenine or guanine in the RNA can be replaced by modified nucleobase(s). The modified nucleobase replacing cytosine can be 5-methylcytosine (m5C). The modified nucleobase 69 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 replacing adenine can be N6-methyladenine (m6A). Any other modified nucleobase known in the art for reducing the immunogenicity of the molecule can be used.

[0378] The modified nucleoside replacing one or more uridine in the RNA can include, but is not limited to, one or more of 3-methyl-uridine (m3U), 5-methoxy-uridine (mo5U), 5-aza-uridine, 6-aza- uridine, 2- thio-5-aza-uridine, 2-thio-uridine (s2U), 4-thio-uridine (s4U), 4-thio-pseudouridine, 2-thio-pseudouridine, 5-hydroxy-uridine (ho5U), 5-aminoallyl-uridine, 5-halo-uridine (e.g., 5- iodo-uridineor 5-bromo-uridine), uridine 5-oxyacetic acid (cmo5U), uridine 5-oxyacetic acid methyl ester (mcmo5U), 5-carboxymethyl- uridine (cm5U), 1-carboxymethyl-pseudouridine, 5-carboxyhydroxymethyl-uridine (chm5U), 5- carboxyhydroxymethyl-uridine methyl ester (mchm5U), 5-methoxycarbonylmethyl-uridine (mcm5U), 5- methoxycarbonylmethyl-2-thio- uridine (mcm5s2U), 5-aminomethyl-2-thio-uridine (nm5s2U), 5- methylaminomethyl-uridine (mnm5U), 1-ethyl-pseudouridine, 5-methylaminomethyl-2-thio-uridine (mnm5s2U), 5- methylaminomethyl-2-seleno-uridine (mnm5se2U), 5-carbamoylmethyl-uridine (ncm5U), 5- carboxymethylaminomethyl-uridine (cmnm5U), 5-carboxymethylaminomethyl-2-thio-uridine (cmnm5s2U), 5-propynyl-uridine, 1-propynyl-pseudouridine, 5-taurinomethyl-uridine (τm5U), 1- taurinomethyl-pseudouridine, 5-taurinomethyl-2-thio-uridine(τm5s2U), 1-taurinomethyl-4- thio- pseudouridine), 5-methyl-2-thio-uridine (m5s2U), 1-methyl-4-thio-pseudouridine (m1s4ψ), 4-thio-1- methyl-pseudouridine, 3-methyl-pseudouridine (m3ψ), 2-thio-1-methyl- pseudouridine, 1-methyl-1-deaza- pseudouridine, 2-thio-1-methyl-1-deaza-pseudouridine, dihydrouridine (D), dihydropseudouridine, 5,6- dihydrouridine, 5-methyl-dihydrouridine (m5D), 2-thio-dihydrouridine, 2-thio-dihydropseudouridine, 2- methoxy-uridine, 2-methoxy- 4-thio-uridine, 4-methoxy-pseudouridine, 4-methoxy-2-thio-pseudouridine, N1-methyl- pseudouridine, 3-(3-amino-3-carboxypropyl)uridine (acp3U), 1-methyl-3-(3-amino-3- carboxypropyl)pseudouridine (acp3ψ), 5-(isopentenylaminomethyl)uridine (inm5U), 5- (isopentenylaminomethyl)-2-thio-uridine (inm5s2U), α-thio-uridine, 2′-O-methyl-uridine (Um), 5,2′-O- dimethyl-uridine (m5Um), 2′-O-methyl-pseudouridine (ψm), 2-thio-2′-O- methyl-uridine (s2Um), 5- methoxycarbonylmethyl-2′-O-methyl-uridine (mcm5Um), 5- carbamoylmethyl-2′-O-methyl-uridine (ncm5Um), 5-carboxymethylaminomethyl-2′-O- methyl-uridine (cmnm5Um), 3,2′-O-dimethyl-uridine (m3Um), 5-(isopentenylaminomethyl)- 2′-O-methyl-uridine (inm5Um), 1-thio-uridine, deoxythymidine, 2′-F-ara-uridine, 2′-F- uridine, 2′-OH-ara-uridine, 5-(2-carbomethoxyvinyl) uridine, 5-[3-(1-E- propenylamino)uridine, or any other modified uridine known in the art.

[0379] The compositions disclosed herein can comprising one or more nucleic acids (e.g. DNA or RNA) as described herein. For example, the composition can comprise one RNA, two RNAs, three RNAs, four RNAs, five RNAs, or more. 70 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 D. Therapeutic Applications

[0380] Also provided herein, are methods and uses for the treatment of a disease, disorder or condition comprising administering to a subject in need thereof an IL-18 polypeptide variants as described herein, or a fusion polypeptide or conjugate as described herein. Diseases, disorders, or conditions include, but are not limited to, cancer, inflammatory disease, an immunological disorder, autoimmune disease, infectious disease (i.e., bacterial, viral, or parasitic disease). Preferably, the disease, disorder, or condition is cancer.

[0381] Any suitable cancer may be treated with the IL-18 polypeptide variants provided herein, or a fusion polypeptide provided herein or conjugates described herein. Illustrative suitable cancers, in particular solid tumors, such as sarcomas and carcinomas. For examples, the methods and compositions disclosed herein can be used to treat acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), adrenocortical carcinoma, anal cancer, appendix cancer, astrocytoma, basal cell carcinoma, brain tumor, bile duct cancer, bladder cancer, bone cancer, breast cancer, bronchial tumor, carcinoma of unknown primary origin, cardiac tumor, cervical cancer, chordoma, colon cancer, colorectal cancer, craniopharyngioma, ductal carcinoma, embryonal tumor, endometrial cancer, ependymoma, esophageal cancer, esthesioneuroblastoma, fibrous histiocytoma, Ewing sarcoma, eye cancer, germ cell tumor, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor, gestational trophoblastic disease, glioma, head and neck cancer, hepatocellular cancer, histiocytosis, Hodgkin lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell tumor, Kaposi sarcoma, kidney cancer, Langerhans cell histiocytosis, laryngeal cancer, lip and oral cavity cancer, liver cancer, lobular carcinoma in situ, lung cancer, macroglobulinemia, malignant fibrous histiocytoma, melanoma, Merkel cell carcinoma, mesothelioma, metastatic squamous neck cancer with occult primary, midline tract carcinoma involving NUT gene, mouth cancer, multiple endocrine neoplasia syndrome, multiple myeloma, mycosis fungoides, myelodysplastic syndrome, myelodysplastic / myeloproliferative neoplasm, nasal cavity and par nasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-small cell lung cancer, oropharyngeal cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papillomatosis, paraganglioma, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytomas, pituitary tumor, pleuropulmonary blastoma, primary central nervous system lymphoma, prostate cancer, rectal cancer, renal cell cancer, renal pelvis and ureter cancer, retinoblastoma, rhabdoid tumor, salivary gland cancer, Sezary syndrome, skin cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, spinal cord tumor, stomach cancer, T-cell lymphoma, teratoid tumor, testicular cancer, throat cancer, thymoma and thymic carcinoma, thyroid cancer, urethral cancer, uterine cancer, vaginal cancer, vulvar cancer, and Wilms tumor. 71 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0382] In certain embodiments, the methods and compositions disclosed herein can be used to treat adrenocortical carcinoma, anal cancer, appendix cancer, astrocytoma, basal cell carcinoma, brain tumor, bile duct cancer, bladder cancer, bone cancer, breast cancer, bronchial tumor, carcinoma of unknown primary origin, cardiac tumor, cervical cancer, chordoma, colon cancer, colorectal cancer, craniopharyngioma, ductal carcinoma, embryonal tumor, endometrial cancer, ependymoma, esophageal cancer, esthesioneuroblastoma, fibrous histiocytoma, Ewing sarcoma, eye cancer, germ cell tumor, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor, gestational trophoblastic disease, glioma, head and neck cancer, hepatocellular cancer, histiocytosis, Hodgkin lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell tumor, Kaposi sarcoma, kidney cancer, Langerhans cell histiocytosis, laryngeal cancer, lip and oral cavity cancer, liver cancer, lobular carcinoma in situ, lung cancer, malignant fibrous histiocytoma, melanoma, Merkel cell carcinoma, mesothelioma, metastatic squamous neck cancer with occult primary, midline tract carcinoma involving NUT gene, mouth cancer, multiple endocrine neoplasia syndrome, mycosis fungoides, nasal cavity and par nasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-small cell lung cancer, oropharyngeal cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papillomatosis, paraganglioma, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytomas, pituitary tumor, pleuropulmonary blastoma, primary central nervous system lymphoma, prostate cancer, rectal cancer, renal cell cancer, renal pelvis and ureter cancer, retinoblastoma, rhabdoid tumor, salivary gland cancer, Sezary syndrome, skin cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, spinal cord tumor, stomach cancer, T-cell lymphoma, teratoid tumor, testicular cancer, throat cancer, thymoma and thymic carcinoma, thyroid cancer, urethral cancer, uterine cancer, vaginal cancer, vulvar cancer, non-Hodgkin lymphoma, squamous carcinoma of the head and neck, malignant pleural mesothelioma, and Wilms tumor.

[0383] In certain preferred embodiments, the methods and compositions disclosed herein are used to treat melanoma, non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), head and neck squamous cell cancer (HNSCC), classical Hodgkin lymphoma (cHL), primary mediastinal large B cell lymphoma (PMBCL), urothelial carcinoma, microsatellite instability high or mismatch repair deficient cancer, microsatellite instability high or mismatch repair deficient colorectal cancer, gastric cancer, esophageal cancer, cervical cancer, hepatocellular carcinoma (HCC), merkel cell carcinoma (MCC), renal cell carcinoma (RCC), endometrial carcinoma, tumor mutational burden high cancer, cutaneous squamous cell carcinoma (cSCC), triple negative breast cancer (TNBC), urothelial carcinoma, colorectal cancer or oesophageal carcinoma. 72 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0384] In certain preferred embodiments, the methods and compositions disclosed herein are used to treat Merkel cell carcinoma (MCC), urothelial carcinoma (UC), renal cell carcinoma (RCC), non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), triple negative breast cancer (TNBC), endometrial cancer, cutaneous squamous cell carcinoma (CSCC), basal cell carcinoma (BCC), melanoma, malignant pleural mesothelioma, classical Hodgkin lymphoma (cHL), squamous cell carcinoma of the head and neck (SCCHN), hepatocellular carcinoma (HCC), esophageal squamous cell carcinoma (ESCC), non-squamous non-small cell lung cancer, or nasopharyngeal carcinoma (NPC).

[0385] Preferably, the methods and compositions disclosed herein are used to treat colon cancer, lung cancer, melanoma, renal cell carcinoma, or breast cancer.

[0386] In certain preferred embodiments, the methods and compositions disclosed herein are used to treat melanoma. As an example, the methods and compositions disclosed herein can be used to treat melanoma in subjects with unresectable or metastatic melanoma. As another example, the methods and compositions disclosed herein can be used for the adjuvant treatment of subjects with melanoma with involvement of lymph node(s) following complete resection.

[0387] In some embodiments, provided herein is a method of enhancing an immune response in a subject in need thereof by administering an effective amount of an IL-18 polypeptide variant as described herein, or a fusion polypeptide or conjugates described herein to the subject. The enhanced immune response may prevent, delay, or treat the onset of cancer, a tumor, or a viral disease. In some embodiments, the methods described herein increase the activity of Natural Killer Cells and T lymphocytes.

[0388] The method can further involve the administration of one or more additional agents to treat cancer, such as chemotherapeutic agents (e.g., Adriamycin, Cerubidine, Bleomycin, Alkeran, Velban, Oncovin, Fluorouracil, Thiotepa, Methotrexate, Bisantrene, Noantrone, Thiguanine, Cytaribine, Procarabizine), immuno-oncology agents (e.g., anti-PD-L1, anti-CTLA4, anti-PD-1, anti-LAG3, anti- CD47, anti-GD2), cellular therapies (e.g., CAR-T, T-cell therapy), oncolytic viruses and the like. Non- limiting examples of anti-cancer agents that can be used include acivicin; aclarubicin; acodazole hydrochloride; acronine; adozelesin; aldesleukin; altretamine; ambomycin; ametantrone acetate; aminoglutethimide; amsacrine; anastrozole; anthramycin; asparaginase; asperlin; azacitidine; azetepa; azotomycin; batimastat; benzodepa; bicalutamide; bisantrene hydrochloride; bisnafide dimesylate; bizelesin; bleomycin sulfate; brequinar sodium; bropirimine; busulfan; cactinomycin; calusterone; caracemide; carbetimer; carboplatin; carmustine; carubicin hydrochloride; carzelesin; cedefingol; chlorambucil; cirolemycin; cisplatin; cladribine; crisnatol mesylate; cyclophosphamide; cytarabine; dacarbazine; dactinomycin; daunorubicin hydrochloride; decitabine; dexormaplatin; dezaguanine; dezaguanine mesylate; diaziquone; docetaxel; doxorubicin; doxorubicin hydrochloride; droloxifene; 73 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 droloxifene citrate; dromostanolone propionate; duazomycin; edatrexate; eflornithine hydrochloride; elsamitrucin; enloplatin; enpromate; epipropidine; epirubicin hydrochloride; erbulozole; esorubicin hydrochloride; estramustine; estramustine phosphate sodium; etanidazole; etoposide; etoposide phosphate; etoprine; fadrozole hydrochloride; fazarabine; fenretinide; floxuridine; fludarabine phosphate; fluorouracil; flurocitabine; fosquidone; fostriecin sodium; gemcitabine; gemcitabine hydrochloride; hydroxyurea; idarubicin hydrochloride; ifosfamide; ilmofosine; interleukin II (including recombinant interleukin II, or rIL2), interferon alpha-2a; interferon alpha-2b; interferon alpha-nl interferon alpha-n3; interferon beta-I; interferon gamma-I b; iproplatin; irinotecan hydrochloride; lanreotide acetate; letrozole; leuprolide acetate; liarozole hydrochloride; lometrexol sodium; lomustine; losoxantrone hydrochloride; masoprocol; maytansine; mechlorethamine hydrochloride; megestrol acetate; melengestrol acetate; melphalan; menogaril; mercaptopurine; methotrexate; methotrexate sodium; metoprine; meturedepa; mitindomide; mitocarcin; mitocromin; mitogillin; mitomalcin; mitomycin; mitosper; mitotane; mitoxantrone hydrochloride; mycophenolic acid; nocodazole; nogalamycin; ormaplatin; oxisuran; paclitaxel; pegaspargase; peliomycin; pentamustine; peplomycin sulfate; perfosfamide; pipobroman; piposulfan; piroxantrone hydrochloride; plicamycin; plomestane; porfimer sodium; porfiromycin; prednimustine; procarbazine hydrochloride; puromycin; puromycin hydrochloride; pyrazofurin; riboprine; rogletimide; safingol; safingol hydrochloride; semustine; simtrazene; sparfosate sodium; sparsomycin; spirogermanium hydrochloride; spiromustine; spiroplatin; streptonigrin; streptozocin; sulofenur; talisomycin; tecogalan sodium; tegafur; teloxantrone hydrochloride; temoporfin; teniposide; teroxirone; testolactone; thiamiprine; thioguanine; thiotepa; tiazofurin; tirapazamine; toremifene citrate; trestolone acetate; triciribine phosphate; trimetrexate; trimetrexate glucuronate; triptorelin; tubulozole hydrochloride; uracil mustard; uredepa; vapreotide; verteporfin; vinblastine sulfate; vincristine sulfate; vindesine; vindesine sulfate; vinepidine sulfate; vinglycinate sulfate; vinleurosine sulfate; vinorelbine tartrate; vinzolidine sulfate; vinzolidine sulfate; vorozole; zeniplatin; zinostatin; zorubicin hydrochloride.

[0389] In some embodiments of the methods described herein, the IL-18 polypeptide variants as described herein, or a fusion polypeptide or conjugates described herein is administered in combination with an agent for the treatment of the particular disease, disorder, or condition. Agents include, but are not limited to, therapies involving antibodies, small molecules (e.g., chemotherapeutics), hormones (steroidal, peptide, and the like), radiotherapies (γ-rays, C-rays, and / or the directed delivery of radioisotopes, microwaves, UV radiation and the like), gene therapies (e.g., antisense, retroviral therapy and the like) and other immunotherapies. In some embodiments, the IL-18 polypeptide variants as described herein, or the fusion polypeptides or conjugates described herein or is administered in combination with anti- diarrheal agents, anti-emetic agents, analgesics and / or non-steroidal anti-inflammatory agents. 74 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0390] This disclosure relates to a therapeutic combination of any of the IL-18 polypeptide variants as described herein, or a fusion polypeptide or conjugate described herein in combination with one or more additional agents to treat cancer (such as lymphoma), such as chemotherapeutic agents (e.g., cyclophosphamide, mechlorethamine, melphalan, chlorambucil, ifosfamide, busulfan, N-Nitroso-N- methylurea (MNU), carmustine (BCNU), lomustine (CCNU), semustine (MeCCNU), fotemustine, streptozotocin, dacarbazine, mitozolomide, temozolomide, thiotepa, mitomycin, diaziquone (AZQ), cisplatin, carboplatin, oxaliplatin, procarbazine, hexamethylmelamine, methotrexate, pemetrexed, fluorouracil (e.g.5-fluorouracil), capecitabine, cytarabine, gemcitabine, decitabine, azacitidine, fludarabine, nelarabine, cladribine, clofarabine, pentostatin, thioguanine, mercaptopurine, vincristine, vinblastine, vinorelbine, vindesine, vinflunine, paclitaxel, docetaxel, etoposide, teniposide, doxorubicin, daunorubicin, epirubicin, idarubicin, pirarubicin, aclarubicin, mitoxantrone, actinomycin, bleomycin, bisantrene, gemcitabine, cytarabine, and the like), immuno-oncology agents and immune checkpoint inhibitors (e.g., anti-PD-L1, anti-CTLA4, anti-PD-1, anti-LAG3, anti-CD47, anti-GD2), oncolytic viruses and the like.

[0391] The IL-18 polypeptide variants as described herein, or a fusion polypeptide or conjugates described herein can be combined with any desired additional anti-cancer agent. The IL-18 polypeptide variants as described herein, or a fusion polypeptide or conjugates described herein can be combined with any desired anti-PD-1 antibody or any desired anti-PD-L1 antibody.

[0392] Exemplary anti-PD-1 antibodies that can be combined with the IL-18 polypeptide variants as described herein, or a fusion polypeptide or conjugates described herein, but are not limited to, AMP-224 (AstraZenica), 609A (3SBio), 704 (3SBio), 705 (3SBio), ABBV-181 (AbbVie), ADU-1503 / bion-004 (Chinook Therapeutics), AGEN2034 / balstilimab (Agenus), AK103 (Akeso), AK104 (Akeso), AK112 (Akeso), AK123 (Akeso), AMG 256 (Amgen), AMG 404 (Amgen), ANB030 (AnaptysBio), ANKEBIO Anti-PD1 product (Anhui Anke Biotechnology), Anti PD-1 / Anti-CD47 (DiNonA), ASKG915 (Ask Gene Pharmaceuticals), AV-MEL-1 (Aivita Biomedical), BCD-100 (Biocad CJSC), BI 754091 (Boehringer Ingelheim), BiCKI-IL-7 (OSE Immunotherapeutics), Boehringer-PD-1-unknown (Boehringer Ingelheim), BSK-050K01 (Biosion), Camrelizumab (Jiangsu Hengrui Medicine), CB201 (Crescendo Biologics), CB213 (Crescendo Biologics), CC-90006 (AnaptsBio), cetrelimab (J&J), chPD1 (Kiromic Biopharma), CMAB819 (Mabpharm), CS1003 (CStone Pharmaceuticals), CS17938 (Shenzhen Chipscreen Biosciences), CTX-8371 (Compass Therapeutics), CX-072 (CytomX Therapeutics), CX-188 (CytomX Therapeutics), cypalizumab (Harbin Gloria Pharmaceuticals), DB004 (DotBio), EMB02 (EpimAb Biotherapeutics), Geptanblimab / genolimzumab (Apollomics), GS19 (Suzhou Zelgen Biopharmaceuticals), HLX10 (Shanghai Henlius Biotech), HX008 (Taizhou HanZhong Pharmaceuticals), 75 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 HY003 (Juventas Cell Therapy), IBI315 / BH2950 (Innovent Biologics), IBI318 (Innovent Biologics), IBI319 (Innovent Biologics), IMM1802 (ImmuneOnco Biopharma), IMT200 (TrueBinding), Jemperli / dostarlimab (AnaptysBio), JTX-4014 (Jounce Therapeutics), Keytruda / pembrolizumab (Merck), LBL- 006 (Nanjing Leads Biolabs), Libtayo / cemiplimab-rwlc (Regeneron Pharmaceuticals), LVGN3616 (Lyvgen Biopharma), LXF821 (Novartis), LY01015 (Luye Pharma Group), LY3462817 (Eli Lilly), MCLA-134 (Merus N.V.), MEDI5752 (AstraZenica), NIR178 (Novartis), ONCR-177 (Oncorus), ONO- 4685 (Ono Pharmaceutical), Opdivo / nivolumab (Ono Pharmaceutical), MGD019 (MacroGenius), PD1- GDT CAR-T (Kiromic Biopharma), penpulimab (Akeso), PSB205 (Qilu Puget Sound Biotherapeutics), PT-001 (Merck), PT627 (Merck), RB-M1 (Refuge Biotechnologies), Retifanlimab (MacroGenics), RG6139 (Roche), RG6279 (Roche), RTX-002 (RubrYc Therapeutics), sasanlimab (Pfizer), Servier- PD1xLAG3-unknown (Servier), SL-279252 / TAK-252 (Shattuck Labs), Sofusa anti-PD1 (Sorrento Therapeutics), spartalizumab (Novartis), SSI-361 (Lyvgen Biopharma), Sym021 (Servier), Tebotelimab (MacroGenics), tislelizumab (BeiGene), TSR-075 (AnaptsBio), Tuhura-DO / PD-1-unknown (Tuhura Biopharma), toripalimab (Shanghai Junshi Biosciences), sintilimab (Innovent Biologics), Unicar-CAR- T&PD-1-unknown (Shanghai Unicar-Therapy Bio-Medicine Technology), Xdivane (Xbrane Biopharma), XmAb20717 (Xencor), XmAb23104 (Xencor), YBL-006 (Y-Biologics), and zimberelimab (Arcus Biosciences).

[0393] The anti-PD-1 antibody that can be combined with the IL-18 polypeptide variants as described herein, or fusion polypeptides or conjugates described herein is typically an approved anti-PD-1 antibody. Approved anti-PD-1 antibodies include, but are not limited to, pembrolizumab (KEYTRUDA), dostarlimab (JEMPERLI), cemiplimab-rwlc (LIBATYO), nivolumab (OPDIVO), camrelizumab, tislelizumab, toripalimab, and sintilimab (TYVYT).

[0394] Exemplary anti-PD-L1 antibodies that can be combined with the IL-18 polypeptide variants as described herein, or fusion polypeptides or conjugates described herein include, but are not limited to, A167 (Sichuan Kelun), ABL501 (ABL Bio), ABL503 (ABL Bio), ABSK041 (Abbisko Therapeutics), ACE1708 (Acepodia), ACE-NK-PDL1 (Acepodia), ADG104 (Adagene), AK106 (Akeso), ALPN-202 (Alpine Immune Sciences), AN4005 (Adlai Nortye Biopharma), BMS-936559 / MDX-1105 (BMS), APL-502 / TQB2450 (Apollomics), Arbutus-PD-L1-unknown (Arbutus Biopharma), ASC22 (Ascletis Pharma), ATG-101 (Antengene), AVA-004 (Avacta Group), AVA021 (Avacta Group), AVA027 (Avacta Group), AVA-040-100 (Avacta Group), AVA04-Vbp (Avacta Group), Bavencio / avelumab (Merck), BCD-135 (Biocad CJSC), BGB-A333 (BeiGene), Bintrafusp alfa / GSK4045154 (Merck), CA-170 / aupm-170 (Dr. Reddy’s Laboratories), CCX559 (ChemoCentryx), CDR101 (CDR-Life), cosibelimab (Checkpoint Therapeutics), CTX-8371 (Compass Therapeutics), DiNonA-Solid Tumors-unknown 76 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 (DiNonA), DR30207 (Zhejiang Doer Biologics), DuoBody-PD-L1x4-1BB (Ligand Pharmaceuticals), envafolimab (Alphamab Oncology), EPIM-001 (Elpis Biopharmaceuticals), ES101 (Elpiscience Biopharma), INBRX-105 (Inhibrx), FAZ053 (Novartis), FS118 (F-star Therapeutics), GB262 (Genor Biopharma), GS-4224 (Gilead), GT900008 (Kintor Pharmaceuticals), GX-P2 (Genexine), Hamni-PS- L1 / CD47-Unknown (Hanmi Pharmaceutical), HBM7015 (HBM Holdings), HBM9167 (HBM Holdings), HLX20 (Shanghai Henlius Biotech), HTI-1088 (Jiangsu Hengrui Medicine), IBI318 (Innovent Biologics), IBI322 (Innovent Biologics), IBI323 (Innovent Biologics), IGM-7354 (IGM Biosciences), IMC-001 (Sorrento Therapeutics), Imfinzi / durvalumab (AstraZenica), IMM25 (ImmuneOnco Biopharma), IMM2502 (ImmuneOnco Biopharma), IMM2503 (ImmuneOnco Biopharma), IMM2504 (ImmuneOnco Biopharma), INCB86550 (Incyte), IO103 (IO Biotech), JS003 (Shanghai Junshi Biosciences), Jubilant- PD-L1-unknown (Jubilant Therapeutics), KD033 (Kadmon Holdings), KN046 (Alphamab Oncology), KY1003 (Sanofi), KY1043 (Sanofi), LY3300054 (Eli Lilly), LY3415244 (Eli Lilly), MRNA-6981 (Moderna), MSB2311 (Transcenta Holding), MT-6035 (Molecular Templates), ND021 / NM21-1480 (Numab Therapeutics), OX001R (Oxford BioTherapeutics), PD-L1 based BsAbs (I-Mab), PD-L1 Boltbody ISAC (Bolt Biotherapeutics), PDL-GEX (Glycotope GmbH), PMC-122 (PharmAbcine), PMI06 (D&D Pharmatech), Protheragen-RV-scFv-PDL1-unknown (Protheragen), PRS-344 (Pieris Pharmaceuticals), Q-1802 (Merck), RC98 (Yantai Rongchang Pharmaceutical), RV-scFv-PDL1 (Protheragen), SenI_TAAx22P (Hebei Senlang Biotechnology), SHC020 (Nanjing Sanhome Pharmaceutical), sugemalimab (Ligand Pharmaceuticals), atezolizumab (Roche), TST005 (Transcenta Holding), TT-01 (Topmunnity Therapeutics), TTX-siPDL1 (TransCode Therapeutics), UniCAR-T-PD- L1 (GEMoaB monoclonals), Vaximm (VXM10), and YBL-013 (Y-Biologics).

[0395] The anti-PD-L1 antibody that can be combined with the IL-18 polypeptide variants as described herein, or a fusion polypeptide or conjugate described herein is typically an approved anti-PD-L1 antibody. Approved anti-PD-1 antibodies include, but are not limited to, avelumab (BAVENCIO), durvalumab (IMFINZI), and atezolizumab (TECENTRIQ).

[0396] The anti-CTLA4 antibody that can be combined with the IL-18 polypeptide variants as described herein, or a fusion polypeptide or conjugate as described herein is typically an approved anti-CTLA4 antibody. Approved anti-CTLA4 antibodies include, but are not limited to, ipilimumab (YERVOY) and tremelimumab (IMJUDO).

[0397] The anti-LAG3 antibody that can be combined with the IL-18 polypeptide variants as described herein, or a fusion polypeptide or conjugate is typically an approved anti-LAG3 antibody. An approved anti-LAG3 antibody includes, relatlimab (OPDUALAG).   77 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0398] The IL-18 polypeptide variants as described herein, or a fusion polypeptide or conjugates described herein can be combined with any desired additional cytokine. Suitable cytokines, include for example, IFN-alpha, IFN-beta, IFN-gamma, IL-1, IL-2, IL-4, IL-5, IL-6, IL-10, IL-18, IL-22, IL-23p19, IL-11, IL-12, IL-13, IL-15, IL-12p35, IL-21, IL-30 (IL27p28), IL-34, IL-35, IL-36 and the like. Preferable cytokines include IL-2, IL-12 (IL-12 p35 and / or IL-12p40), IL-10, IL-18, and IL-21. For example, the preferred cytokine can be IL-2. For example, the preferred cytokine can be IL-12. For example, the preferred cytokine can be IL-2. If desired, the IL-18 polypeptide variants as disclosed herein can be combined with an inducible cytokine prodrug. For example, the inducible cytokine prodrug can be an inducible IL-2 prodrug, an inducible IL-12 prodrug, a inducible IL-10 prodrug, an inducible IL-21 prodrug, an inducible IL-21 prodrug, or an inducible IFN prodrug. Inducible IL-12, IL-2, and interferon prodrugs have been described in International Publication Nos.: WO2019 / 222294, WO2019 / 222295, WO2019 / 222296, WO2021 / 097376.    Definitions

[0399] All publications and patents cited in this disclosure are incorporated by reference in their entirety. To the extent the material incorporated by reference contradicts or is inconsistent with this specification, the specification will supersede any such material. The citation of any references herein is not an admission that such references are prior art to the present disclosure. When a range of values is expressed, it includes embodiments using any particular value within the range. Further, reference to values stated in ranges includes each and every value within that range. All ranges are inclusive of their endpoints and combinable. When values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms another embodiment. Reference to a particular numerical value includes at least that particular value, unless the context clearly dictates otherwise. The use of “or” will mean “and / or” unless the specific context of its use dictates otherwise.

[0400] Various terms relating to aspects of the description are used throughout the specification and claims. Such terms are to be given their ordinary meaning in the art unless otherwise indicated. Other specifically defined terms are to be construed in a manner consistent with the definitions provided herein. The techniques and procedures described or referenced herein are generally well understood and commonly employed using conventional methodologies by those skilled in the art, such as, for example, the widely utilized molecular cloning methodologies described in Sambrook et al., Molecular Cloning: A Laboratory Manual 4th ed. (2012) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY. As appropriate, procedures involving the use of commercially available kits and reagents are generally carried out in accordance with manufacturer-defined protocols and conditions unless otherwise noted. 78 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320

[0401] As used herein, the singular forms “a,” “an,” and “the” include plural forms unless the context clearly indicates otherwise. The terms “include,” “such as,” and the like are intended to convey inclusion without limitation, unless otherwise specifically indicated.

[0402] Unless otherwise indicated, the terms "at least," "less than," and "about," or similar terms preceding a series of elements or a range are to be understood to refer to every element in the series or range. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims.

[0403] As used herein, the term “operably linked” in the context of a polypeptide (e.g., a inducible cytokine prodrug) refers to the orientation of the components of a polypeptide that permits the components to function in their intended manner.

[0404] As used throughout, “subject” can be a vertebrate, more specifically a mammal (e.g. a human, horse, cat, dog, cow, pig, sheep, goat, mouse, rabbit, rat, and guinea pig), birds, reptiles, amphibians, fish, and any other animal. The term does not denote a particular age or sex. Thus, adult and newborn subjects, whether male or female, are intended to be covered. As used herein, “patient” or “subject” may be used interchangeably and can refer to a subject with a disease or disorder (e.g. cancer). The term patient or subject includes human and veterinary subjects.

[0405] As used herein, the term “therapeutically effective amount” refers to an amount of a compound described herein (e.g., a inducible cytokine prodrug) that is sufficient to achieve a desired pharmacological or physiological effect under the conditions of administration. For example, a “therapeutically effective amount” can be an amount that is sufficient to reduce the signs or symptoms of a disease or condition (e.g., a tumor). Those skilled in the art will appreciate that the therapeutic effects need not be complete or curative, as long as some benefit is provided to the subject. A therapeutically effective amount of a pharmaceutical composition can vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the pharmaceutical composition to elicit a desired response in the individual. An ordinarily skilled clinician can determine appropriate amounts to administer to achieve the desired therapeutic benefit based on these and other considerations.

[0406] As used herein the terms “treatment”, “treat”, or “treating” refers to a method of reducing the effects of a disease or condition or symptom of the disease or condition. Thus, in the disclosed methods, treatment can refer to at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or substantially complete reduction in the severity of an established disease or condition or symptom of the disease or condition, such as reduction in tumor volume, reduction in tumor burden, reduction in death. 79 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 For example, a method for treating a disease is considered to be a treatment if there is a 10% reduction in one or more symptoms of the disease in a subject as compared to a control. Thus, the reduction can be a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or any percent reduction in between 10% and 100% as compared to native or control levels. It is understood that treatment does not necessarily refer to a cure or complete ablation of the disease, condition, or symptoms of the disease or condition.

[0407] As used herein, the terms “prevent”, “preventing”, and “prevention” of a disease or disorder refers to an action, for example, administration of the chimeric polypeptide or nucleic acid sequence encoding the chimeric polypeptide, that occurs before or at about the same time a subject begins to show one or more symptoms of the disease or disorder, which inhibits or delays onset or exacerbation of one or more symptoms of the disease or disorder. 5. EXAMPLES

[0408] The present invention is further described by the following examples, which are not intended to be limiting in any way. Example 1. Discovery of IL-18 variant polypeptides 1.1. Library designs

[0409] Structure-based optimization libraries were designed by analyzing complexes of wildtype human IL-18 binding to human IL-18BP and human IL-18 Receptor α (IL-18Rα) and human IL-18 Receptor β (IL-18Rβ) from the Protein Data Bank coordinates 7AL7 and 3WO4, respectively. Sixty-one IL-18 residues were identified and divided into five libraries based on their contacts to IL-18BP only (Library A), a combination of IL-18BP, IL-18Rα, and / or IL-18Rβ (Library B or E), IL-18Rα only (Library C), and IL-18Rβ only (Library D). The five libraries were further sub-divided into a total of 15 sub-libraries (Libraries A, B, C.1-C.5, D.1-D.2, E.1-E.6) containing up to 5 NNK-diversified codons apiece for a maximum combinatorial diversity of up to 3.2 x 106per sub-library and between 1.7 x 105to 1.3 x 107maximum diversity per library. Oligos were ordered from IDT and recombined into a single-chain Fc (scFc) vector and propagated in a proprietary yeast strain prior to selections. 1.2. Cycle 1 affinity maturation selections

[0410] Five IL-18 variant yeast libraries were propagated as described previously described (see, e.g., Y. Xu et al, PEDS 26(10), 663-70 (2013); WO2009036379; WO2010105256; and WO2012009568.) Libraries of IL-18 variants were subjected to four to five rounds of selection using flow cytometry (FACS). Yeast were pelleted, washed twice with wash buffer (phosphate-buffered saline (PBS) / 0.1% 80 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 bovine serum albumin (BSA)), and incubated at 24 °C with one of the following: decreasing concentrations of biotinylated monomeric human IL-18Rα / Rβ (complexed on ice for 20 minutes at a molar ratio of 1:2 for IL-18Rα:IL-18Rβ; 100 nM to 1 nM) to pressure for higher affinity; increasing concentrations of biotinylated monomeric IL-18BP (0.1 nM to 10 nM); or, increasing concentrations of biotinylated dimeric IL-18BP (10 nM to 100 nM). Yeast were washed twice with wash buffer and resuspended in anti-heavy chain 488 goat F(ab’)2 anti-human IgG1 Fc fragment specific (HC488) diluted at 1:200, propidium iodide (PI) diluted at 1:100, and streptavidin-AF633 (SA-633) diluted at 1:500 for 15 minutes at 24 °C. Yeast were washed three times with wash buffer and resuspended in 200-500 µl wash buffer and transferred into strainer-capped sort tubes. Sorting was carried out using a FACS ARIA (BD Biosciences) and sort gates were drawn to select for IL-18 variants with desired expression and binding properties. Selection rounds were repeated iteratively until final profiles were reached for each library. Throughout the FACS selections, the libraries were evaluated for species cross-reactivity and affinity pressure via antigen titration. Following the terminal round of selections, yeast were plated and individual colonies were picked for sequencing and characterization. 1.3. Cycle 2 affinity maturation selections

[0411] Eleven clones were nominated for further affinity maturation and were divided into nine individual libraries and combined into four discrete library pools numbered 1 through 4. For the library pools 1-3, NNK-based diversification was carried out at up to five residues per sub-library contacting either IL-18Rα or IL-18Rβ. For library pool 4, NNK-based diversification was performed at up to five residues per sub-library restricted to IL-18BP binding or to the shared interface between IL-18BP as well as IL-18Rα and / or IL-18Rβ. Library pools 1-4 had maximum theoretical diversities between 1.3 x 107and 3.9 x 107. Selections were carried out on FACS and were performed iteratively as in Cycle 1 until the final desired expression and antigen binding profiles were obtained. 1.4 Antibody production and purification

[0412] Yeast clones were grown to saturation and then induced for 48 h at 30 °C with shaking. After induction, yeast cells were pelleted and the supernatants were harvested for purification. IL-18 muteins having a single-chain Fc tag were purified using a Protein A column and eluted with phosphoric acid, pH 3.0 and neutralized with sodium phosphate, pH 12. 81 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 1.5 Developability analyses

[0413] The various developability assessments, including HIC, LC-MS, PSR, SEC, DSF Tm and biophysical and biochemical analyses, were performed as described previously (see, Jain T, et al. (2017) PNAS 114(5): 944-949; and Lu X, et al. (2018) MAbs 11(1): 45-57). Example 2. Measurement of Binding Affinities to IL-18 Binding Protein and IL-18Rα / β

[0414] Biolayer interferometry (BLI) kinetic measurements were acquired at 25 °C with an Octet HTX instrument (Sartorius, Bohemia, NY, USA) at an orbital shaking speed of 1000 rpm. Reagents were formulated in a running buffer of phosphate buffered saline with 0.1% IgG-free bovine serum albumin (PBSF). The IL-18 Ra and IL-18 Rb antigens (human and cyno) were precomplexed on ice for 20-30 minutes at a 1:2 (a:b) molar ratio prior to analysis. For each binding assessment the IL-18 scFc samples were formulated to 100 nM and loaded (300 s) to human IgG Fc capture (AHC) sensors (Sartorius, Bohemia, NY, USA) to a response level of 0.6-1.3 nm.

[0415] For the binding assessment to the Fc-fused antigen (human IL-18BP dimer), the loaded sensor tips were exposed (10 min) to a solution of adalimumab (3.3 M) to block unoccupied human Fc binding sites on the sensor tips. This was followed by a stand (15 min) in PBSF buffer before proceeding to the kinetic measurements. The kinetic measurements begin with a short (60 s) baseline dip into PBSF before exposing (180 s) the IL-18 scFc-loaded and blocked sensors to the Fc-fused antigen (100 nM). This was immediately followed by a dip (300 s) into PBSF to measure the rate of dissociation of the sample / antigen complex.

[0416] For monovalent binding assessments (human and cyno IL-18 Ra / b; mouse IL-18R1; human, cyno and mouse IL-18 BP) the loaded sensor tips were first equilibrated (15 min) in PBSF before proceeding to the kinetic measurements. The kinetic measurements begin with a short (60 s) baseline dip into PBSF before exposing (180 s) the loaded sensor tips to the monovalent antigen(s) (100 nM). This was immediately followed by a dip (180-300 s) into PBSF to measure the rate of dissociation of the sample / antigen complex.

[0417] All kinetic data were y-axis aligned, inter-step corrected and then fit to a 1:1 binding model using the ForteBio Data Analysis Software version 11.1.3.10. For interactions where the decay in response during the dissociation phase was less than 3.55%, a koffvalue of 1.20E-4 (s-1) was assigned and used to determine the approximate KDof the interaction. 82 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 Table 1. Binding Affinities for IL-18 polypeptide variants IL-18 Polypeptide Octet IL-18BP KD (M) Octet IL-18Rα / β KD (M) Variant Monovalent Monovalent CTL 88716 ( ild t \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 ADI-88703 9.73E-10 8.82E-09 ADI-88704 8.64E-10 7.16E-09 Table 2. Binding Affinities for IL-18 polypeptide variants IL-18 Polypeptide Octet Human Octet Human Octet Human \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 CTL-88717 1.34E-08 N.B. N.B. ADI-88623 4.49E-09 5.70E-10 5.69E-10 85 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 Example 3. HEK Blue Assay 3.1. IL-18

[0418] HEK-Blue IL-18 cells (InvivoGen) were plated at a concentration of 54,000 cells / well in culture media (without selection antibiotics) and stimulated with a dilution series of either recombinant hIL-18 (R&D Systems, Cat. No.9124-IL / CF) or hIL-18 variants (scFc fusions) for 20 hours at 37 °C and 5% CO2. IL-18 activity was assessed by quantification of Secreted Alkaline Phosphatase (SEAP) activity by mixing 20 µL of culture supernatant with 180 µL of reagent QUANTI-Blue (InvivoGen) and incubating at 37°C for 30 minutes before measuring absorbance at 620 nm. Results are shown in FIGs.3A-3D. 3.2. IL-18 Binding Protein

[0419] HEK-Blue IL-18 cells (InvivoGen) were plated at a concentration of 54,000 cells / well in culture media (without selection antibiotics) and stimulated with either recombinant hIL-18 (R&D Systems, Cat. No.9124-IL / CF) or hIL-18 variants (scFc fusions) at their respective EC70 concentrations and a dose titration of hIL-18BP-Fc for 20 hours at 37 °C and 5% CO2. IL-18 activity was assessed by quantification of Secreted Alkaline Phosphatase (SEAP) activity by mixing 20 µL of culture supernatant with 180 µL of reagent QUANTI-Blue (InvivoGen) and incubating at 37 °C for 30 minutes before measuring absorbance at 620 nm. Results are shown in FIGs.4A-4B. Example 4. Expression and purification of recombinant IL-18 polypeptide variants

[0420] C-terminally His-tagged IL-18 molecules were expressed in the Escherichia coli strain BL21(DE3) as an N-terminal SUMO fusion protein. The cells were grown in Terrific Broth with 100 µg / mL carbenicillin at 37°C until its OD600reached 0.8. Expression was induced by the addition of 0.5 mM IPTG and incubation at 15°C for 17 hours. The cells were collected by centrifugation and resuspended in lysis buffer (1XPBS, 20 mM imidazole) with protease inhibitor cocktail tablets (Roche, 11836170001). Cleared lysate was prepared by sonication (60s on, 20s off, 10 minutes total at 70% strength) followed by centrifugation. The lysate was incubated with Ni Sepharose 6 fast flow resin (Cytiva, 17531802) for at least 2 hours at 4°C, and the resin was collected in a gravity column. The resin was washed with 1X PBS, followed by wash buffer (20 mM sodium phosphate pH7, 500 mM NaCl, 30 mM imidazole), 150 mM imidazole buffer (20 mM sodium phosphate pH7, 500 mM NaCl, 150 mM imidazole), and 500 mM imidazole buffer (20 mM sodium phosphate pH7, 500 mM NaCl, 500 mM imidazole). Fractions with SUMO-IL-18 molecules were identified by SDS-PAGE and combined for 86 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 subsequent dialysis into 1XPBS, 1 mM DTT, 1 mM EDTA. SUMO tag was cleaved by incubation with SUMO protease (Millipore Sigma, SAE0067) for 3 days at 4°C, and cleavage efficiency was monitored by SDS-PAGE. Cleaved IL-18 molecules were captured with Ni Sepharose 6 fast flow resin, and the resin was washed in the same order as described above. Fractions containing IL-18 molecules were pooled and passed through Mustang E syringe filters (Cytiva, MSTG25E3) until the endotoxin level was below 5 EU / mg. Proteins were further purified on gel filtration column (Cytiva, 29148721) equilibrated with 1XPBS 1 mM DTT 1 mM EDTA. Elution fractions were concentrated, filtered sterile, aliquoted, and stored at -80 °C for further use. Example 5. IL-18 variants single chain Fc fusion protein production and SDS-PAGE analysis

[0421] IL-18 single chain Fc fusion (scFc) variants were produced. The proteins were expressed using the Expi293F transient expression system (Life Technologies, A14635) according to the manufacturer's protocol. Proteins were purified by Protein A chromatography. Briefly, clarified culture supernatants were left to rotate overnight in the presence of MabSelect Sure LX protein A resin (Cytiva, 17547402). The following day, the culture / resin mixture was applied to gravity columns. The resin was washed with 1× PBS, and the protein was eluted with 200 mM acetic acid pH 3.5, 50 mM sodium chloride and quenched with 1 M Tris pH 8. Elutions were dialyzed against 1× PBS, concentrated, aliquoted, and stored for future use at −80°C. Theoretical extinction coefficients were determined for each protein, and concentration was determined by A280. To determine the quality of each variant, proteins were analyzed by SDS-PAGE to characterize purity. Briefly, 3 μg of protein was loaded on a 4% to 12% Bis-Tris gel under nonreducing and reducing conditions. Gels were stained using the Pierce Mini Gel Staining Kit (Thermo Scientific) and the Pierce Power stain cassette (Thermo Scientific) and visualized using a ChemiDoc Imaging system (BioRad). Example 6. IL-18 Human PBMC Assay

[0422] Human PBMCs (from Donor HMN1139657) were used to assess the activity of IL-18 proteins. PBMCs were isolated from human leukopaks and cryopreserved prior to use. Frozen PBMCs were thawed and plated at 100,000 cells per well in X-Vivo 15 Media (Lonza). Cells were stimulated with 10 ng / mL IL-12 (WW0814 / WW0636), while concurrently being treated with titrated amounts of recombinant BPR IL-18 protein WW50991or WW50993, 10 nM, 5x dilutions over 9 points) for 48 hours at 37°C / 5% CO2. See Example 8 regarding expression and purification of the human BPR IL-18 proteins used in this assay. IL-18 activity was accessed by measuring IFNγ secretion with a human IFNγ alphaLISA (Revvity) according to the manufacturer’s protocols. EC50 values for each protein were 87 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 determined using a 4-parameter nonlinear curve fit (Graphpad Prism) and are shown in FIGs.5A-5B and 8A and 8B.

[0423] In order to measure resistance to IL-18BP, a similar assay was used. Frozen PBMCs were thawed and plated at 100,000 cells per well in X-Vivo 15 Media (Lonza). Cells were stimulated with 10 ng / mL IL-12 (WW0814 / WW0636), while concurrently being treated with titrated amounts of recombinant IL- 18BP protein (WW50300, 10 nM, 5x dilutions over 9 points) in the presence of 0.1 nM BPR IL-18 protein (approximate EC70 of BPR IL-18) for 48 hours at 37°C / 5% CO2. IL-18 activity was accessed by measuring IFNγ secretion with a human IFNγ alphaLISA (Revvity) according to the manufacturer’s protocols. Example 7. IL-18 HEK Blue Assay

[0424] HEK-Blue IL-18 cells (InvivoGen) were plated at a concentration of 54,000 cells / well in culture media (without selection antibiotics) and stimulated with a dilution series of either recombinant hIL-18- scFc (CTL-88716, 1.5nM, 5x dilutions over 9 points) or human BPR IL-18 proteins (WW50991 or WW50993, 1.3nM, 5x dilutions over 9 points) or (WW50992, 1.0nM, 25x dilutions over 5 points, or WW50994, 1.0nM, 5x dilutions over 9 points) for 20 hours at 37°C and 5% CO2. See Example 8 regarding expression and purification of the human BPR IL-18 proteins used in this assay. IL-18 activity was assessed by quantification of Secreted Alkaline Phosphatase (SEAP) activity by mixing 20 µL of culture supernatant with 180 µL of reagent QUANTI-Blue (InvivoGen) and incubating at 37°C for 30 minutes before measuring absorbance at 620 nm. EC50 values for each protein were determined using a 4-parameter nonlinear curve fit (GraphPad Prism). Results are shown in FIGs.6A-6E.

[0425] In order to measure resistance to IL-18BP, a similar assay was used. HEK-Blue IL-18 cells (InvivoGen) were plated at a concentration of 54,000 cells / well in culture media (without selection antibiotics) and stimulated with a constant concentration of human wildtype IL-18 [12.5pM] or human BPR IL-18 sumo proteins (WW50991 [8pM], WW50993 [2.5pM], WW50992 [4.5pM], or WW50994 [5pM]) and a dose titration of hIL-18BP-Fc (3000nM, 5x dilutions over 9 points) for 20 hours at 37 °C and 5% CO2. The approximate EC70 of wild-type IL-18 and BPR IL-18 was used. IL-18 activity was assessed by quantification of Secreted Alkaline Phosphatase (SEAP) activity by mixing 20 µL of culture supernatant with 180 µL of reagent QUANTI-Blue (InvivoGen) and incubating at 37 °C for 30 minutes before measuring absorbance at 620 nm. 88 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 Example 8. Expression and purification of recombinant SUMO tagged IL-18 molecules using mammalian cells

[0426] IL-18 molecules were expressed using the Expi293F transient expression system (Life Technologies, A14635) according to the manufacturer's protocol as an N-terminal human SUMO fusion and C-terminal His-tagged protein. During protein expression, the SUMO tag is removed by endogenous proteases resulting in IL-18 with a native N-terminus. The cells were collected by centrifugation and resuspended in hypotonic lysis buffer (50 mM Tris-HCl, pH 7.5) with protease inhibitor cocktail tablets (Roche, 05892953001). Lysate was stirred at 4°C for 1 hour. Salt concentration was adjusted to 250 mM NaCl prior to sonication. Clarified lysate was prepared by sonication (10 seconds on, 10 seconds off, 1 minute total at 70% strength) followed by centrifugation. The lysate was incubated with Ni Sepharose 6 fast flow resin (Cytiva, 17531802) for at least 1 hour at 4°C. The resin was collected in a gravity column and washed with wash buffer (50 mM Tris-HCl, 250 mM NaCl, pH 7.5), followed by wash buffer containing 20 mM imidazole buffer. Protein was eluted with wash buffer containing 500 mM imidazole buffer. Fractions with IL-18 molecules were identified by SDS-PAGE, then combined and concentrated. Proteins were further purified on a size-exclusion chromatography column (Cytiva, 28990944) equilibrated with 1X PBS. Elution fractions were concentrated, filtered sterile, aliquoted, and stored at -80 °C for further use. Example 9. Activation Of Murine Splenocytes For IL-18 Activity Assays

[0427] Murine splenocytes were used to assess the activity of IL-18 proteins. Murine splenocytes were isolated from C57BL / 6 spleens and rested in a T75 flask for 1 hour in complete RPMI (RPMI-1640 (Gibco), 10% heat inactivated FBS (Gibco), 100 U / mL Penicillin / Streptomycin (Gibco), 1X 2- mercaptoethanol (Gibco)). Suspension cells were then diluted to 1x106cells / mL in complete RPMI and stimulated with 5 µg / mL anti-CD28 (Biolegend) in T75 flasks coated with anti-CD3 (Biolegend) for 72 hours at 37°C / 5%CO2. Flasks were coated for 3 hours with 10 ug / mL anti-CD3 in 1X PBS (Gibco) and washed twice with 1X PBS prior to addition of cells. Cells were then harvested and resuspended at 1x106cells / mL in complete RPMI containing 2 ng / mL rmIL-12 (R&D Systems) and incubated for 24 hours at 37°C / 5%CO2. Cells were washed twice in complete X-VIVO media (X-VIVO 15 media (Lonza) with 10% heat inactivated FBS (Gibco)) and used in IL-18 activity assays or cryopreserved in Recovery Cell Culture Freezing media (Gibco) for future use. Results are shown in FIGs.7A-7C. 89 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 Example 10. IL-18 Murine Splenocyte Assay

[0428] Freshly activated murine splenocytes were plated at 40,000 cells per well. Cells were then stimulated with 0.1 ng / mL of rmIL-12, while concurrently being treated with titrated amounts of either recombinant mIL-18 protein or activatable IL-18 proteins for 24 hours at 37°C / 5% CO2. Cleaved inducible IL-18 was generated by incubation for 17 hours at 22°C with porcine pancreatic elastase (Sigma) in 1X PBS pH 7.4 (Gibco). IL-18 activity was assessed by measuring proliferation with Cell Titer Glo (Promega) according to the manufacturer’s protocol, and by IFNγ secretion using a mouse IFNγ alphaLISA kit (Revvity) according to the manufacturer’s protocol. Results are shown in FIGs.7A-7C. Example 11. Inhibition of IL-18 activity by IL-18BP

[0429] Frozen activated murine splenocytes were thawed in complete X-VIVO media and plated at 40,000 cells per well. A titration of IL-18BP was prepared with a constant concentration of 1, 0.1, or 0.01 nM IL-18 recombinant protein and incubated for approximately 30 minutes prior to addition to cells. Cells were stimulated with 0.1 ng / mL of rmIL-12, while concurrently being treated with titrated amounts of either recombinant mIL-18 protein or prepared titration of IL-18BP with a fixed concentration of IL-18 protein for 24 hours at 37°C / 5% CO2. IL-18 activity was assessed by measuring proliferation with Cell Titer Glo (Promega) according to the manufacturer’s protocol, and by IFNγ secretion using a mouse IFNγ alphaLISA kit (Revvity) according to the manufacturer’s protocol. Results are shown in FIGs.7A-7C. 6. SEQUENCE DISCLOSURE SEQ ID NO Description Sequence 1 Wildt e Human IL-18 (CTL- YFGKLESKLSVIRNLNDQVLFIDQGNRPLFEDMTDSDCRDNAPRTIFI KS G FI V V FI K L FI KS G FI K \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 SDIIFFQRSVPGHDNKMQFESSSYEGYFLACEKERDLFKLILKKEDEL GDRSIMFTVQNED FI K L FI KS G FI KS G FI K L FI KS G FI S G FI KS G FI S G FI K L I K L FI KS G FI K L FI KS G FI KS G 91 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 20 ADI-88685 YFGKLESKLSVIRNLNDQVLFIDQGNRPLFEDMTDSDCRDNAPRTIFI ISMYKDSQPRGMAVTISVKCEKISTLSCENKIISFKEMNPPDWIKDTK SDIIFFQRSVPGHDNKMQFESSSYEGYFLACEKERDLFKLILKKEDEL FI K L FI K L FI K L FI S G FI K L FI KS G FI K L FI K L FI KS G FI S G FI K L FI S G FI K L I K L \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 35 ADI-88701 YFGKLESKLSVIRNLNDQVLFIDQGNRPLFEDMTDSDCRDNAPRTIFI ISMYKDSQPRGMAVTISVKCEKISTLSCENKIISFKEMNPPDMIKDTK SDIIFFQRSVPGHDNKMQFESSSYEGYFLACEKERDLFKLILKKEDEL FI KS G FI KS G FI KS G FI S G FI S G FI KS G FI K L FI K L I K L FI S G FI K L II S G FI S G FI S G \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 50 ADI-88633 YFPKLRSKLSVIRNLNDQVLFIDQGNRPLFEDMTDSDCRDNAPRTIFII SMYKDSQPRGMAVTISVKCEKISTLSCENKIISFKEMNPPDNIKDTKS DIIFFQRSVPGHDNKMQFESSSYEGYFLACEKERDLFKLILKKEDELG FII S G II S G FI KS G FII S G II S G IF K L FI KS G FII S G FI KS G FI KS FI KS G FI KS G FI KS G FII S G \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 65 ADI-88642 WFGKLESKLSVIRNLNDQVLFIDQGNRPLFEDMTDSDCRDNAPRTIFI ISRYSDSQPRGMAVTISVKCEKISTLSCENKIISFKEMNPPDNIKDTKS DIIFFQRSVPGHDNKMQFESSSYEGYFLACEKERDLFKLILKKEDELG FI S G FI S G FI S G FI S G FIr S G Ir S G FII S G II S G FII DI D II DI D FII DI D II DI D FIr S G Ir S G \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 80 hIL18_S10K_I49R_P57S_S36E YFGKLESKLkVIRNLNDQVLFIDQGNRPLFEDMTDeDCRnNAPRTIFIr _D40N-L-scFc_(L = G4S (SEQ SMYKDSQsRGMAVTISVKCEKISTLSCENKIISFKEMNPPDNIKDTKS ID NO: 246)) DIIFFQRSVPGHDNKMQFESSSYEGYFLACEKERDLFKLILKKEDELG IS D D IS D D S D D FI SD D FII DI D II DI D FI S G FIr S G Ir S G II S G Ir S G Ir S G rS DI D II S G \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 95 hIL18_ADI- YFGKLESKLSVIRNLNDQVLFIDQGNRPLFEDMTDeDsRDNAPRTIFII 88631_(54P)_36E_C38S SMYKpSQPRGMAVTISVKCEKISTLSCENKIISFKEMNPPDNIKDTKS DIIFFQRSVPGHDNKMQFESSSYEGYFLACEKERDLFKLILKKEDELG IS DI D II DI D II DI D IS II R II DI D II DI D IS I R Ir S G Ir S G rS DI D II S G II S G IS D D IS D D \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 110 hIL18_ADI-88629_(36E)_C38S YFGKLESKLSVIRNLNDQVLFIDQGNRPLFEDMTDeDsRDNAPRTIFII SMYKDSQPRGMAVTISVKCEKISTLSCENKIISFKEMNPPDNIKDTKS DIIFFQRSVPGHDNKMQFESSSYEGYFLACEKERDLFKLILKKEDELG II DI D II DI D IS II R Ir S G Ir S G rS DI D FI KS G FI S G FIr S G Ir S G FI S G FII S G II S G FI D D \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 125 hIL18_ADI- YFGKLESKLSVIRNLNDQVLFIDQGNRPLFEDMTDeDCRDNAPRTIFII 88644_(51R_53S)_36E_A162C SrYsDSQPRGMAVTISVKCEKISTLSCENKIISFKEMNPPDNIKDTKSDI _I185C IFFQRSVPGHDNKMQFESSSYEGYFLcCEKERDLFKLILKKEDELGDR II DI R FI D D FII DI R II DI R FIr S G Ir S G Ir S G IS D D IS D D S D D II S G FII S G FI SD D FII DI R \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 140 hIL18_51R_57S_36E_40N_A16 YFGKLESKLSVIRNLNDQVLFIDQGNRPLFEDMTDeDCRnNAPRTIFII 2C_I185C SrYKDSQsRGMAVTISVKCEKISTLSCENKIISFKEMNPPDNIKDTKSDI IFFQRSVPGHDNKMQFESSSYEGYFLcCEKERDLFKLILKKEDELGDR FI S G FIr S G Ir S G II S G Ir S G Ir S G rS DI R II S G II S G IS DI R II DI R II DI R IS II R II DI R \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 155 hIL18_ADI- YFGKLESKLSVIRNLNDQVLFIDQGNRPLFEDMTDeDsRDNAPRTIFII 88651_(51R_53T)_36E_C38S_ SrYtDSQPRGMAVTISVKCEKISTLSCENKIISFKEMNPPDNIKDTKSDI A162C_I185C IFFQRSVPGHDNKMQFESSSYEGYFLcCEKERDLFKLILKKEDELGDR IS I R Ir S G Ir S G rS DI R II S G II S G IS D D IS D D II S G II DI R II DI R IS II R Ir S G Ir S G \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 170 hIL18_49R_53S_36E_40N_C38 YFGKLESKLSVIRNLNDQVLFIDQGNRPLFEDMTDeDsRnNAPRTIFIrS S_A162C_I185C MYsDSQPRGMAVTISVKCEKISTLSCENKIISFKEMNPPDNIKDTKSDI IFFQRSVPGHDNKMQFESSSYEGYFLcCEKERDLFKLILKKEDELGDR FI S G FI S G C L P L S PS H A A FS FI S G C L P L S PS H A A FS FI D D V T PS S G VF A IE E S FI D D V T PS S G VF A IE \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSPGK II S G C L P L S PS H A A FS FII S G C L P L S PS H A A FS FI SD D V T PS S G VF A IE E S FII DI D V T PS S G VF A IE E S 103 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 180 WW50865 YFGKLESKLSVIRNLNDQVLFIDQGNRPLFEDMTDeDCRnNAPRTIFII SrYKDSQsRGMAVTISVKCEKISTLSCENKIISFKEMNPPDNIKDTKSDI IFFQRSVPGHDNKMQFESSSYEGYFLACEKERDLFKLILKKEDELGD CV T PS S G VF A IE E S FI S G C L P L S PS H A A FS FIr S G C L P L S PS H A A FS Ir S G C L P L S PS H A A FS FII S G C L \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 TVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLP PSRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PS H A A FS II S G C L P L S PS H A A FS FII DI D V T PS S G VF A IE E S II DI D V T PS S G VF A IE E S FII DI D V T PS S G VF A \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 KTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCS II DI D V T PS S G VF A IE E S FIr S G C L P L S PS H A A FS Ir S G C L P L S PS H A A FS Ir S G C L P L S PS H A A FS 106 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 193 WW50858 FGKLESKLSVIRNLNDQVLFIDQGNRPLFEDMTDSDCRDNAPRTIFIIS MYKDSQPRGMAVTISVKCEKISTLSCENKIISFKEMNPPDNIKDTKSD IIFFQRSVPGHDNKMQFESSSYEGYFLACEKERDLFKLILKKEDELGD CV T PS S G VF A IE E S IS D D V T PS S G VF A IE E S S D D V T PS S G VF A IE E S FIr S G C L P L S PS H A A FS Ir S G C L \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 TVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLP PSRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PS H A A FS ne de ch sd ik na FI S G FI D D FI KS G II D D Ir S G II Q N Ir S G II DI D II D D Ir S G 108 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 210 scFc GGGGSAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE VKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNG KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQV KL G TL Y Q N H FII S G FII S G FII S G FII S G FII S G FII S L FII S G FII S G FI KS G FI KS G FI KS G FI KS G 109 \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 223 ADI-92594 YFGKLESKLSVIRNLNDQVLFIDQGNRPLFEDMTDQECRDNAPRTIFI ISMYKDSQVRGKAVTISVKCEKISTLSCENKIISFKEMNPPDNIKDTKS DIIFFQRSVPGHDNKMQFESSSYEGYFLACEKERDLFKLILKKEDELG FI KS G FI K L FI KS G FI KS G FI S G FI S G FI S G FI S G FI S R FI S G FI S G FI S G FI K L FI K L \\4149-1379-6182 v1 Attorney Docket No.: 761146.272320 238 ADI-92563 YFGKLESKLSVIRNLNDQVLFIDQGNRPLFEDMTDSECRDNAPRTIFI RSMYKDSQSRGMAVTISVKCEKISTLSCENKIISFKEMNPPDNILDTK SDIIFFQRSVPGHDNKMQFESSSYEGYFLACEKERDLSKLILKKEDEL FI K L FI K L FI K L FI K L FI K EL FII S G FI K L 111 \\4149-1379-6182 v1

Claims

Attorney Docket No.: 761146.272320 CLAIMS 1. A human interleukin-18 (IL-18) polypeptide comprising SEQ ID NO: 1 or an amino acid sequence that has at least about 95% identity to SEQ ID NO: 1, wherein a) the amino acid at position 1 is tryptophan; b) the amino acid at position 3 is not glycine, the amino acid at position 6 is not glutamic acid, the amino acid at position 10 is not serine, the amino acid at position 49 is not isolucine, the amino acid at position 54 is not aspartic acid, the amino acid at position 62 is not valine, and / or the amino acid at position 91 is not asparagine; the amino acid at position 40 is asparagine or glutamic acid; the amino acid at position 56 is proline; or the amino acid at position 57 is histidine; and wherein the IL-18 polypeptide has binding specificity for the human IL-18 receptor.

2. The IL-18 polypeptide of claim 1, wherein the amino acid at position 1 is tryptophan.

3. The IL-18 polypeptide of claim 1 or 2, wherein the amino acid at position 3 is selected from the group consisting of proline, aspartic acid, arginine, glutamine, and valine.

4. The IL-18 polypeptide of any one of the preceding claims, wherein the amino acid at position 6 is selected from the group consisting of arginine, glutamine, asparagine, tryptophan, and lysine.

5. The IL-18 polypeptide of any one of the preceding claims, wherein the amino acid at position 10 is selected from the group consisting of S10L, S10N, S10A, S10T, S10Q, S10H, S10L, S10I, S10H, S10A, S10K, S10R, S10G, S10F, and S10V.

6. The IL-18 polypeptide of any one of the preceding claims, wherein the amino acid substitution at position 49 is selected from the group consisting of I49L, I49T, I49Q, I49K, I49R, and I49S.

7. The IL-18 polypeptide of any one of the preceding claims, wherein the amino acid substitution at position 54 is selected from the group consisting of D54F, D54P, and D54H.

8. The IL-18 polypeptide of any one of the preceding claims, wherein the amino acid substitution at position 56 is Q56P. 112 \\4149-1379-6182 v1Attorney Docket No.: 761146.272320 u) D40X.

13. The IL-18 polypeptide of claim 12, wherein the IL-18 polypeptide comprises: a) S10L, I49L, and N91R; b) S10L, I49Q, and N91Y; c) S10I, I49K, and N91K; d) Q56P, P57H, and M60K; e) S10A, I49L, and V62Y; f) Y1W, M51R, K53S; and g) S10T, I49T, and N91L91.

14. The IL-18 polypeptide of any one of claims 1-12, wherein the IL-18 polypeptide comprises: a) S10A and I49L; b) S10R and I49T; c) S10I, I49K; d) V62A and N91M; e) I49L and N91A; f) I49T and N91R; g) I49I and N91K; h) I49R and N91Y; i) I49S and N91R; j) S10H and N91Y; k) S10H and N91L; l) S10H and N91R; m) S10H and N91T; n) S10T and N91L; o) S10T and N91R; p) S10F and N91M; q) G3P and D45P; r) G3D and D54F; s) G3P and 6EQ; t) G3Q and 6EW; u) E6K and D40N; 114 \\4149-1379-6182 v1Attorney Docket No.: 761146.272320 v) K53S and M51R; w) K53S and M51K; x) K53T and M51R; y) M60K and N91L; z) G3P and D54H; or za) G3R and E6N.

15. The IL-18 polypeptide of claim 14, wherein the IL-18 polypeptide comprises S10R and I49T, S10H and N91T, or G3R and E6N.

16. The IL-18 polypeptide of any one of claims 1-12, wherein the IL-18 polypeptide comprises: a) S10N; b) S10H; c) S10G; d) S10Q; e) S10K; f) S10R; g) V62S; h) N91G; i) N91R; j) N91W; k) N91L; l) N91T; m) N91I; n) N91V; o) N91S; p) N91M; q) N91K; r) N91Y; s) N91K; t) N91A; u) I49R; v) D54P; 115 \\4149-1379-6182 v1Attorney Docket No.: 761146.272320 w) E6Q; x) G3R; y) G3V; z) D40N; and za) D40E.

17. The IL-18 variant polypeptide of claim 16, wherein the IL-18 polypeptide variant comprises one amino acid substitutions at positions S10K, N91V, or I49R.

18. An IL-18 polypeptide comprising an amino acid sequence that is at least about 95% identical to the amino acid sequence of any one of SEQ ID NO: 2-170, 200-209, and 211-245.

19. The IL-18 polypeptide of claim 18, wherein the IL-18 polypeptide comprises the amino acid sequence of SEQ ID NO: 2 or an amino acid sequence that is at least about 95% identical to the amino acid sequence of SEQ ID NO:

2.

20. The IL-18 polypeptide of claim 18, wherein the IL-18 polypeptide comprises the amino acid sequence of SEQ ID NO: 13 or an amino acid sequence that is at least about 95% identical to the amino acid sequence of SEQ ID NO:

13.

21. The IL-18 polypeptide of claim 18, wherein the IL-18 polypeptide comprises the amino acid sequence of SEQ ID NO: 27 or an amino acid sequence that is at least about 95% identical to the amino acid sequence of SEQ ID NO:

27.

22. The IL-18 polypeptide of claim 18, wherein the IL-18 polypeptide comprises the amino acid sequence of SEQ ID NO: 66 or an amino acid sequence that is at least about 95% identical to the amino acid sequence of SEQ ID NO:

66.

23. The IL-18 polypeptide of claim 18, wherein the IL-18 polypeptide comprises the amino acid sequence of SEQ ID NO: 71 or an amino acid sequence that is at least about 95% identical to the amino acid sequence of SEQ ID NO:

71. 116 \\4149-1379-6182 v1Attorney Docket No.: 761146.272320 24. The IL-18 polypeptide of claim 18, wherein the IL-18 polypeptide comprises the amino acid sequence of SEQ ID NO: 75 or an amino acid sequence that is at least about 95% identical to the amino acid sequence of SEQ ID NO:

75.

25. The IL-18 variant polypeptide of claim 18, wherein the IL-18 polypeptide comprises the amino acid sequence of SEQ ID NO: 69 or an amino acid sequence that is at least about 95% identical to the amino acid sequence of SEQ ID NO:

69.

26. The IL-18 polypeptide of any one of claims 1-25, wherein the IL-18 polypeptide has substantially reduced binding to the IL-18 binding protein compared to wildtype human IL-18.

27. The IL-18 variant polypeptide of any one of claims 1-26, wherein the IL-18 polypeptide has binding specificity for IL-18Rα.

28. The IL-18 variant polypeptide of any one of claims 1-27, wherein the IL-18 polypeptide is synthetic.

29. A nucleic acid encoding the IL-18 polypeptide of any one of the preceding claims.

30. A vector comprising the nucleic acid of claim 29.

31. A host cell comprising the vector of claim 30 or nucleic acid of claim 29.

32. A method of making an IL-18 polypeptide of any one of claims 1-28, comprising culturing the host cell of claim 31 under suitable conditions for expression of the IL-18 polypeptide.

33. The method of claim 32, further comprising isolating the IL-18 polypeptide.

34. A fusion polypeptide comprising: a) the IL-18 polypeptide of any one of claims 1-28; and b) an amino acid sequence of interest. 117 \\4149-1379-6182 v1Attorney Docket No.: 761146.272320 35. The fusion polypeptide of claim 34, wherein the amino acid sequence of interest is an antibody or antibody binding fragment thereof.

36. The fusion polypeptide of claim 35, wherein the antibody or antibody binding fragment thereof is a Fc domain.

37. The fusion polypeptide of claim 34, wherein the amino acid sequence of interest is a polypeptide with biological activity.

38. The fusion polypeptide of claim 37, wherein the polypeptide with biological activity comprises a cytokine, a chemokine, a growth factor, a soluble receptor, a tumor antigen binding domain, a peptide hormone, a hormone receptor agonist, or muteins, variants or combinations of any of the foregoing.

39. A pharmaceutical composition comprising the IL-18 polypeptide of any one of claims 1-28 or the fusion protein of any one of claims 34-38, and a pharmaceutically acceptable carrier.

40. A method for treating cancer in a subject in need thereof, the method comprising administering to the subject an effective amount of the IL-18 variant polypeptide of any one of claims 1-28, the fusion protein of any one of claims 34-38, or the pharmaceutical composition of claim 39. 118 \\4149-1379-6182 v135. The fusion polypeptide of claim 34, wherein the amino acid sequence of interest is an antibody or antibody binding fragment thereof.

36. Tire fusion polypeptide of claim 35, wherein the antibody or antibody binding fragment thereof is a Fc domain.

37. The fusion polypeptide of claim 34, wherein the amino acid sequence of interest is a polypeptide with biological activity.

38. Tire fusion polypeptide of claim 37, wherein the polypeptide with biological activity comprises a cytokine, a chemokine, a growth factor, a soluble receptor, a tumor antigen binding domain, a peptide hormone, a hormone receptor agonist, or muteins. variants or combinations of any of the foregoing.

39. A pharmaceutical composition comprising the IL-18 polypeptide of any one of claims 1-28 or the fusion protein of any one of claims 34-38, and a pharmaceutically acceptable carrier.

40. A method for treating cancer in a subject in need thereof, the method comprising administering to the subject an effective amount of the IL-18 variant polypeptide of any one of claims 1-28, the fusion protein of any one of claims 34-38, orthe pharmaceutical composition of claim 39.

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