Membrane-binding IL-15, CD8 polypeptides, cells, compositions, and methods of use thereof

Abstract

The present disclosure relates to cells capable of co-expressing T cell receptors ("TCRs") with membrane-binding IL-15 polypeptides and / or CD8 polypeptides and their use in adoptive cell therapy. The disclosure further provides modified IL-15, IL-15R [alpha], IL-15 / IL-15R [alpha] fusion polypeptides, and IL-15R [alpha] / IL-15 fusion polypeptide sequences, vectors, and related methods of making and using the same. The disclosure further provides modified CD8 sequences, vectors, and related methods of making and using the same.

Description

[0001] Cross-reference to related applications

[0002] 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 contents of which are hereby incorporated by reference for all purposes.

[0003] Reference to a sequence listing submitted electronically

[0004] An official copy of the sequence listing is submitted simultaneously via EFS-Web as a sequence listing in ASCII format, with the file name "3000011-029977_Sequence-Listing_ST26", created on April 27, 2023 and 722,707 bytes in size, and submitted simultaneously with this specification. The sequence listing contained in this ASCII formatted document is part of this specification and is incorporated herein by reference in its entirety. Technical Field

[0005] The present disclosure relates to cells capable of co-expressing 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, vectors, compositions, transformed cells and related methods thereof. Background Art

[0006] CD8 and CD4 are transmembrane glycoproteins specific to different populations of T lymphocytes, whose antigenic responses are restricted by class I and class II MHC molecules, respectively. They play a major role in both the differentiation and selection of T cells during thymic development and the activation of mature T lymphocytes that respond to antigen presenting cells. Both CD8 and CD4 are immunoglobulin superfamily proteins. They determine antigenic restriction by binding to MHC molecules at an interface different from the region presenting the antigenic peptide, but the structural basis of their similar functions appears to be very different. Their sequence similarity is very low, and while CD4 is expressed on the cell surface as a monomer, CD8 is expressed as an αα homodimer (e.g., Fig.55C ) or αβ heterodimer expression (e.g., Fig.55A ). In humans, this CD8αα homodimer may functionally replace the CD8αβ heterodimer. CD8 contacts the acidic loop in the α3 domain of class I MHC, thereby increasing the avidity of T cells for their targets. CD8 also binds to the tyrosine kinase p56 through its α chain cytoplasmic tail. lck The association of β-actin with β-actin involves phosphorylation events leading to CTL activation.

[0007] The pleiotropic cytokine interleukin-15 ("IL-15" or "IL15") is a member of the 4α-helical bundle cytokine family. (Waldmann TA and Tagaya Y, Ann. Rev. Immunol. 17:19-49, 1999, the contents of which are incorporated herein by reference). The 14-15 kDa glycoprotein wild-type IL-15 shares partial structural homology with IL-2. (Ibid.). Wild-type IL-15 can be expressed in two isoforms, one with a signal peptide of 48 amino acids and the other with a signal peptide of 21 amino acids. (Ibid.). The mature form of wild-type IL-15 consists of 114 amino acids. (Ibid.). Wild-type IL-15 expression is regulated at the transcription, translation, and intracellular trafficking levels. (Ibid.). Wild-type IL-15 utilizes a private receptor, IL-15Rα (or “IL15Rα”), which binds IL-15 with high affinity in lymphocytes and binds to IL-2Rβ (also known as IL-2 / IL-15Rβ) and IL-2Rγ (also known as γ c ) trimerization. (Same as above; Okada S et al., Immunol. and Cell Biol. 93:461-471, 2015, the contents of which are incorporated herein by reference). Wild-type IL-15Rα comprises a signal peptide, an extracellular domain, a transmembrane domain, and a cytoplasmic domain. (Waldman and Tagaya). The extracellular domain of wild-type IL-15Rα comprises a sushi domain (also known as a GP-1 motif). (Same as above).

[0008] Adoptive cell therapy (ACT) is a promising method for treating diseases such as cancer. T cell therapy has successfully treated various cancers. Li et al. Signal Transduction and Targeted Therapy 4(35):(2019), which is incorporated by reference in its entirety. However, the cells used in ACT often fail to persist in the tumor microenvironment and quickly lose their ability to kill tumor cells. Therefore, T cells and natural killer cells that exhibit longer persistence and / or sustained ability to kill tumor cells in the tumor microenvironment are needed. It is also desirable to develop methods for preparing T cells and natural killer cells with enhanced specific cytotoxic activity for immunotherapy. Summary of the invention

[0009] In an embodiment, a membrane-bound IL-15 polypeptide (membrane-bound IL-15 or mbIL-15) may be provided. In an embodiment, a nucleic acid described herein may comprise and / or encode a membrane-bound IL-15 polypeptide. In an embodiment, a vector described herein may comprise and / or encode a membrane-bound IL-15 polypeptide. In an embodiment, a cell described herein may comprise and / or express a membrane-bound IL-15 polypeptide. In an embodiment, a composition described herein may comprise a membrane-bound IL-15 polypeptide or may comprise a cell comprising and / or expressing a membrane-bound IL-15 polypeptide. In an embodiment, IL-15 may be membrane-bound by expressing an IL-15 polypeptide and an IL-15Rα polypeptide in an IL-15 / IL-15Rα fusion polypeptide (IL-15 / IL-15Rα). Other membrane-bound forms of IL-15 / IL-15Rα fusion polypeptides and IL-15 may be referred to as membrane-bound IL-15 (mbIL-15).

[0010] In embodiments, isolated membrane-bound IL-15 polypeptides may be provided. Isolated nucleic acid sequences may be provided, comprising one or more nucleic acid sequences encoding one or more membrane-bound IL-15 polypeptides. In embodiments, isolated vectors may be provided, comprising one or more nucleic acid sequences encoding one or more membrane-bound IL-15 polypeptides. In embodiments, cells may be provided, comprising and / or expressing one or more membrane-bound IL-15 polypeptides. In embodiments, cells may be provided, comprising or expressing one or more nucleic acid sequences encoding one or more membrane-bound IL-15 polypeptides. In embodiments, cells may be provided, comprising or expressing one or more vectors comprising one or more nucleic acid sequences encoding one or more membrane-bound IL-15 polypeptides. In embodiments, compositions may be provided, comprising such polypeptides, nucleic acids, vectors and / or cells.

[0011] In an embodiment, the IL-15 polypeptide may be located at the N-terminus of the IL-15Rα polypeptide in the membrane-bound IL-15 polypeptide. Fig.67A In an embodiment, the IL-15 polypeptide may be located at the C-terminus of the IL-15Rα polypeptide in the membrane-bound IL-15 polypeptide. Fig.67B ). Fig.67A and 67B The IL-15 polypeptide in can be immature wild type ("wt"), immature mutated, mature wild type or mature mutated. Fig.67A and67B The IL-15Rα polypeptide in 67A and 67B may be immature wild type, immature mutated, mature wild type, or mature mutated. In embodiments, the IL-15 polypeptide in 67A and 67B is mature and may or may not be mutated, and the IL-15Rα polypeptide in 67A and 67B is mature and may or may not be mutated. In embodiments, the IL-15 polypeptide in 67A and 67B is mature and may or may not be mutated, and the IL-15Rα polypeptide in 67A and 67B is mature and mutated. Although a linker is depicted in Figures: 67A and 67B, the mbIL-15 polypeptide may or may not include a linker.

[0012] In an embodiment, the IL-15 polypeptide and the IL-15Rα polypeptide may be linked by one or more linkers. In an embodiment, the membrane-bound IL-15 may comprise Fig.67A or Fig.67B The structure shown and / or encoded thereby. Fig.67A and 67B In the figure, the lines connecting IL-15 to the one or more linkers (L) and connecting the one or more linkers (L) to IL-15Rα can represent a direct connection without intervening sequences, or can represent intervening sequences, such as but not limited to linkers, untranslated sequences (in the case of nucleic acid sequences), translated sequences, sequences comprising one or more restriction endonuclease sites (in the case of nucleic acid sequences), or combinations thereof.

[0013] In embodiments, the IL-15 / IL-15Rα polypeptide may comprise one or more signal peptides. In embodiments, the membrane-bound IL-15 comprising one or more signal peptides and optionally one or more linkers may comprise Fig.68A or Fig.68B The structure shown and / or encoded thereby. Fig.68A and 68B The IL-15 polypeptide in the can be immature wild type, immature mutated, mature wild type or mature mutated. Fig.68A and 68B The IL-15Rα polypeptide in can be immature wild type, immature mutated, mature wild type or mature mutated. In an embodiment, Fig.68A and Fig.68B The IL-15 polypeptide in is mature and may or may not be mutated, and Fig.68A and Fig.68B The IL-15Rα polypeptide in is mature and may or may not be mutated. In an embodiment, Fig.68A and Fig.68B The IL-15 polypeptide in is mature and may or may not be mutated, and Fig.68A and Fig.68B The IL-15Rα polypeptide in is mature and mutated. Although a linker is depicted in Figures 68A and 68B, the mbIL-15 polypeptide comprising a signal peptide may or may not comprise a linker. Fig.68A and 68B In the method, (a) the one or more signal peptides (SP) are linked to IL-15, IL-15 is linked to the one or more linkers (L), and the one or more linkers are linked to IL-15Rα (e.g. Fig.68A (a) linking the one or more signal peptides (SP) to IL-15α, linking IL-15α to the one or more linkers (L), and linking the one or more linkers to IL-15 (as shown in Fig.68B A line (shown in the figure) may represent a direct connection without intervening sequences, or may represent intervening sequences, such as, but not limited to, linkers, untranslated sequences (in the case of nucleic acid sequences), translated sequences, sequences comprising one or more restriction endonuclease sites (in the case of nucleic acid sequences), or combinations thereof.

[0014] In embodiments, the CD8 polypeptide described herein may comprise a CD8 alpha immunoglobulin (Ig)-like domain, a CD8 β region, a CD8 alpha transmembrane domain, and a CD8 alpha cytoplasmic domain. In embodiments, the CD8 β region may be a CD8 β stem region or domain.

[0015] In embodiments, the CD8 polypeptides described herein can comprise (a) an immunoglobulin (Ig)-like domain comprising 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 CD8 beta region comprising 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 CD8 beta region comprising 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: NO:3's amino acid sequence comprises a transmembrane domain 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, and (d) a cytoplasmic domain 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:4.

[0016] In 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.

[0017] In 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.

[0018] In embodiments, the CD8 polypeptide described herein may comprise 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 the amino acid sequence of any one of SEQ ID NO:6, SEQ ID NO:293 or SEQ ID NO:294, wherein the one or more signal peptides are fused directly or indirectly to the N-terminus or C-terminus of the CD8 polypeptide described herein.

[0019] In embodiments, the CD8 polypeptides described herein may comprise (a) SEQ ID NO: 1 comprising one, two, three, four or five amino acid substitutions; (b) SEQ ID NO: 2 comprising one, two, three, four or five amino acid substitutions; (c) SEQ ID NO: 3 comprising one, two, three, four or five amino acid substitutions, and (d) SEQ ID NO: 4 comprising one, two, three, four or five amino acid substitutions. In embodiments, the amino acid substitutions may be conservative or non-conservative. In embodiments, the amino acid substitutions may be conservative amino acid substitutions.

[0020] In embodiments, the CD8 polypeptide described herein may be CD8α or a modified CD8α polypeptide.

[0021] In embodiments, the CD8 polypeptide described herein may be CD8αβ or a modified CD8α polypeptide.

[0022] In an embodiment, the CD8 β polypeptide may comprise the amino acid sequence of any one of SEQ ID NO: 8, 9, 10, 11, 12, 13 or 14.

[0023] In an embodiment, the TCR α chain and the TCR β chain can be selected from 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; and 91 and 92.

[0024] In an embodiment, the isolated nucleic acid may include a nucleic acid sequence encoding a T cell receptor comprising an alpha chain and a beta chain and a CD8 polypeptide comprising an alpha chain and a beta chain. In an embodiment, the CD8 polypeptide may include a CD8 alpha chain and / or a CD8 beta chain, and the CD8 alpha chain and / or the CD8 beta chain may be independently modified or unmodified. The isolated nucleic acid may include a nucleic acid at least about 80% identical to a 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.

[0025] In one aspect, the polypeptide sequences and / or nucleic acid sequences described herein may be isolated and / or recombinant sequences.

[0026] In one aspect, the cells described herein can be isolated and / or recombinant cells.

[0027] In embodiments, the isolated nucleic acid comprises the nucleic acid sequence of SEQ ID NO:267.

[0028] In embodiments, the isolated nucleic acid comprises the nucleic acid sequence of SEQ ID NO:279.

[0029] In embodiments, an isolated polypeptide can be encoded by a nucleic acid described herein, or, due to, for example, codon degeneracy, by a nucleic acid encoding the same polypeptide.

[0030] In embodiments, the isolated polypeptide may 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 may 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 comprises 1, 2, 3, 4, 5, 10, 15 or 20 or more amino acid substitutions or deletions. 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 comprises at most 1, 2, 3, 4, 5, 10, 15 or 20 amino acid substitutions or deletions.

[0031] In an embodiment, the isolated polypeptide may comprise the amino acid sequence of SEQ ID NO:268.

[0032] In an embodiment, the isolated polypeptide may comprise the amino acid sequence of SEQ ID NO:280.

[0033] In an embodiment, a nucleic acid may be provided, the nucleic acid encoding the fusion polypeptide of formula I:

[0034] N-terminus-P6-PL-P7-C-terminus [I]

[0035] wherein P6 and P7 are each independently a first and a second polypeptide, and PL is a linker, wherein PL comprises SEQ ID NO: 387 or 389 or can provide 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.

[0036] In an embodiment, a nucleic acid may be provided, the nucleic acid comprising Formula II:

[0037] 5'-N6-NL-N7-3'[II]

[0038] wherein N6 and N7 each independently encode a first and a second polypeptide, and NL encodes a linker, wherein NL comprises SEQ ID NO: 388 or 390 or can provide 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.

[0039] In an embodiment, a nucleic acid can be provided that encodes 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.

[0040] In an embodiment, a nucleic acid can be provided that encodes 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.

[0041] In an embodiment, a nucleic acid can be provided, comprising SEQ ID NO:310, 312, 314 or 316 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:310, 312, 314 or 316.

[0042] In an embodiment, a nucleic acid can be provided, comprising SEQ ID NO:312, 314 or 316 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:312, 314 or 316.

[0043] In an embodiment, a nucleic acid can be provided, which encodes (i) a polypeptide comprising SEQ ID NO: 307, which is directly or indirectly fused to the N-terminus of a polypeptide comprising any one of SEQ ID NOs: 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, which is 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 one of SEQ ID NOs: 309, 311, 313 or 315.

[0044] In an embodiment, a nucleic acid can be provided, which encodes (i) a polypeptide comprising SEQ ID NO:307, which is directly or indirectly fused to the N-terminus of a polypeptide comprising any one of SEQ ID NOs: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, which is 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 one of SEQ ID NOs:311, 313 or 315.

[0045] In an embodiment, the nucleic acid may further comprise a nucleic acid encoding a signal peptide, the signal peptide being fused directly or indirectly to the N-terminus of SEQ ID NO: 307 or to the N-terminus of a sequence 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 an embodiment, the signal peptide may be derived from an IgE polypeptide. In an embodiment, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367 or a sequence 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.

[0046] In an embodiment, a nucleic acid can be provided, comprising (i) SEQ ID NO:308, which is directly or indirectly fused to the 5' end of any one of SEQ ID NO: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, which is directly or indirectly fused to the 5' end of any one of the sequences that are 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 one of SEQ ID NO:310, 312, 314 or 316.

[0047] In an embodiment, a nucleic acid can be provided, comprising (i) SEQ ID NO:308, which is directly or indirectly fused to the 5' end of any one 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, which is directly or indirectly fused to the 5' end of any one of the sequences that are 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 one of SEQ ID NO:312, 314 or 316.

[0048] In an embodiment, the nucleic acid may further comprise a nucleic acid encoding a signal peptide, the signal peptide being fused directly or indirectly to the 5' end of SEQ ID NO: 308 or to the 5' end of a sequence 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 an embodiment, the signal peptide may be derived from an IgE polypeptide. In an embodiment, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 368 or a sequence 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 thereto.

[0049] In an embodiment, 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.

[0050] In an embodiment, a nucleic acid can be provided that encodes 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.

[0051] In an embodiment, the nucleic acid may further comprise a nucleic acid encoding a signal peptide, the signal peptide being fused directly or indirectly to the N-terminus of SEQ ID NO: 317, 321, 325, 327, 329, 331 or 333 or to the N-terminus of a sequence 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 an embodiment, the signal peptide may be derived from an IgE polypeptide. In an embodiment, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367 or a sequence 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.

[0052] In an embodiment, a nucleic acid can be provided, comprising SEQ ID NO:318, 322, 326, 328, 330, 332, 334 or 336 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:318, 322, 326, 328, 330, 332, 334 or 336.

[0053] In an embodiment, a nucleic acid can be provided, comprising SEQ ID NO:318, 322, 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 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.

[0054] In embodiments, the nucleic acid may further comprise a nucleic acid encoding a signal peptide, the signal peptide being fused directly or indirectly 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 embodiments, the signal peptide may be derived from an IgE polypeptide. In embodiments, the signal peptide derived from an IgE polypeptide may 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 thereto.

[0055] In an embodiment, a nucleic acid can be provided that encodes 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.

[0056] In an embodiment, a nucleic acid can be provided that encodes 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.

[0057] In an embodiment, a nucleic acid can be provided, comprising SEQ ID NO:338, 342, 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 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.

[0058] In an embodiment, a nucleic acid can be provided, comprising 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 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.

[0059] In an embodiment, the nucleic acid described herein may further comprise a nucleic acid encoding: (a) at least one TCR polypeptide, the at least one TCR polypeptide comprising an α chain and a β chain, (b) at least one CD8 polypeptide, the 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 and at least one CD8 polypeptide, the at least one TCR polypeptide comprising an α chain and a β chain, the at least one CD8 polypeptide comprising (i) an α chain, (ii) a β chain, or (iii) both an α chain and a β chain.

[0060] In an embodiment, a polypeptide, a plurality of polypeptides or a fusion polypeptide may be provided, wherein the polypeptide, a plurality of polypeptides or a fusion polypeptide is encoded by a nucleic acid described herein.

[0061] In an embodiment, a polypeptide or fusion polypeptide can be provided, which 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.

[0062] In an embodiment, a polypeptide or fusion polypeptide can be provided, which 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.

[0063] In an embodiment, a fusion polypeptide can be provided, comprising 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, and the polypeptide is directly or indirectly fused to the N-terminus of any of the polypeptides that are 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.

[0064] In an embodiment, a fusion polypeptide can be provided, comprising 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 the polypeptides that are 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.

[0065] In embodiments, the fusion polypeptide may further comprise a signal peptide, the signal peptide being fused directly or indirectly 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 embodiments, the signal peptide may be derived from an IgE polypeptide. In 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 thereto.

[0066] In an embodiment, a nucleic acid may be provided, comprising: (a) a nucleic acid encoding: (i) a T cell receptor (TCR) and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain and a β chain, or (ii) a TCR and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain without a β chain, and (b) a nucleic acid encoding an IL-15 / IL-15Rα fusion polypeptide, wherein the TCRα chain and the TCRβ chain are selected from 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 and 91 and 92; wherein the CD8 alpha chain is SEQ ID NO:7, 258, 259, 262 or a variant thereof; wherein the CD8 β chain, if present, is SEQ ID NO:8, 9, 10, 11, 12, 13 or 14; and wherein the IL-15 / IL-15Rα fusion polypeptide is selected from (i) SEQ ID NO:307 or a sequence 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, fused directly or indirectly to the N-terminus of SEQ ID NO:309 or a sequence 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; (ii) SEQ ID NO:307 or a sequence 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 NO:311, or a sequence 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, fused directly or indirectly to the N-terminus of the sequence; (iii) SEQ ID NO:307, or a sequence 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, fused directly or indirectly to the N-terminus of SEQ ID NO:313, or a sequence 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;or (iv) SEQ ID NO:307, or a sequence 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, fused directly or indirectly to the N-terminus of SEQ ID NO:315, or a sequence 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.;

[0067] In an embodiment, the IL-15 / IL-15Rα fusion polypeptide may further comprise a signal peptide, which is 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).

[0068] In an embodiment, a nucleic acid may be provided, comprising: (a) a nucleic acid encoding: (i) a T cell receptor (TCR) and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain and a β chain, or (ii) a TCR and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain without a β chain, and (b) a nucleic acid encoding an IL-15 / IL-15Rα fusion polypeptide, wherein the TCRα chain and the TCRβ chain are selected from 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 and 91 and 92; wherein the CD8 alpha chain is SEQ ID NO:7, 258, 259, 262 or a variant thereof; wherein the CD8 β chain, if present, is SEQ ID NO:8, 9, 10, 11, 12, 13 or 14; and wherein the IL-15 / IL-15Rα fusion polypeptide is selected from (i) SEQ ID NO:307 or a sequence 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, fused directly or indirectly to the N-terminus of SEQ ID NO:311 or a sequence 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; (ii) SEQ ID NO:307 or a sequence 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 NO:313, or a sequence 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 the sequence; or (iii) SEQ ID NO:307, or a sequence 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 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.

[0069] In an embodiment, the IL-15 / IL-15Rα fusion polypeptide may further comprise a signal peptide, which is 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).

[0070] In embodiments, the signal peptide may be derived from an IgE polypeptide. In embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367, or a sequence 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.

[0071] In an embodiment, a nucleic acid may be provided, comprising: (a) a nucleic acid encoding: (i) a T cell receptor (TCR) and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain and a β chain, or (ii) a TCR and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain without a β chain, and (b) a nucleic acid encoding an IL-15 / IL-15Rα fusion polypeptide, wherein the TCRα chain and the TCRβ chain are selected from 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 and 91 and 92; wherein the CD8 alpha chain is SEQ ID NO:7, 258, 259, 262 or a variant thereof; wherein the CD8 β chain, if present, is SEQ ID NO:8, 9, 10, 11, 12, 13 or 14; and wherein 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.

[0072] In an embodiment, the IL-15 / IL-15Rα fusion polypeptide may further comprise a signal peptide, which is fused directly or indirectly to the N-terminus of SEQ ID NO: SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, 333 or 335, or directly or indirectly 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.

[0073] In an embodiment, a nucleic acid may be provided, comprising: (a) a nucleic acid encoding: (i) a T cell receptor (TCR) and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain and a β chain, or (ii) a TCR and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain without a β chain, and (b) a nucleic acid encoding an IL-15 / IL-15Rα fusion polypeptide, wherein the TCRα chain and the TCRβ chain are selected from 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 and 91 and 92; wherein the CD8 alpha chain is SEQ ID NO:7, 258, 259, 262 or a variant thereof; wherein the CD8 β chain, if present, is SEQ ID NO:8, 9, 10, 11, 12, 13 or 14; and wherein the IL-15 / IL-15Rα fusion polypeptide is selected from SEQ ID NO: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 NO:317, 319, 321, 323, 325, 327, 329, 331 or 333.

[0074] In an embodiment, the IL-15 / IL-15Rα fusion polypeptide may further comprise a signal peptide, which is directly or indirectly fused to the N-terminus of SEQ ID NO: SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331 or 333, or 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.

[0075] In embodiments, the signal peptide may be derived from an IgE polypeptide. In embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367, or a sequence 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.

[0076] In an embodiment, a nucleic acid may be provided, comprising: (a) a nucleic acid encoding: (i) a T cell receptor (TCR) and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain and a β chain, or (ii) a TCR and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain without a β chain, and (b) a nucleic acid encoding a fusion polypeptide, wherein the fusion polypeptide comprises (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, and (ii) the N-terminus of an IL-15 / IL-15Rα fusion polypeptide, wherein the TCR α chain and the TCR β chain are selected from SEQ ID NO: 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 and 91 and 92; wherein the CD8 alpha chain is SEQ ID NO:7, 258, 259, 262 or a variant thereof; wherein the CD8 β chain, if present, is SEQ ID NO:8, 9, 10, 11, 12, 13 or 14; and wherein the nucleic acid of (b) encodes a fusion polypeptide selected from SEQ ID NO: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 NO:337, 339, 341, 343, 345, 347, 349, 351, 353 or 355.

[0077] In an embodiment, a nucleic acid may be provided, comprising: (a) a nucleic acid encoding: (i) a T cell receptor (TCR) and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain and a β chain, or (ii) a TCR and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain without a β chain, and (b) a nucleic acid encoding a fusion polypeptide, wherein the fusion polypeptide comprises (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, and (ii) the N-terminus of an IL-15 / IL-15Rα fusion polypeptide, wherein the TCR α chain and the TCR β chain are selected from SEQ ID NO: 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 and 91 and 92; wherein the CD8 alpha chain is SEQ ID NO:7, 258, 259, 262 or a variant thereof; wherein the CD8 β chain, if present, is SEQ ID NO:8, 9, 10, 11, 12, 13 or 14; and wherein the nucleic acid of (b) encodes a fusion polypeptide selected from SEQ ID NO: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 NO:337, 339, 341, 343, 345, 347, 349, 351 or 353.

[0078] In an embodiment, a nucleic acid may be provided, comprising: (a) a nucleic acid encoding: (i) a T cell receptor (TCR) and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain and a β chain, or (ii) a TCR and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain without a β chain, and (b) a nucleic acid encoding an IL-15 / IL-15Rα fusion polypeptide, wherein the TCRα chain and the TCRβ chain are selected from 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 and 91 and 92; wherein the CD8 alpha chain is SEQ ID NO:7, 258, 259, 262 or a variant thereof; wherein the CD8 β chain, if present, is SEQ ID NO:8, 9, 10, 11, 12, 13 or 14; and wherein 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.

[0079] In an embodiment, the IL-15 / IL-15Rα fusion polypeptide may further comprise a signal peptide, which is fused directly or indirectly to the N-terminus of SEQ ID NO: 317, 321, 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, 321, 325, 327, 329, 331, 333 or 335.

[0080] In an embodiment, a nucleic acid may be provided, comprising: (a) a nucleic acid encoding: (i) a T cell receptor (TCR) and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain and a β chain, or (ii) a TCR and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain without a β chain, and (b) a nucleic acid encoding an IL-15 / IL-15Rα fusion polypeptide, wherein the TCRα chain and the TCRβ chain are selected from 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 and 91 and 92; wherein the CD8 alpha chain is SEQ ID NO:7, 258, 259, 262 or a variant thereof; wherein the CD8 β chain, if present, is SEQ ID NO:8, 9, 10, 11, 12, 13 or 14; and wherein 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.

[0081] In an embodiment, the IL-15 / IL-15Rα fusion polypeptide may further comprise a signal peptide, which 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.

[0082] In embodiments, the signal peptide may be derived from an IgE polypeptide. In embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367, or a sequence 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.

[0083] In an embodiment, a nucleic acid may be provided, comprising: (a) a nucleic acid encoding: (i) a T cell receptor (TCR) and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain and a β chain, or (ii) a TCR and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain without a β chain, and (b) a nucleic acid encoding a fusion polypeptide, wherein the fusion polypeptide comprises (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, and (ii) the N-terminus of an IL-15 / IL-15Rα fusion polypeptide, wherein the TCR α chain and the TCR β chain are selected from SEQ ID NO: 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 and 91 and 92; wherein the CD8 alpha chain is SEQ ID NO:7, 258, 259, 262 or a variant thereof; wherein the CD8 β chain, if present, is SEQ ID NO:8, 9, 10, 11, 12, 13 or 14; and wherein the nucleic acid of (b) encodes a fusion polypeptide selected from SEQ ID NO: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 NO:337, 339, 341, 345, 347, 349, 351, 353 or 355.

[0084] In an embodiment, a nucleic acid may be provided, comprising: (a) a nucleic acid encoding: (i) a T cell receptor (TCR) and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain and a β chain, or (ii) a TCR and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain without a β chain, and (b) a nucleic acid encoding a fusion polypeptide, wherein the fusion polypeptide comprises (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, and (ii) the N-terminus of an IL-15 / IL-15Rα fusion polypeptide, wherein the TCR α chain and the TCR β chain are selected from SEQ ID NO: 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 and 91 and 92; wherein the CD8 alpha chain is SEQ ID NO:7, 258, 259, 262 or a variant thereof; wherein the CD8 β chain, if present, is SEQ ID NO:8, 9, 10, 11, 12, 13 or 14; and wherein the nucleic acid of (b) encodes a fusion polypeptide selected from SEQ ID NO: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 NO:337, 341, 345, 347, 349, 351 or 353.

[0085] In an embodiment, a nucleic acid may be provided, comprising: (a) a nucleic acid encoding: (i) a T cell receptor (TCR) and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain and a β chain, or (ii) a TCR and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain without a β chain, and (b) a nucleic acid encoding an IL-15 / IL-15Rα fusion polypeptide, wherein 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; wherein the CD8α chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof; and wherein the CD8β chain, if present, is SEQ ID NO: 8, 9, 10, 11, 12, 13 or 14.

[0086] In embodiments, the IL-15 / IL-15Rα fusion polypeptide may 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 thereto.

[0087] In embodiments, the IL-15 / IL-15Rα fusion polypeptide may 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 thereto.

[0088] In embodiments, the IL-15 / IL-15Rα fusion polypeptide may 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 thereto.

[0089] In embodiments, the IL-15 / IL-15Rα fusion polypeptide may comprise SEQ ID NO: 323, 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 thereto.

[0090] In embodiments, the IL-15 / IL-15Rα fusion polypeptide may comprise SEQ ID NO: 325, 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 thereto.

[0091] In embodiments, the IL-15 / IL-15Rα fusion polypeptide may comprise SEQ ID NO: 327, 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 thereto.

[0092] In embodiments, the IL-15 / IL-15Rα fusion polypeptide may comprise SEQ ID NO: 329, 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 thereto.

[0093] In embodiments, the IL-15 / IL-15Rα fusion polypeptide may comprise SEQ ID NO: 331, 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 thereto.

[0094] In embodiments, the IL-15 / IL-15Rα fusion polypeptide may comprise SEQ ID NO: 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 thereto.

[0095] In embodiments, the IL-15 / IL-15Rα fusion polypeptide may comprise 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 thereto.

[0096] In an embodiment, the IL-15 / IL-15Rα fusion polypeptide may further comprise a signal peptide, which is directly or indirectly fused to the N-terminus of the IL-15 / IL-15Rα fusion polypeptide. In an embodiment, the signal peptide may be derived from an IgE polypeptide. In an embodiment, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367 or a sequence 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.

[0097] In an embodiment, a nucleic acid may be provided, comprising: (a) a nucleic acid encoding: (i) a T cell receptor (TCR) and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain and a β chain, or (ii) a TCR and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain without a β chain, and (b) a nucleic acid encoding a fusion polypeptide, wherein the fusion polypeptide comprises (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, and (ii) the N-terminus of an IL-15 / IL-15Rα fusion polypeptide, wherein the TCR α chain and the TCR β chain are selected from SEQ ID NO: NO: 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; wherein the CD8 alpha chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof; and wherein the CD8 beta chain, if present, is SEQ ID NO: 8, 9, 10, 11, 12, 13 or 14.

[0098] In an embodiment, the nucleic acid of (b) can encode SEQ ID NO:337, 339, 341, 343, 345, 347, 349, 351, 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 any of the foregoing SEQ ID NOs.

[0099] In an embodiment, a nucleic acid may be provided, comprising: (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 encoding an IL-15 / IL-15Rα fusion polypeptide.

[0100] In an embodiment, a nucleic acid can be provided, comprising: (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 encoding an IL-15 / IL-15Rα fusion polypeptide.

[0101] In an embodiment, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide can be selected from (i) SEQ ID NO: 308 or a sequence at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical thereto, which is fused directly or indirectly to the 5' end of SEQ ID NO: 310 or a sequence at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical thereto, with or without a nucleic acid encoding a linker therebetween; (ii) SEQ ID NO: 308 or a sequence at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical thereto, with or without a nucleic acid encoding a linker therebetween ... NO:312, or a sequence at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical thereto, fused directly or indirectly to the 5' end thereof; (iii) SEQ ID NO:308, or a sequence at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical thereto, fused directly or indirectly to the 5' end thereof, with or without a nucleic acid encoding a linker, and SEQ ID NO:314, or a sequence at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical thereto; or (iv) SEQ ID NO:315, or a sequence at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical thereto, fused directly or indirectly to the 5' end thereof. NO:308, or a sequence at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical thereto, which is fused directly or indirectly to the 5' end of SEQ ID NO:316, or a sequence at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical thereto, with or without a nucleic acid encoding a linker therebetween.

[0102] In an embodiment, 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 SEQ ID NO: 308 of any one of (i), (ii), (iii) or (iv) 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 SEQ ID NO: 308 of any one of (i), (ii), (iii) or (iv).

[0103] In an embodiment, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide can be selected from (i) SEQ ID NO: 308 or a sequence at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical thereto, which is fused directly or indirectly to the 5' end of SEQ ID NO: 312 or a sequence at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical thereto, with or without a nucleic acid encoding a linker therebetween; (ii) SEQ ID NO: 308 or a sequence at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical thereto, with or without a nucleic acid encoding a linker therebetween ... NO:314, or a sequence at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical thereto, directly or indirectly fused to the 5' end of the sequence; or (iii) SEQ ID NO:308, or a sequence at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical thereto, with or without a nucleic acid encoding a linker therebetween, directly or indirectly fused to the 5' end of SEQ ID NO:316, or a sequence at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical thereto.

[0104] In an embodiment, 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 SEQ ID NO: 308 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 SEQ ID NO: 308 of any one of (i), (ii) or (iii).

[0105] In embodiments, the signal peptide may be derived from an IgE polypeptide. In embodiments, the nucleic acid encoding the signal peptide may comprise SEQ ID NO: 368, or a sequence 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.

[0106] In an embodiment, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide can be selected from 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 acid of SEQ ID NO:318, 320, 322, 324, 326, 328, 330, 332, 334 or 336.

[0107] In an embodiment, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide may further comprise a nucleic acid encoding a signal peptide, wherein 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 a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to a nucleic acid of SEQ ID NO: 318, 320, 322, 324, 326, 328, 330, 332, 334 or 336.

[0108] In an embodiment, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide can 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.

[0109] In an embodiment, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide may further comprise a nucleic acid encoding a signal peptide, wherein 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 a sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to a nucleic acid of SEQ ID NO: 318, 320, 322, 324, 326, 328, 330, 332 or 334.

[0110] In embodiments, the signal peptide may be derived from an IgE polypeptide. In embodiments, the nucleic acid encoding the signal peptide may comprise SEQ ID NO: 368, or a sequence 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.

[0111] In an embodiment, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide (i) may further comprise a nucleic acid encoding a signal peptide derived from an IgE polypeptide, and (ii) may be selected from SEQ ID NO: 338, 340, 342, 344, 346, 348, 350, 352, 354 or 356 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: 338, 340, 342, 344, 346, 348, 350, 352, 354 or 356.

[0112] In an embodiment, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide (i) may further comprise a nucleic acid encoding a signal peptide derived from an IgE polypeptide, and (ii) may be selected from SEQ ID NO: 338, 340, 342, 344, 346, 348, 350, 352 or 354 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: 338, 340, 342, 344, 346, 348, 350, 352 or 354.

[0113] In an embodiment, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide can be selected from SEQ ID NO:318, 322, 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 acid of SEQ ID NO:318, 322, 326, 328, 330, 332, 334 or 336.

[0114] In an embodiment, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide may further comprise a nucleic acid encoding a signal peptide, wherein the nucleic acid encoding the signal peptide is fused directly or indirectly to the 5' end of SEQ ID NO: 318, 322, 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 a nucleic acid of SEQ ID NO: 318, 322, 326, 328, 330, 332, 334 or 336.

[0115] In an embodiment, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide can 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.

[0116] In an embodiment, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide may further comprise a nucleic acid encoding a signal peptide, wherein the nucleic acid encoding the signal peptide is fused directly or indirectly to the 5' end of 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 a nucleic acid of SEQ ID NO: 318, 322, 326, 328, 330, 332 or 334.

[0117] In embodiments, the signal peptide may be derived from an IgE polypeptide. In embodiments, the nucleic acid encoding the signal peptide may comprise SEQ ID NO: 368, or a sequence 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.

[0118] In an embodiment, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide (i) may further comprise a nucleic acid encoding a signal peptide derived from an IgE polypeptide, and (ii) may be selected from SEQ ID NO: 338, 342, 346, 348, 350, 352, 354 or 356 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: 338, 342, 346, 348, 350, 352, 354 or 356.

[0119] In an embodiment, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide (i) may further comprise a nucleic acid encoding a signal peptide derived from an IgE polypeptide, and (ii) may be selected from SEQ ID NO: 338, 342, 346, 348, 350, 352 or 354 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: 338, 342, 346, 348, 350, 352 or 354.

[0120] In an embodiment, a vector may be provided, comprising a nucleic acid encoding at least one CD8 α chain, at least one TCR α chain, at least one TCR β chain, at least one IL-15 / IL-15Rα fusion polypeptide, and optionally at least one CD8 β chain.

[0121] In an embodiment, a vector may be provided, comprising N1, N2, N3, N4, N5, L1, L2, L3 and L4 in any order, wherein N1 comprises a nucleic acid encoding a CD8 β chain and is present or absent, N2 comprises a nucleic acid encoding a CD8 α chain, N3 comprises a nucleic acid encoding a TCR β chain, N4 comprises a nucleic acid encoding a TCR α chain, N5 comprises a nucleic acid encoding an IL-15 / IL-15Rα fusion polypeptide; and wherein L1-L4 each comprises a nucleic acid encoding at least one linker, wherein each of L1-L4 is independently the same or different, and wherein each of L1-L4 is independently present or absent.

[0122] In an embodiment, the vector may comprise Formula III or Formula IV:

[0123] 5'-N1-L1-N2-L2-N3-L3-N4-L4-N5-3' [III]

[0124] 5'-N5-L1-N1-L2-N2-L3-N3-L4-N4-3'[IV].

[0125] In an embodiment, N1 may comprise a nucleic acid encoding SEQ ID NO: 8, 9, 10, 11, 12, 13 or 14.

[0126] In an embodiment, N2 comprises a nucleic acid encoding SEQ ID NO: 7, 258, 259, 262 or a variant thereof.

[0127] In an embodiment, N4 and N3 may contain a sequence encoding SEQ ID 83 and 84, 85 and 86, 87 and 88, 89 and 90, 91 and 92, 93 and 94, 95 and 96, 97 and 98, 99 and 100, 101 and 102, 103 and 104, 105 and 106, 107 and 108, 109 and 110, 111 and 112, 112 and 113, 113 and 114, 115 and 116, 117 and 118, 119 and 120, 121 and 122, 123 and 124, 125 and 126, 127 and 128, 129 and 130, 131 and 132, 133 and 134, 135 and 136, 137 and 138, 139 and 140

[0128] In an embodiment, N5 may comprise a nucleic acid encoding: (i) SEQ ID NO: 307, or a sequence 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, fused directly or indirectly to the N-terminus of SEQ ID NO: 309, or a sequence 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, with or without a linker therebetween; (ii) SEQ ID NO: 307, or a sequence 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, fused directly or indirectly to the N-terminus of SEQ ID NO: 311, or a sequence 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, with or without a linker therebetween 313 or a sequence 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, fused directly or indirectly to the N-terminus of the sequence; (iii) SEQ ID NO: 307 or a sequence 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, fused directly or indirectly to the N-terminus of SEQ ID NO: 313 or a sequence 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, with or without a linker therebetween; or (iv) SEQ ID NO: 307 or a sequence 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 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.

[0129] In an embodiment, N5 may further comprise a nucleic acid encoding a signal peptide, which is directly or indirectly fused to the 5' end of the nucleic acid encoding SEQ ID NO: 307 of any one 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 SEQ ID NO: 307 of any one of (i), (ii), (iii) or (iv).

[0130] In an embodiment, N5 may comprise a nucleic acid encoding: (i) SEQ ID NO: 307, or a sequence 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, fused directly or indirectly to the N-terminus of SEQ ID NO: 311, or a sequence 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, with or without a linker therebetween; (ii) SEQ ID NO: 307, or a sequence 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, fused directly or indirectly to the N-terminus of SEQ ID NO: 313, or a sequence 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, with or without a linker therebetween. 315 or a sequence 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 the sequence; or (iii) SEQ ID NO: 307 or a sequence 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 SEQ ID NO: 315 or a sequence 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, with or without a linker in between.

[0131] In an embodiment, N5 may further comprise a nucleic acid encoding a signal peptide, which is directly or indirectly fused to the 5' end of the nucleic acid encoding SEQ ID NO: 307 of any one 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 SEQ ID NO: 307 of any one of (i), (ii) or (iii).

[0132] In embodiments, the signal peptide may be derived from an IgE polypeptide. In embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367, or a sequence 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.

[0133] In an embodiment, N5 may comprise a nucleic acid encoding 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.

[0134] In an embodiment, N5 may further comprise a nucleic acid encoding a signal peptide, which 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, 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, 319, 321, 323, 325, 327, 329, 331, 333 or 335.

[0135] In an embodiment, N5 may comprise a nucleic acid encoding SEQ ID NO: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 NO:317, 319, 321, 323, 325, 327, 329, 331 or 333.

[0136] In an embodiment, N5 may further comprise a nucleic acid encoding a signal peptide, which 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.

[0137] In embodiments, the signal peptide may be derived from an IgE polypeptide. In embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367, or a sequence 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.

[0138] In an embodiment, (i) N5 may further comprise a nucleic acid encoding a signal peptide derived from an IgE polypeptide, and (ii) N5 may encode SEQ ID NO: 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 NO: 337, 339, 341, 343, 345, 347, 349, 351, 353 or 355.

[0139] In an embodiment, (i) N5 may further comprise a nucleic acid encoding a signal peptide derived from an IgE polypeptide, and (ii) N5 may encode SEQ ID NO: 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 NO: 337, 339, 341, 343, 345, 347, 349, 351 or 353.

[0140] In an embodiment, N5 may comprise a nucleic acid encoding 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.

[0141] In an embodiment, N5 may further comprise a nucleic acid encoding a signal peptide, which 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.

[0142] In an embodiment, N5 may comprise a nucleic acid encoding 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.

[0143] In an embodiment, N5 may further comprise a nucleic acid encoding a signal peptide, which 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.

[0144] In embodiments, the signal peptide may be derived from an IgE polypeptide. In embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367, or a sequence 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.

[0145] In an embodiment, (i) N5 may further comprise a nucleic acid encoding a signal peptide derived from an IgE polypeptide, and (ii) N5 may encode SEQ ID NO: 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 NO: 337, 341, 345, 347, 349, 351 or 353.

[0146] In an embodiment, (i) the vector may further encode a 2A peptide or an internal ribosome entry site (IRES) located 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) located 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.

[0147] In an embodiment, (i) the vector may further encode a furin located 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 furin located 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.

[0148] In an embodiment, 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).

[0149] In an embodiment, the IRES may be selected from the group consisting of an IRES from a picornavirus, an IRES from a flavivirus, an IRES from a pestivirus, an IRES from a retrovirus, an IRES from a lentivirus, an IRES from an insect RNA virus, and an IRES from a cellular mRNA.

[0150] In an embodiment, a T cell and / or natural killer cell may be provided, wherein the T cell and / or natural killer cell comprises: (a) (i) a T cell receptor (TCR) and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain and a β chain, or (ii) a TCR and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain without a β chain, and (b) an IL-15 / IL-15Rα fusion polypeptide, wherein the TCRα chain and the TCRβ chain are selected from 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 and 91 and 92; wherein the CD8 alpha chain is SEQ ID NO:7, 258, 259, 262 or a variant thereof; wherein the CD8 β chain, if present, is SEQ ID NO:8, 9, 10, 11, 12, 13 or 14; and wherein the IL-15 / IL-15Rα fusion polypeptide is selected from (i) 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, which is fused directly or indirectly 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, with or without a linker therebetween; (ii) 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:309; NO:307 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, which is fused directly or indirectly to the N-terminus of SEQ ID NO:311, or a sequence 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, with or without a linker therebetween;(iii) SEQ ID NO:307, or a sequence 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, fused directly or indirectly to the N-terminus of SEQ ID NO:313, or a sequence 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, with or without a linker therebetween; or (iv) SEQ ID NO:307, or a sequence 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, fused, with or without a linker therebetween, to the N-terminus of SEQ ID NO:315, or a sequence 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:316. NO:315 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 the sequence. ;

[0151] In an embodiment, the IL-15 / IL-15Rα fusion polypeptide may further comprise a signal peptide, which is 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).

[0152] In an embodiment, a T cell and / or natural killer cell may be provided, wherein the T cell and / or natural killer cell comprises: (a) (i) a T cell receptor (TCR) and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain and a β chain, or (ii) a TCR and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain without a β chain, and (b) an IL-15 / IL-15Rα fusion polypeptide, wherein the TCRα chain and the TCRβ chain are selected from 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 and 91 and 92; wherein the CD8 alpha chain is SEQ ID NO:7, 258, 259, 262 or a variant thereof; wherein the CD8 β chain, if present, is SEQ ID NO:8, 9, 10, 11, 12, 13 or 14; and wherein the IL-15 / IL-15Rα fusion polypeptide is selected from (i) SEQ ID NO:307 or a sequence 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, fused directly or indirectly to the N-terminus of SEQ ID NO:311 or a sequence 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, with or without a linker therebetween; (ii) SEQ ID NO:307 or a sequence 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; NO:307 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, which is fused directly or indirectly to the N-terminus of SEQ ID NO:313, or a sequence 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, with or without a linker therebetween;or (iii) 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, fused directly or indirectly to 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, with or without a linker therebetween.;

[0153] In an embodiment, the IL-15 / IL-15Rα fusion polypeptide may further comprise a signal peptide, which is 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).

[0154] In embodiments, the signal peptide may be derived from an IgE polypeptide. In embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367, or a sequence 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.

[0155] In an embodiment, a T cell and / or natural killer cell may be provided, wherein the T cell and / or natural killer cell comprises: (a) (i) a T cell receptor (TCR) and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain and a β chain, or (ii) a TCR and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain without a β chain, and (b) an IL-15 / IL-15Rα fusion polypeptide, wherein the TCRα chain and the TCRβ chain are selected from 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 and 91 and 92; wherein the CD8 alpha chain is SEQ ID NO:7, 258, 259, 262 or a variant thereof; wherein the CD8 β chain, if present, is SEQ ID NO:8, 9, 10, 11, 12, 13 or 14; and wherein 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.

[0156] In an embodiment, the IL-15 / IL-15Rα fusion polypeptide may further comprise a signal peptide, which is fused directly or indirectly to the N-terminus of SEQ ID NO: SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, 333 or 335, or directly or indirectly 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.

[0157] In an embodiment, a T cell and / or natural killer cell may be provided, wherein the T cell and / or natural killer cell comprises: (a) (i) a T cell receptor (TCR) and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain and a β chain, or (ii) a TCR and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain without a β chain, and (b) an IL-15 / IL-15Rα fusion polypeptide, wherein the TCRα chain and the TCRβ chain are selected from 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 and 91 and 92; wherein the CD8 alpha chain is SEQ ID NO:7, 258, 259, 262 or a variant thereof; wherein the CD8 β chain, if present, is SEQ ID NO:8, 9, 10, 11, 12, 13 or 14; and wherein the IL-15 / IL-15Rα fusion polypeptide is selected from SEQ ID NO: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 NO:317, 319, 321, 323, 325, 327, 329, 331 or 333.

[0158] In an embodiment, the IL-15 / IL-15Rα fusion polypeptide may further comprise a signal peptide, which is directly or indirectly fused to the N-terminus of SEQ ID NO: SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331 or 333, or 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.

[0159] In embodiments, the signal peptide may be derived from an IgE polypeptide. In embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367, or a sequence 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.

[0160] In an embodiment, a T cell and / or natural killer cell may be provided, comprising: (a) (i) a T cell receptor (TCR) and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain and a β chain, or (ii) a TCR and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain without a β chain, and (b) a fusion polypeptide, wherein the fusion polypeptide comprises (i) a signal peptide, wherein the signal peptide comprises a fused 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) an IL-15 / IL-15Rα fusion polypeptide, wherein the TCRα chain and the TCRβ chain are selected from SEQ ID NO: 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 and 91 and 92; wherein the CD8 alpha chain is SEQ ID NO:7, 258, 259, 262 or a variant thereof; wherein the CD8 β chain, if present, is SEQ ID NO:8, 9, 10, 11, 12, 13 or 14; and wherein the fusion polypeptide of (b) is selected from SEQ ID NO: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 NO:337, 339, 341, 343, 345, 347, 349, 351, 353 or 355.

[0161] In an embodiment, a T cell and / or natural killer cell may be provided, comprising: (a) (i) a T cell receptor (TCR) and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain and a β chain, or (ii) a TCR and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain without a β chain, and (b) a fusion polypeptide, wherein the fusion polypeptide comprises (i) a signal peptide, wherein the signal peptide comprises a fused 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) an IL-15 / IL-15Rα fusion polypeptide, wherein the TCRα chain and the TCRβ chain are selected from SEQ ID NO: 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 and 91 and 92; wherein the CD8 alpha chain is SEQ ID NO:7, 258, 259, 262 or a variant thereof; wherein the CD8 β chain, if present, is SEQ ID NO:8, 9, 10, 11, 12, 13 or 14; and wherein the fusion polypeptide of (b) is selected from SEQ ID NO: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 NO:337, 339, 341, 343, 345, 347, 349, 351 or 353.

[0162] In an embodiment, a T cell and / or natural killer cell may be provided, wherein the T cell and / or natural killer cell is transduced to express (a) (i) a T cell receptor (TCR) and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain and a β chain, or (ii) a TCR and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain without a β chain, and (b) an IL-15 / IL-15Rα fusion polypeptide, wherein the TCRα chain and the TCRβ chain are selected from 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 and 91 and 92; wherein the CD8 alpha chain is SEQ ID NO:7, 258, 259, 262 or a variant thereof; wherein the CD8 β chain, if present, is SEQ ID NO:8, 9, 10, 11, 12, 13 or 14; and wherein 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.

[0163] In an embodiment, the IL-15 / IL-15Rα fusion polypeptide may further comprise a signal peptide, which is directly or indirectly fused to the N-terminus of SEQ ID NO: SEQ ID NO: 317, 321, 325, 327, 329, 331, 333 or 335, or 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.

[0164] In an embodiment, a T cell and / or natural killer cell may be provided, wherein the T cell and / or natural killer cell is transduced to express (a) (i) a T cell receptor (TCR) and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain and a β chain, or (ii) a TCR and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain without a β chain, and (b) an IL-15 / IL-15Rα fusion polypeptide, wherein the TCRα chain and the TCRβ chain are selected from 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 and 91 and 92; wherein the CD8 alpha chain is SEQ ID NO:7, 258, 259, 262 or a variant thereof; wherein the CD8 β chain, if present, is SEQ ID NO:8, 9, 10, 11, 12, 13 or 14; and wherein 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.

[0165] In an embodiment, the IL-15 / IL-15Rα fusion polypeptide may further comprise a signal peptide, which is directly or indirectly fused to the N-terminus of SEQ ID NO: SEQ ID NO: 317, 321, 325, 327, 329, 331 or 333, or 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.

[0166] In embodiments, the signal peptide may be derived from an IgE polypeptide. In embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367, or a sequence 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.

[0167] In an embodiment, a T cell and / or natural killer cell may be provided, comprising: (a) (i) a T cell receptor (TCR) and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain and a β chain, or (ii) a TCR and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain without a β chain, and (b) a fusion polypeptide, wherein the fusion polypeptide comprises (i) a signal peptide 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 fused directly or indirectly, and (ii) the N-terminus of an IL-15 / IL-15Rα fusion polypeptide, wherein the TCR α chain and the TCR β chain are selected from SEQ ID NO: 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 and 91 and 92; wherein the CD8 alpha chain is SEQ ID NO:7, 258, 259, 262 or a variant thereof; wherein the CD8 β chain, if present, is SEQ ID NO:8, 9, 10, 11, 12, 13 or 14; and wherein the fusion polypeptide of (b) is selected from SEQ ID NO: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 NO:337, 341, 345, 347, 349, 351 or 353.

[0168] In an embodiment, a T cell and / or natural killer cell, comprising: (a) (i) a T cell receptor (TCR) and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain and a β chain, or (ii) a TCR and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain without a β chain, and (b) an IL-15 / IL-15Rα fusion polypeptide, wherein the TCR α chain and the 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; wherein the CD8 α chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof; and wherein the CD8 β chain, if present, is SEQ ID NO: 8, 9, 10, 11, 12, 13 or 14.

[0169] In embodiments, the IL-15 / IL-15Rα fusion polypeptide may 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 thereto.

[0170] In embodiments, the IL-15 / IL-15Rα fusion polypeptide may 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 thereto.

[0171] In embodiments, the IL-15 / IL-15Rα fusion polypeptide may 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 thereto.

[0172] In embodiments, the IL-15 / IL-15Rα fusion polypeptide may comprise SEQ ID NO: 323, 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 thereto.

[0173] In embodiments, the IL-15 / IL-15Rα fusion polypeptide may comprise SEQ ID NO: 325, 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 thereto.

[0174] In embodiments, the IL-15 / IL-15Rα fusion polypeptide may comprise SEQ ID NO: 327, 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 thereto.

[0175] In embodiments, the IL-15 / IL-15Rα fusion polypeptide may comprise SEQ ID NO: 329, 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 thereto.

[0176] In embodiments, the IL-15 / IL-15Rα fusion polypeptide may comprise SEQ ID NO: 331, 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 thereto.

[0177] In embodiments, the IL-15 / IL-15Rα fusion polypeptide may comprise SEQ ID NO: 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 thereto.

[0178] In embodiments, the IL-15 / IL-15Rα fusion polypeptide may comprise 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 thereto.

[0179] In an embodiment, the IL-15 / IL-15Rα fusion polypeptide may further comprise a signal peptide, which is directly or indirectly fused to the N-terminus of the IL-15 / IL-15Rα fusion polypeptide. In an embodiment, the signal peptide may be derived from an IgE polypeptide. In an embodiment, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367 or a sequence 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.

[0180] In embodiments, the T cell and / or natural killer cell comprises: (a) (i) a T cell receptor (TCR) and a CD8 polypeptide, wherein the TCR comprises an alpha chain and a beta chain, and the CD8 polypeptide comprises an alpha chain and a beta chain, or (ii) a TCR and a CD8 polypeptide, wherein the TCR comprises an alpha chain and a beta chain, and the CD8 polypeptide comprises an alpha chain without a beta chain, and (b) a fusion polypeptide comprising (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, and (ii) the N-terminus of an IL-15 / IL-15Rα fusion polypeptide, wherein the TCR alpha chain and the TCR beta chain are selected from SEQ ID NO: NO: 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; wherein the CD8 alpha chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof; and wherein the CD8 beta chain, if present, is SEQ ID NO: 8, 9, 10, 11, 12, 13 or 14.

[0181] In an embodiment, the fusion polypeptide of (b) may comprise SEQ ID NO: 337, 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 thereto.

[0182] In an embodiment, the fusion polypeptide of (b) may comprise SEQ ID NO: 339, 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 thereto.

[0183] In an embodiment, the fusion polypeptide of (b) may comprise SEQ ID NO: 341, 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 thereto.

[0184] In an embodiment, the fusion polypeptide of (b) may comprise SEQ ID NO: 343, 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 thereto.

[0185] In an embodiment, the fusion polypeptide of (b) may comprise SEQ ID NO: 345, 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 thereto.

[0186] In an embodiment, the fusion polypeptide of (b) may comprise SEQ ID NO: 347, 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 thereto.

[0187] In an embodiment, the fusion polypeptide of (b) may comprise SEQ ID NO: 349, 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 thereto.

[0188] In an embodiment, the fusion polypeptide of (b) may comprise SEQ ID NO: 351, 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 thereto.

[0189] In an embodiment, the fusion polypeptide of (b) may comprise SEQ ID NO: 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 thereto.

[0190] In an embodiment, the fusion polypeptide of (b) may comprise SEQ ID NO: 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 thereto.

[0191] In an embodiment, the T cell may be an αβT cell, a γδT cell, a natural killer T cell, or any combination thereof. In an embodiment, the αβT cell may be a CD4+T cell. In an embodiment, the αβT cell may be a CD8+T cell. In an embodiment, the γδT cell may be a Vγ9Vδ2+T cell.

[0192] In an embodiment, a composition may be provided, comprising T cells and / or natural killer cells as described herein. In an embodiment, the composition may be a pharmaceutical composition. In an embodiment, the composition may further comprise an adjuvant, an excipient, a carrier, a diluent, a buffer, a stabilizer, or a combination thereof.

[0193] In an embodiment, the adjuvant can be an anti-CD40 antibody, imiquimod, resiquimod, GM-CSF, cyclophosphamide, sunitinib, bevacizumab, atezolizumab, interferon alpha, interferon beta, CpG oligonucleotides and derivatives, poly(I:C) and 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 an embodiment, the adjuvant can be IL-2, IL-7, IL-12, IL-15, IL-21 or any combination thereof.

[0194] In an embodiment, a method for preparing T cells and / or natural killer cells for immunotherapy may 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 an embodiment, the method further comprises 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 an embodiment, the blood sample may comprise peripheral blood mononuclear cells (PMBC). In an embodiment, the activation may comprise contacting the T cells and / or natural killer cells with anti-CD3 and anti-CD28 antibodies. In an embodiment, the T cells may be CD4+T cells. In an embodiment, the T cells may be CD8+T cells. In an embodiment, the T cells may be γδT cells or αβT cells. In embodiments, said activation, said amplification, or both may be performed in the presence of a combination of IL-2 and IL-15 and optionally zoledronate.

[0195] In an embodiment, a method of increasing the persistence, functionality, naive state, lifespan, ability to kill antigen presenting cells, or a combination thereof of T cells and / or natural killer cells may 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, a vector described herein, or a combination thereof to obtain transduced T cells and / or natural killer cells, and obtaining transduced T cells and / or natural killer cells, wherein the persistence, lifespan, naive state, ability to kill antigen presenting cells, or a combination thereof of the transduced T cells and / or natural killer cells are increased compared to the persistence, lifespan, naive state, ability to kill antigen presenting cells, or a combination thereof of a control cell. In an embodiment, the method may further comprise expanding the transduced T cells and / or natural killer cells.

[0196] In embodiments, the control cells may comprise untransduced T cells and / or natural killer cells, T cells and / or natural killer cells transduced with only TCR, or a combination thereof. In embodiments, the control cells may comprise untransduced T cells and / or natural killer cells, T cells and / or natural killer cells transduced with only TCR, T cells and / or natural killer cells transduced with only TCR and CD8, or a combination thereof. In embodiments, the persistence, lifespan, functionality, naive state, ability to kill antigen presenting cells, or a combination thereof of the transduced T cells and / or natural killer cells and control cells may be determined after stimulating with antigen presenting cells once, twice, three times, four times, five times, six times, seven times, or more times with antigen presenting cells. The persistence, lifespan, functionality, naive state, ability to kill antigen presenting cells, or a combination thereof, of the transduced T cells and / or natural killer cells and control cells can be determined after five or more stimulations with antigen presenting cells or multiple stimulations with antigen presenting cells.

[0197] In an embodiment, a method for increasing the secretion of interferon gamma (IFNγ) by T cells and / or natural killer cells may 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, a vector described herein, or a combination thereof to obtain transduced T cells and / or natural killer cells, and obtaining 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 the IFNγ secretion of control cells. In an embodiment, the method may further comprise amplifying the transduced T cells and / or natural killer cells. In an embodiment, the control cells may comprise untransduced T cells and / or natural killer cells, T cells and / or natural killer cells transduced with only TCR, or a combination thereof.

[0198] In embodiments, the control cells may comprise untransduced T cells and / or natural killer cells, T cells and / or natural killer cells transduced with only TCR, T cells and / or natural killer cells transduced with only TCR and CD8, or a combination thereof. In embodiments, IFNγ secretion by transduced T cells and / or natural killer cells and control cells may be determined after stimulating with antigen presenting cells once, twice, three times, four times, five times, six times, seven times, or more times with antigen presenting cells. In embodiments, IFNγ secretion by transduced T cells and / or natural killer cells and control cells may be determined after stimulating with antigen presenting cells five or more times or multiple times with antigen presenting cells.

[0199] In an embodiment, the antigen presenting cell can present an antigen on the cell surface, and the transduced T cells and / or natural killer cells and control cells can kill the antigen presenting cell. In an embodiment, the antigen can comprise a peptide. In an embodiment, the peptide can be complexed with an MHC molecule on the cell surface.

[0200] In an embodiment, a polypeptide, a plurality of polypeptides or a fusion polypeptide may be provided, wherein the polypeptide, a plurality of polypeptides or a fusion polypeptide is encoded by a nucleic acid described herein.

[0201] In embodiments, the nucleic acids described herein may be isolated, recombinant, or both isolated and recombinant.

[0202] In embodiments, the vectors described herein may be isolated, recombinant, or both isolated and recombinant.

[0203] In embodiments, the T cells and / or natural killer cells described herein may be isolated, recombinant, engineered, or any combination thereof.

[0204] In embodiments, a polypeptide, polypeptides, or fusion polypeptides described herein may be isolated, recombinant, or both isolated and recombinant.

[0205] In an embodiment, a vector may be provided, the vector comprising a nucleic acid described herein. In an embodiment, the vector described herein may further comprise a nucleic acid encoding a 2A peptide or an internal ribosome entry site (IRES) between a nucleic acid encoding a CD8 alpha chain and a nucleic acid encoding a CD8 beta chain. In an embodiment, the vector may further comprise a nucleic acid encoding a 2A peptide or an IRES between a nucleic acid encoding a TCR alpha chain and a nucleic acid encoding a TCR beta chain. In an embodiment, the vector may further comprise a nucleic acid encoding a 2A peptide or an IRES 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 an embodiment, 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 an embodiment, the IRES may be selected from the group consisting of: an IRES from a picornavirus, an IRES from a flavivirus, an IRES from a pestivirus, an IRES from a retrovirus, an IRES from a lentivirus, an IRES from an insect RNA virus, and an IRES from a cellular mRNA. In an embodiment, the vector may further comprise a post-transcriptional regulatory element (PRE) sequence selected from woodchuck PRE (WPRE) (SEQ ID NO: 264), woodchuck PRE (WPRE) mutant 1 (SEQ ID NO: 256), woodchuck PRE (WPRE) mutant 2 (SEQ ID NO: 257), or hepatitis B virus (HBV) PRE (HPRE) (SEQ ID NO: 437). In an embodiment, the post-transcriptional regulatory element (PRE) sequence may be woodchuck PRE (WPRE) mutant 1, the WPRE mutant comprising the nucleic acid sequence of SEQ ID NO: 256. In an embodiment, the post-transcriptional regulatory element (PRE) sequence may be a woodchuck PRE (WPRE) mutant 2, the WPRE mutant comprising the nucleic acid sequence of SEQ ID NO: 257. In an embodiment, 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 acid protein (GFAP) promoter, a modified MoMuLV LTR comprising a myeloproliferative sarcoma virus enhancer (MNDU3), a ubiquitin C promoter, an EF-1α promoter, or a murine stem cell virus (MSCV) promoter. In an embodiment, the promoter may be a murine stem cell virus (MSCV) promoter. In an embodiment, the vector may be a viral vector or a non-viral vector. In an embodiment, the vector may be a viral vector.In an embodiment, the viral vector can be selected from adenovirus, poxvirus, alphavirus, arenavirus, flavivirus, rhabdovirus, retrovirus, lentivirus, herpesvirus, paramyxovirus, picornavirus and any combination thereof. In an embodiment, the viral vector can be selected from the natural cat endogenous virus (RD114), a version of RD114 (RD114TR), gibbon ape leukemia virus (GALV), a version of GALV (GALV-TR), amphoteric murine leukemia virus (MLV4070A), baculovirus (GP64), vesicular stomatitis virus (VSV-G), fowl plague virus (FPV), Ebola virus (EboV), or baboon retrovirus envelope glycoprotein (BaEV) and lymphocytic choriomeningitis virus (LCMV) The envelope protein of the virus is pseudotyped. In an embodiment, the vector can be a lentiviral vector. In an embodiment, the vector may further include a nucleic acid encoding a chimeric antigen receptor (CAR).

[0206] In an embodiment, a T cell and / or a natural killer cell may be provided, the T cell and / or the natural killer cell expressing a polypeptide as described herein and / or comprising a vector as described herein and / or produced by a method as described herein. In an embodiment, the T cell may be an αβT cell, a γδT cell, a natural killer T cell, or any combination thereof. In an embodiment, the αβT cell may be a CD4+T cell. In an embodiment, the αβT cell may be a CD8+T cell. In an embodiment, the γδT cell may be a Vγ9Vδ2+T cell.

[0207] In an embodiment, a composition may be provided, comprising T cells and / or natural killer cells as described herein. In an embodiment, the composition may be a pharmaceutical composition. In an embodiment, the composition may further comprise an adjuvant, an excipient, a carrier, a diluent, a buffer, a stabilizer, or a combination thereof. In embodiments, the adjuvant may be an anti-CD40 antibody, imiquimod, resiquimod, GM-CSF, cyclophosphamide, sunitinib, bevacizumab, atezolizumab, interferon alpha, interferon beta, CpG oligonucleotides and derivatives, poly(I:C) and derivatives, RNA, sildenafil, poly(lactide-co-glycolide) (PLG)-containing particle formulations, 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 embodiments, the adjuvant may be IL-2, IL-7, IL-12, IL-15, IL-21, or any combination thereof.

[0208] In an embodiment, a method of treating a patient with cancer may be provided, the method comprising 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 cancer, gastric cancer, and prostate cancer. In an embodiment, a method of eliciting an immune response in a patient with cancer may be provided, the method comprising 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 cancer, gastric cancer, and prostate cancer. In an embodiment, the T cells and / or natural killer cells can kill cancer cells that present peptides complexed with MHC molecules on the cell surface.

[0209] In an embodiment, the nucleic acid sequences disclosed herein may be mutated so that the encoded amino acid remains unchanged, but the nucleic acid codons are changed to maintain improved expression in the target cell and / or by the target vector. In an embodiment, the nucleic acid disclosed herein may be codon optimized. In an embodiment, the nucleic acid sequences shown in this article are codon optimized. In an embodiment, the nucleic acid sequences shown in this article may be codon optimized, and the nucleic acid sequences encoding the polypeptides shown in this article may be codon optimized. In an embodiment, the mutations of the nucleic acid sequences shown in this article may encompass codon optimization.

[0210] In embodiments, the expression of membrane-bound IL-15 can improve immune cells, such as but not limited to T cells and / or natural killer cells, persistence, functionality, growth, viability, expansion, or any combination thereof, compared to cells that do not express membrane-bound IL-15. In embodiments, the expression of membrane-bound IL-15 can improve immune cells, such as but not limited to T cells and / or natural killer cells, persistence, functionality, growth, viability, expansion, or any combination thereof in a tumor microenvironment, compared to cells that do not express membrane-bound IL-15. In embodiments, the expression of membrane-bound IL-15 can increase the efficacy of immune cells, such as but not limited to T cells and / or natural killer cells, in killing tumor cells, compared to cells that do not express membrane-bound IL-15. In embodiments, the expression of membrane-bound IL-15 can increase the ability of immune cells, such as but not limited to T cells and / or natural killer cells, to survive in a tumor microenvironment, to persist in killing tumor cells, or any combination thereof, compared to cells that do not express membrane-bound IL-15. In embodiments, expression of membrane-bound IL-15 can increase the ability of immune cells, such as but not limited to T cells and / or natural killer cells, to maintain a naive phenotype.

[0211] As a non-limiting example, persistence can be assessed by the length of time of cells detectable in an individual (e.g., patient) after infusion. As a non-limiting example, persistence can be measured in a few days, weeks, months, or years after infusion, as a non-limiting example, 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. As a non-limiting example, persistence can be assessed by PCR of peripheral blood samples, flow cytometry of peripheral blood samples, and / or by analysis of tumor biopsy samples. As a non-limiting example, the persistence of cells expressing membrane-bound IL-15 can be compared with the typical persistence of infused ACT cells or the persistence of similar cells that do not express membrane-bound IL-15.

[0212] As non-limiting examples, the sustained ability to kill tumor cells can be measured by: (i) using the IncuCyte's serial killing assay (wherein the ability to kill / impair tumor growth as measured by growth fold over a period of time is assessed), and / or (ii) by cytokine / effector molecule production (IFNγ by ELISA and other proinflammatory cytokines by Luminex (as non-limiting examples, the cytokines measured can include IFNγ, TNFα, granzyme B, perforin, IL-2, IL-6, MIP-1β, MIP-1α, GM-CSF, RANTES, IL-18, IL-4, IL-10, and IP10)). As a non-limiting example, the sustained ability of cells expressing membrane-bound IL-15 to kill tumor cells can be compared to the sustained ability of similar cells that do not express membrane-bound IL-15, or the sustained ability of other control cells to kill tumor cells.

[0213] As a non-limiting example, the original state of the phenotype can be assessed by flow cytometry through the Tmem set assay. Typically, flow cytometer gating excludes CD8+TCR+ cells. Typically, the higher frequencies of T memory subsets T original / scm (CD45RA+CCR7+) and Tcm (CD45RA-CCR7+) and the increase or retention of CD39-CD69- and CD27+CD28+ colonies may indicate a more original phenotype. Low CD57 expression can also be desirable.

[0214] When assessing the persistence, functionality, growth, viability, expansion, tumor killing efficacy, naive state or other properties of cells expressing dnTGFRβRII, cells such as, by way of non-limiting example, untransduced cells, cells transduced with TCR only, cells transduced with CD8 and TCR, or a combination thereof, can serve as control cells.

[0215] In embodiments, membrane-bound IL-15 can act in a cis manner (e.g., affecting cells that express it), in a trans manner (e.g., affecting cells that do not express it), or any combination thereof. In embodiments where membrane-bound IL-15 acts in trans, cells that are adjacent to or near cells expressing membrane-bound IL-15 (e.g., within a tumor microenvironment) can exhibit any improvement or combination of improvements that are the same or similar to the described cells expressing membrane-bound IL-15 compared to cells that are not adjacent to or near cells expressing membrane-bound IL-15.

[0216] In an embodiment, the present disclosure provides nucleic acids encoding polypeptides described herein. In an embodiment, the present disclosure provides vectors comprising nucleic acids encoding polypeptides described herein. In an embodiment, one or more vectors may comprise nucleic acids encoding IL-15 / IL-15Rα fusion polypeptides. In an embodiment, one or more vectors may comprise nucleic acids encoding CD8 polypeptides. In an embodiment, one or more vectors may comprise nucleic acids encoding CD8α polypeptides. In an embodiment, one or more vectors may comprise nucleic acids encoding CD8β polypeptides.

[0217] In an embodiment, one or more vectors may include one or more nucleic acids encoding a T cell receptor (TCR) comprising an alpha chain and a beta chain. In an embodiment, one or more vectors may include one or more nucleic acids encoding a T cell receptor (TCR) comprising a gamma chain and a delta chain. In an embodiment, one or more vectors may include one or more nucleic acids encoding a chimeric antigen receptor (CAR).

[0218] In an embodiment, one or more vectors may be provided, the one or more vectors comprising one or more nucleic acids encoding a TCR comprising an alpha chain and a beta chain, a TCR comprising a gamma chain and a delta chain, a CAR, an IL-15 polypeptide, an IL-15Rα polypeptide, an IL-15 / IL-15Rα fusion polypeptide, and / or a CD8 polypeptide, or any combination thereof. In an embodiment, the CD8 polypeptide may comprise a CD8α chain and / or a CD8β chain, and the CD8α chain and / or the CD8β chain may be independently modified or unmodified.

[0219] In an embodiment, one or more vectors may be provided, comprising one or more nucleic acids encoding one or more TCRs comprising α chains and β chains, IL-15 / IL-15Rα fusion polypeptides, and / or CD8 polypeptides or any combination thereof. In an embodiment, one or more vectors may be provided, comprising one or more nucleic acids encoding one or more TCRs comprising γ chains and δ chains, IL-15 / IL-15Rα fusion polypeptides, and / or CD8 polypeptides or any combination thereof. In an embodiment, one or more vectors may be provided, comprising one or more nucleic acids encoding one or more CARs, IL-15 / IL-15Rα fusion polypeptides, and / or CD8 polypeptides or any combination thereof. In an embodiment, a CD8 polypeptide may comprise a CD8 α chain and / or a CD8 β chain, and the CD8 α chain and / or the CD8 β chain may be independently modified or unmodified.

[0220] In an embodiment, one or more vectors may be provided, comprising one or more nucleic acids encoding TCRs comprising α chains and β chains, IL-15 / IL-15Rα fusion polypeptides, and CD8 polypeptides. In an embodiment, one or more vectors may be provided, comprising one or more nucleic acids encoding TCRs comprising γ chains and δ chains, IL-15 / IL-15Rα fusion polypeptides, and CD8 polypeptides. In an embodiment, one or more vectors may be provided, comprising one or more nucleic acids encoding CARs, IL-15 / IL-15Rα fusion polypeptides, and CD8 polypeptides. In an embodiment, a CD8 polypeptide may include a CD8α chain and / or a CD8β chain, and the CD8α chain and / or the CD8β chain may be independently modified or unmodified.

[0221] In an embodiment, one or more vectors may be provided, comprising one or more nucleic acids encoding TCR and IL-15 / IL-15Rα fusion polypeptides comprising α chain and β chain. In an embodiment, one or more vectors may be provided, comprising one or more nucleic acids encoding TCR and IL-15 / IL-15Rα fusion polypeptides comprising γ chain and δ chain. In an embodiment, one or more vectors may be provided, comprising one or more nucleic acids encoding CAR and IL-15 / IL-15Rα fusion polypeptides.

[0222] In an embodiment, one or more vectors may be provided, comprising one or more nucleic acids, the one or more nucleic acids encoding TCR and CD8 polypeptides comprising α chains and β chains. In an embodiment, one or more vectors may be provided, the one or more vectors comprising one or more nucleic acids, the one or more nucleic acids encoding TCR and CD8 polypeptides comprising γ chains and δ chains. In an embodiment, one or more cells may be provided, the one or more cells comprising one or more nucleic acids, the one or more nucleic acids encoding CAR and CD8 polypeptides. In an embodiment, the CD8 polypeptide may include a CD8 α chain and / or a CD8 β chain, and the CD8 α chain and / or the CD8 β chain may be independently modified or unmodified.

[0223] In embodiments, more than one vector may be co-transduced into one or more cells, co-expressed in one or more cells, or any combination thereof. In embodiments, the one or more cells may comprise αβT cells, γδT cells, natural killer (NK) cells, natural killer T cells, CD4+T cells, CD8+T cells, CD4+ / CD8+ cells, or any combination thereof.

[0224] In an embodiment, more than one vector may comprise one or more nucleic acids encoding 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, or any combination thereof. In an embodiment, a CD8 polypeptide may comprise a CD8α chain and / or a CD8β chain, and the CD8α chain and / or the CD8β chain may be independently modified or unmodified.

[0225] In an embodiment, a single vector may comprise one or more nucleic acids encoding 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, or any combination thereof. In an embodiment, a CD8 polypeptide may comprise a CD8α chain and / or a CD8β chain, and the CD8α chain and / or the CD8β chain may be independently modified or unmodified.

[0226] In an embodiment, the nucleic acid can be polycistronic, and one or more polycistronic nucleic acids can be utilized. Expression of multiple (e.g., 2, 3, 4, 5 or more) polypeptides from a polycistronic nucleic acid can also be achieved by any suitable method, such as i) pre-mRNA splicing; ii) proteolytic cleavage sites; iii) fusion proteins; iv) including one or more nucleic acids encoding 2A peptides (e.g., but not limited to P2A, T2A, E2A, and F2A); v) including one or more internal ribosome entry sites (IRES), or other mechanisms. Each of these methods has some advantages and disadvantages for providing multiple transcription units. Among the five methods, the most widely used are self-cleaving 2A peptides and IRES. In an embodiment, the nucleic acid can be monocistronic, and one or more monocistronic nucleic acids can be utilized.

[0227] In an embodiment, the 2A peptide may be selected from P2A (SEQ ID NO: 93), T2A (SEQ ID NO: 94), E2A (SEQ ID NO: 95) or F2A (SEQ ID NO: 96).

[0228] In an embodiment, the IRES may be selected from the group consisting of an IRES from a picornavirus, an IRES from a flavivirus, an IRES from a pestivirus, an IRES from a retrovirus, an IRES from a lentivirus, an IRES from an insect RNA virus, and an IRES from a cellular mRNA.

[0229] In embodiments, the vector may comprise a nucleic acid encoding a 2A peptide or an internal ribosome entry site (IRES) located between the nucleic acid encoding the modified CD8 alpha polypeptide and the nucleic acid encoding the CD8 beta polypeptide.

[0230] In an embodiment, the vector may comprise a nucleic acid encoding a 2A peptide located between a nucleic acid encoding a TCR alpha chain and a nucleic acid encoding a TCR beta chain.

[0231] In an embodiment, the vector may comprise a nucleic acid encoding a 2A peptide or an internal ribosome entry site (IRES) located between a nucleic acid encoding a modified CD8α polypeptide, a nucleic acid encoding a CD8β polypeptide, a nucleic acid encoding a TCRα chain, or a nucleic acid encoding a TCRβ chain and a nucleic acid encoding membrane-bound IL-15.

[0232] In an embodiment, a single vector may comprise one or more nucleic acids encoding 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, or any combination thereof, and the vector may comprise a nucleic acid encoding a 2A peptide or an internal ribosome entry site (IRES) between any or each of the nucleic acids encoding the polypeptide or fusion polypeptide. In an embodiment, a CD8 polypeptide may comprise a CD8α chain and / or a CD8β chain, and the CD8α chain and / or the CD8β chain may be independently modified or unmodified.

[0233] In an embodiment, the vector may further comprise a post-transcriptional regulatory element (PRE) sequence. In an embodiment, the post-transcriptional regulatory element (PRE) sequence may be selected from woodchuck hepatitis virus PRE (WPRE) (such as but not limited to wild-type WPRE, such as but not limited to SEQ ID NO: 264, or mutated WPRE, such as but not limited to WPREmut1 (SEQ ID NO: 256) or WPREmut2 (SEQ ID NO: 257)) or hepatitis B virus (HBV) PRE (HPRE) (SEQ ID NO: 437), variants thereof, or any combination thereof.

[0234] In an embodiment, the vector may further comprise one or more promoters. In an embodiment, the promoter may be selected from a cytomegalovirus (CMV) promoter, a phosphoglycerate kinase (PGK) promoter, a myelin basic protein (MBP) promoter, a glial fibrillary acid protein (GFAP) promoter, a modified MoMuLV LTR comprising a myeloproliferative sarcoma virus enhancer (MNDU3), a ubiquitin C promoter, an EF-1α promoter, a murine stem cell virus (MSCV) promoter, a promoter from CD69, a nuclear factor of activated T cells (NFAT) promoter, an IL-2 promoter, a minimal IL-2 promoter, or a combination thereof.

[0235] In embodiments, the vector can be a viral vector or a non-viral vector.

[0236] In an embodiment, 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.

[0237] In an embodiment, the vector can be pseudotyped with an envelope protein of a virus selected from the natural feline endogenous virus (RD114), a chimeric version of RD114 (RD114TR), Gibbon Ape Leukemia Virus (GALV), a chimeric version of GALV (GALV-TR), amphotropic murine leukemia virus (MLV 4070A), baculovirus (GP64), vesicular stomatitis virus (VSV-G), fowl plague virus (FPV), Ebola virus (EboV), or baboon retrovirus envelope glycoprotein (BaEV), lymphocytic choriomeningitis virus (LCMV), or a combination thereof.

[0238] In an embodiment, the vector may comprise one or more Kozak sequences. In an embodiment, the Kozak sequence may initiate, increase or promote translation or a combination thereof. In an embodiment, the Kozak sequence may be GCCACC. In an embodiment, the Kozak sequence may be ACCATGG. In an embodiment, the Kozak sequence may be GCCNCCATGG, wherein N is a purine (A or G) (SEQ ID NO: 382).

[0239] In embodiments, the vector may comprise one or more Factor Xa sites.

[0240] In an embodiment, the vector may comprise one or more enhancers. In an embodiment, the enhancer may comprise conserved non-coding sequence (CNS) 0, CNS 1, CNS 2, CNS 3, CNS 4 or a portion or any combination thereof.

[0241] In an embodiment, the present disclosure provides one or more cells transduced with and / or expressing one or more vectors comprising a nucleic acid encoding a polypeptide.

[0242] In embodiments, the cells may comprise 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.

[0243] In an embodiment, the T cell may be a CD4+T cell. In an embodiment, the T cell may be a CD8+T cell. In an embodiment, the T cell may be a CD4+ / CD8+T cell. In an embodiment, the T cell may be an αβT cell. In an embodiment, the T cell may be a γδT cell.

[0244] In an embodiment, the T cell may be an αβT cell and may express a CD8 polypeptide described herein. In an embodiment, the CD8 polypeptide may comprise a CD8 α chain and / or a CD8 β chain, and the CD8 α chain and / or the CD8 β chain may be independently modified or unmodified. In an embodiment, the T cell may be an αβT cell and may express a modified CD8 polypeptide described herein, such as a modified CD8 α polypeptide or a modified CD8 α polypeptide having a CD8 β stem region, such as m1CD8α in constructs #11 and #12 ( Figure 4 ) and CD8α*( Fig.55B ). In an embodiment, the T cell may be an αβT cell and may 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.

[0245] In embodiments, the T cell may be a γδT cell and may express a CD8 polypeptide described herein. In embodiments, the CD8 polypeptide may comprise a CD8 α chain and / or a CD8 β chain, and the CD8 α chain and / or the CD8 β chain may be independently modified or unmodified. In embodiments, the T cell may be a γδT cell and may express a modified CD8 polypeptide described herein, such as a modified CD8 α polypeptide or a modified CD8 α polypeptide having a CD8 β stem region, such as m1CD8α in constructs #11 and #12 ( Figure 4 ) and CD8α*( Fig.55B). In an embodiment, the T cell may be a γδT cell and may 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 an embodiment, the CD8 polypeptide may comprise a CD8α chain and / or a CD8β chain, and the CD8α chain and / or the CD8β chain may be independently modified or unmodified.

[0246] In an embodiment, one or more cells may be provided, the one or more cells comprising or comprising one or more nucleic acids encoding a TCR comprising an alpha chain and a beta chain, a TCR comprising a gamma chain and a delta chain, a CAR, an IL-15 polypeptide, an IL-15Rα polypeptide, an IL-15 / IL-15Rα fusion polypeptide, and / or a CD8 polypeptide, or any combination thereof. In an embodiment, the CD8 polypeptide may comprise a CD8 alpha chain and / or a CD8 beta chain, and the CD8 alpha chain and / or the CD8 beta chain may be independently modified or unmodified.

[0247] In an embodiment, one or more cells may be provided, the one or more cells comprising or comprising one or more nucleic acids, the one or more nucleic acids encoding one or any combination of TCRs comprising α chains and β chains, IL-15 / IL-15Rα fusion polypeptides, and / or CD8 polypeptides. In an embodiment, one or more cells may be provided, the one or more cells comprising or comprising one or more nucleic acids, the one or more nucleic acids encoding one or any combination of TCRs comprising γ chains and δ chains, IL-15 / IL-15Rα fusion polypeptides, and / or CD8 polypeptides. In an embodiment, one or more cells may be provided, the one or more cells comprising or comprising one or more nucleic acids, the one or more nucleic acids encoding CARs, IL-15 / IL-15Rα fusion polypeptides, and / or CD8 polypeptides. In an embodiment, a CD8 polypeptide may comprise a CD8 α chain and / or a CD8 β chain, and the CD8 α chain and / or the CD8 β chain may be independently modified or unmodified.

[0248] In an embodiment, one or more cells may be provided, the one or more cells comprising or comprising one or more nucleic acids, the one or more nucleic acids encoding TCR comprising α chain and β chain, IL-15 / IL-15Rα fusion polypeptide and CD8 polypeptide. In an embodiment, one or more cells may be provided, the one or more cells comprising or comprising one or more nucleic acids, the one or more nucleic acids encoding TCR comprising γ chain and δ chain, IL-15 / IL-15Rα fusion polypeptide and CD8 polypeptide. In an embodiment, one or more cells may be provided, the one or more cells comprising or comprising one or more nucleic acids, the one or more nucleic acids encoding CAR, IL-15 / IL-15Rα fusion polypeptide and CD8 polypeptide. In an embodiment, the CD8 polypeptide may comprise a CD8 α chain and / or a CD8 β chain, and the CD8 α chain and / or the CD8 β chain may be independently modified or unmodified.

[0249] In an embodiment, one or more cells may be provided, the one or more cells comprising or comprising one or more nucleic acids, the one or more nucleic acids encoding TCR and IL-15 / IL-15Rα fusion polypeptides comprising α chain and β chain. In an embodiment, one or more cells may be provided, the one or more cells comprising or comprising one or more nucleic acids, the one or more nucleic acids encoding TCR and IL-15 / IL-15Rα fusion polypeptides comprising γ chain and δ chain. In an embodiment, one or more cells may be provided, the one or more cells comprising or comprising one or more nucleic acids, the one or more nucleic acids encoding CAR and IL-15 / IL-15Rα fusion polypeptides.

[0250] In an embodiment, one or more cells may be provided, the one or more cells comprising or comprising one or more nucleic acids, the one or more nucleic acids encoding TCR and CD8 polypeptides comprising α chain and β chain. In an embodiment, one or more cells may be provided, the one or more cells comprising or comprising one or more nucleic acids, the one or more nucleic acids encoding TCR and CD8 polypeptides comprising γ chain and δ chain. In an embodiment, one or more cells may be provided, the one or more cells comprising or comprising one or more nucleic acids, the one or more nucleic acids encoding CAR and CD8 polypeptides. In an embodiment, the CD8 polypeptide may comprise a CD8 α chain and / or a CD8 β chain, and the CD8 α chain and / or the CD8 β chain may be independently modified or unmodified.

[0251] In embodiments, one or more nucleic acids may be contained in and / or expressed from one or more vectors.

[0252] In embodiments, the one or more cells may comprise αβ T cells, γδ T cells, natural killer cells, CD4+ T cells, CD8+ T cells, CD4+ / CD8+ cells, or any combination thereof.

[0253] In an embodiment, a cell population as described herein can be provided. As a non-limiting example, the present disclosure provides a modified cell population, the modified cell population comprising or comprising one or more nucleic acids encoding one or any combination of the following: an exogenous CD8 co-receptor, the exogenous CD8 co-receptor comprising a polypeptide described herein, e.g., 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 (e.g., an IL-15 / IL-15Rα fusion polypeptide) as described herein; and / or a T cell receptor. In an embodiment, the cell population may comprise 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.

[0254] In one aspect, the polypeptide sequences and / or nucleic acid sequences described herein may be isolated and / or recombinant sequences.

[0255] In one aspect, the cells described herein can be isolated and / or recombinant cells.

[0256] In an embodiment, a method of preparing cells for immunotherapy may 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 expanding the transduced cells. In an embodiment, the cells may include αβT cells, γδT cells, natural killer cells, natural killer T cells, CD4+T cells, CD8+T cells, CD4+ / CD8+ cells, or any combination thereof.

[0257] In an embodiment, a method of treating a patient with cancer may comprise administering to the patient a composition comprising an expanded cell population, wherein the cells kill cancer cells presenting a peptide complexed with an MHC molecule on the surface, wherein the peptide is selected from SEQ ID NO: 98-255, 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 cancer, gastric cancer, prostate cancer, or a combination thereof. In an embodiment, the cell may comprise an αβT cell, a γδT 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. In an embodiment, the composition may further comprise an adjuvant. In an embodiment, the adjuvant can be selected from anti-CD40 antibodies, imiquimod, resiquimod, GM-CSF, cyclophosphamide, sunitinib, bevacizumab, atezolizumab, interferon alpha, interferon beta, CpG oligonucleotides and derivatives, poly (I: C) and derivatives, RNA, sildenafil, poly (lactide-co-glycolide) (PLG)-containing particle formulations, virosomes, interleukin (IL) -1, IL-2, IL-4, IL-7, IL-12, IL-13, IL-15, IL-21, IL-23 or any combination thereof.

[0258] In an embodiment, a method of eliciting an immune response in a patient with cancer may comprise administering to the patient a composition comprising an expanded population of cells, wherein the cells kill cancer cells presenting a peptide complexed with an MHC molecule on the surface, wherein the peptide is selected from SEQ ID NO: 98-255, 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 cancer, gastric cancer, prostate cancer, or a combination thereof. In an embodiment, the cell may comprise an αβT cell, a γδT 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. BRIEF DESCRIPTION OF THE DRAWINGS

[0259] Figure 1 A representative CD8α subunit is shown, for example, SEQ ID NO: 258 (CD8α1). CD8α1 includes five domains: (1) a signal peptide; (2) an Ig-like domain-1; (3) a stem region; (4) a transmembrane (TM) domain; and (5) a cytoplasmic tail (Cyto) containing an lck binding motif.

[0260] Figure 2Shown is a sequence alignment between CD8α1 (SEQ ID NO: 258) and m1CD8α (SEQ ID NO: 7).

[0261] Figure 3 A sequence alignment between CD8α2 (SEQ ID NO: 259) and m2CD8α (SEQ ID NO: 262) is shown, with the cysteine ​​substitution at position 112 indicated by an arrow.

[0262] Figure 4 An exemplary vector according to aspects of the present disclosure is shown. In an embodiment, the vector may further comprise additional elements, such as those described herein, such as but not limited to one or more promoters or one or more post-transcriptional regulatory elements. Figure 4 In the figure, the line can represent a direct connection without intervening sequences, or can represent intervening sequences, such as but not limited to linkers, furin, sequences encoding 2A polypeptides, factor Xa sites, untranslated sequences, translated sequences, sequences comprising one or more restriction endonuclease sites, or combinations thereof.

[0263] Figure 5A Shows Figure 4 Titers of viral vectors are indicated.

[0264] Figure 5B Titers of additional viral vectors according to the present disclosure are shown. 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 NO:15 and SEQ ID NO:16, which can be encoded by SEQ ID NO:72 and SEQ ID NO:73, respectively) (Construct #8), which is combined with 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 different batches of the same construct (SEQ ID NOs:285 and 286).

[0265] Figure 6 T cell preparation is shown.

[0266] Fig. 7A Shown is the expression of activation markers before and after activation in CD3+CD8+ cells.

[0267] Figure 7B Shown is the expression of activation markers before and after activation in CD3+CD4+ cells.

[0268] Fig. 8A The fold expansion of cells transduced with various constructs from donor #1 is shown. 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); NT = non-transduced T cells (as negative control). Note that constructs #9 and #9b are different batches of the same construct (SEQ ID NOs: 287 and 288).

[0269] Figure 8B The fold expansion of cells transduced with various constructs from donor #2 is shown. 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 (as negative control).

[0270] Fig. 9A Shown are flow cytometry plots of cells transduced with construct #9.

[0271] Fig. 9B Shown is a flow cytometric plot of cells transduced with construct #10 according to one embodiment of the present disclosure.

[0272] Fig. 9C Flow cytometry of cells transduced with construct #11 is shown.

[0273] Fig.9D Shown are flow cytometry plots of cells transduced with construct #12.

[0274] Fig.10 Shown are the CD8+CD4+% in cells transduced with various constructs from donor #1 and donor #2. 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); NT = non-transduced T cells (as negative control).

[0275] Fig.11 Shown are the % Tet relative to CD8+CD4+ in cells transduced with various constructs. 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); NT = non-transduced T cells (as negative control).

[0276] Fig.12 Tet MFI (CD8+CD4+Tet+) in cells transduced with various constructs are shown. 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); NT = non-transduced T cells (as negative control).

[0277] Fig.13 CD8α MFI (CD8+CD4+Tet+) in cells transduced with various constructs are shown. 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); NT = non-transduced T cells (as negative control).

[0278] Fig.14 The CD8+CD4% (relative to CD3+) in cells transduced with various constructs are shown. 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); NT = non-transduced T cells (as negative control).

[0279] Fig.15 The CD8+Tet+% (relative to CD3+) in cells transduced with various constructs are shown. 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); NT = non-transduced T cells (as negative control).

[0280] Fig.16 Tet MFI (CD8+Tet+) in cells transduced with various constructs are shown. 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); NT = non-transduced T cells (as negative control).

[0281] Fig.17CD8α MFI (CD8+Tet+) in cells transduced with various constructs are shown. 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); NT = non-transduced T cells (as negative control).

[0282] Fig.18 The % Tet+ (relative to CD3+) in cells transduced with various constructs are shown. 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); NT = non-transduced T cells (as negative control).

[0283] Fig.19 VCN (upper panel) and CD3+Tet+ / VCN (lower panel) of cells transduced with various constructs are shown. 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); NT = non-transduced T cells (as negative control).

[0284] Figures 20A-20C Depicted are data showing that constructs (#10, #11, and #12) are comparable to TCR alone in mediating cytotoxicity against target positive cell lines expressing antigen at varying levels (1081 copies per cell for UACC257 and 50 copies per cell for A375).

[0285] Figures 21A-21B Depicted are data showing that IFNγ secretion in response to UACC257 is comparable among constructs, however for A375, #10 expression is the highest among all constructs. However, comparing #9 and #11 expressing wild-type and modified CD8 co-receptor sequences, respectively, T cells transduced with #11 induced a stronger cytokine response as measured by quantitative IFNγ in the supernatant from Incucyte plates. Construct #9; Construct #10; Construct #11; Construct #12; Construct #1; Construct #2; Construct #8 = R11KEATCR only.

[0286] Fig. 22 Depicted is an exemplary experimental design for evaluating DC maturation and cytokine secretion of PBMC-derived products in response to UACC257 and A375 targets. N=2.

[0287] Figures 23A-23B Depicted are data showing increased IFNγ secretion in response to A375 in the presence of iDCs. In triple culture with iDCs, IFNγ secretion was higher in construct #10 compared to the other constructs. However, construct #9 expressing wild-type and modified CD8 co-receptor sequences, respectively, was compared to construct #11, and T cells transduced with #11 induced a stronger cytokine response as measured by quantitative IFNγ in the culture supernatant of the three-way co-culture using donor D600115, E:T:iDC::1:1 / 10:1 / 4. Construct #9; Construct #10; Construct #11; Construct #12; Construct #1; Construct #2; Construct #8 = R11KEATCR only.

[0288] Figures 24A-24B Depicted are data showing increased IFNγ secretion in response to A375 in the presence of iDCs. In triple culture with iDCs, IFNγ secretion was higher in construct #10 compared to the other constructs. IFNγ was quantified in culture supernatants of three-way co-cultures using donor D150081, E:T:iDC::1:1 / 10:1 / 4. Construct #9; Construct #10; Construct #11; Construct #12; Construct #1; Construct #2; Construct #8 = R11KEATCR only.

[0289] Figures 25A-25B Depict data showing increased IFNγ secretion in response to UACC257 in the presence of iDC. In triple culture with iDC, IFNγ secretion in construct #10 was higher than in other constructs. However, construct #9 and construct #11, which express wild-type and modified CD8 co-receptor sequences, respectively, were compared, as measured by quantitative IFNγ in the culture supernatant of three-way co-cultures using donor D600115, E:T:iDC::1:1 / 10:1 / 4, and T cells transduced with construct #11 induced a stronger cytokine response. Construct #9; construct #10; construct #11; construct #12; construct #1; construct #2; construct #8 = R11KEA TCR only.

[0290] Fig.26 T cell preparation according to one embodiment of the present disclosure is shown.

[0291] Fig.27A Shown is the expression of activation markers before and after activation in CD3+CD8+ cells.

[0292] Fig.27BShown is the expression of activation markers before and after activation in CD3+CD4+ cells according to one embodiment of the present disclosure.

[0293] Fig.28 The fold expansion of cells transduced with various constructs is shown.

[0294] Figures 29A-29B Shown are the CD8+CD4+ % in cells transduced with various constructs according to one embodiment of the present disclosure.

[0295] Figures 30A-30B Shown is the % CD8+CD4+ of Tet relative to in cells transduced with various constructs according to one embodiment of the present disclosure.

[0296] Figures 31A-31B Shown is Tet MFI (relative to CD8+CD4+Tet+) in cells transduced with various constructs according to one embodiment of the present disclosure.

[0297] Figures 32A-32B Shown is the CD8+CD4- % (relative to CD3+) in cells transduced with various constructs according to one embodiment of the present disclosure.

[0298] Figures 33A-33B Shown is the CD8+Tet+ % (relative to CD3+) in cells transduced with various constructs according to one embodiment of the present disclosure.

[0299] Figures 34A-34B Shown is Tet MFI (CD8+Tet+) in cells transduced with various constructs according to one embodiment of the present disclosure.

[0300] Figures 35A-35B Shown is the % Tet+ (relative to CD3+) in cells transduced with various constructs according to one embodiment of the present disclosure.

[0301] Figures 36A-36B Shown are VCNs of cells transduced with various constructs according to one embodiment of the present disclosure.

[0302] Fig.37 T cell preparation according to one embodiment of the present disclosure is shown.

[0303] Fig.38 Shown are the % of Tet relative to CD8+CD4+ in cells transduced with various constructs.

[0304] Fig.39 Shown are Tet MFI of CD8+CD4+Tet+ in cells transduced with various constructs.

[0305] Fig.40 Shown are Tet MFI of CD8+Tet+ in cells transduced with various constructs.

[0306] Fig.41 Shown are the % of Tet+ versus CD3+ cells transduced with various constructs.

[0307] Fig.42 The vector copy number (VCN) of cells transduced with various constructs is shown.

[0308] Fig.43 The % T cell subsets in cells transduced with various constructs are shown. FACS analysis was gated on CD3+TCR+.

[0309] Figures 44A-44B The % T cell subsets in cells transduced with various constructs are shown. FACS analysis for Fig.44A Gating was performed on CD4+CD8+ and for Fig.44B Gate on CD4-CD8+TCR+.

[0310] Figures 45A-45B Depict display construct #13 and #10 in mediation for the cytotoxicity of the UACC257 target positive cell line expressing high level antigen with only TCR suitable data (1081 copies per cell).Construct #15 is also effective, but killing is slower than construct #13 and #10.Effector for producing these results: target ratio is 4:1.

[0311] Fig.46 IFNγ secretion in response in the UACC257 cell line is shown, with higher secretion for construct #13 compared to construct #10. Quantified IFNγ in supernatant from Incucyte plates. The effector:target ratio used to generate these results was 4:1.

[0312] Fig.47 ICI marker frequencies are shown (2B4, 41BB, LAG3, PD-1, TIGIT, TIM3, CD39+CD69+, and CD39-CD69-).

[0313] Figures 48A-48G Shown is increased expression of IFNγ, IL-2 and TNFα by CD4+CD8+ cells transduced with construct #10 (WT signal peptide, CD8β1) compared to other constructs.FACS analysis was gated on CD3+CD4+CD8+ cells for UACC257, 4:1 E:T.

[0314] Figures 49A-49GShown is increased expression of IFNγ, IL-2, MIP-1β and TNFα by CD4-CD8+ cells transduced with construct #10 (WT signal peptide, CD8β1) compared to other constructs.FACS analysis was gated on CD3+CD4-CD8+ cells for UACC257, 4:1E:T.

[0315] Figures 50A-50G Shown is increased expression of IL-2 and TNFα by CD3+TCR+ cells transduced with construct #10 (WT signal peptide, CD8β1) compared to the other constructs.FACS analysis CD3+TCR+ cells were gated for UACC257, 4:1 E:T.

[0316] Figures 51A-51C Shown are results from FACS analysis gating CD4+CD8+ cells for A375, 4:1 E:T.

[0317] Figures 52A-52C Shown are results from FACS analysis gating CD4-CD8+ cells for A375, 4:1E:T.

[0318] Figures 53A-53C Shown are results from FACS analysis gating CD3+TCR+ cells for A375, 4:1E:T.

[0319] Fig.54 T cell preparation according to one embodiment of the present disclosure is shown.

[0320] Figures 55A-55C Figure 2 shows that antigen presenting cells (APCs) bind to peptide / MHC complexes of T cells by binding to the complex of TCR and CD8αβ heterodimers ( Fig.55A , e.g., produced by transducing T cells with construct #2, #3, #4, #10, #13, #14, #15, #16, #17, #18, or #21), a complex of a TCR whose stem region is replaced by the CD8β stem region (CD8αα*) and homodimeric CD8α ( Fig.55B , e.g., produced by transducing T cells with constructs #11, #12, or #19), and complexes of TCR and CD8α homodimers ( Fig.55C , e.g., generated by transducing T cells with construct #1, #5, #6, #7, or #9).

[0321] Fig.56 Shown is the level of IL-12 secretion by dendritic cells (DCs) in the presence of CD4+ T cells and immature dendritic cells (iDCs) transduced with construct #10 or #11, according to one embodiment of the present disclosure.

[0322] Fig.57 Shown is the level of TNF-α secretion by dendritic cells (DCs) in the presence of CD4+ T cells and immature dendritic cells (iDCs) transduced with construct #10 or #11, according to one embodiment of the present disclosure.

[0323] Fig.58 Shown is the level of IL-6 secretion by dendritic cells (DCs) in the presence of CD4+ T cells and immature dendritic cells (iDCs) transduced with construct #10 or #11, according to one embodiment of the present disclosure.

[0324] Fig.59 Shown is a protocol for determining the level of cytokine secretion by dendritic cells (DCs) in the presence of PBMCs and target cells transduced with various constructs according to one embodiment of the present disclosure.

[0325] Fig.60 Shown is the level of IL-12 secretion by dendritic cells (DCs) in the presence of PBMCs and target cells transduced with various constructs according to one embodiment of the present disclosure.

[0326] Fig.61 Shown is the level of TNF-α secretion by dendritic cells (DCs) in the presence of PBMCs and target cells transduced with various constructs according to one embodiment of the present disclosure.

[0327] Fig.62 Shown is the level of IL-6 secretion by dendritic cells (DCs) in the presence of PBMCs and target cells transduced with various constructs according to one embodiment of the present disclosure.

[0328] Figures 63A-63C According to one embodiment of the present disclosure, Fig.63A ), Donor #2( Fig.63B ) and donor #3( Fig.63C ) IFNγ production by transduced CD4+ selected T cells obtained.

[0329] Fig.63D Shows Figures 63A-63C EC50 value (ng / ml).

[0330] Figures 64A-64C According to one embodiment of the present disclosure, Fig.64A ), Donor #1( Fig.64B ) and donor #3( Fig.64C ) IFNγ production of transduced PBMCs and their corresponding EC50 values ​​(ng / ml).

[0331] Fig.64D Shows Figures 64A-64C Comparison of EC50 values ​​(ng / ml) among different donors in.

[0332] Figures 65A-65C According to one embodiment of the present disclosure, a transduced PBMC ( Fig.65A ), CD8+ selected T cells ( Fig.65B ) and CD4+ selected T cells ( Fig.65C ) of IFNγ production and its corresponding EC50 value (ng / ml).

[0333] Fig.66 An exemplary membrane-bound IL-15 that can be provided in the Examples and that is bound to a T cell membrane is schematically depicted. In the Examples, as a non-limiting example, the membrane-bound IL-15 can signal through the intermediary IL-2 / IL-15 receptor.

[0334] Fig.67A Exemplary membrane-bound IL-15 polypeptides that may be provided in the Examples are depicted, wherein the IL-15 polypeptide is located at the N-terminus of an IL-15Rα polypeptide.

[0335] Fig.67B Exemplary membrane-bound IL-15 polypeptides that can be provided in the Examples are depicted, wherein the IL-15 polypeptide is located at the C-terminus of the IL-15Rα polypeptide. In the Examples, nucleic acids encoding such constructs are also provided. Fig.67A and 67B In the figure, L represents one or more optional linkers, and the line may represent a direct connection without intervening sequences, or may represent intervening sequences, such as but not limited to linkers, untranslated sequences (in the case of nucleic acid sequences), translated sequences, sequences comprising one or more restriction endonuclease sites (in the case of nucleic acid sequences), or combinations thereof.

[0336] Fig.68A An exemplary membrane-bound IL-15 polypeptide that may be provided in the Examples is depicted, wherein the IL-15 polypeptide is located at the N-terminus of an IL-15Rα polypeptide, and a signal peptide (SP) is located at the N-terminus of the IL-15 polypeptide.

[0337] Fig.68B Depicted are exemplary membrane-bound IL-15 polypeptides that may be provided in the Examples, wherein 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. In the Examples, nucleic acids encoding such constructs are also provided. Fig.68A and 68BIn the figure, L represents one or more optional linkers, and the line may represent a direct connection without intervening sequences, or may represent intervening sequences, such as but not limited to linkers, untranslated sequences (in the case of nucleic acid sequences), translated sequences, sequences comprising one or more restriction endonuclease sites (in the case of nucleic acid sequences), or combinations thereof.

[0338] Fig.69A Exemplary polypeptide constructs that may be provided in the Examples are depicted.

[0339] Fig.69B Depicted are exemplary nucleic acid constructs that may be provided in the Examples. Fig.69A and 69B In the figure, the line can represent a direct connection without intervening sequences, or can represent intervening sequences, such as but not limited to linkers, untranslated sequences (in the case of nucleic acid sequences), translated sequences, sequences comprising one or more restriction endonuclease sites (in the case of nucleic acid sequences), or combinations thereof.

[0340] Fig.70 Depicted are exemplary vector constructs that may be provided in the Examples. In the Examples, the vector may further comprise additional elements, such as those described herein, such as but not limited to one or more promoters or one or more post-transcriptional regulatory elements. Fig.70 In the figure, the line can represent a direct connection without intervening sequences, or can represent intervening sequences, such as but not limited to linkers, furin, sequences encoding 2A polypeptides, factor Xa sites, untranslated sequences, translated sequences, sequences comprising one or more restriction endonuclease sites, or combinations thereof.

[0341] Figures 71A-71D Shown in an exemplary assay, TCR+% (A), IL15Rα+TCR+% (B), expansion fold (C) and cell viability (D) of different vector constructs (Alt v1-v4, v7) co-expressing TCR and mbIL15 compared to TCR only ("TCR") and untransduced cells ("NT") as controls. Flow cytometry was gated for CD8+. Data were grouped (n=2) and expressed as mean values.

[0342] Figures 72A-72D Shown are TCR+% (A), IL15Rα+TCR+% (B), expansion fold (C), and total TCR+ cells (D) of different vector constructs (Alt v1-v4, v7) co-expressing TCR and mbIL15 compared to TCR only ("TCR") and untransduced cells ("NT") as controls in an exemplary assay. Flow cytometry gating excludes CD8+. Data are grouped (n=4) and expressed as mean values.

[0343] Figures 73A-73C Shown are exemplary kinetic killing (A), tumor growth index (B) and IFNγ release (C) in co-culture assays of UACC257 tumor cells ("UACC257-RFP") expressing red fluorescent protein (RFP) and untransduced ("NT") cells, cells transduced with only TCR ("TCR"), or cells transduced with various constructs (Alt v1-v4, v7) co-expressing TCR and mbIL15. Within 9-10 days, cells were stimulated four times with UACC257 cells at a 1:1 effector: target ratio at about 0 hours, about 70 hours, about 120 hours, and about 170 hours. Individual UACC257-RFP cells were determined as controls. Data were grouped (n=4), expressed as mean values, and normalized relative to TCR+. Data were collected using IncuCyte and ELISA.

[0344] Figures 74A-74C Exemplary kinetic killing (A), tumor growth index (B), and IFNγ release (C) in co-culture assays of hs695T tumor cells ("hs695T-RFP") expressing red fluorescent protein (RFP) with non-transduced ("NT") cells, cells transduced with TCR alone ("TCR"), or cells transduced with various constructs (Alt v1-v4, v7) co-expressing TCR and mbIL15. Cells were stimulated four times with hs695T cells at an effector: target ratio of 2: 1 at about 0 hours, about 70 hours, about 120 hours, and about 170 hours over 9-10 days. hs695T-RFP cells alone were assayed as controls. Data were grouped (n=4), expressed as mean values, and normalized to TCR+. Data were collected using IncuCyte and ELISA.

[0345] Figures 75A-75C Shown are exemplary kinetic killing (A), tumor growth index (B), and IFNγ release (C) in co-culture assays of A375 tumor cells ("A375-RFP") expressing red fluorescent protein (RFP) with untransduced ("NT") cells, cells transduced with only TCR ("TCR"), or cells transduced with various constructs. Cells were stimulated four times with A375 cells at an effector: target ratio of 4:1 at about 0 hours, about 70 hours, about 120 hours, and about 170 hours over 9-10 days. A375-RFP cells alone were determined as controls. Data were grouped (n=4), expressed as mean values, and normalized relative to TCR+. Data were collected using IncuCyte and ELISA.

[0346] Figures 76A-76DThe percentages of TemRA, Tem, Tnaive / scm, and Tcm cells in cells transduced with TCR alone ("TCR") and various constructs in the Examples are shown. Non-transduced cells ("NT") were determined as controls. Fig.76A The panels in are performed on cells that were not exposed to antigen presenting tumor cells (pre-antigen). Fig.76B Panels in -D were taken after four tumor stimulations with UACC257 cells (B), hs695T cells (C), and A375 cells (D) over 9-10 days. Flow cytometry was gated on CD3+CD8+ cells. Data are presented as mean values.

[0347] Figures 77A-77D The percentage of CD8+ cells positive for each of LAG-3, PD-1, TIGIT, TIM-3, CD39 and CD69 is shown in the examples before (A) or after cells are exposed to UACC257 cells (B), hs695T cells (C) or A375 cells (D) carrying antigens. Cells are stimulated four times with tumor cells in 9-10 days. Untransduced ("NT") cells and cells transduced with only TCR ("TCR"), as well as each of cells transduced with various constructs show expression percentages. Data are grouped (n=4) and expressed as mean values.

[0348] Figures 78A-78F Flow cytometry plots of cells transduced with TCR only ("TCR") and various constructs in the Examples are shown. Non-transduced cells ("NT") were determined as controls. The X-axis shows staining for cell viability markers (Helix NP) and the Y-axis shows staining for apoptosis markers (ApoTracker TM ) staining. Flow cytometry plots of cells after four antigen challenges with antigen presenting UACC257 tumor cells over 9-10 days are shown.

[0349] Figure 78G-78H The frequencies of live and dead apoptotic cells are shown, respectively. Cells with similar results were obtained after stimulation with hs695T and A375 tumor cells (data not shown). Flow cytometry was gated on CD8+ cells. Data were grouped (n=4) and expressed as mean values.

[0350] Figures 79A-79C Proliferation index of cells transduced with TCR only ("TCR") and various constructs is shown. Cells were stimulated twice with UACC257 tumor cells (A), hs695T tumor cells (B), or A375 tumor cells (C) within 6 days. Flow cytometry was gated for CD8+ cells. Data were grouped (n=4) and expressed as mean values.

[0351] Figures 80A-80C Total cell counts (A), expansion folds (B) and cell viability (C) of cells transduced with only TCR ("TCR") and various constructs in the absence or presence of exogenous interleukin-7 and interleukin-15 ("IL7 / 15") and in the absence of antigenic stimulation are shown. The conditions using exogenous interleukin-7 and interleukin-15 ("IL7 / 15") terminated at day 17, while the conditions without exogenous interleukin-7 and interleukin-15 ("IL7 / 15") were cultured for up to 31 days. Data were grouped (n=4) and expressed as mean values.

[0352] Figures 81A-81B Shown is an exemplary kinetic killing of cells transduced with various constructs after 31 days of culture in the absence of exogenous cytokine addition or antigen stimulation. To assess killing ability, transduced cells were co-cultured with UACC257 tumor cells at an effector: target ratio of 1: 1 (A) or hs695T tumor cells at an effector: target ratio of 2: 1 (B). Data were grouped (n = 4), expressed as mean values, and normalized relative to TCR+. DETAILED DESCRIPTION

[0353] Membrane-bound IL-15

[0354] In embodiments, membrane-bound IL-15 polypeptides (membrane-bound IL-15 or mbIL-15) are provided. In embodiments, the nucleic acids described herein comprise and / or encode membrane-bound IL-15 polypeptides. In embodiments, the vectors described herein comprise and / or encode membrane-bound IL-15 polypeptides. In embodiments, the cells described herein comprise and / or express membrane-bound IL-15 polypeptides. In embodiments, the compositions described herein comprise membrane-bound IL-15 polypeptides or comprise cells comprising and / or expressing membrane-bound IL-15 polypeptides. In embodiments, IL-15 is membrane-bound by expressing an IL-15 polypeptide and an IL-15Rα polypeptide in an IL-15 / IL-15Rα fusion polypeptide (IL-15 / IL-15Rα).

[0355] A membrane-bound IL-15 polypeptide is provided. An isolated nucleic acid sequence is provided, the isolated nucleic acid sequence comprising one or more nucleic acid sequences, the one or more nucleic acid sequences encoding one or more membrane-bound IL-15 polypeptides. A vector is provided, the vector comprising one or more nucleic acid sequences comprising one or more nucleic acid sequences, the one or more nucleic acid sequences encoding one or more membrane-bound IL-15 polypeptides. A cell comprising and / or expressing one or more membrane-bound IL-15 polypeptides is provided. A cell is provided, the cell comprising or expressing one or more nucleic acid sequences comprising one or more nucleic acid sequences, the one or more nucleic acid sequences encoding one or more membrane-bound IL-15 polypeptides. A cell is provided, the cell comprising or expressing one or more vectors, the one or more vectors comprising one or more nucleic acid sequences comprising one or more nucleic acid sequences, the one or more nucleic acid sequences encoding one or more membrane-bound IL-15 polypeptides. In an embodiment, the cell described herein may comprise a membrane-bound IL-15 polypeptide, a CD8 polypeptide, a cell receptor (TCR) comprising an alpha chain and a beta chain, a TCR comprising a gamma chain and a delta chain, a chimeric antigen receptor (CAR), or any combination thereof. In an embodiment, the cell may comprise an αβT cell, a γδT 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 an embodiment, such polypeptides, nucleic acids, vectors and / or cells may be isolated, recombinant and / or engineered. Compositions comprising such polypeptides, nucleic acids, vectors and / or cells are provided.

[0356] In one aspect, the polypeptide sequences and / or nucleic acid sequences described herein may be isolated and / or recombinant sequences.

[0357] In one aspect, the cells described herein can be isolated and / or recombinant cells.

[0358] The membrane-bound IL-15 may comprise, for example, an IL-15 / IL-15Rα fusion polypeptide and / or an IL-15Rα / IL-15 fusion polypeptide. One or more linkers may be located between IL-15 and IL-15Rα or between IL-15Rα and IL-15. In an embodiment, 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 an embodiment, the IL-15 polypeptide is located at the C-terminus of the IL-15Rα polypeptide in the membrane-bound IL-15 polypeptide. Fig.67B ). Fig.67A and 67B The IL-15 polypeptide in the can be immature wild type, immature mutated, mature wild type or mature mutated. Fig.67A and 67BThe IL-15Rα polypeptide in can be immature wild type, immature mutated, mature wild type or mature mutated. In an embodiment, Fig.67A and Fig.67B The IL-15 polypeptide in is mature and may or may not be mutated, and Fig.67A and Fig.67B The IL-15Rα polypeptide in is mature and may or may not be mutated. In an embodiment, Fig.67A and Fig.67B The IL-15 polypeptide in is mature and may or may not be mutated, and Fig.67A and Fig.67B The IL-15Rα polypeptide in is mature and mutated. Fig.67A and 67B A linker is depicted in , but mbIL-15 may or may not include a linker.

[0359] In an embodiment, 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 further comprise one or more linkers. In an embodiment, the membrane-bound IL-15 comprises Fig.67A or Fig.67B The structure shown and / or encoded thereby. Fig.67A and 67B In the figure, the lines connecting IL-15 to the one or more linkers (L) and connecting the one or more linkers (L) to IL-15Rα can represent a direct connection without intervening sequences, or can represent intervening sequences, such as but not limited to linkers, untranslated sequences (in the case of nucleic acid sequences), translated sequences, sequences comprising one or more restriction endonuclease sites (in the case of nucleic acid sequences), or combinations thereof.

[0360] In embodiments, the IL-15 / IL-15Rα fusion polypeptide and / or IL-15Rα / IL-15 fusion polypeptide may comprise one or more signal peptides. In embodiments, the membrane-bound IL-15 comprising one or more signal peptides and optionally one or more linkers may comprise Fig.68A or Fig.68B The structure shown and / or encoded thereby. Fig.68A An exemplary IL-15 / IL-15Rα fusion polypeptide comprising, optionally, at least one linker and at least one signal peptide is depicted in . Fig.68B An exemplary 15Rα / IL-15 fusion polypeptide comprising, optionally, at least one linker and at least one signal peptide is depicted in . Fig.68A and 68BThe IL-15 polypeptide in the can be immature wild type, immature mutated, mature wild type or mature mutated. Fig.68A and 68B The IL-15Rα polypeptide in can be immature wild type, immature mutated, mature wild type or mature mutated. In an embodiment, Fig.68A and Fig.68B The IL-15 polypeptide in is mature and may or may not be mutated, and Fig.68A and Fig.68B The IL-15Rα polypeptide in is mature and may or may not be mutated. In an embodiment, Fig.68A and 68B The IL-15 polypeptide in is mature and may or may not be mutated, and Fig.68A and 68B The IL-15Rα polypeptide in is mature and mutated. Fig.68A and 68B In the method, (a) the one or more signal peptides (SP) are linked to IL-15, IL-15 is linked to the one or more linkers (L), and the one or more linkers are linked to IL-15Rα (e.g. Fig.68A (a) linking the one or more signal peptides (SP) to IL-15α, linking IL-15α to the one or more linkers (L), and linking the one or more linkers to IL-15 (as shown in Fig.68B A line (shown in the figure) may represent a direct connection without intervening sequences, or may represent intervening sequences, such as, but not limited to, linkers, untranslated sequences (in the case of nucleic acid sequences), translated sequences, sequences comprising one or more restriction endonuclease sites (in the case of nucleic acid sequences), or combinations thereof.

[0361] In embodiments, the IL-15 / IL-15Rα fusion polypeptide comprises an intact IL-15 polypeptide, an intact IL-15Rα polypeptide, or both. In embodiments, the intact or full wild-type IL-15 polypeptide may comprise SEQ ID NO: 305. In embodiments, the intact or full wild-type IL-15Rα polypeptide may comprise SEQ ID NO: 306.

[0362] In embodiments, the IL-15 / IL-15Rα fusion polypeptide comprises a mature IL-15 polypeptide (e.g., SEQ ID NO: 307), a mature IL-15Rα polypeptide (e.g., SEQ ID NO: 309), or both, which may be mutated (e.g., SEQ ID NO: 311, 313, 315). In 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 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 embodiments, the mature wild-type IL-15 polypeptide is encoded by a nucleic acid comprising or consisting of a nucleic acid as set forth in SEQ ID NO: 308. In embodiments, the mature wild-type IL-15Rα polypeptide is encoded by a nucleic acid comprising or consisting of a nucleic acid as set forth in SEQ ID NO: 310. However, in embodiments, the IL-15 / IL-15Rα fusion polypeptide does not comprise the mature wild-type IL-15Rα as in SEQ ID NO: 309, or a sequence having about 95% or more sequence identity thereto. In embodiments, the IL-15 / IL-15Rα fusion polypeptide does not comprise the mature wild-type IL-15Rα encoded by SEQ ID NO: 310, or a sequence having about 80%, about 85%, about 90%, or about 95% or more sequence identity thereto.

[0363] In embodiments, the IL-15 polypeptide is mutated and / or truncated, the IL-15Rα polypeptide is mutated and / or truncated, or both are mutated and / or truncated.

[0364] In an embodiment, the IL-15 polypeptide may comprise or may lack a native signal peptide (which may have a sequence comprising SEQ ID NO: 369), may comprise or may lack a native propeptide (which may have a sequence comprising SEQ ID NO: 371), or any combination thereof.

[0365] In embodiments, the IL-15Rα polypeptide may comprise or may lack a native signal sequence (which may have a sequence comprising SEQ ID NO: 370).

[0366] In an embodiment, the IL-15Rα polypeptide, which may be a mature IL-15Rα polypeptide (e.g., SEQ ID NO: 309), may be mutated. In an embodiment, the IL-15Rα polypeptide IL-15Rα polypeptide may comprise a mutated transmembrane domain. In an embodiment, the transmembrane domain of the IL-15Rα polypeptide may comprise or consist of SEQ ID NO: 376 or SEQ ID NO: 378. In an embodiment, the transmembrane domain of the IL-15Rα polypeptide may be encoded by a nucleic acid comprising or consisting of a sequence shown in SEQ ID NO: 377 or SEQ ID NO: 379. In an embodiment, the mutant IL-15Rα may comprise a heterologous transmembrane domain. In an embodiment, the heterologous transmembrane domain may be derived from CD25. In embodiments, the transmembrane domain derived from CD25 comprises or consists 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 thereof. In embodiments, the transmembrane domain derived from CD25 is encoded by a nucleic acid comprising or consisting of the 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 thereof. In embodiments, the IL-15Rα polypeptide comprising a CD25 transmembrane domain comprises or consists of the 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 thereof. In embodiments, the IL-15Rα polypeptide comprising a CD25 transmembrane domain is encoded by a nucleic acid comprising or consisting of the 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 thereof. In embodiments, the heterologous transmembrane domain may be derived from CD28.In embodiments, the transmembrane domain derived from CD28 comprises or consists of SEQ ID NO: 374, 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 thereof. In embodiments, the transmembrane domain derived from CD28 is encoded by a nucleic acid comprising or consisting of the 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 thereof. In embodiments, the IL-15Rα polypeptide comprising a CD28 transmembrane domain comprises or consists of the 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 thereof. In embodiments, the IL-15Rα polypeptide comprising a CD28 transmembrane domain is encoded by a nucleic acid comprising or consisting of the 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 thereof. In embodiments, IL-15Rα can be mutated by deleting exon 3 of human IL-15Rα genomic DNA. In embodiments, the IL-15Rα polypeptide comprising an exon 3 deletion comprises or consists of a sequence as 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 thereto. In embodiments, an IL-15Rα polypeptide comprising an exon 3 deletion is encoded by a nucleic acid comprising or consisting of a sequence as set forth in SEQ ID NO:316, or a sequence 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.In embodiments, the functions of IL-15Rα, such as but not limited to the ability of IL-15Rα to be membrane bound and one or more signaling functions of IL-15α are retained and / or enhanced in an IL-15Rα polypeptide having a heterologous transmembrane domain or deleted exon 3.

[0367] In an embodiment, the disclosure provides nucleic acids encoding the polypeptides described herein.

[0368] In one aspect, the polypeptide sequences and / or nucleic acid sequences described herein may be isolated and / or recombinant sequences.

[0369] In one aspect, the cells described herein can be isolated and / or recombinant cells.

[0370] In embodiments, the IL-15 polypeptide has a sequence comprising 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 embodiments, the function of IL-15, such as but not limited to one or more signaling functions of IL-15, is retained and / or enhanced in the mutated IL-15 polypeptide.

[0371] In embodiments, the IL-15 polypeptide has a sequence comprising 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 embodiments, the function of IL-15, such as but not limited to one or more signaling functions of IL-15, is retained and / or enhanced in the mutated IL-15 polypeptide.

[0372] In embodiments, the IL-15 polypeptide comprises (a) SEQ ID NO: 305 comprising one, two, three, four or five amino acid substitutions, or (b) SEQ ID NO: 307 comprising one, two, three, four or five amino acid substitutions. In embodiments, the amnion substitutions are conservative or non-conservative. In embodiments, the amino acid substitutions are conservative amino acid substitutions. In embodiments, the function of IL-15, such as but not limited to one or more signaling functions of IL-15, is retained and / or enhanced in the mutated IL-15 polypeptide.

[0373] In embodiments, the IL-15 polypeptide is encoded by a nucleic acid comprising 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 of SEQ ID NO: 308. In embodiments, the function of IL-15, such as but not limited to one or more signaling functions of IL-15, is retained and / or enhanced in the IL-15 polypeptide encoded by the mutated nucleic acid sequence.

[0374] In an embodiment, the IL-15 polypeptide is encoded by a nucleic acid comprising (a) SEQ ID NO: 308 comprising one, two, three, four or five nucleic acid substitutions. In an embodiment, one or more nucleic acid substitutions in a codon may generate a codon encoding the same amino acid or may generate a codon encoding a different amino acid. In an embodiment, one or more nucleic acid substitutions in a codon may generate a codon encoding a conservative amino acid substitution. In an embodiment, one or more nucleic acid substitutions in a codon may generate a codon encoding the same amino acid. In an embodiment, the function of IL-15, such as but not limited to one or more signaling functions of IL-15, is retained and / or enhanced in the IL-15 polypeptide encoded by the mutated nucleic acid sequence.

[0375] In an embodiment, the nucleic acid encoding the IL-15 polypeptide may comprise a stop codon (such as TAA, TAG or TGA), as a non-limiting example, located at the 3' end of the nucleotides encoding the IL-15 polypeptide.

[0376] In embodiments, the IL-15Rα polypeptide has a sequence comprising 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 embodiments, the IL-15Rα polypeptide has a sequence comprising 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 embodiments, the IL-15Rα polypeptide does not have a sequence comprising, consisting of, or having about 95% or more sequence identity with SEQ ID NO: 309. In embodiments, the IL-15Rα polypeptide has a sequence comprising 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 with the amino acid sequence of SEQ ID NO: 311. In embodiments, the IL-15Rα polypeptide has a sequence comprising 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 embodiments, the IL-15Rα polypeptide has a sequence comprising 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 embodiments, the functions of IL-15Rα, such as but not limited to the ability of IL-15Rα to be membrane bound and one or more signaling functions of IL-15α are retained and / or enhanced in the mutated IL-15Rα polypeptide.

[0377] In embodiments, the IL-15Rα polypeptide may comprise (a) SEQ ID NO:306 comprising one, two, three, four or five amino acid substitutions; (b) SEQ ID NO:309 comprising one, two, three, four or five amino acid substitutions; (c) SEQ ID NO:311 comprising one, two, three, four or five amino acid substitutions; (d) SEQ ID NO:313 comprising one, two, three, four or five amino acid substitutions; or (d) SEQ ID NO:315 comprising one, two, three, four or five amino acid substitutions. However, in embodiments, the IL-15Rα polypeptide does not have a sequence comprising or consisting of SEQ ID NO:309 or a sequence having about 95% or more sequence identity thereto. In embodiments, the amnion substitutions are conservative or non-conservative. In embodiments, the amino acid substitutions are conservative amino acid substitutions. In embodiments, the functions of IL-15Rα, such as but not limited to the ability of IL-15Rα to be membrane bound and one or more signaling functions of IL-15α are retained and / or enhanced in the mutated IL-15Rα polypeptide.

[0378] In embodiments, the IL-15Rα polypeptide is encoded by a nucleic acid comprising 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 of SEQ ID NO: 310. However, in embodiments, the IL-15Rα polypeptide is not encoded by a nucleic acid comprising SEQ ID NO: 310, or having about 85%, about 90%, about 95%, or more sequence identity thereto. In embodiments, the IL-15Rα polypeptide is encoded by a nucleic acid comprising 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 of SEQ ID NO: 312. In embodiments, the IL-15Rα polypeptide is encoded by a nucleic acid comprising 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 of SEQ ID NO: 314. In embodiments, the IL-15Rα polypeptide is encoded by a nucleic acid comprising 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 of SEQ ID NO: 316. In embodiments, the function of IL-15Rα, such as but not limited to the ability of IL-15Rα to be membrane-bound and one or more signaling functions of IL-15α, is retained and / or enhanced in the IL-15α polypeptide encoded by the mutated nucleic acid sequence.

[0379] In embodiments, the IL-15Rα polypeptide is encoded by a nucleic acid comprising (a) SEQ ID NO: 310 comprising one, two, three, four or five nucleic acid substitutions; (b) SEQ ID NO: 312 comprising one, two, three, four or five nucleic acid substitutions; (c) SEQ ID NO: 314 comprising one, two, three, four or five nucleic acid substitutions, and (d) SEQ ID NO: 316 comprising one, two, three, four or five nucleic acid substitutions. However, in embodiments, the IL-15Rα polypeptide is not encoded by a nucleic acid comprising SEQ ID NO: 310 or having about 85%, about 90%, about 95% or more sequence identity thereto. In embodiments, one or more nucleic acid substitutions in a codon may produce a codon encoding the same amino acid or may produce a codon encoding a different amino acid. In embodiments, one or more nucleic acid substitutions in a codon may produce a codon encoding a conservative amino acid substitution. In embodiments, one or more nucleic acid substitutions in a codon may produce a codon encoding the same amino acid. In embodiments, the functions of IL-15Rα, such as but not limited to the ability of IL-15Rα to be membrane bound and one or more signaling functions of IL-15α are retained and / or enhanced in the IL-15α polypeptide encoded by the mutated nucleic acid sequence.

[0380] In embodiments, the nucleic acid encoding the IL-15Rα polypeptide may comprise a stop codon (such as TAA, TAG or TGA), as a non-limiting example, located at the 3' end of the nucleotides encoding the IL-15Rα polypeptide.

[0381] In embodiments, the IL-15 polypeptide and the IL-15Rα polypeptide are connected by one or more linkers. In embodiments, the linker is a peptide linker. In embodiments, the peptide linker is rigid or flexible. In embodiments, the linker is cleavable. In embodiments, the linker can promote the stability or proper folding of the fusion polypeptide, can increase the expression of the fusion polypeptide, can improve the biological activity of the fusion polypeptide, can promote the targeting of the fusion polypeptide, can change the PK of the fusion polypeptide, or any combination thereof.

[0382] In embodiments, the linker comprises about 2-40 amniotic acids, about 4-38 amino acids, about 6-34 amino acids, about 8-32 amino acids, about 10-30 amino acids, about 10 amino acids, about 11 amino acids, about 12 amino acids, about 12-28 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, about 14-26 amino acids, about 12-24 amino acids, about 10-22 amino acids, about 10-20 amino acids, about 12-18 amino acids, about 14-16 amino acids, about 8-22 amino acids, about 6-24 amino acids, about 4-26 amino acids, or about 2-28 amino acids.

[0383] In embodiments, the one or more linkers of the IL-15 / IL-15Rα fusion polypeptide independently comprise or consist of any of GSG, LE, SEQ ID NO: 266, 383, 385, 387, 389, 391, or 393 or 395-432, 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% sequence identity to any of SEQ ID NO: 266, 383, 385, 387, 389, 391, or 393 or 395-432. However, in embodiments, the 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 embodiments, one or more linkers of the IL-15 / IL-15Rα fusion polypeptide independently comprise or consist of GSG, LE, SEQ ID NO: 266, 383, 385, 387, 389, 393 or 396-432, 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% sequence identity to any of SEQ ID NO: 266, 383, 385, 387, 389, 393 or 396-432. In embodiments, one or more linkers of the IL-15 / IL-15Rα fusion polypeptide independently comprise or consist of any of SEQ ID NO: 383, 385, 387 or 389, 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% sequence identity to any of SEQ ID NO: 383, 385, 387 or 389.

[0384] In 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 one of SEQ ID NOs: 384, 386, 388, 390, or 392, having 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 any one of SEQ ID NOs: 384, 386, 388, 390, or 392, one or more nucleic acids encoding any linker comprising or consisting of GSG, LE, or one or more linkers shown in SEQ ID NOs: 266 or 393-432, or one or more nucleic acids encoding any linker that is identical to any one of SEQ ID NOs: 384, 386, 388, 390, or 392. NO: 266 or any of 393-432 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% identity. However, in 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.

[0385] In 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 one of SEQ ID NOs: 384, 386, 388, or 390, having 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 any one of SEQ ID NOs: 384, 386, 388, or 390, one or more nucleic acids encoding any linker comprising or consisting of GSG or one or more linkers shown in SEQ ID NOs: 266, 383, 385, 387, 389, 393, or 396-432, or one or more nucleic acids encoding any linker that is identical to any one of SEQ ID NOs: Any of NO: 266, 383, 385, 387, 389, 393 or 396-432 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% identity.

[0386] In embodiments, one or more linkers of the IL-15 / IL-15Rα fusion polypeptide are independently encoded by any one of SEQ ID NOs: 384, 386, 388 or 390, or by 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 any one of SEQ ID NOs: 384, 386, 388 or 390.

[0387] In embodiments, the linker has a sequence comprising 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 embodiments, the linker has a sequence comprising 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 embodiments, the linker has a sequence comprising 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 embodiments, the linker has a sequence comprising 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 an embodiment, one or more functions of a linker, such as but not limited to the flexibility, rigidity, cleavability of the linker, the ability to promote stability or proper folding of a fusion polypeptide, the ability to increase expression of a fusion polypeptide, the ability to improve the biological activity of a fusion polypeptide, the ability to promote targeting of a fusion polypeptide, the ability to alter the PK of a fusion polypeptide, or a combination thereof, are retained and / or enhanced in the mutated linker.

[0388] In an embodiment, the linker comprises (a) SEQ ID NO: 383 comprising one, two, three, four or five amino acid substitutions; (b) SEQ ID NO: 385 comprising one, two, three, four or five amino acid substitutions; (c) SEQ ID NO: 387 comprising one, two, three, four or five amino acid substitutions; (d) SEQ ID NO: 389 comprising one, two, three, four or five amino acid substitutions; or (d) SEQ ID NO: 391 comprising one, two, three, four or five amino acid substitutions. In an embodiment, the amnion substitutions may be conservative or non-conservative. In an embodiment, the amino acid substitutions may be conservative amino acid substitutions. In an embodiment, one or more functions of the linker, such as but not limited to the flexibility, rigidity, cleavage of the linker, the ability to promote 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 one or more of a combination thereof, are retained and / or enhanced in the mutated linker.

[0389] In embodiments, the linker is encoded by a nucleic acid comprising 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 of SEQ ID NO: 384. In embodiments, the linker is encoded by a nucleic acid comprising 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 of SEQ ID NO: 386. In embodiments, the linker is encoded by a nucleic acid comprising 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 of SEQ ID NO: 388. In embodiments, the linker is encoded by a nucleic acid comprising 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 of SEQ ID NO: 390. In an embodiment, one or more functions of a linker, such as but not limited to the flexibility, rigidity, cleavability of the linker, the ability to promote stability or proper folding of a fusion polypeptide, the ability to increase expression of a fusion polypeptide, the ability to improve the biological activity of a fusion polypeptide, the ability to promote targeting of a fusion polypeptide, the ability to alter the PK of a fusion polypeptide, or a combination thereof, are retained and / or enhanced in the mutated linker.

[0390] In an embodiment, the linker is encoded by a nucleic acid comprising (a) SEQ ID NO:384 comprising one, two, three, four or five nucleic acid substitutions; (b) SEQ ID NO:386 comprising one, two, three, four or five nucleic acid substitutions; (c) SEQ ID NO:388 comprising one, two, three, four or five nucleic acid substitutions; (d) SEQ ID NO:390 comprising one, two, three, four or five nucleic acid substitutions; or (d) SEQ ID NO:392 comprising one, two, three, four or five nucleic acid substitutions. In an embodiment, one or more nucleic acid substitutions in a codon can produce codons encoding the same amino acid or can produce codons encoding different amino acids. In an embodiment, one or more nucleic acid substitutions in a codon can produce codons encoding conservative amino acid substitutions. In an embodiment, one or more nucleic acid substitutions in a codon can produce codons encoding the same amino acid. In an embodiment, one or more functions of a linker, such as but not limited to the flexibility, rigidity, cleavability of the linker, the ability to promote stability or proper folding of a fusion polypeptide, the ability to increase expression of a fusion polypeptide, the ability to improve the biological activity of a fusion polypeptide, the ability to promote targeting of a fusion polypeptide, the ability to alter the PK of a fusion polypeptide, or a combination thereof, are retained and / or enhanced in the mutated linker.

[0391] In 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 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 embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker does not comprise or consist of SEQ ID NO: 335, or a sequence having about 95% or more sequence identity thereof. In 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, or a sequence having about 80%, about 85%, about 90%, or about 95% or more sequence identity thereof. In 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 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 embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker comprises SEQ ID NO: 317, 321, 325, 327, 329, 331, or 333. In 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.

[0392] In an embodiment, the IL-15 / IL-15Rα fusion polypeptide comprising a linker may comprise (a) SEQ ID NO: 317 comprising one, two, three, four or five amino acid substitutions; (b) SEQ ID NO: 319 comprising one, two, three, four or five amino acid substitutions; (c) SEQ ID NO: 321 comprising one, two, three, four or five amino acid substitutions; (d) SEQ ID NO: 323 comprising one, two, three, four or five amino acid substitutions; (e) SEQ ID NO: 325 comprising one, two, three, four or five amino acid substitutions; (f) SEQ ID NO: 327 comprising one, two, three, four or five amino acid substitutions; (g) SEQ ID NO: 329 comprising one, two, three, four or five amino acid substitutions; (h) SEQ ID NO: 331 comprising one, two, three, four or five amino acid substitutions; (i) SEQ ID NO: 332 comprising one, two, three, four or five amino acid substitutions. NO:333; or (j) SEQ ID NO:335 comprising one, two, three, four or five amino acid substitutions. However, in embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker does not comprise or consists of SEQ ID NO:335 or a sequence having about 95% or more sequence identity thereto. In embodiments, amniotic acid substitutions may be conservative or non-conservative. In embodiments, amino acid substitutions may be conservative amino acid substitutions. In 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 be membrane-bound and the signaling function of IL-15Rα, or (iii) both (i) and (ii) are retained and / or enhanced in the mutated IL-15 / IL-15Rα fusion polypeptide.

[0393] In embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker 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: 317. In embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker 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: 319. In embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker 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: 321. In embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker 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: 323. In embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker 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: 325. In embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker 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: 327. In embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker 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:329.In embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker 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: 331. In embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker 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: 333. In embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker 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%, or at least about 94% sequence identity to SEQ ID NO: 335. In embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker does not comprise or consist of SEQ ID NO: 335 or a sequence having about 95% or more sequence identity thereto. In 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 be membrane-bound and the signaling function of IL-15Rα, or (iii) both (i) and (ii) are retained and / or enhanced in the IL-15 / IL-15Rα fusion polypeptide encoded by the mutated nucleic acid sequence.

[0394] In embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker may be encoded by a nucleic acid comprising 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 of SEQ ID NO: 318. In embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker may be encoded by a nucleic acid comprising 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 of SEQ ID NO: 320. In embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker may be encoded by a nucleic acid comprising 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 of SEQ ID NO: 322. In embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker may be encoded by a nucleic acid comprising 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 of SEQ ID NO: 324. In embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker may be encoded by a nucleic acid comprising 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 of SEQ ID NO: 326. In embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker may be encoded by a nucleic acid comprising 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 of SEQ ID NO: 328.In embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker may be encoded by a nucleic acid comprising 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 of SEQ ID NO: 330. In embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker may be encoded by a nucleic acid comprising 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 of SEQ ID NO: 332. In embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker may be encoded by a nucleic acid comprising 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 of SEQ ID NO: 334. In embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker may be encoded by a nucleic acid comprising 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 embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker is not encoded by a nucleic acid encoding a polypeptide having about 95% or more sequence identity to SEQ ID NO: 335. In embodiments, (i) a function of IL-15, such as but not limited to one or more signaling functions of IL-15, (ii) a function of IL-15Rα, such as but not limited to the ability of IL-15Rα to be membrane bound and one or more signaling functions of IL-15Rα, or (iii) both (i) and (ii) are retained and / or enhanced in the IL-15 / IL-15Rα fusion polypeptide encoded by the mutated nucleic acid sequence.

[0395] In an embodiment, an IL-15 / IL-15Rα fusion polypeptide comprising a linker can be encoded by a nucleic acid comprising (a) SEQ ID NO:318 comprising one, two, three, four or five nucleic acid substitutions; (b) SEQ ID NO:320 comprising one, two, three, four or five nucleic acid substitutions; (c) SEQ ID NO:322 comprising one, two, three, four or five nucleic acid substitutions; (d) SEQ ID NO:324 comprising one, two, three, four or five nucleic acid substitutions; (e) SEQ ID NO:326 comprising one, two, three, four or five nucleic acid substitutions; (f) SEQ ID NO:328 comprising one, two, three, four or five nucleic acid substitutions; (g) SEQ ID NO:330 comprising one, two, three, four or five nucleic acid substitutions; (h) SEQ ID NO:332 comprising one, two, three, four or five nucleic acid substitutions; or (i) SEQ ID NO:334 comprising one, two, three, four or five nucleic acid substitutions. In an embodiment, one or more nucleic acid substitutions in a codon may generate a codon encoding the same amino acid or may generate a codon encoding a different amino acid. In an embodiment, one or more nucleic acid substitutions in a codon may generate a codon encoding a conservative amino acid substitution. In an embodiment, one or more nucleic acid substitutions in a codon may generate a codon encoding the same amino acid. In an embodiment, (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 be membrane-bound and one or more signaling functions of IL-15Rα or (iii) (i) and (ii) are retained and / or enhanced in an IL-15 / IL-15Rα fusion polypeptide encoded by a mutated nucleic acid sequence.

[0396] In embodiments, an IL-15 / IL-15Rα fusion polypeptide optionally comprising one or more linkers comprises or consists of, e.g., SEQ ID NO: 307 (or a sequence 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) fused directly or indirectly to the N-terminus of SEQ ID NO: 309 (or a sequence 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; NO:307 (or a sequence 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 fused directly or indirectly to the N-terminus of SEQ ID NO:311 (or a sequence 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:312 (or a sequence 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); NO:307 (or a sequence 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 fused directly or indirectly to the N-terminus of SEQ ID NO:313 (or a sequence 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;or SEQ ID NO: 307 (or a sequence 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) fused directly or indirectly to the N-terminus of SEQ ID NO: 315 (or a sequence 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; or SEQ ID NO: 316 (or a sequence 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); NO:317, 319, 321, 323, 325, 327, 329, 331, 333 or 335 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, 333 or 335. However, in embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker does not comprise or consist of: (i) SEQ ID NO: 307, which is fused directly or indirectly to the N-terminus of SEQ ID NO: 309 with a linker therebetween; (ii) SEQ ID NO: 335, (iii) a sequence having about 95% or more sequence identity to SEQ ID NO: 307, which is fused directly or indirectly to the N-terminus of SEQ ID NO: 309 with a linker therebetween; or (iv) a sequence having about 95% or more sequence identity to SEQ ID NO: 335. In embodiments, one or more linkers comprise or consist of a linker sequence as set forth herein. ;

[0397] In embodiments, an IL-15 / IL-15Rα fusion polypeptide optionally comprising one or more linkers comprises or consists of, e.g., SEQ ID NO: 307 (or a sequence 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) fused directly or indirectly to the N-terminus of SEQ ID NO: 311 (or a sequence 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; NO:307 (or a sequence 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 fused directly or indirectly to the N-terminus of SEQ ID NO:313 (or a sequence 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; or SEQ ID NO:314 (or a sequence 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); NO:307 (or a sequence 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) fused directly or indirectly to the N-terminus of SEQ ID NO:315 (or a sequence 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; or SEQ ID NO:317, 319, 321, 323, 325, 327, 329, 331 or 333 or to SEQ ID NO: NO:317, 319, 321, 323, 325, 327, 329, 331 or 333 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 sequence.In embodiments, one or more linkers comprise or consist of a linker sequence set forth herein.

[0398] In embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising one or more linkers comprises, e.g., 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.

[0399] In embodiments, an IL-15 / IL-15Rα fusion polypeptide optionally comprising one or more linkers is encoded by a nucleic acid comprising or consisting of, e.g., SEQ ID NO: 308 (or a sequence 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), fused directly or indirectly to the 5' end of SEQ ID NO: 310 (or a sequence 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; NO:308 (or a sequence 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 fused directly or indirectly to the 5' end of SEQ ID NO:312 (or a sequence 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; SEQ ID NO:314 (or a sequence 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); NO:308 (or a sequence 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 fused directly or indirectly to the 5' end of SEQ ID NO:314 (or a sequence 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;or SEQ ID NO:308 (or a sequence 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 fused directly or indirectly to the 5' end of SEQ ID NO:316 (or a sequence 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; or SEQ ID NO:317 (or a sequence 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); NO:318, 320, 322, 324, 326, 328, 330, 332, 334 or 336 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, 334 or 336. However, in 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 about 80%, about 85%, about 90%, or about 95% or more 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 about 80%, about 85%, about 90%, or about 95% or more sequence identity to SEQ ID NO: 336. In embodiments, one or more linkers are encoded by one or more nucleic acids comprising or consisting of nucleic acids encoding linkers as set forth herein. ;

[0400] In embodiments, an IL-15 / IL-15Rα fusion polypeptide, optionally comprising one or more linkers, is encoded by a nucleic acid comprising or consisting of, e.g., SEQ ID NO: 308 (or a sequence 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), fused directly or indirectly to the 5' end of SEQ ID NO: 312 (or a sequence 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; NO:308 (or a sequence 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 fused directly or indirectly to the 5' end of SEQ ID NO:314 (or a sequence 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; or SEQ ID NO:315 (or a sequence 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); NO:308 (or a sequence 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 fused directly or indirectly to the 5' end of SEQ ID NO:316 (or a sequence 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;or 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 embodiments, the one or more linkers are encoded by one or more nucleic acids comprising or consisting of a nucleic acid encoding a linker as set forth herein. ;

[0401] In embodiments, the IL-15 / IL-15Rα fusion polypeptide is encoded by a nucleic acid comprising, e.g., SEQ ID NO: 318, 322, 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, 322, 326, 328, 330, 332, or 334.

[0402] In an embodiment, the IL-15 / IL-15Rα fusion polypeptide comprising a linker (L) comprises Fig.69A In an embodiment, the IL-15 / IL-15Rα fusion polypeptide comprising a linker (L) comprises: Fig.69A In an embodiment, the IL-15 / IL-15Rα fusion polypeptide comprising a linker comprises: Fig.69A In an embodiment, the IL-15 / IL-15Rα fusion polypeptide comprising a linker is encoded by one or more nucleic acids comprising: Fig.69B In an embodiment, the IL-15 / IL-15Rα fusion polypeptide comprising a linker is encoded by one or more nucleic acids, wherein the one or more nucleic acids comprise Fig.69B In an embodiment, the IL-15 / IL-15Rα fusion polypeptide comprising a linker is encoded by one or more nucleic acids, wherein the one or more nucleic acids comprise Fig.69B Any construct A', C' or E'-I' shown in or consisting of it. In the embodiments, also provided with Fig.69A and 69BAny of the sequences shown in comprises 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. Fig.69A and 69B In the example, the lines connecting IL-15 to the linker and the linker to IL-15Rα can represent direct connections, without intervening sequences, or can represent intervening sequences, such as, but not limited to, linkers, untranslated sequences (in Fig.69B ), translated sequences, sequences containing one or more restriction endonuclease sites (in Fig.69B ) or a combination thereof.

[0403] In an embodiment, the nucleic acid encoding the IL-15 / IL-15Rα fusion polypeptide may comprise a stop codon (e.g., TAA, TAG, or TGA), as a non-limiting example, located at the 3' end of the nucleotides encoding the IL-15Rα polypeptide, such as where the encoded fusion polypeptide is at Fig.67A Or the orientation shown in Figure 668A, or at the 3' end of the IL-15 polypeptide, such as where the encoded fusion polypeptide is in Fig.67B Or the orientation shown in Figure 668B.

[0404] In an embodiment, the IL-15 / IL-15Rα fusion polypeptide and / or the IL-15Rα / IL-15 fusion polypeptide may comprise one or more signal peptides. In an embodiment, the fusion polypeptide may comprise 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 an embodiment, the complete signal peptide or part of the signal peptide of IL-15, IL-15Rα or both may be mutated or deleted. In an embodiment, the fusion polypeptide may comprise one or more heterologous signal peptides, i.e., all or part of the signal peptide from molecules other than IL-15 and IL-15Rα. In an embodiment, the heterologous signal peptide may be derived from IL-2, CD33, IgVκ or IgE. In an embodiment, the signal peptide may be a signal peptide derived from IgE. In an embodiment, the signal peptide derived from IgE may comprise SEQ ID NO: 367 or consist thereof. In an embodiment, the signal peptide derived from IgE may be encoded by a nucleic acid comprising or consisting of the sequence shown in SEQ ID NO: 368.

[0405] In embodiments, the signal peptide can be cleaved or otherwise removed from the IL-15 / IL-15Rα fusion polypeptide.

[0406] In an embodiment, the signal peptide can increase or promote transcription, translation, translocation, or a combination thereof of the fusion polypeptide compared to the native IL-15Rα signal peptide, the native IL-15 signal peptide, or both. In an embodiment, the signal peptide can be fused directly or indirectly to the N-terminus or C-terminus of the IL-15 / IL-15Rα fusion polypeptide.

[0407] In embodiments, the signal peptide has a sequence comprising 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 embodiments, the function of the signal peptide, such as but not limited to one or more signaling functions of the signal peptide, is retained and / or enhanced in the mutated signal peptide.

[0408] In embodiments, the signal peptide may comprise SEQ ID NO: 367 comprising one, two, three, four or five amino acid substitutions. In embodiments, the function of the signal peptide, such as but not limited to one or more signal transduction functions of the signal peptide, is retained and / or enhanced in the mutated signal peptide.

[0409] In embodiments, the signal peptide may be encoded by a nucleic acid comprising 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 of SEQ ID NO: 368. In embodiments, the function of the signal peptide, such as but not limited to one or more signaling functions of the signal peptide, is retained and / or enhanced in the signal peptide encoded by the mutated nucleic acid sequence.

[0410] In an embodiment, the signal peptide can be encoded by a nucleic acid comprising SEQ ID NO:368 including one, two, three, four or five nucleic acid substitutions. In an embodiment, one or more nucleic acid substitutions in a codon can produce a codon encoding the same amino acid or can produce a codon encoding a different amino acid. In an embodiment, one or more nucleic acid substitutions in a codon can produce a codon encoding a conservative amino acid substitution. In an embodiment, one or more nucleic acid substitutions in a codon can produce a codon encoding the same amino acid. In an embodiment, the function of a signal peptide, such as but not limited to one or more signal transduction functions of a signal peptide, is retained and / or enhanced in a signal peptide encoded by a mutated nucleic acid sequence.

[0411] In embodiments, an 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 embodiments, an 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 embodiments, an 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 embodiments, an 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 embodiments, an 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: 345. In embodiments, an 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: 347.In embodiments, an 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: 349. In embodiments, an 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: 351. In embodiments, an 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: 353. In embodiments, an 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: 355. However, in embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE does not comprise or consist of SEQ ID NO: 355 or a sequence having about 95% or more sequence identity thereto. In 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 be membrane-bound and one or more signaling functions of IL-15Rα, (iii) the function of the signal peptide derived from IgE, such as but not limited to one or more signaling functions of the signal peptide, or (iv) all of (i), (ii) and (iii) are retained and / or enhanced in a mutated IL-15 / IL-15Rα fusion polypeptide comprising a signal peptide derived from IgE.

[0412] In an embodiment, an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE may comprise (a) SEQ ID NO: 337 comprising one, two, three, four or five amino acid substitutions; (b) SEQ ID NO: 339 comprising one, two, three, four or five amino acid substitutions; (c) SEQ ID NO: 341 comprising one, two, three, four or five amino acid substitutions; (d) SEQ ID NO: 343 comprising one, two, three, four or five amino acid substitutions; (e) SEQ ID NO: 345 comprising one, two, three, four or five amino acid substitutions; (f) SEQ ID NO: 347 comprising one, two, three, four or five amino acid substitutions; (g) SEQ ID NO: 349 comprising one, two, three, four or five amino acid substitutions; (h) SEQ ID NO: 351 comprising one, two, three, four or five amino acid substitutions; (i) SEQ ID NO: 352 comprising one, two, three, four or five amino acid substitutions. NO:353; or (j) SEQ ID NO:355 comprising one, two, three, four or five amino acid substitutions. However, in an embodiment, the IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE does not comprise SEQ ID NO:355 or a sequence having about 95% or more sequence identity therewith or consists thereof. In an embodiment, the amnion substitution may be conservative or non-conservative. In an embodiment, the amino acid substitution may be a conservative amino acid substitution. In an embodiment, (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 be membrane-bound and one or more signaling functions of IL-15Rα, (iii) the function of a signal peptide derived from IgE, such as but not limited to one or more signaling functions of a signal peptide or (iv) all of (i), (ii) and (iii) are retained and / or enhanced in a mutated IL-15 / IL-15Rα fusion polypeptide comprising a signal peptide derived from IgE.

[0413] In an embodiment, 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 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 of SEQ ID NO:338. In an embodiment, 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 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 of SEQ ID NO:340. In an embodiment, 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 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 of SEQ ID NO:342. In an embodiment, 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 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 of SEQ ID NO:344. In an embodiment, 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 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 of SEQ ID NO:346. In an embodiment, 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 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 of SEQ ID NO:348.In an embodiment, 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 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 of SEQ ID NO:350. In an embodiment, 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 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 of SEQ ID NO:352. In an embodiment, 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 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 of SEQ ID NO:354. In embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE may be encoded by a nucleic acid comprising 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 of SEQ ID NO: 356. However, in 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 about 80%, about 85%, about 90%, or about 95% or more sequence identity thereto. In an embodiment, (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 be membrane bound and one or more signaling functions of IL-15Rα, (iii) the function of the signal peptide derived from IgE, such as but not limited to one or more signaling functions of the signal peptide, or (iv) all of (i), (ii) and (iii) are retained and / or enhanced in the IL-15 / IL-15Rα fusion polypeptide encoded by the mutated nucleic acid sequence comprising a signal peptide derived from IgE.

[0414] In an embodiment, 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 comprising one, two, three, four or five nucleic acid substitutions; (b) SEQ ID NO: 340 comprising one, two, three, four or five nucleic acid substitutions; (c) SEQ ID NO: 342 comprising one, two, three, four or five nucleic acid substitutions; (d) SEQ ID NO: 344 comprising one, two, three, four or five nucleic acid substitutions; (e) SEQ ID NO: 346 comprising one, two, three, four or five nucleic acid substitutions; (f) SEQ ID NO: 348 comprising one, two, three, four or five nucleic acid substitutions; (g) SEQ ID NO: 350 comprising one, two, three, four or five nucleic acid substitutions; (h) SEQ ID NO: 352 comprising one, two, three, four or five nucleic acid substitutions; (i) SEQ ID NO: 353 comprising one, two, three, four or five nucleic acid substitutions. NO:354; or (j) SEQ ID NO:356 comprising one, two, three, four or five nucleic acid substitutions. However, in an embodiment, the 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 about 80%, about 85%, about 90%, or about 95% or more sequence identity thereto. In an embodiment, one or more nucleic acid substitutions in a codon can produce a codon encoding the same amino acid or can produce a codon encoding a different amino acid. In an embodiment, one or more nucleic acid substitutions in a codon can produce a codon encoding a conservative amino acid substitution. In an embodiment, (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 be membrane bound and one or more signaling functions of IL-15Rα, (iii) the function of the signal peptide derived from IgE, such as but not limited to one or more signaling functions of the signal peptide, or (iv) all of (i), (ii) and (iii) are retained and / or enhanced in the IL-15 / IL-15Rα fusion polypeptide encoded by the mutated nucleic acid sequence comprising a signal peptide derived from IgE.

[0415] In embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a signal peptide derived from IgE and optionally comprising one or more linkers comprises or consists of, e.g., SEQ ID NO: 367 (or a sequence 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) fused directly or indirectly to the N-terminus of SEQ ID NO: 307 (or a sequence 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. NO:309 (or a sequence 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) is fused directly or indirectly to the N-terminus of SEQ ID NO:309 (or a sequence 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:367 (or a sequence 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 fused directly or indirectly to the N-terminus of SEQ ID NO:367 NO:307 (or a sequence 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) is fused directly or indirectly to the N-terminus of SEQ ID NO:311 (or a sequence 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 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), fused directly or indirectly to the N-terminus of SEQ ID NO:307 (or a sequence 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:313 (or a sequence 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) is fused directly or indirectly to the N-terminus of SEQ ID NO:367 (or a sequence 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 fused directly or indirectly to the N-terminus of SEQ ID NO:313 (or a sequence 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), NO:307 (or a sequence 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) is fused directly or indirectly to the N-terminus of SEQ ID NO:315 (or a sequence 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;or SEQ ID NO:337, 339, 341, 343, 345, 347, 349, 351, 353 or 355, 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, 353 or 355. However, in embodiments, the IL-15 / IL-15Rα fusion polypeptide comprising a signal peptide derived from IgE and optionally comprising one or more linkers does not comprise or consist of: (i) SEQ ID NO: 307, which is fused directly or indirectly to the N-terminus of SEQ ID NO: 309 with a linker therebetween; (ii) SEQ ID NO: 335 or SEQ ID NO: 355; (iii) a sequence having about 95% or more sequence identity to SEQ ID NO: 307, which is fused directly or indirectly to the N-terminus of SEQ ID NO: 309 with a linker therebetween; or (iv) a sequence having about 95% or more sequence identity to SEQ ID NO: 335 or SEQ ID NO: 355. In embodiments, the one or more linkers comprise or consist of a linker sequence as set forth herein. ;

[0416] In embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a signal peptide derived from IgE and optionally comprising one or more linkers comprises or consists of, e.g., SEQ ID NO: 367 (or a sequence 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) fused directly or indirectly to the N-terminus of SEQ ID NO: 307 (or a sequence 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. NO:311 (or a sequence 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) is fused directly or indirectly to the N-terminus of SEQ ID NO:367 (or a sequence 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 fused directly or indirectly to the N-terminus of SEQ ID NO:311 (or a sequence 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), NO:307 (or a sequence 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) is fused directly or indirectly to the N-terminus of SEQ ID NO:313 (or a sequence 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;or SEQ ID NO: 367 (or a sequence 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 fused directly or indirectly to the N-terminus of SEQ ID NO: 307 (or a sequence 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. NO:315 (or a sequence 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:337, 339, 341, 343, 345, 347, 349, 351 or 353 or a sequence with SEQ ID NO:354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370, 371, 372, 373, 374, 375, 376, 377, 378, 379, 380, 381, 382, ​​383, 384, 385, 386, 387, 388, 389, 390, 401, 402, 403, 414, 415, 416, 417, 418, 420, 421, 422, 423, 424, 425 NO:337, 339, 341, 343, 345, 347, 349, 351 or 353 are 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. In embodiments, one or more linkers comprise or consist of a linker sequence as set forth herein. ;

[0417] In embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a signal peptide derived from IgE and comprising a linker comprises, e.g., 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.

[0418] In embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a signal peptide derived from IgE and optionally comprising one or more linkers is encoded by a nucleic acid comprising or consisting of, e.g., SEQ ID NO: 368 (or a sequence 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 identical to SEQ ID NO: 369. NO:308 (or a sequence 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) is fused directly or indirectly to the 5' end of SEQ ID NO:308 (or a sequence 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 fused directly or indirectly to the 5' end of SEQ ID NO:310 (or a sequence 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; NO:368 (or a sequence 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 fused directly or indirectly to the 5' end of SEQ ID NO:308 (or a sequence 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 the use of a nucleic acid encoding a linker therebetween. NO:312 (or a sequence 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) is fused directly or indirectly to the 5' end;SEQ ID NO:368 (or a sequence 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), fused directly or indirectly to the 5' end of SEQ ID NO:308 (or a sequence 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. NO:314 (or a sequence 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) is fused directly or indirectly to the 5' end; or SEQ ID NO:368 (or a sequence 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 fused directly or indirectly to the 5' end of SEQ ID NO:314 (or a sequence 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), 308 (or a sequence 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) is fused directly or indirectly to the 5' end of SEQ ID NO:308 (or a sequence 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 fused directly or indirectly to the 5' end of SEQ ID NO:316 (or a sequence 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, 354, or 356 or a sequence at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, NO:338, 340, 342, 344, 346, 348, 350, 352, 354 or 356 are 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 sequences.;

[0419] However, in 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 (i) SEQ ID NO: 356 or consisting thereof, or a sequence having about 80%, about 85%, about 90%, or about 95% or more sequence identity thereto, or (ii) SEQ ID NO: 368, fused directly or indirectly to the 5' end of SEQ ID NO: 308, fused directly or indirectly to the 5' end of SEQ ID NO: 310, or a sequence having about 80%, about 85%, about 90%, or about 95% or more sequence identity thereto. In embodiments, one or more linkers are encoded by one or more nucleic acids comprising or consisting of a nucleic acid encoding a linker as set forth herein.

[0420] In embodiments, an IL-15 / IL-15Rα fusion polypeptide comprising a signal peptide derived from IgE and optionally comprising one or more linkers is encoded by a nucleic acid comprising or consisting of, e.g., SEQ ID NO: 368 (or a sequence 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 identical to SEQ ID NO: 369. NO:308 (or a sequence 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) is fused directly or indirectly to the 5' end of SEQ ID NO:308 (or a sequence 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 fused directly or indirectly to the 5' end of SEQ ID NO:312 (or a sequence 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; NO:368 (or a sequence 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 fused directly or indirectly to the 5' end of SEQ ID NO:308 (or a sequence 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 the use of a nucleic acid encoding a linker therebetween. NO:314 (or a sequence 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) fused directly or indirectly to the 5' end;or SEQ ID NO:368 (or a sequence 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 fused directly or indirectly to the 5' end of SEQ ID NO:308 (or a sequence 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 fused directly or indirectly to the 5' end of SEQ ID NO:308 (or a sequence 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), NO:316 (or a sequence 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) fused directly or indirectly to the 5' end; or SEQ ID NO:338, 340, 342, 344, 346, 348, 350, 352 or 354 or a sequence with SEQ ID NO:338, 340, 342, 344, 346, 348, 350, 352 or 354 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 sequence. In embodiments, one or more linkers are encoded by one or more nucleic acids that comprise or consist of nucleic acids encoding linkers as set forth herein. ;

[0421] In an embodiment, an IL-15 / IL-15Rα fusion polypeptide comprising a signal peptide derived from IgE and comprising one or more linkers is encoded by a nucleic acid comprising, for example, 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, or consists of.

[0422] In an embodiment, the vector may further comprise a post-transcriptional regulatory element (PRE) sequence. In an embodiment, the post-transcriptional regulatory element (PRE) sequence may be selected from woodchuck hepatitis virus PRE (WPRE) (such as but not limited to wild-type WPRE, such as but not limited to SEQ ID NO: 264, or mutated WPRE, such as but not limited to WPREmut1 (SEQ ID NO: 256) or WPREmut2 (SEQ ID NO: 257)) or hepatitis B virus (HBV) PRE (HPRE) (SEQ ID NO: 437), variants thereof, or any combination thereof.

[0423] In an embodiment, (i) an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE (ii) followed by WPREmut2 can be encoded by a nucleic acid comprising 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 of SEQ ID NO:357. In an embodiment, (i) an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE (ii) followed by WPREmut2 can be encoded by a nucleic acid comprising 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 of SEQ ID NO:358. In an embodiment, (i) an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE (ii) followed by WPREmut2 can be encoded by a nucleic acid comprising 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 of SEQ ID NO:359. In an embodiment, (i) an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE (ii) followed by WPREmut2 can be encoded by a nucleic acid comprising 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 of SEQ ID NO:360. In an embodiment, (i) an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE (ii) followed by WPREmut2 can be encoded by a nucleic acid comprising 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 of SEQ ID NO:361.In an embodiment, (i) an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE (ii) followed by WPREmut2 can be encoded by a nucleic acid comprising 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 of SEQ ID NO:362. In an embodiment, (i) an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE (ii) followed by WPREmut2 can be encoded by a nucleic acid comprising 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 of SEQ ID NO: 363. In an embodiment, (i) an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE (ii) followed by WPREmut2 can be encoded by a nucleic acid comprising 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 of SEQ ID NO:364. In an embodiment, (i) an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE (ii) followed by WPREmut2 can be encoded by a nucleic acid comprising 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 of SEQ ID NO:365. In an embodiment, (i) an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE (ii) followed by WPRE can be encoded by a nucleic acid comprising 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 of SEQ ID NO:366.However, in embodiments, (i) the IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE, (ii) followed by a mutant or wild-type WPRE is not encoded by SEQ ID NO: 366 or a sequence having about 80%, about 85%, about 90%, or about 95% or more sequence identity thereto. In 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 be membrane-bound and the signaling function of IL-15Rα, (iii) the function of the signal peptide derived from IgE, such as but not limited to one or more signaling functions of the signal peptide, (iv) the post-transcriptional regulatory function of the wild-type or mutant WPRE, or (v) all of (i), (ii), (iii) and (iv) are retained and / or enhanced in an IL-15 / IL-15Rα fusion polypeptide encoded by a mutated nucleic acid sequence comprising a signal peptide derived from IgE.

[0424] In an embodiment, (i) an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE (ii) followed by WPREmut2 or wild-type WPRE (where wt is indicated) can be encoded by a nucleic acid comprising (a) SEQ ID NO: 357 comprising one, two, three, four or five nucleic acid substitutions; (b) SEQ ID NO: 358 comprising one, two, three, four or five nucleic acid substitutions; (c) SEQ ID NO: 359 comprising one, two, three, four or five nucleic acid substitutions; (d) SEQ ID NO: 360 comprising one, two, three, four or five nucleic acid substitutions; (e) SEQ ID NO: 361 comprising one, two, three, four or five nucleic acid substitutions; (f) SEQ ID NO: 362 comprising one, two, three, four or five nucleic acid substitutions; (g) SEQ ID NO: 363 comprising one, two, three, four or five nucleic acid substitutions; (h) SEQ ID NO: 364 comprising one, two, three, four or five nucleic acid substitutions. NO:364; (i) SEQ ID NO:365 comprising one, two, three, four or five nucleic acid substitutions; or (j) SEQ ID NO:366 (wt WPRE) comprising one, two, three, four or five nucleic acid substitutions. However, in embodiments, (i) an IL-15 / IL-15Rα fusion polypeptide comprising a linker and a signal peptide derived from IgE, (ii) followed by a wild-type or mutant WPRE is not encoded by SEQ ID NO:366 or a sequence having about 80%, about 85%, about 90%, or about 95% or more sequence identity thereto. In embodiments, one or more nucleic acid substitutions in a codon may produce a codon encoding the same amino acid or may produce a codon encoding a differ...

Claims

1. A nucleic acid encoding a polypeptide comprising SEQ ID NO:311, 313 or 315, or a polypeptide 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, wherein the nucleic acid optionally comprises SEQ ID NO:312, 314 or 316, or a sequence 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.

2. The nucleic acid of claim 1, encoding (i) a polypeptide comprising SEQ ID NO: 307 fused directly or indirectly to the N-terminus of a polypeptide comprising any one of SEQ ID NOs: 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 fused directly or indirectly 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 one of SEQ ID NOs: 311, 313, or 315.

3. The nucleic acid of claim 1 or 2, encoding a polypeptide comprising SEQ ID NO: 317, 321, 325, 327, 329, 331 or 333, or a polypeptide 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, optionally comprising SEQ ID NO: 318, 320, 322, 324, 326, 328, 330, 332 or 334, or a polypeptide 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 ... A nucleic acid of NO:318, 320, 322, 324, 326, 328, 330, 332 or 334 that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid of NO:318, 320, 322, 324, 326, 328, 330, 332 or 334.

4. The nucleic acid of claim 3, further comprising a nucleic acid encoding a signal peptide, which is fused directly or indirectly 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.

5. The nucleic acid of claim 4, wherein the signal peptide is derived from an IgE polypeptide and optionally comprises 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 thereto, and the nucleic acid optionally comprises 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 thereto.

6. A nucleic acid according to claim 4 or 5, which encodes a polypeptide comprising SEQ ID NO:337, 341, 345, 347, 349, 351 or 353, or a polypeptide 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, and the nucleic acid optionally comprises SEQ ID NO:338, 342, 346, 348, 350, 352 or 354, or a sequence 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.

7. A nucleic acid according to any one of claims 1 to 6, further comprising a nucleic acid encoding (a) at least one TCR polypeptide, the 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 and at least one CD8 polypeptide, wherein the at least one TCR polypeptide comprises an α chain and a β chain, and the at least one CD8 polypeptide comprises (i) an α chain, (ii) a β chain, or (iii) both an α chain and a β chain.

8. The nucleic acid of claim 7, comprising a nucleic acid that is at least about 80%, 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, 301, 438, 439, 440, 441, 442, 443, 444, 445, 446 or 447.

9. A nucleic acid according to any one of the preceding claims, comprising: (a) Nucleic acid encoding (i) a T cell receptor (TCR) and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, The CD8 polypeptide comprises an α chain and a β chain, or (ii) a TCR and a CD8 polypeptide, wherein the TCR comprises an α chain and a β chain, and the CD8 polypeptide comprises an α chain without a β chain, and (b) a nucleic acid encoding a fusion polypeptide according to any one of claims 3 or 6, wherein the TCR α chain and the TCR β chain are selected from 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, 91 and 92; wherein the CD8 α chain is SEQ ID NO: 7, 258, 259, 262 or a variant thereof; wherein the CD8 beta chain, if present, is SEQ ID NO: 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 according to any one of claims 1 to 9.

12. The vector according to claim 11, comprising 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], in N1 comprises a nucleic acid encoding the CD8 β chain and is present or absent, N2 contains nucleic acid encoding the CD8 alpha chain, N3 contains nucleic acid encoding the TCR β chain, N4 comprises nucleic acid encoding a TCR alpha 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; and wherein L1-L4 each comprises a nucleic acid encoding at least one linker, wherein each of L1-L4 is independently the same or different, and wherein each of L1-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 of 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 of any one of claims 11 to 14, wherein the vector further comprises a nucleic acid encoding a chimeric antigen receptor (CAR).

16. A method for preparing T cells and / or natural killer cells for immunotherapy, the method comprising: isolating T cells and / or natural killer cells from a blood sample of a human subject, activating isolated T cells and / or natural killer cells, transducing activated T cells and / or natural killer cells with a nucleic acid according to any one of claims 1 to 9 or a vector according to any one of claims 11 to 15, Expanding transduced T cells and / or natural killer cells.

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

18. The method of claim 16 or 17, wherein (i) the activation comprises contacting the T cells and / or natural killer cells with anti-CD3 and anti-CD28 antibodies; and / or (ii) the activation, the expansion, or both are performed in the presence of a combination of IL-2 and IL-15 and optionally zoledronate.

19. A T cell and / or natural killer cell (i) transduced with a nucleic acid according to any one of claims 1 to 9, or (ii) comprising a vector according to any one of claims 11 to 15, or (iii) obtained by a method according to 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 cells and / or natural killer cells according to claim 19 or 20, wherein the αβ T cells are CD4+ or CD8+ T cells.

22. A composition comprising the T cells and / or natural killer cells according to claim 19.

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

24. The composition of 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 of claim 24, wherein the adjuvant is an anti-CD40 antibody, imiquimod, resiquimod, GM-CSF, cyclophosphamide, sunitinib, bevacizumab, atezolizumab, interferon alpha, interferon beta, CpG oligonucleotides and derivatives, poly(I:C) and derivatives, RN A. sildenafil, a particle formulation 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.

26. A method of treating a patient with cancer and / or eliciting an immune response in a patient with cancer, the method comprising administering to the patient a composition according to any one of claims 23 to 25, 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 cancer, stomach cancer and prostate cancer.

27. 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 a natural killer cell according to any one of claims 19 to 21 or a composition according to any one of claims 22 to 25 for use as a medicament.

28. 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 a natural killer cell according to any one of claims 19 to 21, or a composition according to any one of claims 22 to 25, for use in a method of treating cancer, 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 cancer, stomach cancer and prostate cancer.

29. 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 a natural killer cell according to any one of claims 19 to 21, or a composition according to any one of claims 22 to 26, for the preparation of a medicament for treating cancer, the cancer optionally being 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 cancer, stomach cancer and prostate cancer.

30. 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 a natural killer cell according to any one of claims 19 to 21 or a composition according to any one of claims 22 to 26 as a medicament.

31. 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 a natural killer cell according to any one of claims 19 to 21, or a composition according to any one of claims 22 to 26 for treating cancer, 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 cancer, stomach cancer and prostate cancer.

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

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