Membrane-bound IL-15, CD8 polypeptide, cell, composition, and methods of using the same

Abstract

The present disclosure relates to cells that can co-express a T cell receptor (“TCR”) together with a membrane-bound IL-15 polypeptide and / or a CD8 polypeptide, and to their use in adoptive cell therapy. The present disclosure further provides modified IL-15, IL-15Rα, IL-15 / IL-15Rα fusion polypeptide and IL-15Rα / IL-15 fusion polypeptide sequences, vectors, and related methods for making and using them. The present disclosure further provides modified CD8 sequences, vectors, and related methods for making and using them.

Description

Technical Field

[0001] Related Applications This application is an international application claiming priority to U.S. Provisional Patent Application No. 63 / 336,025, filed on April 28, 2022, the entire content of which is hereby incorporated by reference in its entirety for all purposes.

[0002] Reference to Electronically Submitted Sequence Listing An official copy of the sequence listing was created on April 27, 2023, and simultaneously submitted via EFS-Web as an ASCII format sequence listing with the file name "3000011-029977_Sequence-Listing_ST26" and a size of 722,707 bytes, and is filed simultaneously with this specification. The sequence listing contained in this ASCII format document is part of this specification and is hereby incorporated by reference in its entirety.

[0003] The present disclosure relates to cells that can co-express one or any combination of a T cell receptor (TCR), a CD8 polypeptide, and / or a membrane-bound interleukin 15 (IL-15), and their use in adoptive cell therapy (ACT). The present disclosure further provides modified CD8 sequences, IL-15 sequences, IL-15 receptor alpha (IL-15-Rα) sequences, IL-15 / IL-15Rα fusion polypeptides, their vectors, compositions, transformed cells, and related methods.

Background Art

[0004] CD8 and CD4 are transmembrane glycoproteins characteristic of different populations of T lymphocytes, and the antigen responses of those T lymphocytes are restricted by class I MHC molecules and class II MHC molecules, respectively. Both play major roles in the differentiation and selection of T cells during thymic development and in the activation of mature T lymphocytes in response to antigen-presenting cells. CD8 and CD4 are both immunoglobulin superfamily proteins. They determine antigen restriction by binding to MHC molecules at an interface quite distinct from the region presenting the antigenic peptide, although their structural bases for similar functions would appear to be quite different. Their sequence similarity is low. CD4 is expressed on the cell surface as a monomer, while CD8 is expressed as an αα homodimer (e.g., Figure 55C) or as an αβ heterodimer (e.g., Figure 55A). In humans, this CD8αα homodimer can be functionally replaced by the CD8αβ heterodimer. CD8 contacts an acidic loop in the α3 domain of class I MHC, thereby increasing the avidity of the T cell for its target. CD8 is also involved in phosphorylation, by which its α-chain cytoplasmic tail associates with tyrosine kinase p56 lck and CTL activation occurs.

[0005] Interleukin-15 (IL-15 or IL15), a multifaceted cytokine, is a member of the 4α-helix bundle type cytokine family. (Waldmann TA and Tagaya Y, Ann. Rev. Immunol. 17:19-49, 1999. The content is incorporated herein by reference). Wild-type IL-15, a glycoprotein of 14-15 kDa, shares partial structural homology with IL-2. (Ibid.). Wild-type IL-15 can be expressed in two isoforms. One has a 48-amino acid signal peptide and the other has a 21-amino acid signal peptide. (Ibid.). The mature form of wild-type IL-15 consists of 114 amino acids. (Ibid.). The expression of wild-type IL-15 is regulated during transcription, translation, and intracellular transport. (Ibid.). Wild-type IL-15 utilizes a unique receptor, IL-15Rα (or "IL15Rα"). In lymphocytes, this receptor binds to IL-15 with high affinity and forms a trimer with IL-2Rβ (also called IL-2 / IL-15Rβ) and IL-2Rγ (γ c also called). (Ibid., Okada S at al., Immunol. and Cell Biol. 93:461-471, 2015. The content is incorporated herein by reference). Wild-type IL-15Rα contains a signal peptide, an extracellular domain, a transmembrane domain, and a cytoplasmic domain. (Waldman and Tagaya). The extracellular domain of wild-type IL-15Rα contains a sushi domain (also called GP-1 motif). (Ibid.).

[0006] Adoptive cell therapy (ACT) is a promising approach for the treatment of diseases such as cancer. T cell therapy has been successful in treating various cancers. Li et al., Signal Transduction and Targeted Therapy 4(35):(2019). The content of which is incorporated herein by reference. However, the cells used in ACT often do not persist in the tumor microenvironment and rapidly lose their ability to kill tumor cells. Therefore, there is a need for T cells and natural killer cells that can persist in the tumor microenvironment and / or exhibit a sustained ability to kill tumor cells. There is also a desire to develop methods for producing T cells and natural killer cells with enhanced specific cytotoxicity for immunotherapy. SUMMARY OF THE INVENTION

[0007] In multiple embodiments, a membrane-bound IL-15 polypeptide (membrane-bound IL-15 or mbIL-15) can be provided. In multiple embodiments, the nucleic acids described herein can include and / or encode a membrane-bound IL-15 polypeptide. In multiple embodiments, the vectors described herein can include and / or encode a membrane-bound IL-15 polypeptide. In multiple embodiments, the cells described herein can include and / or express a membrane-bound IL-15 polypeptide. In multiple embodiments, the compositions described herein can include a membrane-bound IL-15 polypeptide, or can include cells that include and / or express a membrane-bound IL-15 polypeptide. In multiple embodiments, IL-15 can be in a membrane-bound state by expressing the IL-15 polypeptide and the IL-15Rα polypeptide in an IL-15 / IL-15Rα fusion polypeptide (IL-15 / IL-15Rα). The IL-15 / IL-15Rα fusion polypeptide, and other membrane-bound forms of IL-15, can be referred to as membrane-bound IL-15 (mbIL-15).

[0008] In multiple embodiments, an isolated membrane-bound IL-15 polypeptide may be provided. An isolated nucleic acid sequence may be provided that includes one or more nucleic acid sequences encoding one or more membrane-bound IL-15 polypeptides. In multiple embodiments, an isolated vector may be provided that includes one or more nucleic acid sequences encoding one or more membrane-bound IL-15 polypeptides. In multiple embodiments, cells may be provided that contain and / or express one or more membrane-bound IL-15 polypeptides. In multiple embodiments, cells may be provided that contain, express, or include one or more nucleic acid sequences that include one or more nucleic acid sequences encoding one or more membrane-bound IL-15 polypeptides. In multiple embodiments, cells may be provided that contain, express, or include one or more vectors that include one or more nucleic acid sequences that include one or more nucleic acid sequences encoding one or more membrane-bound IL-15 polypeptides. In multiple embodiments, a composition may be provided that includes such polypeptides, nucleic acids, vectors, and / or cells.

[0009] In multiple embodiments, the IL-15 polypeptide can be located at the N-terminus of the IL-15Rα polypeptide in the membrane-bound IL-15 polypeptide. (Figure 67A). In multiple embodiments, the IL-15 polypeptide can be located at the C-terminus of the IL-15Rα polypeptide in the membrane-bound IL-15 polypeptide. (Figure 67B). The IL-15 polypeptide in Figures 67A and 67B can be an immature wild-type (wt), immature mutant, mature wild-type, or mature mutant. The IL-15Rα polypeptide in Figures 67A and 67B can be an immature wild-type, immature mutant, mature wild-type, or mature mutant. In multiple embodiments, the IL-15 polypeptides in 67A and 67B are mature, with or without mutations, and the IL-15Rα polypeptides in 67A and 67B are mature, with or without mutations. In multiple embodiments, the IL-15 polypeptides in 67A and 67B are mature, with or without mutations, and the IL-15Rα polypeptides in 67A and 67B are mature and mutant. A linker is shown in Figures 67A and 67B, but the mbIL-15 polypeptide may or may not contain a linker.

[0010] In multiple embodiments, the IL-15 polypeptide and the IL-15Rα polypeptide can be linked by one or more linkers. In multiple embodiments, the membrane-bound IL-15 can be included and / or encoded by the structure shown in Figure 67A or Figure 67B. In Figures 67A and 67B, the line connecting IL-15 to one or more linkers and connecting the one or more linkers to IL-15Rα may represent a direct bond without intervening sequences, or may represent intervening sequences such as, for example, but not limited to, a linker, an untranslated sequence (in the case of a nucleic acid sequence), a translated sequence, a sequence containing one or more restriction endonuclease sites (in the case of a nucleic acid sequence), or a combination thereof.

[0011] In multiple embodiments, the IL-15 / IL-15Rα polypeptide may include one or more signal peptides. In multiple embodiments, membrane-bound IL-15, which includes one or more signal peptides and optionally one or more linkers, may be included and / or encoded by the structures shown in FIGS. 68A or 68B. In FIGS. 68A and 68B, the IL-15 polypeptide can be an immature wild-type, immature mutant, mature wild-type, or mature mutant. In FIGS. 68A and 68B, the IL-15Rα polypeptide can be an immature wild-type, immature mutant, mature wild-type, or mature mutant. In multiple embodiments, the IL-15 polypeptides of FIGS. 68A and 68B are mature, with or without mutations, and the IL-15Rα polypeptides of FIGS. 68A and 68B are mature, with or without mutations. In multiple embodiments, the IL-15 polypeptides of FIGS. 68A and 68B are mature, with or without mutations, and the IL-15Rα polypeptides of FIGS. 68A and 68B are mature and mutant. Linkers are shown in FIGS. 68A and 68B, but the mbIL-15 polypeptide containing a signal peptide may or may not include a linker. As shown in FIGS. 68A and 68B, (a) a line connecting one or more signal peptides (SP) to IL-15, the IL-15 to one or more linkers (L), and the one or more linkers to IL-15Rα (FIG. 68A), or (b) a line connecting one or more signal peptides (SP) to IL-15α, the IL-15α to one or more linkers (L), and the one or more linkers to IL-15 (FIG. 68B) can represent a direct bond without intervening sequences or can represent intervening sequences such as, for example, but not limited to, linkers, untranslated sequences (in the case of nucleic acid sequences), translated sequences, sequences containing one or more restriction endonuclease sites (in the case of nucleic acid sequences), or combinations thereof.

[0012] In multiple embodiments, the CD8 polypeptide described herein may include a CD8α immunoglobulin (Ig)-like domain, a CD8β region, a CD8α transmembrane domain, and a CD8α cytoplasmic domain. In multiple embodiments, the CD8β region can be a CD8β stalk region or domain.

[0013] In multiple embodiments, the CD8 polypeptide described herein includes: (a) an immunoglobulin (Ig)-like domain having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the amino acid sequence of SEQ ID NO: 1; (b) a CD8β region having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the amino acid sequence of SEQ ID NO: 2; (c) a transmembrane domain having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the amino acid sequence of SEQ ID NO: 3; and (d) a cytoplasmic domain having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the amino acid sequence of SEQ ID NO: 4.

[0014] In multiple embodiments, the CD8 polypeptide described herein has at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the amino acid sequence of SEQ ID NO: 5.

[0015] In multiple embodiments, the CD8 polypeptide described herein has at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the amino acid sequence of SEQ ID NO: 7.

[0016] In multiple embodiments, the CD8 polypeptide described herein may include one or more signal peptides having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to any one of the amino acid sequences of SEQ ID NO: 6, SEQ ID NO: 293 or SEQ ID NO: 294, which are directly or indirectly fused to the N-terminus or C-terminus of the CD8 polypeptide described herein.

[0017] In multiple embodiments, the CD8 polypeptide described herein can include (a) SEQ ID NO: 1 containing 1, 2, 3, 4, or 5 amino acid substitutions, (b) SEQ ID NO: 2 containing 1, 2, 3, 4, or 5 amino acid substitutions, (c) SEQ ID NO: 3 containing 1, 2, 3, 4, or 5 amino acid substitutions, and (d) SEQ ID NO: 4 containing 1, 2, 3, 4, or 5 amino acid substitutions. In multiple embodiments, the amino acid substitutions can be conservative or non-conservative. In multiple embodiments, the amino acid substitutions can be conservative amino acid substitutions.

[0018] In multiple embodiments, the CD8 polypeptide described herein can be a CD8α polypeptide or a modified CD8α polypeptide.

[0019] In multiple embodiments, the CD8 polypeptide described herein can be a CD8αβ polypeptide or a modified CD8α polypeptide.

[0020] In multiple embodiments, the CD8β polypeptide can include any one of the amino acid sequences of SEQ ID NOs: 8, 9, 10, 11, 12, 13, or 14.

[0021] In multiple embodiments, the TCRα chain and the TCRβ chain can be selected from SEQ ID NOs: 15 and 16, 17 and 18, 19 and 20, 21 and 22, 23 and 24, 25 and 26, 27 and 28, 29 and 30, 31 and 32, 33 and 34, 35 and 36, 37 and 38, 39 and 40, 41 and 42, 43 and 44, 45 and 46, 47 and 48, 49 and 50, 51 and 52, 53 and 54, 55 and 56, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, 71 and 303, 304 and 74, 75 and 76, 77 and 78, 79 and 80, 81 and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90, and 91 and 92.

[0022] In multiple embodiments, the isolated nucleic acid can include a nucleic acid sequence encoding a T cell receptor comprising an α-chain and a β-chain, as well as a CD8 polypeptide comprising an α-chain and a β-chain. In multiple embodiments, the CD8 polypeptide can include a CD8α-chain and / or a CD8β-chain, and the CD8α-chain and / or the CD8β-chain can independently be modified or unmodified. The isolated nucleic acid can include a nucleic acid that is at least about 80% identical to the nucleic acid sequence of SEQ ID NO: 267, 269, 271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291, 295, 297, 299, or 301. The isolated nucleic acid can be at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 267, 269, 271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291, 295, 297, 299, or 301.

[0023] In certain aspects, the polypeptide sequences and / or nucleic acid sequences described herein can be isolated sequences and / or recombinant sequences.

[0024] In certain aspects, the cells described herein can be isolated cells and / or recombinant cells.

[0025] In multiple embodiments, the isolated nucleic acid includes the nucleic acid sequence of SEQ ID NO: 267.

[0026] In multiple embodiments, the isolated nucleic acid includes the nucleic acid sequence of SEQ ID NO: 279.

[0027] In multiple embodiments, the isolated polypeptide can be encoded by the nucleic acids described herein or, for example, by nucleic acids that encode the same polypeptide due to codon degeneracy.

[0028] In multiple embodiments, the isolated polypeptide can comprise an amino acid sequence that is at least about 80% identical to the amino acid sequence of SEQ ID NO: 268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 296, 298, 300, or 302. The amino acid sequence can be at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 296, 298, 300, or 302. In another aspect, SEQ ID NO: 268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 296, 298, 300, or 302 can include substitutions or deletions of 1, 2, 3, 4, 5, 10, 15 or 20 or more amino acids. In yet another aspect, SEQ ID NO: 268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 296, 298, 300, or 302 can include substitutions or deletions of up to 1, 2, 3, 4, 5, 10, 15 or 20 amino acids.

[0029] In multiple embodiments. The isolated polypeptide can comprise the amino acid sequence of SEQ ID NO: 268.

[0030] In multiple embodiments. The isolated polypeptide can comprise the amino acid sequence of SEQ ID NO: 280.

[0031] In multiple embodiments, a nucleic acid encoding a fusion polypeptide of Formula I below can be provided: N-terminal - P6 - PL - P7 - C-terminal [I] Wherein P6 and P7 are each independently a first polypeptide and a second polypeptide, PL is a linker, and PL comprises a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 387 or 389.

[0032] In multiple embodiments, a nucleic acid comprising the following formula II can be provided: 5’-N6-NL-N7-3’ [II] Wherein N6 and N7 each independently encode a first polypeptide and a second polypeptide, NL encodes a linker, and NL comprises a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 388 or 390, or SEQ ID NO: 388 or 390.

[0033] In multiple embodiments, a nucleic acid encoding a polypeptide comprising SEQ ID NO: 309, 311, 313 or 315, or a polypeptide that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 309, 311, 313 or 315, can be provided.

[0034] In multiple embodiments, a nucleic acid encoding a polypeptide comprising SEQ ID NO: 311, 313 or 315, or a polypeptide that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 311, 313 or 315, can be provided.

[0035] In multiple embodiments, a nucleic acid comprising SEQ ID NO: 310, 312, 314 or 316, or a nucleic acid comprising a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 310, 312, 314 or 316, can be provided.

[0036] In some embodiments, a nucleic acid comprising SEQ ID NO: 312, 314, or 316, or a nucleic acid comprising a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 312, 314, or 316, can be provided.

[0037] In some embodiments, a nucleic acid encoding a polypeptide comprising: (i) SEQ ID NO: 307 directly or indirectly fused to the N-terminus of a polypeptide comprising any of SEQ ID NO: 309, 311, 313, or 315; or (ii) a polypeptide that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 307 directly or indirectly fused to a polypeptide that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to any of SEQ ID NO: 309, 311, 313, or 315, can be provided.

[0038] In some embodiments, a nucleic acid encoding a polypeptide comprising: (i) SEQ ID NO: 307 directly or indirectly fused to the N-terminus of a polypeptide comprising any of SEQ ID NO: 311, 313, or 315; or (ii) a polypeptide that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 307 directly or indirectly fused to a polypeptide that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to any of SEQ ID NO: 311, 313, or 315, can be provided.

[0039] In multiple embodiments, the nucleic acid may further comprise a nucleic acid encoding a signal peptide that is directly or indirectly fused to the N-terminus of SEQ ID NO: 307, or to the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 307. In multiple embodiments, the signal peptide may be derived from an IgE polypeptide. In multiple embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 367.

[0040] In multiple embodiments, there is provided a nucleic acid comprising (i) SEQ ID NO: 308 directly or indirectly fused to the 5' end of any of SEQ ID NOs: 310, 312, 314 or 316, or (ii) a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 308, directly or indirectly fused to the 5' end of a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to any of SEQ ID NOs: 310, 312, 314 or 316.

[0041] In multiple embodiments, a nucleic acid can be provided that comprises (i) SEQ ID NO: 308 fused directly or indirectly to the 5' end of any of SEQ ID NO: 312, 314, or 316, or (ii) a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 308 and fused directly or indirectly to the 5' end of any sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to any of SEQ ID NO: 312, 314, or 316.

[0042] In multiple embodiments, the nucleic acid can further comprise a nucleic acid encoding a signal peptide fused directly or indirectly to the 5' end of SEQ ID NO: 308 or to the 5' end of a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 308. In multiple embodiments, the signal peptide can be derived from an IgE polypeptide. In multiple embodiments, the signal peptide derived from an IgE polypeptide can comprise SEQ ID NO: 368 or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 368.

[0043] In multiple embodiments, a nucleic acid can be provided that encodes a polypeptide comprising SEQ ID NO: 317, 321, 325, 327, 329, 331, 333, or 335, or a polypeptide that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 317, 321, 325, 327, 329, 331, 333, or 335.

[0044] In multiple embodiments, a nucleic acid encoding a polypeptide comprising SEQ ID NO: 317, 321, 325, 327, 329, 331, or 333, or a polypeptide that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 317, 321, 325, 327, 329, 331, or 333, may be provided.

[0045] In multiple embodiments, the nucleic acid may further comprise a nucleic acid encoding a signal peptide that is directly or indirectly fused to the N-terminus of SEQ ID NO: 317, 321, 325, 327, 329, 331, or 333, or to the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 317, 321, 325, 327, 329, 331, or 333. In multiple embodiments, the signal peptide may be derived from an IgE polypeptide. In multiple embodiments, the signal peptide derived from an IgE polypeptide may comprise a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 367.

[0046] In multiple embodiments, a nucleic acid comprising SEQ ID NO: 318, 322, 326, 328, 330, 332, 334, or 336, or a nucleic acid comprising a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 318, 322, 326, 328, 330, 332, 334, or 336, may be provided.

[0047] In some embodiments, a nucleic acid comprising SEQ ID NO: 318, 322, 326, 328, 330, 332, or 334, or a nucleic acid comprising a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 318, 322, 326, 328, 330, 332, or 334, can be provided.

[0048] In some embodiments, the nucleic acid can further comprise a nucleic acid encoding a signal peptide that is directly or indirectly fused to the 5' end of SEQ ID NO: 318, 322, 326, 328, 330, 332, or 334, or to the 5' end of a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 318, 322, 326, 328, 330, 332, or 334. In some embodiments, the signal peptide can be derived from an IgE polypeptide. In some embodiments, the signal peptide derived from an IgE polypeptide can comprise a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 368.

[0049] In some embodiments, a nucleic acid encoding a polypeptide comprising SEQ ID NO: 337, 341, 345, 347, 349, 351, 353, or 355, or a polypeptide that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 337, 341, 345, 347, 349, 351, 353, or 355, can be provided.

[0050] In multiple embodiments, a nucleic acid encoding a polypeptide comprising SEQ ID NO: 337, 341, 345, 347, 349, 351, or 353, or a polypeptide that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 337, 341, 345, 347, 349, 351, or 353, may be provided.

[0051] In multiple embodiments, a nucleic acid comprising SEQ ID NO: 338, 342, 346, 348, 350, 352, 354, or 356, or a nucleic acid comprising a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 338, 342, 346, 348, 350, 352, 354, or 356, may be provided.

[0052] In multiple embodiments, a nucleic acid comprising SEQ ID NO: 338, 342, 346, 348, 350, 352, or 354, or a nucleic acid comprising a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 338, 342, 346, 348, 350, 352, or 354, may be provided.

[0053] In multiple embodiments, the nucleic acids described herein may further comprise a nucleic acid encoding (a) at least one TCR polypeptide comprising an α-chain and a β-chain, (b) at least one CD8 polypeptide comprising (i) an α-chain, (ii) a β-chain, or (iii) both an α-chain and a β-chain, or (c) at least one TCR polypeptide comprising an α-chain and a β-chain and at least one CD8 polypeptide comprising (i) an α-chain, (ii) a β-chain, or (iii) both an α-chain and a β-chain.

[0054] In some embodiments, a polypeptide, a plurality of polypeptides, or a fusion polypeptide encoded by the nucleic acids described herein may be provided.

[0055] In some embodiments, a polypeptide or fusion polypeptide may be provided that comprises an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 307, 309, 311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331, 333, 335, 337, 339, 341, 343, 345, 347, 349, 351, 353, or 355.

[0056] In some embodiments, a polypeptide or fusion polypeptide may be provided that comprises an amino acid sequence that is at least about 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331, 333, 337, 339, 341, 343, 345, 347, 349, 351, or 353.

[0057] In some embodiments, a fusion polypeptide may be provided that comprises a polypeptide that is at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 307, directly or indirectly fused to the N-terminus of any of a polypeptide that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to any of SEQ ID NO: 309, 311, 313, or 315.

[0058] In some embodiments, a fusion polypeptide can be provided that comprises a polypeptide that is at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 307, directly or indirectly fused to the N-terminus of any polypeptide that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 311, 313, or 315.

[0059] In some embodiments, the fusion polypeptide can further comprise a signal peptide directly or indirectly fused to the N-terminus of a polypeptide that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 307. In some embodiments, the signal peptide can be derived from an IgE polypeptide. In some embodiments, the signal peptide derived from an IgE polypeptide can comprise SEQ ID NO: 367, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 367.

[0060] In multiple embodiments, there can be provided a nucleic acid comprising: (a) a nucleic acid encoding (i) a T cell receptor (TCR) comprising an α-chain and a β-chain and a CD8 polypeptide comprising an α-chain and a β-chain, or (ii) a TCR comprising an α-chain and a β-chain and a CD8 polypeptide comprising an α-chain and not comprising a β-chain; and (b) a nucleic acid encoding an IL-15 / IL-15Rα fusion polypeptide, wherein the TCR α-chain and TCR β-chain are selected from SEQ ID NOs: 15 and 16, 17 and 18, 19 and 20, 21 and 22, 23 and 24, 25 and 26, 27 and 28, 29 and 30, 31 and 32, 33 and 34, 35 and 36, 37 and 38, 39 and 40, 41 and 42, 43 and 44, 45 and 46, 47 and 48, 49 and 50, 51 and 52, 53 and 54, 55 and 56, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, 71 and 303, 304 and 74, 75 and 76, 77 and 78, 79 and 80, 81 and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90, and 91 and 92; the CD8 α-chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof; and if present, the CD8 β-chain is SEQ ID NO: 8, 9, 10, 11, 12, 13 or 14; and the IL-15 / IL-15Rα fusion polypeptide is (i) a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 307, directly or indirectly fused to the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to the N-terminus of SEQ ID NO: 309, with or without a linker therebetween; or (ii) a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 307, directly or indirectly fused to the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to the N-terminus of SEQ ID NO: 311, with or without a linker therebetween.(iii) the N-terminus of SEQ ID NO: 313 or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 313, directly or indirectly fused to the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 307, with or without a linker in between, or (iv) the N-terminus of SEQ ID NO: 315 or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 315, directly or indirectly fused to the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 307, with or without a linker in between, and is selected therefrom.,

[0061] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide may further comprise a signal peptide directly or indirectly fused to the N-terminus of SEQ ID NO: 307 of any one of (i), (ii), (iii) or (iv), or to the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 307 of any one of (i), (ii), (iii) or (iv).

[0062] In some embodiments, a nucleic acid can be provided that comprises: (a) a nucleic acid encoding (i) a T cell receptor (TCR) comprising an α chain and a β chain and a CD8 polypeptide comprising an α chain and a β chain, or (ii) a TCR comprising an α chain and a β chain and a CD8 polypeptide comprising an α chain and not comprising a β chain; and (b) a nucleic acid encoding an IL-15 / IL-15Rα fusion polypeptide, wherein the TCR α chain and TCR β chain are selected from SEQ ID NOs: 15 and 16, 17 and 18, 19 and 20, 21 and 22, 23 and 24, 25 and 26, 27 and 28, 29 and 30, 31 and 32, 33 and 34, 35 and 36, 37 and 38, 39 and 40, 41 and 42, 43 and 44, 45 and 46, 47 and 48, 49 and 50, 51 and 52, 53 and 54, 55 and 56, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, 71 and 303, 304 and 74, 75 and 76, 77 and 78, 79 and 80, 81 and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90, and 91 and 92; the CD8 α chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof; and if present, the CD8 β chain is SEQ ID NO: 8, 9, 10, 11, 12, 13 or 14; and the IL-15 / IL-15Rα fusion polypeptide is (i) a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 307, directly or indirectly fused to the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to the N-terminus of SEQ ID NO: 311, with or without a linker therebetween; or (ii) a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 307, directly or indirectly fused to the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to the N-terminus of SEQ ID NO: 313, with or without a linker therebetween.or (iii) the N-terminus of SEQ ID NO: 315 or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 315, directly or indirectly fused to the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 307, with or without a linker therebetween, and is selected from these.

[0063] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide may further comprise a signal peptide directly or indirectly fused to the N-terminus of SEQ ID NO: 307 of any of (i), (ii) or (iii), or to the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 307 of any of (i), (ii) or (iii).

[0064] In multiple embodiments, the signal peptide may be derived from an IgE polypeptide. In multiple embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367 or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 367.

[0065] In multiple embodiments, a nucleic acid can be provided that comprises (a) a nucleic acid encoding (i) a T cell receptor (TCR) comprising an α-chain and a β-chain, and a CD8 polypeptide comprising an α-chain and a β-chain, or (ii) a TCR comprising an α-chain and a β-chain, and a CD8 polypeptide comprising an α-chain and not comprising a β-chain, and (b) a nucleic acid encoding an IL-15 / IL15Rα fusion polypeptide, wherein the TCR α-chain and TCR β-chain are selected from SEQ ID NOs: 15 and 16, 17 and 18, 19 and 20, 21 and 22, 23 and 24, 25 and 26, 27 and 28, 29 and 30, 31 and 32, 33 and 34, 35 and 36, 37 and 38, 39 and 40, 41 and 42, 43 and 44, 45 and 46, 47 and 48, 49 and 50, 51 and 52, 53 and 54, 55 and 56, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, 71 and 303, 304 and 74, 75 and 76, 77 and 78, 79 and 80, 81 and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90, 91 and 92; the CD8 α-chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof; and if present, the CD8 β-chain is SEQ ID NO: 8, 9, 10, 11, 12, 13 or 14; and the IL-15 / IL-15Rα fusion polypeptide is selected from SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, 333 or 335, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, 333 or 335.

[0066] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide can further comprise a signal peptide that is directly or indirectly fused to the N-terminus of SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, 333 or 335, or to the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, 333 or 335.

[0067] In multiple embodiments, a nucleic acid can be provided that comprises (a) (i) a nucleic acid encoding a T cell receptor (TCR) comprising an α-chain and a β-chain, and a CD8 polypeptide comprising an α-chain and a β-chain, or (ii) a nucleic acid encoding a TCR comprising an α-chain and a β-chain, and a CD8 polypeptide comprising an α-chain and not comprising a β-chain, and (b) a nucleic acid encoding an IL-15 / IL15Rα fusion polypeptide, wherein the TCR α-chain and TCR β-chain are selected from SEQ ID NOs: 15 and 16, 17 and 18, 19 and 20, 21 and 22, 23 and 24, 25 and 26, 27 and 28, 29 and 30, 31 and 32, 33 and 34, 35 and 36, 37 and 38, 39 and 40, 41 and 42, 43 and 44, 45 and 46, 47 and 48, 49 and 50, 51 and 52, 53 and 54, 55 and 56, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, 71 and 303, 304 and 74, 75 and 76, 77 and 78, 79 and 80, 81 and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90 and 91 and 92; the CD8 α-chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof; and if present, the CD8 β-chain is SEQ ID NO: 8, 9, 10, 11, 12, 13 or 14; and the IL-15 / IL-15Rα fusion polypeptide is selected from SEQ ID NOs: 317, 319, 321, 323, 325, 327, 329, 331 or 333, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NOs: 317, 319, 321, 323, 325, 327, 329, 331 or 333.

[0068] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide may further comprise a signal peptide that is directly or indirectly fused to the N-terminus of SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331 or 333, or is directly or indirectly fused to the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331 or 333.

[0069] In multiple embodiments, the signal peptide may be derived from an IgE polypeptide. In multiple embodiments, the signal peptide derived from an IgE polypeptide may comprise a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 367.

[0070] In multiple embodiments, a nucleic acid is provided that can include: (a) (i) a nucleic acid encoding a T cell receptor (TCR) comprising an α-chain and a β-chain, and a CD8 polypeptide comprising an α-chain and a β-chain, or (ii) a nucleic acid encoding a TCR comprising an α-chain and a β-chain, and a CD8 polypeptide comprising an α-chain and not comprising a β-chain; and (b) (i) a nucleic acid encoding a fusion polypeptide comprising SEQ ID NO: 367 or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 367, and (ii) that is fused to the N-terminus of an IL-15 / IL15Rα fusion polypeptide, wherein the TCR α-chain and TCR β-chain are selected from SEQ ID NOs: 15 and 16, 17 and 18, 19 and 20, 21 and 22, 23 and 24, 25 and 26, 27 and 28, 29 and 30, 31 and 32, 33 and 34, 35 and 36, 37 and 38, 39 and 40, 41 and 42, 43 and 44, 45 and 46, 47 and 48, 49 and 50, 51 and 52, 53 and 54, 55 and 56, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, 71 and 303, 304 and 74, 75 and 76, 77 and 78, 79 and 80, 81 and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90 and 91 and 92; the CD8 α-chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof; and if present, the CD8 β-chain is SEQ ID NOs: 8, 9, 10, 11, 12, 13 or 14; and the nucleic acid of (b) encodes a fusion polypeptide selected from SEQ ID NOs: 337, 339, 341, 343, 345, 347, 349, 351, 353 or 355, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NOs: 337, 339, 341, 343, 345, 347, 349, 351, 353 or 355.

[0071] In multiple embodiments, a nucleic acid is provided that can include: (a) (i) a nucleic acid encoding a T cell receptor (TCR) comprising an α-chain and a β-chain, and a CD8 polypeptide comprising an α-chain and a β-chain, or (ii) a nucleic acid encoding a TCR comprising an α-chain and a β-chain, and a CD8 polypeptide comprising an α-chain and not comprising a β-chain; and (b) (i) SEQ ID NO: 367, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 367, which are (ii) fused to the N-terminus of an IL-15 / IL15Rα fusion polypeptide, wherein the TCR α-chain and TCR β-chain are selected from SEQ ID NOs: 15 and 16, 17 and 18, 19 and 20, 21 and 22, 23 and 24, 25 and 26, 27 and 28, 29 and 30, 31 and 32, 33 and 34, 35 and 36, 37 and 38, 39 and 40, 41 and 42, 43 and 44, 45 and 46, 47 and 48, 49 and 50, 51 and 52, 53 and 54, 55 and 56, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, 71 and 303, 304 and 74, 75 and 76, 77 and 78, 79 and 80, 81 and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90 and 91 and 92; the CD8 α-chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof; and if present, the CD8 β-chain is SEQ ID NOs: 8, 9, 10, 11, 12, 13 or 14; and the nucleic acid of (b) encodes a fusion polypeptide selected from SEQ ID NOs: 337, 339, 341, 343, 345, 347, 349, 351 or 353, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NOs: 337, 339, 341, 343, 345, 347, 349, 351 or 353.

[0072] In multiple embodiments, a nucleic acid can be provided that comprises (a) a nucleic acid encoding (i) a T cell receptor (TCR) comprising an α-chain and a β-chain, and a CD8 polypeptide comprising an α-chain and a β-chain, or (ii) a TCR comprising an α-chain and a β-chain, and a CD8 polypeptide comprising an α-chain and not comprising a β-chain, and (b) a nucleic acid encoding an IL-15 / IL15Rα fusion polypeptide, wherein the TCR α-chain and TCR β-chain are selected from SEQ ID NOs: 15 and 16, 17 and 18, 19 and 20, 21 and 22, 23 and 24, 25 and 26, 27 and 28, 29 and 30, 31 and 32, 33 and 34, 35 and 36, 37 and 38, 39 and 40, 41 and 42, 43 and 44, 45 and 46, 47 and 48, 49 and 50, 51 and 52, 53 and 54, 55 and 56, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, 71 and 303, 304 and 74, 75 and 76, 77 and 78, 79 and 80, 81 and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90, 91 and 92; the CD8 α-chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof; and if present, the CD8 β-chain is SEQ ID NO: 8, 9, 10, 11, 12, 13 or 14; and the IL-15 / IL-15Rα fusion polypeptide is selected from SEQ ID NO: 317, 321, 325, 327, 329, 331, 333 or 335, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 317, 321, 325, 327, 329, 331, 333 or 335.

[0073] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide can further comprise a signal peptide that is directly or indirectly fused to the N-terminus of SEQ ID NO: 317, 321, 325, 327, 329, 331, 333 or 335, or that is directly or indirectly fused to the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 317, 321, 325, 327, 329, 331, 333 or 335.

[0074] In multiple embodiments, a nucleic acid can be provided that includes (a) (i) a nucleic acid encoding a T cell receptor (TCR) comprising an α-chain and a β-chain, and a CD8 polypeptide comprising an α-chain and a β-chain, or (ii) a nucleic acid encoding a TCR comprising an α-chain and a β-chain, and a CD8 polypeptide comprising an α-chain and not comprising a β-chain, and (b) a nucleic acid encoding an IL-15 / IL15Rα fusion polypeptide, wherein the TCR α-chain and TCR β-chain are selected from SEQ ID NOs: 15 and 16, 17 and 18, 19 and 20, 21 and 22, 23 and 24, 25 and 26, 27 and 28, 29 and 30, 31 and 32, 33 and 34, 35 and 36, 37 and 38, 39 and 40, 41 and 42, 43 and 44, 45 and 46, 47 and 48, 49 and 50, 51 and 52, 53 and 54, 55 and 56, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, 71 and 303, 304 and 74, 75 and 76, 77 and 78, 79 and 80, 81 and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90 and 91 and 92; the CD8 α-chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof; if present, the CD8 β-chain is SEQ ID NO: 8, 9, 10, 11, 12, 13 or 14; and the IL-15 / IL-15Rα fusion polypeptide is selected from SEQ ID NO: 317, 321, 325, 327, 329, 331 or 333, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 317, 321, 325, 327, 329, 331 or 333.

[0075] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide can further comprise a signal peptide that is directly or indirectly fused to the N-terminus of SEQ ID NO: 317, 321, 325, 327, 329, 331 or 333, or that is directly or indirectly fused to the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 317, 321, 325, 327, 329, 331 or 333.

[0076] In some embodiments, the signal peptide can be derived from an IgE polypeptide. In some embodiments, the signal peptide derived from an IgE polypeptide can include a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 367.

[0077] In multiple embodiments, a nucleic acid is provided that can include (a)(i) a nucleic acid encoding a T cell receptor (TCR) comprising an α chain and a β chain, and a CD8 polypeptide comprising an α chain and a β chain, or (ii) a nucleic acid encoding a TCR comprising an α chain and a β chain, and a CD8 polypeptide comprising an α chain and not comprising a β chain, and (b)(i) SEQ ID NO: 367, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 367, wherein they are (ii) fused to the N-terminus of an IL-15 / IL15Rα fusion polypeptide, the TCR α chain and TCR β chain are selected from SEQ ID NOs: 15 and 16, 17 and 18, 19 and 20, 21 and 22, 23 and 24, 25 and 26, 27 and 28, 29 and 30, 31 and 32, 33 and 34, 35 and 36, 37 and 38, 39 and 40, 41 and 42, 43 and 44, 45 and 46, 47 and 48, 49 and 50, 51 and 52, 53 and 54, 55 and 56, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, 71 and 303, 304 and 74, 75 and 76, 77 and 78, 79 and 80, 81 and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90 and 91 and 92, the CD8α chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof, and if present, the CD8β chain is SEQ ID NOs: 8, 9, 10, 11, 12, 13 or 14, and the nucleic acid of (b) encodes a fusion polypeptide selected from SEQ ID NOs: 337, 339, 341, 343, 345, 347, 349, 351, 353 or 355, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NOs: 337, 339, 341, 345, 347, 349, 351, 353 or 355.

[0078] In multiple embodiments, a nucleic acid is provided that can include: (a) (i) a nucleic acid encoding a T cell receptor (TCR) comprising an α-chain and a β-chain, and a CD8 polypeptide comprising an α-chain and a β-chain, or (ii) a nucleic acid encoding a TCR comprising an α-chain and a β-chain, and a CD8 polypeptide comprising an α-chain and not comprising a β-chain; and (b) (i) a nucleic acid encoding a fusion polypeptide comprising SEQ ID NO: 367 or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 367, and (ii) wherein they are fused to the N-terminus of an IL-15 / IL15Rα fusion polypeptide. In this case, the TCR α-chain and TCR β-chain are selected from SEQ ID NOs: 15 and 16, 17 and 18, 19 and 20, 21 and 22, 23 and 24, 25 and 26, 27 and 28, 29 and 30, 31 and 32, 33 and 34, 35 and 36, 37 and 38, 39 and 40, 41 and 42, 43 and 44, 45 and 46, 47 and 48, 49 and 50, 51 and 52, 53 and 54, 55 and 56, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, 71 and 303, 304 and 74, 75 and 76, 77 and 78, 79 and 80, 81 and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90, 91 and 92. The CD8 α-chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof. If present, the CD8 β-chain is SEQ ID NOs: 8, 9, 10, 11, 12, 13 or 14. The nucleic acid of (b) encodes a fusion polypeptide selected from SEQ ID NOs: 337, 341, 345, 347, 349, 351 or 353, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NOs: 337, 341, 345, 347, 349, 351 or 353.

[0079] In multiple embodiments, a nucleic acid can be provided that includes (a)(i) a nucleic acid encoding a T cell receptor (TCR) comprising an α-chain and a β-chain, and a CD8 polypeptide comprising an α-chain and a β-chain, or (ii) a nucleic acid encoding a TCR comprising an α-chain and a β-chain, and a CD8 polypeptide comprising an α-chain and not comprising a β-chain, and (b) a nucleic acid encoding an IL-15 / IL-15Rα fusion polypeptide, wherein the TCR α-chain and TCR β-chain are selected from SEQ ID NOs: 15 and 16, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, and 71 and 303, the CD8 α-chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof, and if present, the CD8 β-chain is SEQ ID NO: 8, 9, 10, 11, 12, 13 or 14.

[0080] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide can include SEQ ID NO: 317, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 317.

[0081] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide can include SEQ ID NO: 319, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 319.

[0082] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide can include SEQ ID NO: 321, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 321.

[0083] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide may comprise a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 323, or SEQ ID NO: 323.

[0084] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide may comprise a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 325, or SEQ ID NO: 325.

[0085] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide may comprise a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 327, or SEQ ID NO: 327.

[0086] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide may comprise a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 329, or SEQ ID NO: 329.

[0087] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide may comprise a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 331, or SEQ ID NO: 331.

[0088] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide may comprise a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 333, or SEQ ID NO: 333.

[0089] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide can include the sequence of SEQ ID NO: 335, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 335.

[0090] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide can further include a signal peptide that is directly or indirectly fused to the N-terminus of the IL-15 / IL-15Rα fusion polypeptide. In multiple embodiments, the signal peptide can be derived from an IgE polypeptide. In multiple embodiments, the signal peptide derived from an IgE polypeptide can include the sequence of SEQ ID NO: 367, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 367.

[0091] In multiple embodiments, there can be provided a nucleic acid comprising: (a) (i) a nucleic acid encoding a T cell receptor (TCR) comprising an α-chain and a β-chain, and a CD8 polypeptide comprising an α-chain and a β-chain, or (ii) a nucleic acid encoding a TCR comprising an α-chain and a β-chain, and a CD8 polypeptide comprising an α-chain and not comprising a β-chain, and (b) a nucleic acid encoding a fusion polypeptide comprising (i) a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 367, and (ii) a sequence fused to the N-terminus of the IL-15 / IL-15Rα fusion polypeptide, wherein the TCR α-chain and TCR β-chain are selected from SEQ ID NOs: 15 and 16, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, and 71 and 303, the CD8 α-chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof, and when present, the CD8 β-chain is SEQ ID NO: 8, 9, 10, 11, 12, 13 or 14.

[0092] In multiple embodiments, the nucleic acid of (b) can encode a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 337, 339, 341, 343, 345, 347, 349, 351, 353, or any of those sequence numbers.

[0093] In multiple embodiments, a nucleic acid can be provided that comprises (a) a nucleic acid that is at least about 80% identical to the nucleic acid of SEQ ID NO: 267, 269, 271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291, 295, 297, 299 or 301, and (b) a nucleic acid that encodes an IL-15 / IL-15Rα fusion polypeptide.

[0094] In multiple embodiments, a nucleic acid can be provided that comprises (a) a nucleic acid that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid of SEQ ID NO: 267, 269, 271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291, 295, 297, 299 or 301, and (b) a nucleic acid that encodes an IL-15 / IL-15Rα fusion polypeptide.

[0095] In multiple embodiments, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide is (i) SEQ ID NO: 308 directly or indirectly fused to the 5' end of SEQ ID NO: 310, or a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 310, with or without a nucleic acid encoding a linker therebetween; (ii) SEQ ID NO: 308 directly or indirectly fused to the 5' end of SEQ ID NO: 312, or a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 312, with or without a nucleic acid encoding a linker therebetween; (iii) SEQ ID NO: 308 directly or indirectly fused to the 5' end of SEQ ID NO: 314, or a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 314, with or without a nucleic acid encoding a linker therebetween; or (iv) SEQ ID NO: 308 directly or indirectly fused to the 5' end of SEQ ID NO: 316, or a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 316, with or without a nucleic acid encoding a linker therebetween, and can be selected from these.

[0096] In some embodiments, the nucleic acid may further comprise a nucleic acid encoding a signal peptide, wherein the nucleic acid encoding the signal peptide is directly or indirectly fused to the 5' end of any of (i), (ii), (iii) or (iv) of SEQ ID NO: 308, or to the 5' end of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to any of (i), (ii), (iii) or (iv) of SEQ ID NO: 308.

[0097] In multiple embodiments, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide is (i) SEQ ID NO: 308, or a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 308, directly or indirectly fused to the 5' end of SEQ ID NO: 312, or a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 312, regardless of the presence or absence of a nucleic acid encoding a linker therebetween; (ii) SEQ ID NO: 308, or a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 308, directly or indirectly fused to the 5' end of SEQ ID NO: 314, or a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 314, regardless of the presence or absence of a nucleic acid encoding a linker therebetween; or (iii) SEQ ID NO: 308, or a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 308, directly or indirectly fused to the 5' end of SEQ ID NO: 316, or a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 316, regardless of the presence or absence of a nucleic acid encoding a linker therebetween, and can be selected from these.

[0098] In some embodiments, the nucleic acid may further comprise a nucleic acid encoding a signal peptide, wherein the nucleic acid encoding the signal peptide is directly or indirectly fused to the 5' end of any one of (i), (ii), or (iii), or to the 5' end of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to any one of the SEQ ID NO: 308.

[0099] In some embodiments, the signal peptide may be derived from an IgE polypeptide. In some embodiments, the nucleic acid encoding the signal peptide may comprise SEQ ID NO: 368, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 368.

[0100] In some embodiments, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide may be selected from the sequences of SEQ ID NO: 318, 320, 322, 324, 326, 328, 330, 332, 334, or 336, or a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the nucleic acids of SEQ ID NO: 318, 320, 322, 324, 326, 328, 330, 332, 334, or 336.

[0101] In multiple embodiments, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide may further comprise a nucleic acid encoding a signal peptide, in which case the nucleic acid encoding the signal peptide is fused directly or indirectly to the 5' end of SEQ ID NO: 318, 320, 322, 324, 326, 328, 330, 332, 334 or 336, or to the 5' end of a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid of SEQ ID NO: 318, 320, 322, 324, 326, 328, 330, 332, 334 or 336.

[0102] In multiple embodiments, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide may be selected from SEQ ID NO: 318, 320, 322, 324, 326, 328, 330, 332 or 334, or a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid of SEQ ID NO: 318, 320, 322, 324, 326, 328, 330, 332 or 334.

[0103] In multiple embodiments, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide may further comprise a nucleic acid encoding a signal peptide, in which case the nucleic acid encoding the signal peptide is fused directly or indirectly to the 5' end of SEQ ID NO: 318, 320, 322, 324, 326, 328, 330, 332 or 334, or to the 5' end of a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid of SEQ ID NO: 318, 320, 322, 324, 326, 328, 330, 332 or 334.

[0104] In multiple embodiments, the signal peptide can be derived from an IgE polypeptide. In multiple embodiments, the nucleic acid encoding the signal peptide can include a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 368.

[0105] In multiple embodiments, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide can further include (i) a nucleic acid encoding a signal peptide derived from an IgE polypeptide, and (ii) can be selected from a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NOs: 338, 340, 342, 344, 346, 348, 350, 352, 354 or 356, or SEQ ID NOs: 338, 340, 342, 344, 346, 348, 350, 352, 354 or 356.

[0106] In multiple embodiments, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide can further include (i) a nucleic acid encoding a signal peptide derived from an IgE polypeptide, and (ii) can be selected from a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NOs: 338, 340, 342, 344, 346, 348, 350, 352 or 354, or SEQ ID NOs: 338, 340, 342, 344, 346, 348, 350, 352 or 354.

[0107] In multiple embodiments, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide can be selected from a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NOs: 318, 322, 326, 328, 330, 332, 334 or 336, or SEQ ID NOs: 318, 322, 326, 328, 330, 332, 334 or 336.

[0108] In multiple embodiments, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide may further comprise a nucleic acid encoding a signal peptide, in which case the nucleic acid encoding the signal peptide is directly or indirectly fused to the 5' end of SEQ ID NO: 318, 322, 326, 328, 330, 332, 334 or 336, or to the 5' end of a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid of SEQ ID NO: 318, 322, 326, 328, 330, 332, 334 or 336.

[0109] In multiple embodiments, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide may be selected from SEQ ID NO: 318, 322, 326, 328, 330, 332 or 334, or a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid of SEQ ID NO: 318, 322, 326, 328, 330, 332 or 334.

[0110] In multiple embodiments, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide may further comprise a nucleic acid encoding a signal peptide, in which case the nucleic acid encoding the signal peptide is directly or indirectly fused to the 5' end of SEQ ID NO: 318, 322, 326, 328, 330, 332 or 334, or to the 5' end of a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid of SEQ ID NO: 318, 322, 326, 328, 330, 332 or 334.

[0111] In some embodiments, the signal peptide may be derived from an IgE polypeptide. In some embodiments, the nucleic acid encoding the signal peptide may include a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 368.

[0112] In some embodiments, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide may further include (i) a nucleic acid encoding a signal peptide derived from an IgE polypeptide, and (ii) a sequence selected from a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid of SEQ ID NO: 338, 342, 346, 348, 350, 352, 354 or 356.

[0113] In some embodiments, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide may further include (i) a nucleic acid encoding a signal peptide derived from an IgE polypeptide, and (ii) a sequence selected from a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid of SEQ ID NO: 338, 342, 346, 348, 350, 352 or 354.

[0114] In some embodiments, a vector may be provided that includes nucleic acids encoding at least one CD8α, at least one TCRα chain, at least one TCRβ chain, at least one IL-15 / IL-15Rα fusion polypeptide, and optionally at least one CD8β chain.

[0115] In multiple embodiments, a vector comprising N1, N2, N3, N4, N5, L1, L2, L3, and L4 in any order may be provided, where N1 comprises a nucleic acid encoding a CD8β chain, is present or absent, N2 comprises a nucleic acid encoding a CD8α chain, N3 comprises a nucleic acid encoding a TCRβ chain, N4 comprises a nucleic acid encoding a TCRα chain, N5 comprises a nucleic acid encoding an IL-15 / IL-15Rα fusion polypeptide, and L1 to L4 each comprise a nucleic acid encoding at least one linker, L1 to L4 are each independently the same or different, and L1 to L4 are each independently present or absent.

[0116] In multiple embodiments, the vector may comprise Formula III or Formula IV. 5’-N1-L1-N2-L2-N3-L3-N4-L4-N5-3’ [III] 5’-N5-L1-N1-L2-N2-L3-N3-L4-N4 -3’ [IV].

[0117] In multiple embodiments, N1 may comprise a nucleic acid encoding SEQ ID NO: 8, 9, 10, 11, 12, 13, or 14.

[0118] In multiple embodiments, N2 comprises a nucleic acid encoding SEQ ID NO: 7, 258, 259, 262, or a variant thereof.

[0119] In multiple embodiments, N4 and N3 may comprise nucleic acids encoding SEQ ID NO: 15 and 16, 17 and 18, 19 and 20, 21 and 22, 23 and 24, 25 and 26, 27 and 28, 29 and 30, 31 and 32, 33 and 34, 35 and 36, 37 and 38, 39 and 40, 41 and 42, 43 and 44, 45 and 46, 47 and 48, 49 and 50, 51 and 52, 53 and 54, 55 and 56, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, 71 and 303, 304 and 74, 75 and 76, 77 and 78, 79 and 80, 81 and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90, or 91 and 92.

[0120] In multiple embodiments, N5 can include a nucleic acid encoding (i) the N-terminus of SEQ ID NO: 309, or an N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 309, directly or indirectly fused to SEQ ID NO: 307, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 307, with or without a linker therebetween; (ii) the N-terminus of SEQ ID NO: 311, or an N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 311, directly or indirectly fused to SEQ ID NO: 307, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 307, with or without a linker therebetween; (iii) the N-terminus of SEQ ID NO: 313, or an N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 313, directly or indirectly fused to SEQ ID NO: 307, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 307, with or without a linker therebetween; or (iv) the N-terminus of SEQ ID NO: 315, or an N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 315, directly or indirectly fused to SEQ ID NO: 307, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 307, with or without a linker therebetween.

[0121] In multiple embodiments, N5 may further comprise a nucleic acid encoding a signal peptide directly or indirectly fused to the 5' end of a nucleic acid encoding any of the sequence numbers 307 of (i), (ii), (iii), or (iv), or to the 5' end of a nucleic acid encoding a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to any of the sequence numbers 307 of (i), (ii), (iii), or (iv).

[0122] In multiple embodiments, N5 encodes (i) the N-terminus of SEQ ID NO: 311, or the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 311, directly or indirectly fused to SEQ ID NO: 307, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 307, with or without a linker therebetween; (ii) the N-terminus of SEQ ID NO: 313, or the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 313, directly or indirectly fused to SEQ ID NO: 307, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 307, with or without a linker therebetween; or (iii) the N-terminus of SEQ ID NO: 315, or the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 315, directly or indirectly fused to SEQ ID NO: 307, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 307, with or without a linker therebetween.

[0123] In multiple embodiments, N5 may further comprise a nucleic acid encoding a signal peptide that is directly or indirectly fused to the 5' end of a nucleic acid encoding any one of the sequence numbers 307 of (i), (ii), or (iii), or to the 5' end of a nucleic acid encoding a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to any one of the sequence numbers 307 of (i), (ii), or (iii).

[0124] In multiple embodiments, the signal peptide may be derived from an IgE polypeptide. In multiple embodiments, the signal peptide derived from an IgE polypeptide may comprise the sequence number 367, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the sequence number 367.

[0125] In multiple embodiments, N5 may comprise a nucleic acid encoding any one of the sequence numbers 317, 319, 321, 323, 325, 327, 329, 331, 333, or 335, or a nucleic acid encoding a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to any one of the sequence numbers 317, 319, 321, 323, 325, 327, 329, 331, 333, or 335.

[0126] In multiple embodiments, N5 may further comprise a nucleic acid encoding a signal peptide that is directly or indirectly fused to the 5' end of a nucleic acid encoding any one of the sequence numbers 317, 319, 321, 323, 325, 327, 329, 331, 333, or 335, or to the 5' end of a nucleic acid encoding a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to any one of the sequence numbers 317, 319, 321, 323, 325, 327, 329, 331, 333, or 335.

[0127] In multiple embodiments, N5 can include a nucleic acid encoding SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, or 333, or a nucleic acid encoding a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, or 333.

[0128] In multiple embodiments, N5 can further include a nucleic acid encoding a signal peptide that is directly or indirectly fused to the 5'-end of a nucleic acid encoding SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, or 333, or to the 5'-end of a nucleic acid encoding a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, or 333.

[0129] In multiple embodiments, the signal peptide can be derived from an IgE polypeptide. In multiple embodiments, a signal peptide derived from an IgE polypeptide can include SEQ ID NO: 367, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 367.

[0130] In multiple embodiments, (i) N5 can further include a nucleic acid encoding a signal peptide derived from an IgE polypeptide, and (ii) N5 can encode a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 337, 339, 341, 343, 345, 347, 349, 351, 353, or 355.

[0131] In multiple embodiments, (i) N5 may further comprise a nucleic acid encoding a signal peptide derived from an IgE polypeptide, and (ii) N5 may encode a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 337, 339, 341, 343, 345, 347, 349, 351 or 353.

[0132] In multiple embodiments, N5 may comprise a nucleic acid encoding SEQ ID NO: 317, 321, 325, 327, 329, 331, 333 or 335, or a nucleic acid encoding a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 317, 321, 325, 327, 329, 331, 333 or 335.

[0133] In multiple embodiments, N5 may further comprise a nucleic acid encoding a signal peptide that is directly or indirectly fused to the 5' end of a nucleic acid encoding SEQ ID NO: 317, 321, 325, 327, 329, 331, 333 or 335, or to the 5' end of a nucleic acid encoding a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 317, 321, 325, 327, 329, 331, 333 or 335.

[0134] In multiple embodiments, N5 may comprise a nucleic acid encoding SEQ ID NO: 317, 321, 325, 327, 329, 331 or 333, or a nucleic acid encoding a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 317, 321, 325, 327, 329, 331 or 333.

[0135] In multiple embodiments, N5 may further include a nucleic acid encoding a signal peptide that is directly or indirectly fused to the 5' end of a nucleic acid encoding SEQ ID NO: 317, 321, 325, 327, 329, 331, or 333, or to the 5' end of a nucleic acid encoding a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 317, 321, 325, 327, 329, 331, or 333.

[0136] In multiple embodiments, the signal peptide may be derived from an IgE polypeptide. In multiple embodiments, the signal peptide derived from an IgE polypeptide may include a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 367.

[0137] In multiple embodiments, (i) N5 may further include a nucleic acid encoding a signal peptide derived from an IgE polypeptide, and (ii) N5 may encode a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 337, 341, 345, 347, 349, 351, or 353.

[0138] In multiple embodiments, (i) the vector may further encode a 2A peptide or an internal ribosome entry site (IRES) disposed between N1 and L1, between L1 and N2, between N2 and L2, between L2 and N3, between N3 and L3, between L3 and N4, between N4 and L4, between L4 and N5, or any combination thereof, or (ii) the vector may further encode a 2A peptide or an internal ribosome entry site (IRES) disposed between N5 and L1, between L1 and N1, between N1 and L2, between L2 and N2, between N2 and L3, between L3 and N3, between N3 and L4, between L4 and N4, or any combination thereof.

[0139] In multiple embodiments, (i) the vector may further encode Furin disposed between N1 and L1, between L1 and N2, between N2 and L2, between L2 and N3, between N3 and L3, between L3 and N4, between N4 and L4, between L4 and N5, or any combination thereof, or (ii) the vector may further encode Furin disposed between N5 and L1, between L1 and N1, between N1 and L2, between L2 and N2, between N2 and L3, between L3 and N3, between N3 and L4, between L4 and N4, or any combination thereof.

[0140] In multiple embodiments, the 2A peptide can be P2A (SEQ ID NO: 93), T2A (SEQ ID NO: 94), E2A (SEQ ID NO: 95), or F2A (SEQ ID NO: 96).

[0141] In multiple embodiments, the IRES can be selected from the group consisting of an IRES derived from picornavirus, an IRES derived from flavivirus, an IRES derived from pestivirus, an IRES derived from retrovirus, an IRES derived from lentivirus, an IRES derived from insect RNA virus, and an IRES derived from cellular mRNA.

[0142] In multiple embodiments, T cells and / or natural killer cells can be provided that comprise (a) (i) a T cell receptor (TCR) comprising an α-chain and a β-chain and a CD8 polypeptide comprising an α-chain and a β-chain, or (ii) a TCR comprising an α-chain and a β-chain and a CD8 polypeptide comprising an α-chain and not comprising a β-chain, and (b) an IL-15 / IL-15Rα fusion polypeptide, wherein the TCR α-chain and TCR β-chain are selected from SEQ ID NOs: 15 and 16, 17 and 18, 19 and 20, 21 and 22, 23 and 24, 25 and 26, 27 and 28, 29 and 30, 31 and 32, 33 and 34, 35 and 36, 37 and 38, 39 and 40, 41 and 42, 43 and 44, 45 and 46, 47 and 48, 49 and 50, 51 and 52, 53 and 54, 55 and 56, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, 71 and 303, 304 and 74, 75 and 76, 77 and 78, 79 and 80, 81 and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90, and 91 and 92; the CD8 α-chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof; and if present, the CD8 β-chain is SEQ ID NO: 8, 9, 10, 11, 12, 13 or 14; and the IL-15 / IL-15Rα fusion polypeptide is (i) a sequence that is directly or indirectly fused to the N-terminus of SEQ ID NO: 309 or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 309, and is a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 307, with or without a linker therebetween; or (ii) a sequence that is directly or indirectly fused to the N-terminus of SEQ ID NO: 311 or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 311, and is a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 307, with or without a linker therebetween.(iii) the N-terminus of SEQ ID NO: 313 or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 313, directly or indirectly fused to the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 307, with or without a linker therebetween, or (iv) the N-terminus of SEQ ID NO: 315 or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 315, directly or indirectly fused to the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 307, with or without a linker therebetween, selected from.

[0143] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide may further comprise a signal peptide directly or indirectly fused to the N-terminus of SEQ ID NO: 307 of any one of (i), (ii), (iii) or (iv), or to the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 307 of any one of (i), (ii), (iii) or (iv).

[0144] In multiple embodiments, T cells and / or natural killer cells can be provided that include (a) (i) a T cell receptor (TCR) comprising an α chain and a β chain and a CD8 polypeptide comprising an α chain and a β chain, or (ii) a TCR comprising an α chain and a β chain and a CD8 polypeptide comprising an α chain and not comprising a β chain, and (b) an IL-15 / IL-15Rα fusion polypeptide, wherein the TCR α chain and TCR β chain are selected from SEQ ID NOs: 15 and 16, 17 and 18, 19 and 20, 21 and 22, 23 and 24, 25 and 26, 27 and 28, 29 and 30, 31 and 32, 33 and 34, 35 and 36, 37 and 38, 39 and 40, 41 and 42, 43 and 44, 45 and 46, 47 and 48, 49 and 50, 51 and 52, 53 and 54, 55 and 56, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, 71 and 303, 304 and 74, 75 and 76, 77 and 78, 79 and 80, 81 and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90, and 91 and 92; the CD8 α chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof; and if present, the CD8 β chain is SEQ ID NOs: 8, 9, 10, 11, 12, 13 or 14; and the IL-15 / IL-15Rα fusion polypeptide is (i) a sequence that is directly or indirectly fused to the N-terminus of SEQ ID NO: 311 or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 311, and is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 307, with or without a linker therebetween; or (ii) a sequence that is directly or indirectly fused to the N-terminus of SEQ ID NO: 313 or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 313, and is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 307, with or without a linker therebetween.or (iii) the N-terminus of SEQ ID NO: 315 or an N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 315, directly or indirectly fused to a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 307, with or without a linker therebetween, and is selected from the group consisting of:

[0145] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide may further comprise a signal peptide directly or indirectly fused to the N-terminus of SEQ ID NO: 307 of any one of (i), (ii) or (iii), or to the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 307 of any one of (i), (ii) or (iii).

[0146] In multiple embodiments, the signal peptide may be derived from an IgE polypeptide. In multiple embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 367.

[0147] In some embodiments, T cells and / or natural killer cells can be provided that include (a) (i) a T cell receptor (TCR) comprising an α chain and a β chain, and a CD8 polypeptide comprising an α chain and a β chain, or (ii) a TCR comprising an α chain and a β chain, and a CD8 polypeptide comprising an α chain and not comprising a β chain, and (b) an IL-15 / IL15Rα fusion polypeptide, wherein the TCR α chain and the TCR β chain are selected from SEQ ID NOs: 15 and 16, 17 and 18, 19 and 20, 21 and 22, 23 and 24, 25 and 26, 27 and 28, 29 and 30, 31 and 32, 33 and 34, 35 and 36, 37 and 38, 39 and 40, 41 and 42, 43 and 44, 45 and 46, 47 and 48, 49 and 50, 51 and 52, 53 and 54, 55 and 56, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, 71 and 303, 304 and 74, 75 and 76, 77 and 78, 79 and 80, 81 and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90 and 91 and 92; the CD8 α chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof; and if present, the CD8 β chain is SEQ ID NO: 8, 9, 10, 11, 12, 13 or 14; and the IL-15 / IL-15Rα fusion polypeptide is selected from SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, 333 or 335, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, 333 or 335.

[0148] In some embodiments, the IL-15 / IL-15Rα fusion polypeptide can further include a signal peptide that is directly or indirectly fused to the N-terminus of SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, 333 or 335, or that is directly or indirectly fused to the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, 333 or 335.

[0149] In multiple embodiments, T cells and / or natural killer cells can be provided that comprise (a) (i) a T cell receptor (TCR) comprising an α chain and a β chain, and a CD8 polypeptide comprising an α chain and a β chain, or (ii) a TCR comprising an α chain and a β chain, and a CD8 polypeptide comprising an α chain and not comprising a β chain, and (b) an IL-15 / IL15Rα fusion polypeptide, wherein the TCR α chain and TCR β chain are selected from SEQ ID NOs: 15 and 16, 17 and 18, 19 and 20, 21 and 22, 23 and 24, 25 and 26, 27 and 28, 29 and 30, 31 and 32, 33 and 34, 35 and 36, 37 and 38, 39 and 40, 41 and 42, 43 and 44, 45 and 46, 47 and 48, 49 and 50, 51 and 52, 53 and 54, 55 and 56, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, 71 and 303, 304 and 74, 75 and 76, 77 and 78, 79 and 80, 81 and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90 and 91 and 92; the CD8 α chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof; and if present, the CD8 β chain is SEQ ID NO: 8, 9, 10, 11, 12, 13 or 14; and the IL-15 / IL-15Rα fusion polypeptide is selected from SEQ ID NOs: 317, 319, 321, 323, 325, 327, 329, 331 or 333, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NOs: 317, 319, 321, 323, 325, 327, 329, 331 or 333.

[0150] In some embodiments, the IL-15 / IL-15Rα fusion polypeptide may further comprise a signal peptide that is directly or indirectly fused to the N-terminus of SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331 or 333, or that is directly or indirectly fused to the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331 or 333.

[0151] In some embodiments, the signal peptide may be derived from an IgE polypeptide. In some embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 367.

[0152] In multiple embodiments, a T cell and / or natural killer cell is provided that comprises a fusion polypeptide comprising (a) (i) a T cell receptor (TCR) comprising an alpha chain and a beta chain, and a CD8 polypeptide comprising an alpha chain and a beta chain, or (ii) a TCR comprising an alpha chain and a beta chain, and a CD8 polypeptide comprising an alpha chain and not comprising a beta chain, and (b) (i) SEQ ID NO: 367, or a signal peptide comprising a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 367, which is (ii) fused to an IL-15 / IL15Rα fusion polypeptide. In this case, the TCR alpha chain and the TCR beta chain are selected from SEQ ID NOs: 15 and 16, 17 and 18, 19 and 20, 21 and 22, 23 and 24, 25 and 26, 27 and 28, 29 and 30, 31 and 32, 33 and 34, 35 and 36, 37 and 38, 39 and 40, 41 and 42, 43 and 44, 45 and 46, 47 and 48, 49 and 50, 51 and 52, 53 and 54, 55 and 56, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, 71 and 303, 304 and 74, 75 and 76, 77 and 78, 79 and 80, 81 and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90 and 91 and 92; the CD8 alpha chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof; if present, the CD8 beta chain is SEQ ID NOs: 8, 9, 10, 11, 12, 13 or 14; and the fusion polypeptide of (b) is selected from SEQ ID NOs: 337, 339, 341, 343, 345, 347, 349, 351, 353 or 355, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NOs: 337, 339, 341, 343, 345, 347, 349, 351, 353 or 355.

[0153] In multiple embodiments, provided can be T cells and / or natural killer cells comprising a fusion polypeptide comprising: (a) (i) a T cell receptor (TCR) comprising an α chain and a β chain, and a CD8 polypeptide comprising an α chain and a β chain, or (ii) a TCR comprising an α chain and a β chain, and a CD8 polypeptide comprising an α chain and not comprising a β chain, and (b) (i) SEQ ID NO: 367, or a signal peptide comprising a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 367, which are (ii) fused to an IL-15 / IL15Rα fusion polypeptide, wherein the TCR α chain and TCR β chain are selected from SEQ ID NOs: 15 and 16, 17 and 18, 19 and 20, 21 and 22, 23 and 24, 25 and 26, 27 and 28, 29 and 30, 31 and 32, 33 and 34, 35 and 36, 37 and 38, 39 and 40, 41 and 42, 43 and 44, 45 and 46, 47 and 48, 49 and 50, 51 and 52, 53 and 54, 55 and 56, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, 71 and 303, 304 and 74, 75 and 76, 77 and 78, 79 and 80, 81 and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90 and 91 and 92, the CD8 α chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof, and if present, the CD8 β chain is SEQ ID NOs: 8, 9, 10, 11, 12, 13 or 14, and the fusion polypeptide of (b) is selected from SEQ ID NOs: 337, 339, 341, 343, 345, 347, 349, 351 or 353, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NOs: 337, 339, 341, 343, 345, 347, 349, 351 or 353.

[0154] In multiple embodiments, T cells and / or natural killer cells can be provided that are transduced to express (a) (i) a T cell receptor (TCR) comprising an α-chain and a β-chain, and a CD8 polypeptide comprising an α-chain and a β-chain, or (ii) a TCR comprising an α-chain and a β-chain, and a CD8 polypeptide comprising an α-chain and not comprising a β-chain, and (b) an IL-15 / IL15Rα fusion polypeptide, wherein the TCR α-chain and TCR β-chain are selected from SEQ ID NOs: 15 and 16, 17 and 18, 19 and 20, 21 and 22, 23 and 24, 25 and 26, 27 and 28, 29 and 30, 31 and 32, 33 and 34, 35 and 36, 37 and 38, 39 and 40, 41 and 42, 43 and 44, 45 and 46, 47 and 48, 49 and 50, 51 and 52, 53 and 54, 55 and 56, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, 71 and 303, 304 and 74, 75 and 76, 77 and 78, 79 and 80, 81 and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90, 91 and 92, the CD8 α-chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof, and if present, the CD8 β-chain is SEQ ID NO: 8, 9, 10, 11, 12, 13 or 14, and the IL-15 / IL-15Rα fusion polypeptide is selected from SEQ ID NO: 317, 321, 325, 327, 329, 331, 333 or 335, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 317, 321, 325, 327, 329, 331, 333 or 335.

[0155] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide can further comprise a signal peptide that is directly or indirectly fused to the N-terminus of SEQ ID NO: 317, 321, 325, 327, 329, 331, 333 or 335, or that is directly or indirectly fused to the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 317, 321, 325, 327, 329, 331, 333 or 335.

[0156] In multiple embodiments, T cells and / or natural killer cells can be provided that are transduced to express (a) (i) a T cell receptor (TCR) comprising an α chain and a β chain, and a CD8 polypeptide comprising an α chain and a β chain, or (ii) a TCR comprising an α chain and a β chain, and a CD8 polypeptide comprising an α chain and not comprising a β chain, and (b) an IL-15 / IL15Rα fusion polypeptide, wherein the TCR α chain and the TCR β chain are selected from SEQ ID NOs: 15 and 16, 17 and 18, 19 and 20, 21 and 22, 23 and 24, 25 and 26, 27 and 28, 29 and 30, 31 and 32, 33 and 34, 35 and 36, 37 and 38, 39 and 40, 41 and 42, 43 and 44, 45 and 46, 47 and 48, 49 and 50, 51 and 52, 53 and 54, 55 and 56, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, 71 and 303, 304 and 74, 75 and 76, 77 and 78, 79 and 80, 81 and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90 and 91 and 92, the CD8 α chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof, and if present, the CD8 β chain is SEQ ID NO: 8, 9, 10, 11, 12, 13 or 14, and the IL-15 / IL-15Rα fusion polypeptide is selected from SEQ ID NO: 317, 321, 325, 327, 329, 331 or 333, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 317, 321, 325, 327, 329, 331 or 333.

[0157] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide can further comprise a signal peptide that is directly or indirectly fused to the N-terminus of SEQ ID NO: 317, 321, 325, 327, 329, 331 or 333, or that is directly or indirectly fused to the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 317, 321, 325, 327, 329, 331 or 333.

[0158] In multiple embodiments, the signal peptide can be derived from an IgE polypeptide. In multiple embodiments, the signal peptide derived from an IgE polypeptide can include a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 367.

[0159] In multiple embodiments, there are provided T cells and / or natural killer cells comprising a fusion polypeptide comprising: (a) (i) a T cell receptor (TCR) comprising an α-chain and a β-chain, and a CD8 polypeptide comprising an α-chain and a β-chain, or (ii) a TCR comprising an α-chain and a β-chain, and a CD8 polypeptide comprising an α-chain and not comprising a β-chain, and (b) (i) SEQ ID NO: 367, or a signal peptide comprising a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 367, wherein they are (ii) directly or indirectly fused to an IL-15 / IL15Rα fusion polypeptide. In this case, the TCR α-chain and TCR β-chain are selected from SEQ ID NOs: 15 and 16, 17 and 18, 19 and 20, 21 and 22, 23 and 24, 25 and 26, 27 and 28, 29 and 30, 31 and 32, 33 and 34, 35 and 36, 37 and 38, 39 and 40, 41 and 42, 43 and 44, 45 and 46, 47 and 48, 49 and 50, 51 and 52, 53 and 54, 55 and 56, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, 71 and 303, 304 and 74, 75 and 76, 77 and 78, 79 and 80, 81 and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90, 91 and 92; the CD8 α-chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof; if present, the CD8 β-chain is SEQ ID NOs: 8, 9, 10, 11, 12, 13 or 14; and the fusion polypeptide of (b) is selected from SEQ ID NOs: 337, 341, 345, 347, 349, 351 or 353, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NOs: 337, 341, 345, 347, 349, 351 or 353.

[0160] In multiple embodiments, a T cell and / or natural killer cell comprising (a) (i) a T cell receptor (TCR) comprising an α-chain and a β-chain, and a CD8 polypeptide comprising an α-chain and a β-chain, or (ii) a TCR comprising an α-chain and a β-chain, and a CD8 polypeptide comprising an α-chain and not comprising a β-chain, and (b) an IL-15 / IL-15Rα fusion polypeptide, wherein the TCR α-chain and TCR β-chain are selected from SEQ ID NOs: 15 and 16, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, and 71 and 303, the CD8 α-chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof, and if present, the CD8 β-chain is SEQ ID NO: 8, 9, 10, 11, 12, 13 or 14.

[0161] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide can comprise SEQ ID NO: 317, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 317.

[0162] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide can comprise SEQ ID NO: 319, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 319.

[0163] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide can comprise SEQ ID NO: 321, or a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 321.

[0164] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide can comprise a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 323, or SEQ ID NO: 323.

[0165] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide can comprise a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 325, or SEQ ID NO: 325.

[0166] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide can comprise a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 327, or SEQ ID NO: 327.

[0167] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide can comprise a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 329, or SEQ ID NO: 329.

[0168] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide can comprise a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 331, or SEQ ID NO: 331.

[0169] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide can comprise a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 333, or SEQ ID NO: 333.

[0170] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide can include a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 335.

[0171] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide can further include a signal peptide that is directly or indirectly fused to the N-terminus of the IL-15 / IL-15Rα fusion polypeptide. In multiple embodiments, the signal peptide can be derived from an IgE polypeptide. In multiple embodiments, the signal peptide derived from an IgE polypeptide can include a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 367.

[0172] In multiple embodiments, a T cell and / or natural killer cell comprising a fusion polypeptide comprising (a)(i) a T cell receptor (TCR) comprising an α chain and a β chain, and a CD8 polypeptide comprising an α chain and a β chain, or (ii) a TCR comprising an α chain and a β chain, and a CD8 polypeptide comprising an α chain and not comprising a β chain, and (b)(i) a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 367, and (ii) is fused to the N-terminus of the IL-15 / IL-15Rα fusion polypeptide, wherein in this case the TCRα chain and the TCRβ chain are selected from SEQ ID NOs: 15 and 16, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, and 71 and 303, the CD8α chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof, and if present, the CD8β chain is SEQ ID NO: 8, 9, 10, 11, 12, 13 or 14.

[0173] In multiple embodiments, the fusion polypeptide of (b) may comprise a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 337, or SEQ ID NO: 337.

[0174] In multiple embodiments, the fusion polypeptide of (b) may comprise a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 339, or SEQ ID NO: 339.

[0175] In multiple embodiments, the fusion polypeptide of (b) may comprise a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 341, or SEQ ID NO: 341.

[0176] In multiple embodiments, the fusion polypeptide of (b) may comprise a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 343, or SEQ ID NO: 343.

[0177] In multiple embodiments, the fusion polypeptide of (b) may comprise a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 345, or SEQ ID NO: 345.

[0178] In multiple embodiments, the fusion polypeptide of (b) may comprise a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 347, or SEQ ID NO: 347.

[0179] In multiple embodiments, the fusion polypeptide of (b) can comprise a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 349, or SEQ ID NO: 349.

[0180] In multiple embodiments, the fusion polypeptide of (b) can comprise a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 351, or SEQ ID NO: 351.

[0181] In multiple embodiments, the fusion polypeptide of (b) can comprise a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 353, or SEQ ID NO: 353.

[0182] In multiple embodiments, the fusion polypeptide of (b) can comprise a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 355, or SEQ ID NO: 355.

[0183] In multiple embodiments, the T cell can be an αβ T cell, a γδ T cell, a natural killer T cell, or any combination thereof. In multiple embodiments, the αβ T cell can be a CD4+ T cell. In multiple embodiments, the αβ T cell can be a CD8+ T cell. In multiple embodiments, the γδ T cell can be a Vγ9Vδ2+ T cell.

[0184] In multiple embodiments, a composition comprising the T cells and / or natural killer cells described herein can be provided. In multiple embodiments, the composition can be a pharmaceutical composition. In multiple embodiments, the composition can further comprise an adjuvant, an excipient, a carrier, a diluent, a buffer, a stabilizer, or a combination thereof.

[0185] In multiple embodiments, the adjuvant can be an anti-CD40 antibody, imiquimod, resiquimod, GM-CSF, cyclophosphamide, sunitinib, bevacizumab, atezolizumab, interferon alpha, interferon beta, CpG oligonucleotides and their derivatives, poly(I:C) and its derivatives, RNA, sildenafil, particulate formulations containing poly(lactide co-glycolide) (PLG), virosomes, interleukin 1 (IL-1), interleukin 2 (IL-2), interleukin 4 (IL-4), interleukin 7 (IL-7), interleukin 12 (IL-12), interleukin 13 (IL-13), interleukin 15 (IL-15), interleukin 21 (IL-21), interleukin 23 (IL-23), or any combination thereof. In multiple embodiments, the adjuvant can be IL-2, IL-7, IL-12, IL-15, IL-21, or any combination thereof.

[0186] In multiple embodiments, a method for preparing T cells and / or natural killer cells for immunotherapy can be provided, the method comprising isolating T cells and / or natural killer cells from a blood sample of a human subject, activating the isolated T cells and / or natural killer cells, transducing the activated T cells and / or natural killer cells with a nucleic acid described herein or a vector described herein, and amplifying the transduced T cells and / or natural killer cells. In multiple embodiments, the method can further comprise isolating CD4+CD8+ T cells from the transduced T cells and / or natural killer cells, and amplifying the isolated CD4+CD8+ transduced T cells. In multiple embodiments, the blood sample can comprise peripheral blood mononuclear cells (PMBC). In multiple embodiments, activation can comprise contacting the T cells and / or natural killer cells with anti-CD3 antibody and anti-CD28 antibody. In multiple embodiments, the T cells can be CD4+ T cells. In multiple embodiments, the T cells can be CD8+ T cells. In multiple embodiments, the T cells can be γδ T cells or αβ T cells. In multiple embodiments, activation, amplification, or both can be performed in the presence of a combination of IL-2 and IL-15, optionally using zoledronate.

[0187] In some embodiments, methods are provided for increasing the persistence, functionality, naivety, lifespan, ability to kill antigen-presenting cells, or combinations thereof of T cells and / or natural killer cells, the methods including isolating T cells and / or natural killer cells from a blood sample of a human subject, activating the isolated T cells and / or natural killer cells, transducing the activated T cells and / or natural killer cells with a nucleic acid, a vector, or combinations thereof described herein to obtain transduced T cells and / or natural killer cells, and obtaining the transduced T cells and / or natural killer cells, wherein the persistence, lifespan, naivety, ability to kill antigen-presenting cells, or combinations thereof of the transduced T cells and / or natural killer cells are increased compared to those of control cells. In some embodiments, the methods may further include expanding the transduced T cells and / or natural killer cells.

[0188] In multiple embodiments, the control cells can include untransduced T cells and / or natural killer cells, T cells and / or natural killer cells transduced with only the TCR, or combinations thereof. In multiple embodiments, the control cells can include untransduced T cells and / or natural killer cells, T cells and / or natural killer cells transduced with only the TCR, T cells and / or natural killer cells transduced with only the TCR and CD8, or combinations thereof. In multiple embodiments, the persistence, lifespan, functionality, naivety, ability to kill antigen-presenting cells, or combinations thereof of the transduced T cells and / or natural killer cells and the control cells can be determined after one challenge with antigen-presenting cells, two challenges with antigen-presenting cells, three challenges with antigen-presenting cells, four challenges with antigen-presenting cells, five challenges with antigen-presenting cells, six challenges with antigen-presenting cells, seven challenges with antigen-presenting cells, or more than seven challenges with antigen-presenting cells. The persistence, lifespan, functionality, naivety, ability to kill antigen-presenting cells, or combinations thereof of the transduced T cells and / or natural killer cells and the control cells can be determined after five or more challenges with antigen-presenting cells, or more than seven challenges with antigen-presenting cells.

[0189] In multiple embodiments, a method of increasing interferon gamma (IFNγ) secretion by T cells and / or natural killer cells can be provided, the method comprising isolating T cells and / or natural killer cells from a blood sample of a human subject, activating the isolated T cells and / or natural killer cells, transducing the activated T cells and / or natural killer cells with a nucleic acid, a vector described herein, or a combination thereof as described herein to obtain transduced T cells and / or natural killer cells, and obtaining the transduced T cells and / or natural killer cells, wherein the IFNγ secretion of the transduced T cells and / or natural killer cells is increased compared to that of control cells. In multiple embodiments, the method can further comprise amplifying the transduced T cells and / or natural killer cells. In multiple embodiments, the control cells can include untransduced T cells and / or natural killer cells, T cells and / or natural killer cells transduced with only a TCR, or a combination thereof.

[0190] In multiple embodiments, the control cells can include untransduced T cells and / or natural killer cells, T cells and / or natural killer cells transduced with only the TCR, T cells and / or natural killer cells transduced with only the TCR and CD8, or combinations thereof. In multiple embodiments, IFNγ secretion by the transduced T cells and / or natural killer cells and the control cells can be determined after one challenge with antigen-presenting cells, two challenges with antigen-presenting cells, three challenges with antigen-presenting cells, four challenges with antigen-presenting cells, five challenges with antigen-presenting cells, six challenges with antigen-presenting cells, seven challenges with antigen-presenting cells, or more than seven challenges with antigen-presenting cells. In multiple embodiments, IFNγ secretion by the transduced T cells and / or natural killer (NK) cells and the control cells can be determined after five or more challenges with antigen-presenting cells, or more than seven challenges with antigen-presenting cells.

[0191] In multiple embodiments, the antigen-presenting cells can present an antigen on their cell surface, and the transduced T cells and / or natural killer cells, and the control cells can have the ability to kill the antigen-presenting cells. In multiple embodiments, the antigen can include a peptide. In multiple embodiments, the peptide can be in a complex with an MHC molecule on the cell surface.

[0192] In multiple embodiments, a polypeptide, multiple polypeptides, or a fusion polypeptide encoded by the nucleic acids described herein can be provided.

[0193] In multiple embodiments, the nucleic acids described herein can be isolated, recombinant, or both isolated and recombinant.

[0194] In multiple embodiments, the vectors described herein can be isolated, recombinant, or both isolated and recombinant.

[0195] In multiple embodiments, the T cells and / or natural killer cells described herein can be isolated, recombinant, engineered, or any combination thereof.

[0196] In multiple embodiments, the polypeptides, multiple polypeptides, or fusion polypeptides described herein can be isolated, recombinant, or both isolated and recombinant.

[0197] In multiple embodiments, vectors comprising the nucleic acids described herein may be provided. In multiple embodiments, the vectors described herein may further comprise a nucleic acid encoding a 2A peptide or an internal ribosome entry site (IRES) disposed between a nucleic acid encoding a CD8α chain and a nucleic acid encoding a CD8β chain. In multiple embodiments, the vector may further comprise a nucleic acid encoding a 2A peptide or an IRES disposed between a nucleic acid encoding a TCRα chain and a nucleic acid encoding a TCRβ chain. In multiple embodiments, the vector may further comprise a nucleic acid encoding a 2A peptide or an IRES disposed between a nucleic acid encoding a TCR chain or a CD8 chain and a nucleic acid encoding a membrane-bound IL-15, such as an IL-15 / IL-15Rα fusion polypeptide. In multiple embodiments, the 2A peptide may be P2A (SEQ ID NO: 93), T2A (SEQ ID NO: 94), E2A (SEQ ID NO: 95), or F2A (SEQ ID NO: 96). In multiple embodiments, the IRES may be selected from the group consisting of an IRES derived from picornavirus, an IRES derived from flavivirus, an IRES derived from pestivirus, an IRES derived from retrovirus, an IRES derived from lentivirus, an IRES derived from insect RNA virus, and an IRES derived from cellular mRNA. In multiple embodiments, the vector may further comprise a post-transcriptional regulatory element (PRE) sequence selected from woodchuck PRE (WPRE) (SEQ ID NO: 264), woodchuck PRE (WPRE) variant 1 (SEQ ID NO: 256), woodchuck PRE (WPRE) variant 2 (SEQ ID NO: 257), or hepatitis B virus (HBV) PRE (HPRE) (SEQ ID NO: 437). In multiple embodiments, the post-transcriptional regulatory element (PRE) sequence may be woodchuck PRE (WPRE) variant 1 comprising the nucleic acid sequence of SEQ ID NO: 256. In multiple embodiments, the post-transcriptional regulatory element (PRE) sequence may be woodchuck PRE (WPRE) variant 2 comprising the nucleic acid sequence of SEQ ID NO: 257.In some embodiments, the vector may further comprise a promoter selected from a cytomegalovirus (CMV) promoter, a phosphoglycerate kinase (PGK) promoter, a myelin basic protein (MBP) promoter, a glial fibrillary acidic protein (GFAP) promoter, a modified MoMuLV LTR containing the myeloproliferative sarcoma virus enhancer (MNDU3), a ubiquitin C promoter, an EF-1 alpha promoter, or a murine stem cell virus (MSCV) promoter. In some embodiments, the promoter may be a murine stem cell virus (MSCV) promoter. In some embodiments, the vector may be a viral vector or a non-viral vector. In some embodiments, the vector may be a viral vector. In some embodiments, the viral vector may be selected from an adenovirus, a poxvirus, an alphavirus, an arenavirus, a flavivirus, a rhabdovirus, a retrovirus, a lentivirus, a herpesvirus, a paramyxovirus, a picornavirus, and any combination thereof. In some embodiments, the viral vector may be pseudotyped with an envelope protein of a virus selected from endogenous feline virus (RD114), a type of RD114 (RD114TR: a version of RD114), gibbon ape leukemia virus (GALV), a type of GALV (GALV-TR: a version of GALV), amphotropic murine leukemia virus (MLV 4070A), baculovirus (GP64), vesicular stomatitis virus (VSV-G), fowlpox virus (FPV), Ebola virus (EboV), or baboon retrovirus envelope glycoprotein (BaEV), and lymphocytic choriomeningitis virus (LCMV). In some embodiments, the vector may be a lentiviral vector. In some embodiments, the vector may further comprise a nucleic acid encoding a chimeric antigen receptor (CAR).

[0198] In multiple embodiments, T cells and / or natural killer cells that express the polypeptides described herein and / or contain the vectors described herein and / or are produced by the methods described herein can be provided. In multiple embodiments, the T cells can be αβ T cells, γδ T cells, natural killer T cells, or any combination thereof. In multiple embodiments, the αβ T cells can be CD4+ T cells. In multiple embodiments, the αβ T cells can be CD8+ T cells. In multiple embodiments, the γδ T cells can be Vγ9Vδ2+ T cells.

[0199] In multiple embodiments, compositions comprising the T cells and / or natural killer cells described herein can be provided. In multiple embodiments, the compositions can be pharmaceutical compositions. In multiple embodiments, the compositions can further comprise an adjuvant, excipient, carrier, diluent, buffer, stabilizer, or a combination thereof. In multiple embodiments, the adjuvant can be an anti-CD40 antibody, imiquimod, resiquimod, GM-CSF, cyclophosphamide, sunitinib, bevacizumab, atezolizumab, interferon alpha, interferon beta, CpG oligonucleotides and their derivatives, poly(I:C) and its derivatives, RNA, sildenafil, particulate formulations containing poly(lactide co-glycolide) (PLG), virosomes, interleukin 1 (IL-1), interleukin 2 (IL-2), interleukin 4 (IL-4), interleukin 7 (IL-7), interleukin 12 (IL-12), interleukin 13 (IL-13), interleukin 15 (IL-15), interleukin 21 (IL-21), interleukin 23 (IL-23), or any combination thereof. In multiple embodiments, the adjuvant can be IL-2, IL-7, IL-12, IL-15, IL-21, or any combination thereof.

[0200] In multiple embodiments, a method of treating a patient having cancer can be provided, the method including administering to the patient a composition described herein, wherein the cancer is selected from the group consisting of non-small cell lung cancer, small cell lung cancer, melanoma, liver cancer, breast cancer, uterine cancer, Merkel cell carcinoma, pancreatic cancer, gallbladder cancer, bile duct cancer, colorectal cancer, bladder cancer, kidney cancer, leukemia, ovarian cancer, esophageal cancer, brain tumor, stomach cancer, and prostate cancer. In multiple embodiments, a method of eliciting an immune response in a patient having cancer can be provided, the method including administering to the patient a composition described herein, wherein the cancer is selected from the group consisting of non-small cell lung cancer, small cell lung cancer, melanoma, liver cancer, breast cancer, uterine cancer, Merkel cell carcinoma, pancreatic cancer, gallbladder cancer, bile duct cancer, colorectal cancer, bladder cancer, kidney cancer, leukemia, ovarian cancer, esophageal cancer, brain tumor, stomach cancer, and prostate cancer. In multiple embodiments, T cells and / or natural killer cells can kill cancer cells that present peptides in a complex with MHC molecules on the cell surface.

[0201] In multiple embodiments, the nucleic acid sequences disclosed herein can be mutated such that the encoded amino acids remain the same, but the nucleic acid codons are mutated to maintain improved expression in target cells and / or improved expression by the target vector. In multiple embodiments, the nucleic acids disclosed herein can be codon-optimized. In multiple embodiments, the nucleic acid sequences described herein are codon-optimized. In multiple embodiments, the nucleic acid sequences described herein can be codon-optimized, and the nucleic acid sequences encoding the polypeptides described herein can be codon-optimized. In multiple embodiments, the mutations of the nucleic acid sequences described herein can include codon optimization.

[0202] In multiple embodiments, the expression of membrane-bound IL-15 can improve the persistence, functionality, proliferation, survival, amplification, or any combination thereof of immune cells such as, but not limited to, T cells and / or natural killer cells, compared to cells that do not express membrane-bound IL-15. In multiple embodiments, the expression of membrane-bound IL-15 can improve the persistence, functionality, proliferation, survival, amplification, or any combination thereof of immune cells such as, but not limited to, T cells and / or natural killer cells in the tumor microenvironment, compared to cells that do not express membrane-bound IL-15. In multiple embodiments, the expression of membrane-bound IL-15 can increase the effectiveness of immune cells such as, but not limited to, T cells and / or natural killer cells in tumor cell killing, compared to cells that do not express membrane-bound IL-15. In multiple embodiments, the expression of membrane-bound IL-15 can increase the viability of immune cells such as, but not limited to, T cells and / or natural killer cells in the tumor microenvironment, the ability to sustain tumor cell killing, or any combination thereof, compared to cells that do not express membrane-bound IL-15. In multiple embodiments, the expression of membrane-bound IL-15 can increase the ability to maintain the naive phenotype of immune cells such as, but not limited to, T cells and / or natural killer cells.

[0203] Persistence can be evaluated, as a non-limiting example, by the length of time cells are detectable in an individual (e.g., a patient) after infusion. As a non-limiting example, persistence can be measured days, weeks, months, or years after infusion, and as non-limiting examples, can be measured about 1 week, about 2 weeks, about 4 weeks, about 1 month, about 2 months, about 3 months, about 6 months, about 9 months, about 12 months, about 18 months, about 24 months, and / or about 30 months after infusion. Persistence can be evaluated, as a non-limiting example, by PCR of peripheral blood samples, flow cytometry of peripheral blood samples, and / or analysis of tumor biopsy samples. The persistence of cells expressing membrane-bound IL-15 can be compared, as a non-limiting example, to the typical persistence of infused ACT cells or the persistence of similar cells that do not express membrane-bound IL-15.

[0204] The persistence of the ability to kill tumor cells can be measured, as non-limiting examples, through (i) a continuous killing assay using IncuCyte, in which the ability to kill / impair tumor growth is evaluated by the growth rate during repeated tumor stimulations over a period of time, and / or (ii) the production of cytokines / effector molecules (IFNγ via ELISA and other pro-inflammatory cytokines via Luminex (not limited to the cytokines measured, such as IFNγ, TNFα, granzyme B, perforin, IL-2, IL-6, MIP-1β, MIP-1α, GM-CSF, RANTES, IL-18, IL-4, IL-10, and IP10)). The persistence of the ability of cells expressing membrane-bound IL-15 to kill tumor cells can be compared, as non-limiting examples, to the persistence of the ability of similar cells not expressing membrane-bound IL-15 to kill tumor cells, or to the persistence of the ability of other control cells to kill tumor cells.

[0205] The naivety of the phenotype can be evaluated, as non-limiting examples, through a Tmem panel assay by flow cytometry. Typically, the gating of flow cytometry excludes CD8+ TCR+ cells. Typically, a more naive phenotype can be indicated by the high frequency of Tnaive / scm (CD45RA+ CCR7+) and Tcm (CD45RA- CCR7+) of the T memory subset, as well as the increase or retention of the CD39-CD69- and CD27+ CD28+ populations. Low expression of CD57 may also be desirable.

[0206] When evaluating the persistence, functionality, proliferation, survival, amplification, effectiveness of tumor killing, naivety, or other characteristics of cells expressing dnTGFβRII, cells such as, for example, untransduced cells, cells transduced with TCR only, cells transduced with CD8 and TCR, or combinations thereof can be used as control cells, as non-limiting examples.

[0207] In multiple embodiments, membrane-bound IL-15 can act in cis (e.g., affect the cell in which it is expressed), act in trans (e.g., affect cells in which it is not expressed), or act in any combination thereof. In embodiments where membrane-bound IL-15 acts in trans, cells adjacent to or in the vicinity of (e.g., within the tumor microenvironment) cells expressing membrane-bound IL-15 may exhibit any or a combination of the same or equivalent improvements as described for cells expressing membrane-bound IL-15 as compared to cells not adjacent to or in the vicinity of cells expressing membrane-bound IL-15.

[0208] In multiple embodiments, the present disclosure provides nucleic acids encoding the polypeptides described herein. In multiple embodiments, the present disclosure provides vectors comprising nucleic acids encoding the polypeptides described herein. In multiple embodiments, one or more vectors may comprise nucleic acids encoding an IL-15 / IL-15Rα fusion polypeptide. In multiple embodiments, one or more vectors may comprise nucleic acids encoding a CD8 polypeptide. In multiple embodiments, one or more vectors may comprise nucleic acids encoding a CD8α polypeptide. In multiple embodiments, one or more vectors may comprise nucleic acids encoding a CD8β polypeptide.

[0209] In multiple embodiments, one or more vectors may comprise one or more nucleic acids encoding a T cell receptor (TCR) comprising an α chain and a β chain. In multiple embodiments, one or more vectors may comprise one or more nucleic acids encoding a T cell receptor (TCR) comprising a γ chain and a δ chain. In multiple embodiments, one or more vectors may comprise one or more nucleic acids encoding a chimeric antigen receptor (CAR).

[0210] In some embodiments, a vector can be provided that contains one or more nucleic acids encoding one or any combination of a TCR comprising an α-chain and a β-chain, a TCR comprising a γ-chain and a δ-chain, a CAR, an IL-15 polypeptide, an IL-15Rα polypeptide, an IL-15 / IL-15Rα fusion polypeptide, and / or a CD8 polypeptide. In some embodiments, the CD8 polypeptide can include a CD8α-chain and / or a CD8β-chain, and the CD8α-chain and / or the CD8β-chain can independently be modified or unmodified.

[0211] In some embodiments, a vector can be provided that contains one or more nucleic acids encoding one or any combination of a TCR comprising an α-chain and a β-chain, an IL-15 / IL-15Rα fusion polypeptide, and / or a CD8 polypeptide. In some embodiments, a vector can be provided that contains one or more nucleic acids encoding one or any combination of a TCR comprising a γ-chain and a δ-chain, an IL-15 / IL-15Rα fusion polypeptide, and / or a CD8 polypeptide. In some embodiments, a vector can be provided that contains one or more nucleic acids encoding one or any combination of a CAR, an IL-15 / IL-15Rα fusion polypeptide, and / or a CD8 polypeptide. In some embodiments, the CD8 polypeptide can include a CD8α-chain and / or a CD8β-chain, and the CD8α-chain and / or the CD8β-chain can independently be modified or unmodified.

[0212] In some embodiments, a vector can be provided that includes one or more nucleic acids encoding a TCR comprising an α-chain and a β-chain, an IL-15 / IL-15Rα fusion polypeptide, and a CD8 polypeptide. In some embodiments, a vector can be provided that includes one or more nucleic acids encoding a TCR comprising a γ-chain and a δ-chain, an IL-15 / IL-15Rα fusion polypeptide, and a CD8 polypeptide. In some embodiments, a vector can be provided that includes one or more nucleic acids encoding a CAR, an IL-15 / IL-15Rα fusion polypeptide, and a CD8 polypeptide. In some embodiments, the CD8 polypeptide can include a CD8α-chain and / or a CD8β-chain, and the CD8α-chain and / or the CD8β-chain can be independently modified or unmodified.

[0213] In some embodiments, a vector can be provided that includes one or more nucleic acids encoding a TCR comprising an α-chain and a β-chain and an IL-15 / IL-15Rα fusion polypeptide. In some embodiments, a vector can be provided that includes one or more nucleic acids encoding a TCR comprising a γ-chain and a δ-chain and an IL-15 / IL-15Rα fusion polypeptide. In some embodiments, a vector can be provided that includes one or more nucleic acids encoding a CAR and an IL-15 / IL-15Rα fusion polypeptide.

[0214] In some embodiments, a vector can be provided that includes one or more nucleic acids encoding a TCR comprising an α-chain and a β-chain and a CD8 polypeptide. In some embodiments, a vector can be provided that includes one or more nucleic acids encoding a TCR comprising a γ-chain and a δ-chain and a CD8 polypeptide. In some embodiments, a cell can be provided that includes one or more nucleic acids encoding a CAR and a CD8 polypeptide. In some embodiments, the CD8 polypeptide can include a CD8α-chain and / or a CD8β-chain, and the CD8α-chain and / or the CD8β-chain can be independently modified or unmodified.

[0215] In multiple embodiments, two or more vectors can be simultaneously transduced into one or more cells, can be co-expressed in one or more cells, or can be any combination thereof. In multiple embodiments, the cells can include αβ T cells, γδ T cells, natural killer (NK) cells, natural killer T cells, CD4+ T cells, CD8+ T cells, CD4+ / CD8+ cells, or any combination thereof.

[0216] In multiple embodiments, two or more vectors can include nucleic acids encoding one or any combination of an IL-15 polypeptide, an IL-15Rα polypeptide, an IL-15 / IL-15Rα fusion polypeptide, a CD8 polypeptide, a TCR comprising an α chain and a β chain, a TCR comprising a γ chain and a δ chain, and / or a CAR. In multiple embodiments, the CD8 polypeptide can include a CD8α chain and / or a CD8β chain, and the CD8α chain and / or the CD8β chain can independently be modified or unmodified.

[0217] In multiple embodiments, one vector can include nucleic acids encoding one or any combination of an IL-15 polypeptide, an IL-15Rα polypeptide, an IL-15 / IL-15Rα fusion polypeptide, a CD8 polypeptide, a TCR comprising an α chain and a β chain, a TCR comprising a γ chain and a δ chain, and / or a CAR. In multiple embodiments, the CD8 polypeptide can include a CD8α chain and / or a CD8β chain, and the CD8α chain and / or the CD8β chain can independently be modified or unmodified.

[0218] In multiple embodiments, the nucleic acid can be polycistronic, and one or more polycistronic nucleic acids can be utilized. Expressing multiple (e.g., 2, 3, 4, 5, or more) polypeptides from a polycistronic nucleic acid can be achieved, for example, by any suitable method such as i) splicing of pre-mRNA, ii) proteolytic cleavage sites, iii) fusion proteins, iv) inclusion of nucleic acids encoding one or more 2A peptides (e.g., but not limited to, P2A, T2A, E2A, and F2A, etc.), v) inclusion of one or more internal ribosome entry sites (IRES), or other mechanisms. Each of these methods has certain merits and demerits for providing multiple transcription units. The most widely used among the five methods are self-cleaving 2A peptides and IRES. In multiple embodiments, the nucleic acid can be monocistronic, and one or more monocistronic nucleic acids can be utilized.

[0219] In multiple embodiments, the 2A peptide can be selected from P2A (SEQ ID NO: 93), T2A (SEQ ID NO: 94), E2A (SEQ ID NO: 95), or F2A (SEQ ID NO: 96).

[0220] In multiple embodiments, the IRES can be selected from the group consisting of an IRES derived from picornavirus, an IRES derived from flavivirus, an IRES derived from pestivirus, an IRES derived from retrovirus, an IRES derived from lentivirus, an IRES derived from insect RNA virus, and an IRES derived from cellular mRNA.

[0221] In multiple embodiments, the vector can include a nucleic acid encoding a 2A peptide or a nucleic acid encoding an internal ribosome entry site (IRES) disposed between a nucleic acid encoding a modified CD8α polypeptide and a nucleic acid encoding a CD8β polypeptide.

[0222] In multiple embodiments, the vector can include a nucleic acid encoding a 2A peptide disposed between a nucleic acid encoding a TCRα chain and a nucleic acid encoding a TCRβ chain.

[0223] In some embodiments, the vector can include a nucleic acid encoding a modified CD8α polypeptide, a nucleic acid encoding a CD8β polypeptide, a nucleic acid encoding a TCRα chain or a TCRβ chain, and a nucleic acid encoding a 2A peptide or an internal ribosome entry site (IRES) disposed between the nucleic acids encoding a membrane-bound IL-15.

[0224] In some embodiments, one vector can include a nucleic acid encoding one or any combination of an IL-15 polypeptide, an IL-15Rα polypeptide, an IL-15 / IL-15Rα fusion polypeptide, a CD8 polypeptide, a TCR comprising an α chain and a β chain, a TCR comprising a γ chain and a δ chain, and / or a CAR, and the vector can include either a nucleic acid encoding a polypeptide or a fusion polypeptide, or a nucleic acid encoding a 2A peptide or an internal ribosome entry site (IRES) disposed between each. In some embodiments, the CD8 polypeptide can include a CD8α chain and / or a CD8β chain, and the CD8α chain and / or the CD8β chain can be independently modified or unmodified.

[0225] In some embodiments, the vector can further include a post-transcriptional regulatory element (PRE) sequence. In some embodiments, the post-transcriptional regulatory element (PRE) sequence can be selected from a woodchuck hepatitis virus PRE (WPRE) (e.g., but not limited to, wild-type WPRE such as SEQ ID NO: 264, or mutant WPRE such as, but not limited to, WPREmut1 (SEQ ID NO: 256) or WPREmut2 (SEQ ID NO: 257)), or a hepatitis B virus (HBV) PRE (HPRE) (SEQ ID NO: 437), variants thereof, or any combination thereof.

[0226] In some embodiments, the vector may further include one or more promoters. In some embodiments, the promoter may be selected from a cytomegalovirus (CMV) promoter, a phosphoglycerate kinase (PGK) promoter, a myelin basic protein (MBP) promoter, a glial fibrillary acidic protein (GFAP) promoter, a modified MoMuLV LTR containing the myeloproliferative sarcoma virus enhancer (MNDU3), a ubiquitin C promoter, an EF-1 alpha promoter, a murine stem cell virus (MSCV) promoter, a promoter derived from CD69, a nuclear factor of activated T-cells (NFAT) promoter, an IL-2 promoter, a minimal IL-2 promoter, or a combination thereof.

[0227] In some embodiments, the vector can be a viral vector or a non-viral vector.

[0228] In some embodiments, the vector can be selected from an adenovirus, a poxvirus, an alphavirus, an arenavirus, a flavivirus, a rhabdovirus, a retrovirus, a lentivirus, a herpesvirus, a paramyxovirus, a picornavirus, or a combination thereof.

[0229] In some embodiments, the vector can be pseudotyped with an envelope protein of a virus selected from a natural feline endogenous virus (RD114), a chimeric RD114 (RD114TR), a gibbon ape leukemia virus (GALV), a chimeric GALV (GALV-TR), a amphotropic murine leukemia virus (MLV 4070A), a baculovirus (GP64), a vesicular stomatitis virus (VSV-G), a fowlpox virus (FPV), an Ebola virus (EboV), or a baboon endogenous virus envelope glycoprotein (BaEV), and a lymphocytic choriomeningitis virus (LCMV), or a combination thereof.

[0230] In multiple embodiments, the vector may include one or more Kozak sequences. In multiple embodiments, the Kozak sequence may initiate, increase, or enhance translation, or combinations thereof. In multiple embodiments, the Kozak sequence may be GCCACC. In multiple embodiments, the Kozak sequence may be ACCATGG. In multiple embodiments, the Kozak sequence may be GCCNCCATGG, where N is a purine (A or G) (SEQ ID NO: 382).

[0231] In multiple embodiments, the vector may include one or more Factor Xa sites.

[0232] In multiple embodiments, the vector may include one or more enhancers. In multiple embodiments, the enhancer may include conserved non-coding sequence (CNS) 0, CNS1, CNS2, CNS3, CNS4, or a portion thereof, or any combination thereof.

[0233] In multiple embodiments, the present disclosure provides one or more cells transformed with one or more vectors comprising a nucleic acid encoding a polypeptide and / or expressing one or more vectors.

[0234] In multiple embodiments, the cell may include an αβ T cell, a γδ T cell, a natural killer cell, a natural killer T cell, a CD4+ T cell, a CD8+ T cell, a CD4+ / CD8+ cell, or any combination thereof.

[0235] In multiple embodiments, the T cell may be a CD4+ T cell. In multiple embodiments, the T cell may be a CD8+ T cell. In multiple embodiments, the T cell may be a CD4+ / CD8+ T cell. In multiple embodiments, the T cell may be an αβ T cell. In multiple embodiments, the T cell may be a γδ T cell.

[0236] In multiple embodiments, the T cells can be αβ T cells and can express the CD8 polypeptides described herein. In multiple embodiments, the CD8 polypeptide can include the CD8α chain and / or the CD8β chain, and the CD8α chain and / or the CD8β chain can independently be modified or unmodified. In multiple embodiments, the T cells can be αβ T cells and can express a modified CD8 polypeptide described herein, such as a modified CD8α polypeptide, or a modified CD8α polypeptide comprising a CD8β stalk region, such as m1CD8α in constructs #11 and #12 (Figure 4), or CD8α* (Figure 55B). In multiple embodiments, the T cells can be αβ T cells and can express one or any combination of an IL-15 polypeptide, an IL-15Rα polypeptide, an IL-15 / IL-15Rα fusion polypeptide, a modified CD8 polypeptide, and / or a CAR.

[0237] In multiple embodiments, the T cells can be γδ T cells and can express the CD8 polypeptides described herein. In multiple embodiments, the CD8 polypeptide can include the CD8α chain and / or the CD8β chain, and the CD8α chain and / or the CD8β chain can independently be modified or unmodified. In multiple embodiments, the T cells can be γδ T cells and can express a modified CD8 polypeptide described herein, such as a modified CD8α polypeptide, or a modified CD8α polypeptide comprising a CD8β stalk region, such as m1CD8α in constructs #11 and #12 (Figure 4), or CD8α* (Figure 55B). In multiple embodiments, the T cells can be γδ T cells and can express one or any combination of an IL-15 polypeptide, an IL-15Rα polypeptide, an IL-15 / IL-15Rα fusion polypeptide, a CD8 polypeptide, and / or a CAR. In multiple embodiments, the CD8 polypeptide can include the CD8α chain and / or the CD8β chain, and the CD8α chain and / or the CD8β chain can independently be modified or unmodified.

[0238] In some embodiments, a cell can be provided that contains one or more nucleic acids encoding one or any combination of a TCR comprising an α chain and a β chain, a TCR comprising a γ chain and a δ chain, a CAR, an IL-15 polypeptide, an IL-15Rα polypeptide, an IL-15 / IL-15Rα fusion polypeptide, and / or a CD8 polypeptide. In some embodiments, the CD8 polypeptide can comprise a CD8α chain and / or a CD8β chain, and the CD8α chain and / or the CD8β chain can independently be modified or unmodified.

[0239] In some embodiments, a cell can be provided that contains one or more nucleic acids encoding one or any combination of a TCR comprising an α chain and a β chain, an IL-15 / IL-15Rα fusion polypeptide, and / or a CD8 polypeptide. In some embodiments, a cell can be provided that contains one or more nucleic acids encoding one or any combination of a TCR comprising a γ chain and a δ chain, an IL-15 / IL-15Rα fusion polypeptide, and / or a CD8 polypeptide. In some embodiments, a cell can be provided that contains one or more nucleic acids encoding one or any combination of a CAR, an IL-15 / IL-15Rα fusion polypeptide, and / or a CD8 polypeptide. In some embodiments, the CD8 polypeptide can comprise a CD8α chain and / or a CD8β chain, and the CD8α chain and / or the CD8β chain can independently be modified or unmodified.

[0240] In some embodiments, cells can be provided that contain one or more nucleic acids encoding a TCR comprising an α-chain and a β-chain, an IL-15 / IL-15Rα fusion polypeptide, and a CD8 polypeptide. In some embodiments, cells can be provided that contain one or more nucleic acids encoding a TCR comprising a γ-chain and a δ-chain, an IL-15 / IL-15Rα fusion polypeptide, and a CD8 polypeptide. In some embodiments, cells can be provided that contain one or more nucleic acids encoding a CAR, an IL-15 / IL-15Rα fusion polypeptide, and a CD8 polypeptide. In some embodiments, the CD8 polypeptide can comprise a CD8α-chain and / or a CD8β-chain, and the CD8α-chain and / or the CD8β-chain can independently be modified or unmodified.

[0241] In some embodiments, cells can be provided that contain one or more nucleic acids encoding a TCR comprising an α-chain and a β-chain and an IL-15 / IL-15Rα fusion polypeptide. In some embodiments, cells can be provided that contain one or more nucleic acids encoding a TCR comprising a γ-chain and a δ-chain and an IL-15 / IL-15Rα fusion polypeptide. In some embodiments, cells can be provided that contain one or more nucleic acids encoding a CAR and an IL-15 / IL-15Rα fusion polypeptide.

[0242] In some embodiments, cells can be provided that contain one or more nucleic acids encoding a TCR comprising an α-chain and a β-chain and a CD8 polypeptide. In some embodiments, cells can be provided that contain one or more nucleic acids encoding a TCR comprising a γ-chain and a δ-chain and a CD8 polypeptide. In some embodiments, cells can be provided that contain one or more nucleic acids encoding a CAR and a CD8 polypeptide. In some embodiments, the CD8 polypeptide can comprise a CD8α-chain and / or a CD8β-chain, and the CD8α-chain and / or the CD8β-chain can independently be modified or unmodified.

[0243] In some embodiments, the one or more nucleic acids can be contained within and / or expressed from a vector.

[0244] In some embodiments, the cells can include αβ T cells, γδ T cells, natural killer cells, CD4+ T cells, CD8+ T cells, CD4+ / CD8+ cells, or any combination thereof.

[0245] In some embodiments, a population of the cells described herein can be provided. By way of non-limiting example, the disclosure provides a population of modified cells comprising one or more nucleic acids encoding one or a combination of any of the polypeptides described herein, such as an exogenous CD8 co-receptor comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 5, 7, 258, 259, 8, 9, 10, 11, 12, 13, or 14, a membrane-bound IL-15 described herein (e.g., an IL-15 / IL-15Rα fusion polypeptide) and / or a T cell receptor. In some embodiments, the population of cells can include αβ T cells, γδ T cells, natural killer cells, natural killer T cells, CD4+ T cells, CD8+ T cells, CD4+ / CD8+ cells, or any combination thereof.

[0246] In certain aspects, the polypeptide sequences and / or nucleic acid sequences described herein can be isolated sequences and / or recombinant sequences.

[0247] In certain aspects, the cells described herein can be isolated cells and / or recombinant cells.

[0248] In multiple embodiments, a method of preparing cells for immunotherapy can include isolating cells from a blood sample of a human subject, activating the isolated cells, transducing the activated cells with one or more vectors, and amplifying the transduced cells. In multiple embodiments, the cells can include αβ T cells, γδ T cells, natural killer cells, natural killer T cells, CD4+ T cells, CD8+ T cells, CD4+ / CD8+ cells, or any combination thereof.

[0249] In multiple embodiments, a method of treating a patient having cancer can include administering to the patient a composition comprising a population of amplified cells, where the cells kill cancer cells that present a peptide in a complex with MHC molecules on the cell, the peptide is selected from SEQ ID NOs: 98 to 255, and the cancer is selected from the group consisting of non-small cell lung cancer, small cell lung cancer, melanoma, liver cancer, breast cancer, uterine cancer, Merkel cell carcinoma, pancreatic cancer, bile duct cancer, cholangiocarcinoma, colorectal cancer, bladder cancer, kidney cancer, leukemia, ovarian cancer, esophageal cancer, brain tumor, gastric cancer, prostate cancer, or a combination thereof. In multiple embodiments, the cells can include αβ T cells, γδ T cells, natural killer cells, natural killer T cells, CD4+ T cells, CD8+ T cells, CD4+ / CD8+ cells, or any combination thereof. In multiple embodiments, the composition can further include an adjuvant. In multiple embodiments, the adjuvant can be selected from anti-CD40 antibody, imiquimod, resiquimod, GM-CSF, cyclophosphamide, sunitinib, bevacizumab, atezolizumab, interferon alpha, interferon beta, CpG oligonucleotides and derivatives thereof, poly(I:C) and derivatives thereof, RNA, sildenafil, particle formulations comprising poly(lactide co-glycolide) (PLG), virosomes, interleukin (IL)-1, IL-2, IL-4, IL-7, IL-12, IL-13, IL-15, IL-21, IL-23, or any combination thereof.

[0250] In multiple embodiments, a method of eliciting an immune response in a patient having cancer can include administering to the patient a composition comprising a population of amplified cells, where the cells kill cancer cells that present a peptide in a complex with an MHC molecule on the cell surface, the peptide is selected from SEQ ID NOs: 98 - 255, and the cancer is selected from the group consisting of non - small cell lung cancer, small cell lung cancer, melanoma, liver cancer, breast cancer, uterine cancer, Merkel cell carcinoma, pancreatic cancer, gallbladder cancer, bile duct cancer, colorectal cancer, bladder cancer, kidney cancer, leukemia, ovarian cancer, esophageal cancer, brain tumor, stomach cancer, prostate cancer, or combinations thereof. In multiple embodiments, the cells can include αβ T cells, γδ T cells, natural killer cells, natural killer T cells, CD4+ T cells, CD8+ T cells, CD4+ / CD8+ cells, or any combination thereof.

Brief Description of the Drawings

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Figure 1

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Figure 5

[0256] Figure 5B shows the titer of additional viral vectors according to the present disclosure. Construct #13; Construct #14; Construct #15; Construct #16; Construct #17; Construct #18; Construct #19; Construct #21; Construct #10n; Construct #11n; and TCR:R11KEA (SEQ ID NOs: 15 and 16, which may be encoded by SEQ ID NOs: 72 and 73) (Construct #8). This binds to PRAME-004 (SLLQHLIGL) (SEQ ID NO: 147). Note that Constructs #10 and #10n are different batches of the same construct (SEQ ID NOs: 291 and 292), and Constructs #11 and #11n are also different batches of the same construct (SEQ ID NOs: 285 and 286).

[0257]

Figure 6

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Figure 7

[0259] Figure 7B shows the expression of activation markers before and after activation in CD3+CD4+ cells.

[0260]

Figure 8

[0261] Figure 8B shows the expression factors of cells transduced with various constructs derived from donor #2. The constructs are as follows: construct #9b; construct #10; construct #11; construct #12; construct #1; construct #2; TCR = R11KEA.WPRE wt (TCR with wild-type WPRE) (construct #8); NT = non-transduced T cells (negative control).

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[0284] Figure 27B shows the expression of activation markers before and after activation in CD3+CD4+ cells according to one embodiment of the present disclosure.

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Figure 67

[0327] Figure 67B shows an exemplary membrane-bound IL-15 polypeptide that can be provided in multiple embodiments. In the figure, the IL-15 polypeptide is located at the C-terminus of the IL-15Rα polypeptide. Nucleic acids encoding such constructs are also provided in multiple embodiments. In Figures 67A and 67B, L represents one or more optional linkers, and the line may represent a direct bond without intervening sequences, or may represent intervening sequences such as, for example but not limited to, linkers, untranslated sequences (in the case of nucleic acid sequences), translated sequences, sequences containing one or more restriction endonuclease sites (in the case of nucleic acid sequences), or combinations thereof.

[0328]

Figure 68

[0329] Figure 68B shows an exemplary membrane-bound IL-15 polypeptide that can be provided in multiple embodiments. In the figure, the IL-15 polypeptide is located at the C-terminus of the IL-15Rα polypeptide, and the signal peptide (SP) is located at the N-terminus of the IL-15Rα polypeptide. Nucleic acids encoding such constructs are also provided in multiple embodiments. In Figures 68A and 68B, L represents one or more optional linkers, and the line may represent a direct bond without intervening sequences, or may represent intervening sequences such as, for example but not limited to, linkers, untranslated sequences (in the case of nucleic acid sequences), translated sequences, sequences containing one or more restriction endonuclease sites (in the case of nucleic acid sequences), or combinations thereof.

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Mode for Carrying Out the Invention

[0345] Membrane - bound IL - 15 In multiple embodiments, a membrane-bound IL-15 polypeptide (membrane-bound IL-15 or mbIL-15) is provided. In multiple embodiments, the nucleic acids described herein include and / or encode a membrane-bound IL-15 polypeptide. In multiple embodiments, the vectors described herein include and / or encode a membrane-bound IL-15 polypeptide. In multiple embodiments, the cells described herein include and / or express a membrane-bound IL-15 polypeptide. In multiple embodiments, the compositions described herein include a membrane-bound IL-15 polypeptide or include a cell that includes and / or expresses a membrane-bound IL-15 polypeptide. In multiple embodiments, IL-15 becomes membrane-bound by expressing the IL-15 polypeptide and the IL-15Rα polypeptide in an IL-15 / IL-15Rα fusion polypeptide (IL-15 / IL-15Rα).

[0346] A membrane-bound IL-15 polypeptide is provided. An isolated nucleic acid sequence comprising one or more nucleic acid sequences encoding one or more membrane-bound IL-15 polypeptides is provided. A vector comprising one or more nucleic acid sequences encoding one or more membrane-bound IL-15 polypeptides is provided. A cell comprising and / or expressing one or more membrane-bound IL-15 polypeptides is provided. A cell comprising, or expressing, one or more nucleic acid sequences comprising one or more nucleic acid sequences encoding one or more membrane-bound IL-15 polypeptides is provided. A cell comprising, or expressing, one or more vectors comprising one or more nucleic acid sequences comprising one or more nucleic acid sequences encoding one or more membrane-bound IL-15 polypeptides is provided. In multiple embodiments, the cells described herein may comprise a membrane-bound IL-15 polypeptide, a CD8 polypeptide, a cell receptor (TCR) comprising an α chain and a β chain, a TCR comprising a γ chain and a δ chain, a chimeric antigen receptor (CAR), or any combination thereof. In multiple embodiments, the cell may comprise an αβ T cell, a γδ T cell, a natural killer cell, a natural killer T cell, a CD4+ cell, a CD8+ cell, a CD4+ / CD8+ cell, or any combination thereof. In multiple embodiments, such polypeptides, nucleic acids, vectors, and / or cells may be isolated, recombinant, and / or engineered. A composition comprising such polypeptides, nucleic acids, vectors, and / or cells is provided.

[0347] In certain aspects, the polypeptide sequences and / or nucleic acid sequences described herein can be isolated sequences and / or recombinant sequences.

[0348] In certain aspects, the cells described herein can be isolated cells and / or recombinant cells.

[0349] Membrane-bound IL-15 can include, for example, an IL-15 / IL-15Rα fusion polypeptide and / or an IL-15Rα / IL-15 fusion polypeptide. One or more linkers can be disposed between IL-15 and IL-15Rα, or between IL-15Rα and IL-15. In a plurality of embodiments, the IL-15 polypeptide is located at the N-terminus of the IL-15Rα polypeptide in the membrane-bound IL-15 polypeptide. (FIG. 67A). In a plurality of embodiments, the IL-15 polypeptide is located at the C-terminus of the IL-15Rα polypeptide in the membrane-bound IL-15 polypeptide. (FIG. 67B). In FIGS. 67A and 67B, the IL-15 polypeptide can be an immature wild-type, immature mutant, mature wild-type, or mature mutant. In FIGS. 67A and 67B, the IL-15Rα polypeptide can be an immature wild-type, immature mutant, mature wild-type, or mature mutant. In a plurality of embodiments, the IL-15 polypeptides of FIGS. 67A and 67B are mature, with or without mutations, and the IL-15Rα polypeptides of FIGS. 67A and 67B are mature, with or without mutations. In a plurality of embodiments, the IL-15 polypeptides of FIGS. 67A and 67B are mature, with or without mutations, and the IL-15Rα polypeptides of FIGS. 67A and 67B are mature and mutant. Linkers are shown in FIGS. 67A and 67B, but mbIL-15 may or may not include a linker.

[0350] In multiple embodiments, the IL-15 polypeptide and the IL-15Rα polypeptide are linked by one or more linkers. The IL-15 / IL-15Rα fusion polypeptide and / or the IL-15Rα / IL-15 fusion polypeptide may also include one or more linkers. In multiple embodiments, the membrane-bound IL-15 is included and / or encoded by the structure shown in FIG. 67A or FIG. 67B. In FIGS. 67A and 67B, the line connecting IL-15 to one or more linkers and connecting the one or more linkers to IL-15Rα may represent a direct bond without intervening sequences, or may represent intervening sequences such as, for example, but not limited to, linkers, untranslated sequences (in the case of nucleic acid sequences), translated sequences, sequences containing one or more restriction endonuclease sites (in the case of nucleic acid sequences), or combinations thereof.

[0351] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide and / or the IL-15Rα / IL-15 fusion polypeptide can include one or more signal peptides. In multiple embodiments, a membrane-bound IL-15 that includes one or more signal peptides and optionally one or more linkers can be included and / or encoded by the structures shown in FIGS. 68A or 68B. An exemplary IL-15 / IL-15Rα fusion polypeptide that optionally includes at least one linker and at least one signal peptide is shown in FIG. 68A. An exemplary IL-15Rα / IL-15 fusion polypeptide that optionally includes at least one linker and at least one signal peptide is shown in FIG. 68B. In FIGS. 68A and 68B, the IL-15 polypeptide can be an immature wild-type, immature mutant, mature wild-type, or mature mutant. In FIGS. 68A and 68B, the IL-15Rα polypeptide can be an immature wild-type, immature mutant, mature wild-type, or mature mutant. In multiple embodiments, the IL-15 polypeptides of FIGS. 68A and 68B are mature, with or without mutations, and the IL-15Rα polypeptides of FIGS. 68A and 68B are mature, with or without mutations. In multiple embodiments, the IL-15 polypeptides of FIGS. 68A and 68B are mature, with or without mutations, and the IL-15Rα polypeptides of FIGS. 68A and 68B are mature and mutant. As shown in FIGS. 68A and 68B, (a) a line connecting one or more signal peptides (SP) to IL-15, the IL-15 to one or more linkers (L), and the one or more linkers to IL-15Rα (FIG. 68A), or (b) a line connecting one or more signal peptides (SP) to IL-15α, the IL-15α to one or more linkers (L), and the one or more linkers to IL-15 (FIG. 68B) can represent a direct bond without intervening sequences or can represent intervening sequences such as, for example, but not limited to, linkers, untranslated sequences (in the case of nucleic acid sequences), translated sequences, sequences that include one or more restriction endonuclease sites (in the case of nucleic acid sequences), or combinations thereof.

[0352] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide comprises all of the IL-15 polypeptide, all of the IL-15Rα polypeptide, or both. In multiple embodiments, all of the wild-type IL-15 polypeptide, or the complete wild-type IL-15 polypeptide, may comprise SEQ ID NO: 305. In multiple embodiments, all of the wild-type IL-15Rα polypeptide, or the complete wild-type IL-15Rα polypeptide, may comprise SEQ ID NO: 306.

[0353] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide comprises the mature IL-15 polypeptide (e.g., SEQ ID NO: 307), the mature IL-15Rα polypeptide (e.g., SEQ ID NO: 309), which may be mutated (e.g., SEQ ID NO: 311, 313, 315), or both. In multiple embodiments, the mature wild-type IL-15 polypeptide may comprise or consist of SEQ ID NO: 307, or may comprise or consist of amino acids 49-162 of SEQ ID NO: 305. In multiple embodiments, the mature wild-type IL-15Rα polypeptide may comprise or consist of SEQ ID NO: 309, or may comprise or consist of amino acids 31-267 of SEQ ID NO: 306. In multiple embodiments, the mature wild-type IL-15 polypeptide is encoded by a nucleic acid comprising or consisting of the nucleic acid set forth in SEQ ID NO: 308. In multiple embodiments, the mature wild-type IL-15Rα polypeptide is encoded by a nucleic acid comprising or consisting of the nucleic acid set forth in SEQ ID NO: 310. However, in multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide does not comprise mature wild-type IL-15Rα that is in a sequence having at least about 95% sequence identity to SEQ ID NO: 309 or SEQ ID NO: 309. In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide does not comprise mature wild-type IL-15Rα encoded by a sequence having at least about 80%, about 85%, about 90%, or at least about 95% sequence identity to SEQ ID NO: 310 or SEQ ID NO: 310.

[0354] In multiple embodiments, the IL-15 polypeptide is a variant and / or truncated form, the IL-15Rα polypeptide is a variant and / or truncated form, or both are variants and / or truncated forms.

[0355] In multiple embodiments, the IL-15 polypeptide may or may not include a native signal peptide (which may have a sequence including SEQ ID NO: 369), may or may not include a native propeptide (which may have a sequence including SEQ ID NO: 371), or may or may not include any combination thereof.

[0356] In multiple embodiments, the IL-15Rα polypeptide may or may not include a native signal sequence (which may have a sequence including SEQ ID NO: 370).

[0357] In some embodiments, the IL-15Rα polypeptide, which can be a mature IL-15Rα polypeptide (e.g., SEQ ID NO: 309), can be a variant. In some embodiments, the IL-15Rα polypeptide can include a variant transmembrane domain. In some embodiments, the transmembrane domain of the IL-15Rα polypeptide can include or consist of SEQ ID NO: 376 or SEQ ID NO: 378. In some embodiments, the transmembrane domain of the IL-15Rα polypeptide can be encoded by a nucleic acid that includes or consists of the sequence set forth in SEQ ID NO: 377 or SEQ ID NO: 379. In some embodiments, the variant IL-15Rα can include a heterologous transmembrane domain. In some embodiments, the heterologous transmembrane domain can be derived from CD25. In some embodiments, the transmembrane domain derived from CD25 can include or consist of SEQ ID NO: 372 or a sequence having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity to SEQ ID NO: 372. In some embodiments, the transmembrane domain derived from CD25 is encoded by a nucleic acid that includes or consists of a nucleic acid sequence set forth in SEQ ID NO: 373 or a sequence having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity to SEQ ID NO: 373.In some embodiments, an IL-15Rα polypeptide comprising a CD25 transmembrane domain comprises, or consists of, a sequence set forth in SEQ ID NO: 311, or a sequence having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity to SEQ ID NO: 311. In some embodiments, an IL-15Rα polypeptide comprising a CD25 transmembrane domain is encoded by a nucleic acid comprising, or consisting of, a sequence set forth in SEQ ID NO: 312, or a sequence having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity to SEQ ID NO: 312. In some embodiments, the heterologous transmembrane domain can be derived from CD28. In some embodiments, a transmembrane domain derived from CD28 comprises, or consists of, a sequence having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity to SEQ ID NO: 374. In some embodiments, a transmembrane domain derived from CD28 is encoded by a nucleic acid comprising, or consisting of, a nucleic acid sequence set forth in SEQ ID NO: 375, or a sequence having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity to SEQ ID NO: 375.In multiple embodiments, the IL-15Rα polypeptide comprising the CD28 transmembrane domain comprises, or consists of, a sequence set forth in SEQ ID NO: 313, or a sequence having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity to SEQ ID NO: 313. In multiple embodiments, the IL-15Rα polypeptide comprising the CD28 transmembrane domain is encoded by a nucleic acid comprising, or consisting of, a sequence set forth in SEQ ID NO: 314, or a sequence having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity to SEQ ID NO: 314. In multiple embodiments, IL-15Rα can be mutated by deleting exon 3 of the genomic DNA of human IL-15Rα. In multiple embodiments, the IL-15Rα polypeptide comprising the deletion of exon 3 comprises, or consists of, a sequence set forth in SEQ ID NO: 315, or a sequence having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity to SEQ ID NO: 315. In multiple embodiments, the IL-15Rα polypeptide comprising the deletion of exon 3 is encoded by a nucleic acid comprising, or consisting of, a sequence set forth in SEQ ID NO: 316, or a sequence having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity to SEQ ID NO: 316.In multiple embodiments, the function of IL-15Rα, e.g., but not limited to, the ability of IL-15Rα to bind to the membrane, and one or more signaling functions of IL-15α are preserved and / or enhanced in an IL-15Rα polypeptide having a heterologous transmembrane domain or having a deletion of exon 3.

[0358] In multiple embodiments, the present disclosure provides a nucleic acid encoding a polypeptide described herein.

[0359] In one aspect, the polypeptide sequences and / or nucleic acid sequences described herein can be isolated sequences and / or recombinant sequences.

[0360] In one aspect, the cells described herein can be isolated cells and / or recombinant cells.

[0361] In multiple embodiments, the IL-15 polypeptide has a sequence having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the amino acid sequence of SEQ ID NO: 305. In multiple embodiments, the function of IL-15, e.g., but not limited to, one or more signaling functions of IL-15 are preserved and / or enhanced in the mutant IL-15 polypeptide.

[0362] In multiple embodiments, the IL-15 polypeptide has a sequence that has at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the amino acid sequence of SEQ ID NO: 307. In multiple embodiments, one or more signaling functions of IL-15, such as, but not limited to, the signaling functions of IL-15, are conserved and / or enhanced in the mutant IL-15 polypeptide.

[0363] In multiple embodiments, the IL-15 polypeptide comprises (a) SEQ ID NO: 305 that includes 1, 2, 3, 4 or 5 amino acid substitutions, or (b) SEQ ID NO: 307 that includes 1, 2, 3, 4 or 5 amino acid substitutions. In multiple embodiments, the amino acid substitutions are conservative or non-conservative. In multiple embodiments, the amino acid substitutions are conservative amino acid substitutions. In multiple embodiments, one or more signaling functions of IL-15, such as, but not limited to, the signaling functions of IL-15, are conserved and / or enhanced in the mutant IL-15 polypeptide.

[0364] In multiple embodiments, the IL-15 polypeptide is encoded by a nucleic acid that has at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 308. In multiple embodiments, one or more signaling functions of IL-15, such as, but not limited to, the signaling functions of IL-15, are conserved and / or enhanced in the IL-15 polypeptide encoded by the mutant nucleic acid sequence.

[0365] In multiple embodiments, the IL-15 polypeptide is encoded by a nucleic acid comprising (a) SEQ ID NO: 308, which contains 1, 2, 3, 4, or 5 nucleic acid substitutions. In multiple embodiments, one or more nucleic acid substitutions in a codon may result in a codon that encodes the same amino acid or may result in a codon that encodes a different amino acid. In multiple embodiments, one or more nucleic acid substitutions in a codon may result in a codon that encodes a conservative amino acid substitution. In multiple embodiments, one or more nucleic acid substitutions in a codon may result in a codon that encodes the same amino acid. In multiple embodiments, the function of IL-15, for example, but not limited to, one or more signaling functions of IL-15, is conserved and / or enhanced in the IL-15 polypeptide encoded by the variant nucleic acid sequence.

[0366] In multiple embodiments, the nucleic acid encoding the IL-15 polypeptide may contain a stop codon (e.g., TAA, TAG, or TGA), and those stop codons are located, as non-limiting examples, at the 3' end of the nucleotides encoding the IL-15 polypeptide.

[0367] In multiple embodiments, the IL-15Rα polypeptide has a sequence that includes at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the amino acid sequence of SEQ ID NO: 306. In multiple embodiments, the IL-15Rα polypeptide has a sequence that includes at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the amino acid sequence of SEQ ID NO: 309. However, in multiple embodiments, the IL-15Rα polypeptide does not have a sequence that includes or consists of a sequence having at least about 95% sequence identity to SEQ ID NO: 309 or SEQ ID NO: 309. In multiple embodiments, the IL-15Rα polypeptide has a sequence that includes at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the amino acid sequence of SEQ ID NO: 311. In multiple embodiments, the IL-15Rα polypeptide has a sequence that includes at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the amino acid sequence of SEQ ID NO: 313.In multiple embodiments, the IL-15Rα polypeptide has a sequence that has at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the amino acid sequence of SEQ ID NO: 315. In multiple embodiments, the function of IL-15Rα, such as but not limited to, the ability of IL-15Rα to bind to the membrane, and one or more signaling functions of IL-15α are conserved and / or enhanced in the mutant IL-15Rα polypeptide.

[0368] In multiple embodiments, the IL-15Rα polypeptide can comprise (a) SEQ ID NO: 306 comprising 1, 2, 3, 4 or 5 amino acid substitutions, (b) SEQ ID NO: 309 comprising 1, 2, 3, 4 or 5 amino acid substitutions, (c) SEQ ID NO: 311 comprising 1, 2, 3, 4 or 5 amino acid substitutions, (d) SEQ ID NO: 313 comprising 1, 2, 3, 4 or 5 amino acid substitutions, or (e) SEQ ID NO: 315 comprising 1, 2, 3, 4 or 5 amino acid substitutions. However, in multiple embodiments, the IL-15Rα polypeptide does not have a sequence that comprises or consists of a sequence having at least about 95% sequence identity to SEQ ID NO: 309. In multiple embodiments, the amino acid substitutions are conservative or non-conservative. In multiple embodiments, the amino acid substitutions are conservative amino acid substitutions. In multiple embodiments, the function of IL-15Rα, such as but not limited to, the ability of IL-15Rα to bind to the membrane, and one or more signaling functions of IL-15α are conserved and / or enhanced in the mutant IL-15Rα polypeptide.

[0369] In multiple embodiments, the IL-15Rα polypeptide is encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 310. However, in multiple embodiments, the IL-15Rα polypeptide is not encoded by a nucleic acid comprising SEQ ID NO: 310, or a nucleic acid having about 85%, about 90%, about 95% or more sequence identity to SEQ ID NO: 310. In multiple embodiments, the IL-15Rα polypeptide is encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 312. In multiple embodiments, the IL-15Rα polypeptide is encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 314. In multiple embodiments, the IL-15Rα polypeptide is encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 316. In multiple embodiments, the function of IL-15Rα, such as, but not limited to, the ability of IL-15Rα to bind to the membrane, and one or more signaling functions of IL-15α, are conserved and / or enhanced in the IL-15α polypeptide encoded by the mutant nucleic acid sequence.

[0370] In multiple embodiments, the IL-15Rα polypeptide is encoded by a nucleic acid comprising (a) SEQ ID NO: 310 containing 1, 2, 3, 4, or 5 nucleic acid substitutions, (b) SEQ ID NO: 312 containing 1, 2, 3, 4, or 5 nucleic acid substitutions, (c) SEQ ID NO: 314 containing 1, 2, 3, 4, or 5 nucleic acid substitutions, and (d) SEQ ID NO: 316 containing 1, 2, 3, 4, or 5 nucleic acid substitutions. However, in multiple embodiments, the IL-15Rα polypeptide is not encoded by a nucleic acid comprising SEQ ID NO: 310 or a nucleic acid having about 85%, about 90%, about 95% or more sequence identity to SEQ ID NO: 310. In multiple embodiments, one or more nucleic acid substitutions in a codon may result in a codon encoding the same amino acid or may result in a codon encoding a different amino acid. In multiple embodiments, one or more nucleic acid substitutions in a codon may result in a codon encoding a conservative amino acid substitution. In multiple embodiments, one or more nucleic acid substitutions in a codon may result in a codon encoding the same amino acid. In multiple embodiments, the function of IL-15Rα, such as, but not limited to, the ability of IL-15Rα to bind to a membrane and one or more signaling functions of IL-15α, are preserved and / or enhanced in the IL-15α polypeptide encoded by the variant nucleic acid sequence.

[0371] In multiple embodiments, the nucleic acid encoding the IL-15Rα polypeptide may contain a stop codon (e.g., TAA, TAG, or TGA), which, by way of non-limiting example, is located at the 3' end of the nucleotides encoding the IL-15Rα polypeptide.

[0372] In multiple embodiments, the IL-15 polypeptide and the IL-15Rα polypeptide are linked by one or more linkers. In multiple embodiments, the linker is a peptide linker. In multiple embodiments, the peptide linker is rigid or flexible. In multiple embodiments, the linker is cleavable. In multiple embodiments, the linker can promote the stability or proper folding of the fusion polypeptide, increase the expression of the fusion polypeptide, improve the biological activity of the fusion polypeptide, promote the targeting of the fusion polypeptide, change the PK of the fusion polypeptide, or any combination thereof.

[0373] In multiple embodiments, the linker comprises from about 2 to 40 amino acids, from about 4 to 38 amino acids, from about 6 to 34 amino acids, from about 8 to 32 amino acids, from about 10 to 30 amino acids, about 10 amino acids, about 11 amino acids, about 12 amino acids, from about 12 to 28 amino acids, about 13 amino acids, about 14 amino acids, about 15 amino acids, about 16 amino acids, about 17 amino acids, about 18 amino acids, about 19 amino acids, about 20 amino acids, from about 14 to 26 amino acids, from about 12 to 24 amino acids, from about 10 to 22 amino acids, from about 10 to 20 amino acids, from about 12 to 18 amino acids, from about 14 to 16 amino acids, from about 8 to 22 amino acids, from about 6 to 24 amino acids, from about 4 to 26 amino acids, or from about 2 to 28 amino acids.

[0374] In some embodiments, one or more linkers of the IL-15 / IL-15Rα fusion polypeptide independently comprise, or consist of, any sequence having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% sequence identity to GSG, LE, SEQ ID NO: 266, 383, 385, 387, 389, 391, or 393, or 395 - 432, or SEQ ID NO: 266, 383, 385, 387, 389, 391, or 393, or 395 - 432. However, in some embodiments, one or more linkers of the IL-15 / IL-15Rα fusion polypeptide are not SEQ ID NO: 391 and / or SEQ ID NO: 395. In some embodiments, one or more linkers of the IL-15 / IL-15Rα fusion polypeptide independently comprise, or consist of, any sequence having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% sequence identity to GSG, LE, SEQ ID NO: 266, 383, 385, 387, 389, 393, or 396 - 432, or SEQ ID NO: 266, 383, 385, 387, 389, 393, or 396 - 432. In some embodiments, one or more linkers of the IL-15 / IL-15Rα fusion polypeptide independently comprise, or consist of, any sequence having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% sequence identity to SEQ ID NO: 383, 385, 387, or 389, or SEQ ID NO: 383, 385, 387, or 389.

[0375] In multiple embodiments, one or more linkers of the IL-15 / IL-15Rα fusion polypeptide are independently encoded by one or more nucleic acids comprising, or consisting of, any of SEQ ID NO: 384, 386, 388, 390, or 392, or by a sequence having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity to any of SEQ ID NO: 384, 386, 388, 390, or 392, or by one or more nucleic acids encoding any linker comprising, or consisting of, GSG, LE or one or more linkers described in SEQ ID NO: 266 or 393 - 432, or by one or more nucleic acids encoding any linker having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity to any of SEQ ID NO: 266 or 393 - 432. However, in multiple embodiments, one or more linkers of the IL-15 / IL-15Rα fusion polypeptide are not encoded by SEQ ID NO: 392 and are not encoded by a nucleic acid encoding SEQ ID NO: 391 or 395.

[0376] In multiple embodiments, one or more linkers of the IL-15 / IL-15Rα fusion polypeptide are independently encoded by one or more nucleic acids comprising, or consisting of, any of SEQ ID NO: 384, 386, 388, or 390, or by a sequence having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity to any of SEQ ID NO: 384, 386, 388, or 390, encoded by one or more nucleic acids encoding any linker comprising, or consisting of, GSG, or one or more linkers set forth in SEQ ID NO: 266, 383, 385, 387, 389, 393, or 396 - 432, or by one or more nucleic acids encoding any linker having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity to any of SEQ ID NO: 266, 383, 385, 387, 389, 393, or 396 - 432.

[0377] In multiple embodiments, one or more linkers of the IL-15 / IL-15Rα fusion polypeptide are independently encoded by any of SEQ ID NO: 384, 386, 388, or 390, or by a sequence having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity to any of SEQ ID NO: 384, 386, 388, or 390.

[0378] In multiple embodiments, the linker has a sequence that includes at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the amino acid sequence of SEQ ID NO: 383. In multiple embodiments, the linker has a sequence that includes at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the amino acid sequence of SEQ ID NO: 385. In multiple embodiments, the linker has a sequence that includes at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the amino acid sequence of SEQ ID NO: 387. In multiple embodiments, the linker has a sequence that includes at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the amino acid sequence of SEQ ID NO: 389. In multiple embodiments, one or more functions of the linker, such as, but not limited to, the flexibility, rigidity, cleavability of the linker, the ability to promote the stability or proper folding of the fusion polypeptide, the ability to increase the expression of the fusion polypeptide, the ability to improve the biological activity of the fusion polypeptide, the ability to promote the targeting of the fusion polypeptide, the ability to alter the PK of the fusion polypeptide, or any combination thereof, are preserved and / or enhanced in the variant linker.

[0379] In multiple embodiments, the linker comprises (a) SEQ ID NO: 383 comprising 1, 2, 3, 4, or 5 amino acid substitutions, (b) SEQ ID NO: 385 comprising 1, 2, 3, 4, or 5 amino acid substitutions, (c) SEQ ID NO: 387 comprising 1, 2, 3, 4, or 5 amino acid substitutions, (d) SEQ ID NO: 389 comprising 1, 2, 3, 4, or 5 amino acid substitutions, or (e) SEQ ID NO: 391 comprising 1, 2, 3, 4, or 5 amino acid substitutions. In multiple embodiments, the amino acid substitutions can be conservative or non-conservative. In multiple embodiments, the amino acid substitutions can be conservative amino acid substitutions. In multiple embodiments, one or more functions of the linker, such as, but not limited to, the flexibility, rigidity, cleavability of the linker, the ability to promote the stability or proper folding of the fusion polypeptide, the ability to increase the expression of the fusion polypeptide, the ability to improve the biological activity of the fusion polypeptide, the ability to promote the targeting of the fusion polypeptide, the ability to change the PK of the fusion polypeptide, or any combination thereof, are preserved and / or enhanced in the mutant linker.

[0380] In multiple embodiments, the linker is encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 384. In multiple embodiments, the linker is encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 386. In multiple embodiments, the linker is encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 388. In multiple embodiments, the linker is encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 390. In multiple embodiments, one or more functions of the linker, such as, but not limited to, the flexibility, rigidity, cleavability of the linker, the ability to promote the stability or proper folding of the fusion polypeptide, the ability to increase the expression of the fusion polypeptide, the ability to improve the biological activity of the fusion polypeptide, the ability to promote the targeting of the fusion polypeptide, the ability to alter the PK of the fusion polypeptide, or any combination thereof, are conserved and / or enhanced in the variant linker.

[0381] In multiple embodiments, the linker is encoded by a nucleic acid comprising (a) SEQ ID NO: 384 comprising 1, 2, 3, 4, or 5 nucleic acid substitutions, (b) SEQ ID NO: 386 comprising 1, 2, 3, 4, or 5 nucleic acid substitutions, (c) SEQ ID NO: 388 comprising 1, 2, 3, 4, or 5 nucleic acid substitutions, (d) SEQ ID NO: 390 comprising 1, 2, 3, 4, or 5 nucleic acid substitutions, or (e) SEQ ID NO: 392 comprising 1, 2, 3, 4, or 5 nucleic acid substitutions. In multiple embodiments, one or more nucleic acid substitutions in a codon may result in a codon that encodes the same amino acid or may result in a codon that encodes a different amino acid. In multiple embodiments, one or more nucleic acid substitutions in a codon may result in a codon that encodes a conservative amino acid substitution. In multiple embodiments, one or more nucleic acid substitutions in a codon may result in a codon that encodes the same amino acid. In multiple embodiments, one or more functions of the linker, such as, but not limited to, the flexibility, rigidity, cleavability of the linker, the ability to promote the stability or proper folding of the fusion polypeptide, the ability to increase the expression of the fusion polypeptide, the ability to improve the biological activity of the fusion polypeptide, the ability to promote the targeting of the fusion polypeptide, the ability to alter the PK of the fusion polypeptide, or one or more of any combination thereof, are preserved and / or enhanced in the variant linker.

[0382] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker comprises SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, 333, or 335. In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker is encoded by a nucleic acid comprising SEQ ID NO: 318, 320, 322, 324, 326, 328, 330, 332, 334, or 336. However, in multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker does not comprise, or consists of, a sequence having at least about 95% sequence identity to SEQ ID NO: 335. In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker is not encoded by a nucleic acid comprising, or consisting of, SEQ ID NO: 336, and is not encoded by a sequence having at least about 80%, about 85%, about 90%, or about 95% sequence identity to SEQ ID NO: 336. In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker comprises SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, or 333. In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker is encoded by a nucleic acid comprising SEQ ID NO: 318, 320, 322, 324, 326, 328, 330, 332, or 334. In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker comprises SEQ ID NO: 317, 321, 325, 327, 329, 331, or 333. In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker is encoded by a nucleic acid comprising SEQ ID NO: 318, 322, 326, 328, 330, 332, or 334.

[0383] In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker may comprise: (a) SEQ ID NO: 317 comprising 1, 2, 3, 4, or 5 amino acid substitutions; (b) SEQ ID NO: 319 comprising 1, 2, 3, 4, or 5 amino acid substitutions; (c) SEQ ID NO: 321 comprising 1, 2, 3, 4, or 5 amino acid substitutions; (d) SEQ ID NO: 323 comprising 1, 2, 3, 4, or 5 amino acid substitutions; (e) SEQ ID NO: 325 comprising 1, 2, 3, 4, or 5 amino acid substitutions; (f) SEQ ID NO: 327 comprising 1, 2, 3, 4, or 5 amino acid substitutions; (g) SEQ ID NO: 329 comprising 1, 2, 3, 4, or 5 amino acid substitutions; (h) SEQ ID NO: 331 comprising 1, 2, 3, 4, or 5 amino acid substitutions; (i) SEQ ID NO: 333 comprising 1, 2, 3, 4, or 5 amino acid substitutions; or (j) SEQ ID NO: 335 comprising 1, 2, 3, 4, or 5 amino acid substitutions. However, in multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker does not, or does not comprise, a sequence having at least about 95% sequence identity to SEQ ID NO: 335. In multiple embodiments, the amino acid substitutions can be conservative or non-conservative. In multiple embodiments, the amino acid substitutions can be conservative amino acid substitutions. In multiple embodiments, (i) the function of IL-15, such as but not limited to one or more signaling functions of IL-15, (ii) the ability of IL-15Rα to bind to a membrane and the function of IL-15Rα, such as but not limited to the signaling function of IL-15Rα, or (iii) both (i) and (ii) are preserved and / or enhanced in the mutant IL-15 / IL-15Rα fusion polypeptide.

[0384] In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker can have at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to SEQ ID NO: 317. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker can have at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to SEQ ID NO: 319. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker can have at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to SEQ ID NO: 321. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker can have at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to SEQ ID NO: 323. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker can have at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to SEQ ID NO: 325.In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker can comprise at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to SEQ ID NO: 327. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker can comprise at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to SEQ ID NO: 329. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker can comprise at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to SEQ ID NO: 331. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker can comprise at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to SEQ ID NO: 333. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker can comprise at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, or at least about 94% sequence identity to SEQ ID NO: 335.In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker does not include, or consists of, a sequence having at least about 95% sequence identity to SEQ ID NO: 335. In multiple embodiments, (i) the function of IL-15, such as, but not limited to, one or more signaling functions of IL-15, (ii) the ability of IL-15Rα to bind to a membrane and the function of IL-15Rα, such as, but not limited to, the signaling function of IL-15Rα, or (iii) both (i) and (ii) are preserved and / or enhanced in the IL-15 / IL-15Rα fusion polypeptide encoded by the mutant nucleic acid sequence.

[0385] In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 318. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 320. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 322. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 324.In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 326. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 328. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 330. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 332.In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 334. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, or at least about 94% sequence identity to the nucleic acid of SEQ ID NO: 336. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker is not encoded by a nucleic acid encoding a polypeptide having greater than about 95% sequence identity to SEQ ID NO: 335. In multiple embodiments, (i) the function of IL-15, such as, but not limited to, one or more signaling functions of IL-15, (ii) the ability of IL-15Rα to bind to a membrane and the function of IL-15Rα, such as, but not limited to, one or more signaling functions of IL-15Rα, or (iii) both (i) and (ii) are preserved and / or enhanced in an IL-15 / IL-15Rα fusion polypeptide encoded by a mutant nucleic acid sequence.

[0386] In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker can be encoded by a nucleic acid comprising (a) SEQ ID NO: 318 comprising 1, 2, 3, 4, or 5 nucleic acid substitutions, (b) SEQ ID NO: 320 comprising 1, 2, 3, 4, or 5 nucleic acid substitutions, (c) SEQ ID NO: 322 comprising 1, 2, 3, 4, or 5 nucleic acid substitutions, (d) SEQ ID NO: 324 comprising 1, 2, 3, 4, or 5 nucleic acid substitutions, (e) SEQ ID NO: 326 comprising 1, 2, 3, 4, or 5 nucleic acid substitutions, (f) SEQ ID NO: 328 comprising 1, 2, 3, 4, or 5 nucleic acid substitutions, (g) SEQ ID NO: 330 comprising 1, 2, 3, 4, or 5 nucleic acid substitutions, (h) SEQ ID NO: 332 comprising 1, 2, 3, 4, or 5 nucleic acid substitutions, or (i) SEQ ID NO: 334 comprising 1, 2, 3, 4, or 5 nucleic acid substitutions. In multiple embodiments, one or more nucleic acid substitutions in a codon may result in a codon encoding the same amino acid or may result in a codon encoding a different amino acid. In multiple embodiments, one or more nucleic acid substitutions in a codon may result in a codon encoding a conservative amino acid substitution. In multiple embodiments, one or more nucleic acid substitutions in a codon may result in a codon encoding the same amino acid. In multiple embodiments, (i) for example, but not limited to, the function of IL-15 such as one or more signaling functions of IL-15, (ii) for example, but not limited to, the ability of IL-15Rα to bind to a membrane and the function of IL-15Rα such as one or more signaling functions of IL-15Rα, or (iii) both (i) and (ii) are preserved and / or enhanced in an IL-15 / IL-15Rα fusion polypeptide encoded by a mutant nucleic acid sequence.

[0387] In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide optionally containing one or more linkers is, for example, SEQ ID NO: 307 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the N-terminus of SEQ ID NO: 309 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto), regardless of the presence or absence of a linker therebetween; SEQ ID NO: 307 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the N-terminus of SEQ ID NO: 311 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto), regardless of the presence or absence of a linker therebetween;SEQ ID NO: 307 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused, with or without a linker therebetween, to the N-terminus of SEQ ID NO: 313 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto); or SEQ ID NO: 307 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused, with or without a linker therebetween, to the N-terminus of SEQ ID NO: 315 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto);or comprising, or consisting of, a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, 333, or 335. However, in some embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker does not comprise, or consist of, (i) SEQ ID NO: 307 directly or indirectly fused to the N-terminus of SEQ ID NO: 309 with a linker therebetween, (ii) SEQ ID NO: 335, (iii) a sequence having at least about 95% sequence identity to SEQ ID NO: 307 directly or indirectly fused to the N-terminus of SEQ ID NO: 309 with a linker therebetween, or (iv) a sequence having at least about 95% sequence identity to SEQ ID NO: 335. In some embodiments, one or more linkers comprise, or consist of, a linker sequence described herein.;

[0388] In certain embodiments, an IL-15 / IL-15Rα fusion polypeptide optionally comprising one or more linkers is, for example, SEQ ID NO: 307 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused, with or without a linker therebetween, to the N-terminus of SEQ ID NO: 311 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto); SEQ ID NO: 307 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused, with or without a linker therebetween, to the N-terminus of SEQ ID NO: 313 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto);Or SEQ ID NO: 307 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused, with or without a linker therebetween, to the N-terminus of SEQ ID NO: 315 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto); or SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, or 333, or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, or 333, and includes or consists of. In multiple embodiments, one or more linkers include or consist of the linker sequences described herein.;

[0389] In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising one or more linkers includes or consists of, for example, SEQ ID NO: 317, 321, 325, 327, 329, 331, or 333, or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 317, 321, 325, 327, 329, 331, or 333.

[0390] In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide optionally containing one or more linkers is, for example, directly or indirectly fused at the 5' end of SEQ ID NO: 310 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) to SEQ ID NO: 308 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto), regardless of the presence or absence of a nucleic acid encoding the linker therebetween; at the 5' end of SEQ ID NO: 312 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) to SEQ ID NO: 308 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto), regardless of the presence or absence of a nucleic acid encoding the linker therebetween;At the 5' end of SEQ ID NO: 314 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto), SEQ ID NO: 308 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused, regardless of the presence or absence of a nucleic acid encoding the linker therebetween; or at the 5' end of SEQ ID NO: 316 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto), SEQ ID NO: 308 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused, regardless of the presence or absence of a nucleic acid encoding the linker therebetween;Or a nucleic acid encoding, or consisting of, a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 318, 320, 322, 324, 326, 328, 330, 332, 334, or 336. However, in some embodiments, the IL-15 / IL-15Rα fusion polypeptide is not encoded by a nucleic acid comprising, or consisting of, (i) SEQ ID NO: 308 fused to the 5' end of SEQ ID NO: 310 with a linker therebetween, (ii) SEQ ID NO: 336, (iii) a sequence having at least about 80%, about 85%, about 90%, or about 95% sequence identity to SEQ ID NO: 308 fused to the 5' end of SEQ ID NO: 310 with a linker therebetween, or (iv) a sequence having at least about 80%, about 85%, about 90%, or about 95% sequence identity to SEQ ID NO: 336. In some embodiments, one or more linkers are encoded by one or more nucleic acids comprising, or consisting of, nucleic acids encoding the linkers described herein.;

[0391] In certain embodiments, an IL-15 / IL-15Rα fusion polypeptide optionally comprising one or more linkers is, for example, SEQ ID NO: 308 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused, with or without a nucleic acid encoding a linker therebetween, to the 5' end of SEQ ID NO: 312 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto); SEQ ID NO: 308 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused, with or without a nucleic acid encoding a linker therebetween, to the 5' end of SEQ ID NO: 314 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto);or SEQ ID NO: 308 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto), directly or indirectly fused, with or without a nucleic acid encoding a linker therebetween, to the 5' end of SEQ ID NO: 316 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto); or a nucleic acid comprising, or consisting of, SEQ ID NO: 318, 320, 322, 324, 326, 328, 330, 332, or 334, or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 318, 320, 322, 324, 326, 328, 330, 332, or 334. In multiple embodiments, one or more linkers are encoded by one or more nucleic acids comprising, or consisting of, a nucleic acid encoding a linker described herein.;

[0392] In some embodiments, the IL-15 / IL-15Rα fusion polypeptide is encoded by a nucleic acid comprising, or consisting of, for example, a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 318, 322, 326, 328, 330, 332, or 334.

[0393] In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker (L) comprises, or consists of, any of constructs A-J described in FIG. 69A. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker (L) comprises, or consists of, any of constructs A-I described in FIG. 69A. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker comprises, or consists of, any of constructs A, C, or E-I described in FIG. 69A. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker is encoded by one or more nucleic acids comprising, or consisting of, any of constructs A'-J' described in FIG. 69B. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker is encoded by one or more nucleic acids comprising, or consisting of, any of constructs A'-I' described in FIG. 69B. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker is encoded by one or more nucleic acids comprising, or consisting of, any of constructs A', C', or E'-I' described in FIG. 69B. In multiple embodiments, sequences are provided that comprise at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identity to any of the sequences described in FIGS. 69A and 69B. In FIGS. 69A and 69B, the lines connecting IL-15 to the linker and the linker to IL-15Rα may represent a direct bond with no intervening sequence, or may represent an intervening sequence such as, but not limited to, a linker, an untranslated sequence (in the case of FIG. 69B), a translated sequence, a sequence comprising one or more restriction endonuclease sites (in the case of FIG. 69B), or a combination thereof.

[0394] In multiple embodiments, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide may contain a stop codon (e.g., TAA, TAG, or TGA), and these stop codons are, as non-limiting examples, located at the 3' end of the nucleotides encoding the IL-15Rα polypeptide. For example, in this case, the encoded fusion polypeptide is in the orientation shown in Figure 67A or Figure 668A, or at the 3' end of the IL-15 polypeptide. For example, in this case, the encoded fusion polypeptide is in the orientation shown in Figure 67B or Figure 668B.

[0395] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide and / or the IL-15Rα / IL-15 fusion polypeptide may contain one or more signal peptides. In multiple embodiments, the fusion polypeptide may contain all or part of the short or long signal peptide of IL-15, or all or part of the signal peptide of IL-15Rα. In multiple embodiments, all or part of the signal peptides of IL-15, IL-15Rα, or both may be mutated or deleted. In multiple embodiments, the fusion polypeptide may contain all or part of one or more heterologous signal peptides, i.e., signal peptides derived from molecules other than IL-15 and IL-15Rα. In multiple embodiments, the heterologous signal peptide may be derived from IL-2, CD33, IgV K , or IgE. In multiple embodiments, the signal peptide may be a signal peptide derived from IgE. In multiple embodiments, the signal peptide derived from IgE may contain or consist of SEQ ID NO: 367. In multiple embodiments, the signal peptide derived from IgE may be encoded by a nucleic acid containing or consisting of the sequence described in SEQ ID NO: 368.

[0396] In multiple embodiments, the signal peptide may be cleaved from the IL-15 / IL-15Rα fusion polypeptide or removed by other means.

[0397] In multiple embodiments, the signal peptide can increase or facilitate the transcription, translation, translocation, or combinations thereof of the fusion polypeptide as compared to the native IL-15Rα signal peptide, the native IL-15 signal peptide, or both. In multiple embodiments, the signal peptide can be fused directly or indirectly to the N-terminus or C-terminus of the IL-15 / IL-15Rα fusion polypeptide.

[0398] In multiple embodiments, the signal peptide has a sequence that includes at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% sequence identity to the amino acid sequence of SEQ ID NO: 367. In multiple embodiments, one or more signaling functions of the signal peptide, such as, but not limited to, signaling functions of the signal peptide, are conserved and / or enhanced in the variant signal peptide.

[0399] In multiple embodiments, the signal peptide can include SEQ ID NO: 367 and include 1, 2, 3, 4, or 5 amino acid substitutions. In multiple embodiments, one or more signaling functions of the signal peptide, such as, but not limited to, signaling functions of the signal peptide, are conserved and / or enhanced in the variant signal peptide.

[0400] In multiple embodiments, the signal peptide can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 368. In multiple embodiments, the function of the signal peptide, e.g., but not limited to, one or more signaling functions of the signal peptide, is conserved and / or enhanced in the signal peptide encoded by the variant nucleic acid sequence.

[0401] In multiple embodiments, the signal peptide can be encoded by a nucleic acid comprising SEQ ID NO: 368 and containing 1, 2, 3, 4, or 5 nucleic acid substitutions. In multiple embodiments, one or more nucleic acid substitutions in a codon may result in a codon encoding the same amino acid or may result in a codon encoding a different amino acid. In multiple embodiments, one or more nucleic acid substitutions in a codon may result in a codon encoding a conservative amino acid substitution. In multiple embodiments, one or more nucleic acid substitutions in a codon may result in a codon encoding the same amino acid. In multiple embodiments, the function of the signal peptide, e.g., but not limited to, one or more signaling functions of the signal peptide, is conserved and / or enhanced in the signal peptide encoded by the variant nucleic acid sequence.

[0402] In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE may comprise at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to SEQ ID NO: 337. In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE may comprise at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to SEQ ID NO: 339. In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE may comprise at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to SEQ ID NO: 341. In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE may comprise at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to SEQ ID NO: 343.In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE can comprise at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to SEQ ID NO: 345. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE can comprise at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to SEQ ID NO: 347. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE can comprise at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to SEQ ID NO: 349. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE can comprise at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to SEQ ID NO: 351.In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE may have at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to SEQ ID NO: 353. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE may have at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to SEQ ID NO: 355. However, in multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE does not include, or consists of, SEQ ID NO: 355 or a sequence having at least about 95% sequence identity to SEQ ID NO: 355. In multiple embodiments, (i) the function of IL-15, such as, but not limited to, one or more signaling functions of IL-15, (ii) the function of IL-15Rα, such as, but not limited to, the ability of IL-15Rα to bind to a membrane and one or more signaling functions of IL-15Rα, (iii) the function of the signal peptide derived from IgE, such as one or more signaling functions of the signal peptide, or all of (i), (ii), and (iii) are conserved and / or enhanced in a mutant IL-15 / IL-15Rα fusion polypeptide comprising a signal peptide derived from IgE.

[0403] In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE may comprise: (a) SEQ ID NO: 337 with 1, 2, 3, 4, or 5 amino acid substitutions; (b) SEQ ID NO: 339 with 1, 2, 3, 4, or 5 amino acid substitutions; (c) SEQ ID NO: 341 with 1, 2, 3, 4, or 5 amino acid substitutions; (d) SEQ ID NO: 343 with 1, 2, 3, 4, or 5 amino acid substitutions; (e) SEQ ID NO: 345 with 1, 2, 3, 4, or 5 amino acid substitutions; (f) SEQ ID NO: 347 with 1, 2, 3, 4, or 5 amino acid substitutions; (g) SEQ ID NO: 349 with 1, 2, 3, 4, or 5 amino acid substitutions; (h) SEQ ID NO: 351 with 1, 2, 3, 4, or 5 amino acid substitutions; (i) SEQ ID NO: 353 with 1, 2, 3, 4, or 5 amino acid substitutions; or (j) SEQ ID NO: 355 with 1, 2, 3, 4, or 5 amino acid substitutions. However, in multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE does not, or does not comprise, a sequence having at least about 95% sequence identity to SEQ ID NO: 355. In multiple embodiments, the amino acid substitutions can be conservative or non-conservative. In multiple embodiments, the amino acid substitutions can be conservative amino acid substitutions. In multiple embodiments, (i) the function of IL-15, such as, but not limited to, one or more signaling functions of IL-15, (ii) the function of IL-15Rα, such as, but not limited to, the ability of IL-15Rα to bind to the membrane and one or more signaling functions of IL-15Rα, (iii) the function of the signal peptide derived from IgE, such as one or more signaling functions of the signal peptide, or all of (i), (ii), and (iii) are preserved and / or enhanced in a mutant IL-15 / IL-15Rα fusion polypeptide comprising a signal peptide derived from IgE.

[0404] In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 338. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 340. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 342. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 344.In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 346. In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 348. In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 350. In multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 352.In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 354. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 356. However, in multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE is not encoded by SEQ ID NO: 356 or a sequence having at least about 80%, about 85%, about 90% or about 95% or more sequence identity to SEQ ID NO: 356. In multiple embodiments, (i) the function of IL-15, such as but not limited to, one or more signaling functions of IL-15, (ii) the function of IL-15Rα, such as but not limited to, the ability of IL-15Rα to bind to the membrane, and one or more signaling functions of IL-15Rα, (iii) the function of the signal peptide derived from IgE, such as, one or more signaling functions of the signal peptide, or all of (i), (ii), and (iii) are preserved and / or enhanced in an IL-15 / IL-15Rα fusion polypeptide comprising a signal peptide derived from IgE that is encoded by a variant nucleic acid sequence.

[0405] In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE can be encoded by a nucleic acid comprising (a) SEQ ID NO: 338 with 1, 2, 3, 4, or 5 nucleic acid substitutions, (b) SEQ ID NO: 340 with 1, 2, 3, 4, or 5 nucleic acid substitutions, (c) SEQ ID NO: 342 with 1, 2, 3, 4, or 5 nucleic acid substitutions, (d) SEQ ID NO: 344 with 1, 2, 3, 4, or 5 nucleic acid substitutions, (e) SEQ ID NO: 346 with 1, 2, 3, 4, or 5 nucleic acid substitutions, (f) SEQ ID NO: 348 with 1, 2, 3, 4, or 5 nucleic acid substitutions, (g) SEQ ID NO: 350 with 1, 2, 3, 4, or 5 nucleic acid substitutions, (h) SEQ ID NO: 352 with 1, 2, 3, 4, or 5 nucleic acid substitutions, (i) SEQ ID NO: 354 with 1, 2, 3, 4, or 5 nucleic acid substitutions, or (j) SEQ ID NO: 356 with 1, 2, 3, 4, or 5 nucleic acid substitutions. However, in multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE is not encoded by SEQ ID NO: 356 or a sequence having at least about 80%, about 85%, about 90%, or about 95% sequence identity to SEQ ID NO: 356. In multiple embodiments, one or more nucleic acid substitutions in a codon may result in a codon encoding the same amino acid or may result in a codon encoding a different amino acid. In multiple embodiments, one or more nucleic acid substitutions in a codon may result in a codon encoding a conservative amino acid substitution. In multiple embodiments, (i) the function of IL-15, such as, but not limited to, one or more signaling functions of IL-15, (ii) the function of IL-15Rα, such as, but not limited to, the ability of IL-15Rα to bind to a membrane and one or more signaling functions of IL-15Rα, (iii) the function of the signal peptide derived from IgE, such as one or more signaling functions of the signal peptide, or all of (i), (ii), and (iii) are conserved and / or enhanced in an IL-15 / IL-15Rα fusion polypeptide comprising a signal peptide derived from IgE and encoded by a variant nucleic acid sequence.

[0406] In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a signal peptide derived from IgE and optionally one or more linkers, regardless of the presence or absence of a linker therebetween, is directly or indirectly fused to the N-terminus of SEQ ID NO: 307 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) which is directly or indirectly fused to the N-terminus of SEQ ID NO: 367 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto);Regardless of the presence or absence of the linker in between, SEQ ID NO: 367 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the N-terminus of SEQ ID NO: 307 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the N-terminus of SEQ ID NO: 311 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto);Regardless of the presence or absence of the linker in between, SEQ ID NO: 367 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the N-terminus of SEQ ID NO: 307 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the N-terminus of SEQ ID NO: 313 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto);With or without a linker therebetween, SEQ ID NO: 367 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the N-terminus of SEQ ID NO: 307 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the N-terminus of SEQ ID NO: 315 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto);or comprises, or consists of, a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 337, 339, 341, 343, 345, 347, 349, 351, 353, or 355. However, in some embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising an IgE-derived signal peptide and optionally one or more linkers does not comprise, or consist of, (i) SEQ ID NO: 307 fused directly or indirectly to the N-terminus of SEQ ID NO: 309, including the linker therebetween; (ii) SEQ ID NO: 335 or SEQ ID NO: 355; (iii) a sequence having at least about 95% sequence identity to SEQ ID NO: 307 fused directly or indirectly to the N-terminus of SEQ ID NO: 309, including the linker therebetween; or (iv) a sequence having at least about 95% sequence identity to SEQ ID NO: 335 or SEQ ID NO: 355. In some embodiments, one or more linkers comprise, or consist of, a linker sequence described herein.;

[0407] In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide that contains a signal peptide derived from IgE and optionally contains one or more linkers is, for example, directly or indirectly fused to the N-terminus of SEQ ID NO: 307 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto), directly or indirectly fused to the N-terminus of SEQ ID NO: 311 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto), and directly or indirectly fused to the N-terminus of SEQ ID NO: 367 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto), regardless of the presence or absence of a linker therebetween;Regardless of the presence or absence of the linker in between, SEQ ID NO: 367 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the N-terminus of SEQ ID NO: 307 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the N-terminus of SEQ ID NO: 313 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto);With or without a linker therebetween, SEQ ID NO: 367 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the N-terminus of SEQ ID NO: 307 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the N-terminus of SEQ ID NO: 315 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto); or SEQ ID NO: 337, 339, 341, 343, 345, 347, 349, 351, or 353, or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 337, 339, 341, 343, 345, 347, 349, 351, or 353, comprising or consisting of. In a plurality of embodiments, one or more linkers comprise or consist of the linker sequences described herein.;

[0408] In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide that includes a signal peptide derived from IgE and includes a linker comprises, or consists of, for example, SEQ ID NO: 337, 341, 345, 347, 349, 351, or 353, or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 337, 341, 345, 347, 349, 351, or 353.

[0409] In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide that includes a signal peptide derived from IgE and optionally one or more linkers is SEQ ID NO: 368 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the 5' end of SEQ ID NO: 308 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the 5' end of SEQ ID NO: 310 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto), regardless of the presence or absence of nucleic acids encoding linkers therebetween;Regardless of the presence or absence of nucleic acid encoding the linker in between, SEQ ID NO: 368 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the 5' end of SEQ ID NO: 308 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the 5' end of SEQ ID NO: 312 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto);Regardless of the presence or absence of the nucleic acid encoding the linker in between, SEQ ID NO: 368 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the 5' end of SEQ ID NO: 308 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the 5' end of SEQ ID NO: 314 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto);Or SEQ ID NO: 368 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the 5' end of SEQ ID NO: 308 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the 5' end of SEQ ID NO: 316 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto); or a nucleic acid comprising, or consisting of, SEQ ID NO: 338, 340, 342, 344, 346, 348, 350, 352, 354, or 356, or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 338, 340, 342, 344, 346, 348, 350, 352, 354, or 356.

[0410] However, in multiple embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE is not encoded by a nucleic acid comprising, or consisting of, (i) SEQ ID NO: 356, or a sequence having at least about 80%, about 85%, about 90% or about 95% sequence identity thereto, or (ii) SEQ ID NO: 368 directly or indirectly fused to the 5' end of SEQ ID NO: 308 directly or indirectly fused to the 5' end of SEQ ID NO: 310, or a sequence having at least about 80%, about 85%, about 90% or about 95% sequence identity thereto. In multiple embodiments, one or more linkers are encoded by one or more nucleic acids comprising, or consisting of, a nucleic acid encoding a linker described herein.

[0411] In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide that includes a signal peptide derived from IgE and optionally one or more linkers is, for example, SEQ ID NO: 308 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the 5' end of SEQ ID NO: 312 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto), with or without a nucleic acid encoding a linker therebetween; and SEQ ID NO: 368 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the 5' end of SEQ ID NO: 308 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto).Regardless of the presence or absence of nucleic acid encoding the linker therebetween, SEQ ID NO: 368 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the 5' end of SEQ ID NO: 308 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the 5' end of SEQ ID NO: 314 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto);Or SEQ ID NO: 368 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the 5' end of SEQ ID NO: 308 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto), which is directly or indirectly fused to the 5' end of SEQ ID NO: 316 (or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto); or SEQ ID NO: 338, 340, 342, 344, 346, 348, 350, 352, or 354, or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 338, 340, 342, 344, 346, 348, 350, 352, or 354, and is encoded by a nucleic acid comprising or consisting of such a sequence. In a plurality of embodiments, one or more linkers are encoded by one or more nucleic acids comprising or consisting of nucleic acids encoding the linkers described herein.;

[0412] In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide that includes a signal peptide derived from IgE and includes one or more linkers is encoded by a nucleic acid that comprises, or consists of, for example, the sequence of SEQ ID NO: 338, 342, 346, 348, 350, 352, or 354, or a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 338, 342, 346, 348, 350, 352, or 354.

[0413] In multiple embodiments, the vector may further comprise a post-transcriptional regulatory element (PRE) sequence. In multiple embodiments, the post-transcriptional regulatory element (PRE) sequence can be selected from a woodchuck hepatitis virus PRE (WPRE) (e.g., without limitation, wild-type WPRE such as SEQ ID NO: 264, or mutant WPRE such as, without limitation, WPREmut1 (SEQ ID NO: 256) or WPREmut2 (SEQ ID NO: 257)), or a hepatitis B virus (HBV) PRE (HPRE) (SEQ ID NO: 437), variants thereof, or any combination thereof.

[0414] In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising (i) a linker and a signal peptide derived from IgE, and (ii) WPREmut2 following it, can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 357. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising (i) a linker and a signal peptide derived from IgE, and (ii) WPREmut2 following it, can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 358. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising (i) a linker and a signal peptide derived from IgE, and (ii) WPREmut2 following it, can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 359. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising (i) a linker and a signal peptide derived from IgE, and (ii) WPREmut2 following it, can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 360.In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising (i) a linker and a signal peptide derived from IgE, and (ii) WPREmut2 following it, can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 361. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising (i) a linker and a signal peptide derived from IgE, and (ii) WPREmut2 following it, can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 362. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising (i) a linker and a signal peptide derived from IgE, and (ii) WPREmut2 following it, can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 363. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising (i) a linker and a signal peptide derived from IgE, and (ii) WPREmut2 following it, can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 364.In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising (i) a linker and a signal peptide derived from IgE, and (ii) followed by WPREmut2 can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 365. In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising (i) a linker and a signal peptide derived from IgE, and (ii) followed by WPRE can be encoded by a nucleic acid having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% sequence identity to the nucleic acid sequence of SEQ ID NO: 366. However, in multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising (i) a linker and a signal peptide derived from IgE, and (ii) followed by a mutant or wild-type WPRE is not encoded by SEQ ID NO: 366 or a sequence having at least about 80%, about 85%, about 90% or about 95% or more sequence identity to SEQ ID NO: 366. In multiple embodiments, (i) the function of IL-15, such as but not limited to, one or more signaling functions of IL-15, (ii) the function of IL-15Rα, such as but not limited to, the ability of IL-15Rα to bind to the membrane and the signaling function of IL-15Rα, (iii) the function of the signal peptide derived from IgE, such as one or more signaling functions of the signal peptide, (iv) the post-transcriptional regulatory function of wild-type or mutant WPRE, or all of (i), (ii), (iii) and (iv) are preserved and / or enhanced in an IL-15 / IL-15Rα fusion polypeptide encoded by a mutant nucleic acid sequence and comprising a signal peptide derived from IgE.

[0415] In multiple embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising (i) a linker and a signal peptide derived from IgE, and (ii) followed by WPREmut2 or wild-type WPRE (wt if shown) can be encoded by a nucleic acid comprising (a) SEQ ID NO: 357 containing 1, 2, 3, 4, or 5 nucleic acid substitutions, (b) SEQ ID NO: 358 containing 1, 2, 3, 4, or 5 nucleic acid substitutions, (c) SEQ ID NO: 359 containing 1, 2, 3, 4, or 5 nucleic acid substitutions, (d) SEQ ID NO: 360 containing 1, 2, 3, 4, or 5 nucleic acid substitutions, (e) SEQ ID NO: 361 containing 1, 2, 3, 4, or 5 nucleic acid substitutions, (f) SEQ ID NO: 362 containing 1, 2, 3, 4, or 5 nucleic acid substitutions, (g) SEQ ID NO: 363 c...

Claims

1. A nucleic acid encoding a polypeptide containing SEQ ID NO: 313, 311, or 315, or a polypeptide that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 313, 311, or 315, wherein the nucleic acid optionally contains a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 314, 312, or 316.

2. The nucleic acid according to claim 1, encoding (i) a polypeptide comprising SEQ ID NO: 307 directly or indirectly fused to the N-terminus of a polypeptide comprising SEQ ID NO: 313, 311, or 315, or (ii) a polypeptide that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 307, directly or indirectly fused to a polypeptide that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to any of SEQ ID NO: 313, 311, or 315.

3. A polypeptide comprising sequence numbers 329, 317, 321, 325, 327, 331, or 333, or a polypeptide that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to sequence numbers 329, 317, 321, 325, 327, 331, or 333, wherein the nucleic acid is optionally The nucleic acid according to claim 1 or 2, comprising a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the nucleic acid of sequence number 330, 318, 320, 322, 324, 326, 328, 332, or 334.

4. The nucleic acid according to claim 3, further comprising a nucleic acid encoding a signal peptide directly or indirectly fused to the N-terminus of SEQ ID NOs. 329, 317, 321, 325, 327, 331, or 333, or to the N-terminus of a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NOs. 329, 317, 321, 325, 327, 331, or 333.

5. The nucleic acid according to claim 4, wherein the signal peptide is derived from an IgE polypeptide and optionally comprises a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 367, and the nucleic acid comprises a sequence that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO:

368.

6. The nucleic acid according to claim 4 or 5, wherein the nucleic acid encodes a polypeptide comprising sequence number 349, 337, 341, 345, 347, 351 or 353, or a polypeptide that is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to sequence number 349, 337, 341, 345, 347, 351 or 353, and the nucleic acid optionally comprises a sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to sequence number 350, 338, 342, 346, 348, 352, or 354.

7. (a) at least one TCR polypeptide comprising a TCRα chain and a TCRβ chain, (b) At least one CD8 polypeptide comprising (i) a CD8α chain, (ii) a CD8β chain, or (iii) both a CD8α chain and a CD8β chain, (c) The nucleic acid according to any one of claims 1 to 6, further comprising a nucleic acid encoding at least one TCR polypeptide comprising a TCRα chain and a TCRβ chain, and (i) a CD8α chain, (ii) a CD8β chain, or (iii) both a CD8α chain and a CD8β chain.

8. The nucleic acid according to claim 7, comprising nucleic acids that are at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the nucleic acids of sequence numbers 438, 267, 269, 271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291, 295, 297, 299, 301, 439, 440, 441, 442, 443, 444, 445, 446, or 447.

9. (a) Nucleic acids, (i) T cell receptors (TCRs) comprising TCRα chains and TCRβ chains, and CD8 polypeptides comprising CD8α chains and CD8β chains, or (ii) Nucleic acids encoding a TCR including a TCRα chain and a TCRβ chain, and a CD8 polypeptide that does not contain a CD8β chain but contains a CD8α chain, and (b) A nucleic acid comprising a fusion polypeptide according to any one of claims 3 or 6, The α chain and β chain of the TCR are sequence numbers 15 and 16, 17 and 18, 19 and 20, 21 and 22, 23 and 24, 25 and 26, 27 and 28, 29 and 30, 31 and 32, 33 and 34, 35 and 36, 37 and 38, 39 and 40, 41 and 42, 43 and 44, 45 and 46, 47 and 48, 49 and 50, and 51. and 52, 53 and 54, 55 and 56, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, 71 and 303, 304 and 74, 75 and 76, 77 and 78, 79 and 80, 81 and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90, and 91 and 92 are selected, The α chain of CD8 is sequence numbers 7, 258, 259, 262, or a variant thereof. The nucleic acid according to any one of claims 1 to 8, wherein, if present, the β-chain of CD8 is sequence number 8, 9, 10, 11, 12, 13, or 14.

10. A polypeptide or fusion polypeptide encoded by the nucleic acid according to any one of claims 1 to 7.

11. A vector comprising the nucleic acid described in any one of claims 1 to 9.

12. The following formula III or formula IV: 5'-N1-L1-N2-L2-N3-L3-N4-L4-N5-3' [III] 5'-N5-L1-N1-L2-N2-L3-N3-L4-N4-3'[IV], including During the ceremony, N1 contains, and may or may not contain, the nucleic acid encoding the CD8β chain. N2 contains nucleic acid encoding the CD8α chain, N3 contains nucleic acid that encodes the TCRβ chain, N4 contains nucleic acid encoding the TCRα chain, and N5 comprises a nucleic acid encoding an IL-15 / IL-15Rα fusion polypeptide as defined in any one of claims 1 to 3 to 6, The vector according to claim 11, wherein each of L1 to L4 comprises a nucleic acid encoding at least one linker, each of L1 to L4 is independently identical or different, and each of L1 to L4 is independently present or absent.

13. The vector according to any one of claims 11 or 12, wherein the vector is a viral vector or a non-viral vector.

14. The vector according to any one of claims 11 to 13, wherein the viral vector is selected from adenovirus, poxvirus, alphavirus, arenavirus, flavivirus, rhabdovirus, retrovirus, lentivirus, herpesvirus, paramyxovirus, picornavirus, and any combination thereof.

15. The vector according to any one of claims 11 to 14, wherein the vector further comprises a nucleic acid encoding a chimeric antigen receptor (CAR).

16. An in vitro method for preparing T cells and / or natural killer cells for immunotherapy, To prepare isolated T cells and / or natural killer cells derived from human blood samples, Activating the isolated T cells and / or natural killer cells, Transduction of the activated T cells and / or natural killer cells with the nucleic acid according to any one of claims 1 to 9, or the vector according to any one of claims 11 to 15, and A method comprising amplifying the transduced T cells and / or natural killer cells.

17. The method according to claim 16, wherein the blood sample comprises peripheral blood mononuclear cells (PMBCs).

18. The method according to claim 16 or 17, wherein the activation comprises contacting the T cells and / or natural killer cells with an anti-CD3 antibody and an anti-CD28 antibody, and / or (ii) the activation, amplification, or both are optionally carried out using zoledronic acid in the presence of a combination of IL-2 and IL-15.

19. (i) T cells and / or natural killer cells transduced to the nucleic acid described in any one of claims 1 to 9, or (ii) comprising the vector described in any one of claims 11 to 15, or (iii) obtained by the method described in any one of claims 16 to 18.

20. The T cell and / or natural killer cell according to claim 19, wherein the T cell is an αβ T cell, a γδ T cell, a natural killer T cell, or any combination thereof.

21. The T cell and / or natural killer cell according to claim 19 or 20, wherein the αβ T cell is a CD4+ T cell or a CD8+ T cell.

22. A composition comprising T cells and / or natural killer cells as described in claim 19.

23. The composition according to claim 22, wherein the composition is a pharmaceutical composition.

24. The composition according to claim 23, wherein the composition further comprises an adjuvant, an excipient, a carrier, a diluent, a buffer, a stabilizer, or a combination thereof.

25. The composition according to claim 24, wherein the adjuvant is an anti-CD40 antibody, imiquimod, reciquimod, GM-CSF, cyclophosphamide, sunitinib, bevacizumab, atezolizumab, interferon alpha, interferon beta, CpG oligonucleotide and its derivatives, poly(I:C) and its derivatives, RNA, sildenafil, poly(lactidocoglycolide) (PLG) particle formulation, virosom, interleukin 1 (IL-1), interleukin 2 (IL-2), interleukin 4 (IL-4), interleukin 7 (IL-7), interleukin 12 (IL-12), interleukin 13 (IL-13), interleukin 15 (IL-15), interleukin 21 (IL-21), interleukin 23 (IL-23), or any combination thereof.

26. A drug comprising, for use as a pharmaceutical, a nucleic acid according to any one of claims 1 to 9, a polypeptide according to claim 10, a vector according to any one of claims 11 to 15, a T cell and / or natural killer cell according to any one of claims 19 to 21, or a composition according to any one of claims 22 to 25.

27. A drug for treating cancer, comprising a nucleic acid according to any one of claims 1 to 9, a polypeptide according to claim 10, a vector according to any one of claims 11 to 15, a T cell and / or natural killer cell according to any one of claims 19 to 21, or a composition according to any one of claims 22 to 25, wherein the cancer is optionally selected from the group consisting of non-small cell lung cancer, small cell lung cancer, melanoma, liver cancer, breast cancer, uterine cancer, Merkel cell carcinoma, pancreatic cancer, gallbladder cancer, bile duct cancer, colorectal cancer, bladder cancer, kidney cancer, leukemia, ovarian cancer, esophageal cancer, brain tumor, gastric cancer, and prostate cancer.

28. Use of a nucleic acid according to any one of claims 1 to 9, a polypeptide according to claim 10, a vector according to any one of claims 11 to 15, a T cell and / or natural killer cell according to any one of claims 19 to 21, a composition according to any one of claims 22 to 25, or a drug according to claim 26, wherein the cancer is optionally selected from the group consisting of non-small cell lung cancer, small cell lung cancer, melanoma, liver cancer, breast cancer, uterine cancer, Merkel cell carcinoma, pancreatic cancer, gallbladder cancer, bile duct cancer, colorectal cancer, bladder cancer, kidney cancer, leukemia, ovarian cancer, esophageal cancer, brain tumor, gastric cancer, and prostate cancer.

29. Use as a pharmaceutical product of the nucleic acid according to any one of claims 1 to 9, the polypeptide according to claim 10, the vector according to any one of claims 11 to 15, the T cell and / or natural killer cell according to any one of claims 19 to 21, the composition according to any one of claims 22 to 25, or the drug according to claim 26.

30. The nucleic acid, vector, T cells and / or natural killer cells, polypeptide, or fusion polypeptide according to any one of claims 1 to 15 or 19 to 21, wherein the nucleic acid, vector, T cells and / or natural killer cells, polypeptide, or fusion polypeptide is isolated, recombinant, or both isolated and recombinant.

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