Membrane-bound il-15, CD8 polypeptides, cells, compositions, and methods of using thereof

Abstract

The present disclosure relates to cells capable of co-expressing T cell receptors ("TCR") together with membrane-bound IL-15 polypeptides and / or CD8 polypeptides and the use thereof in adoptive cellular therapy. The present disclosure further provides for modified IL-15, IL-15Rα, IL-15 / IL-15Rα fusion polypeptide, and IL-15Rα / IL-15 fusion polypeptide sequences, vectors, and associated methods of making and using the same. The present disclosure further provides for modified CD8 sequences, vectors, and associated methods of making and using the same.

Description

MEMBRANE-BOUND IL-15, CD8 POLYPEPTIDES, CELLS, COMPOSITIONS, AND METHODS OF USING THEREOFCROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application is an International Application claiming priority to U.S. Provisional Application No. 63 / 595,205, filed November 01, 2023, the entire contents of which are hereby incorporated by reference in its entirety.REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY

[0002] A Sequence Listing is submitted herewith as an XML file named “3000011- 035977_Sequence_Listing_ST26” created on 30 October 2024 and having a size of 758,908 bytes as permitted under 37 C.F.R. § 1.821 (c). The material in the aforementioned file is hereby incorporated by reference in its entirety.BACKGROUNDField

[0003] The present disclosure relates to cells capable of co-expressing one or any combination of T cell receptors (“TCR”), CD8 polypeptides, and / or membrane-bound interleukin 15 (IL-15) and the use thereof in adoptive cellular therapy (“ACT”). The present disclosure further provides for modified CD8 sequences, IL- 15 sequences, IL- 15 receptor a (IL-15-Ra) sequences, IL- 15 / IL-15Ra fusion polypeptides, vectors, compositions, transformed cells, and associated methods thereof. Specifically, the present disclosure relates to sequences encoding specific TCRs and IL-15 / IL-15Ra fusion polypeptides and optionally CD8. Further, the present disclosure relates to vectors and cells, specifically T cells such as aP T cells, comprising and capable of co-expressing such sequences.

[0004] CD8 and CD4 are transmembrane glycoproteins characteristic of distinct populations of T lymphocytes whose antigen responses are restricted by class I and class II MHC molecules, respectively. They play major roles both in the differentiation and selection of T cells during thymic development and in the activation of mature T lymphocytes in response to antigen presenting cells. Both CD8 and CD4 are immunoglobulin superfamily proteins. They determine antigen restriction by binding to MHC molecules at an interface distinct from the region presenting the antigenic peptide, but the structural basis for their similar functions appears to be very different. Their sequence similarity is low and, whereas CD4 is expressed on the cell surface as a monomer, CD8 is expressed as an aa homodimer(e.g., FIG. 55C) or an aP heterodimer (e.g., FIG. 55A). In humans, this CD8aa homodimer may functionally substitute for the CD8aP heterodimer. CD8 contacts an acidic loop in the a3 domain of Class I MHC, thereby increasing the avidity of the T cell for its target. CD8 is also involved in the phosphorylation events leading to CTL activation through the association of its a chain cytoplasmic tail with the tyrosine kinase p56 / ot.

[0005] Pleiotropic cytokine interleukin- 15 (“IL- 15” or “IL 15”) is a member of the 4 a- helix bundle cytokine family. (Waldmann TA and Tagaya Y, Ann. Rev. Immunol. 17: 19-49, 1999, the content of which is incorporated herein by reference). A 14-15 kDa glycoprotein, wild type IL-15 shares partial structural homology with IL-2. (Id.). Wild type IL-15 can be expressed in two isoforms, one having a 48 amino acid signal peptide and the other having a 21 amino acid signal peptide. Id.). The mature form of wild type IL-15 consists of 114 amino acids. (Id.). Wild type IL-15 expression is regulated at the transcriptional, translational, and intracellular trafficking levels. (Id.). Wild type IL-15 utilizes a private receptor, IL-15Ra (or “IL15Ra”), which, in lymphocytes, binds IL- 15 with high affinity and trimerizes with IL-2RP (also referred to as IL-2 / IL-15RP) and IL-2Ry (also referred to as yc). (Id Okada S et al., Immunol, and Cell Biol. 93: 461-471, 2015, the content of which is incorporated herein by reference). Wild type IL-15Ra comprises a signal peptide, an extracellular domain, a transmembrane domain, and a cytoplasmic domain. (Waldmann and Tagaya). The extracellular domain of wild type IL-15Ra comprises a sushi domain (also referred to as a GP-1 motif). (Id.).

[0006] Adoptive cell therapy (ACT) is a promising approach to treatment of diseases such as cancer. T-cell therapy has been successful in treating various cancers. Li et al. Signal Transduction and Targeted Therapy 4(35): (2019), the content of which is incorporated by reference in its entirety. However, cells used in ACT often fail to persist in the tumor microenvironment and quickly lose their ability to kill tumor cells. Accordingly, there is a need for T cells and natural killer cells that exhibit longer persistence in the tumor microenvironment and / or sustained capability to kill tumor cells. It is also desirable to develop methods of manufacturing T cells and natural killer cells with enhanced, specific cytotoxic activity for immunotherapy.BRIEF SUMMARY

[0007] In a first aspect, the present disclosure relates to nucleic acids comprising a nucleotide sequence according to any one of SEQ ID NOs: 454, 451, 448, 449, 450, 452 or 453or 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 any one of SEQ ID NOs: 454, 451, 448, 449, 450, 452 or 453. In some embodiments, the nucleic acid of the present disclosure comprises a nucleotide sequence according to any one of SEQ ID NOs: 454, 451, 448 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 any one of SEQ ID NOs: 454, 451, 448. In specific embodiments, the nucleic acid of the present disclosure comprises a nucleotide sequence according to any one of SEQ ID NO: 454 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: 454. The nucleic acid(s) provided herein encode one or more (fusion) polypeptide(s). Specifically, such polypeptides and fusion polypeptides may include a TCR having an alpha chain variable domain as comprised in SEQ ID NO: 15 ,and a TCR beta chain variable domain as comprised in SEQ ID NO: 16. Such polypeptides may also include an IL-15Ra fusion polypeptide having a CD28 or CD25 transmembrane domain. Further, the present disclosure provides vectors comprising the nucleic acids according to said first aspect, T cells and / or natural killer cell transduced with such nucleic acids or vectors and / or expressing the polypeptides encoded by said nucleic acids or vectors. In other words, the T cells and / or natural killer cells of the invention may express a TCR having an alpha chain variable domain as comprised in SEQ ID NO: 15, and a beta chain variable domain as comprised in SEQ ID NO: 16, and further an IL-15 / IL-15Ra fusion polypeptide having a CD28 or CD25 transmembrane domain as comprised in SEQ ID NO: 329, 327, 325, 323 or 321. Specifically, the TCR alpha chain variable domain may have an amino acid sequence comprising the sequence of SEQ ID NO: 455, the TCR beta chain variable domain may have an amino acid sequence comprising SEQ ID NO: 456, and the IL-15 / IL-15 Receptor alpha fusion polypeptide may have an amino acid sequence as comprised in SEQ ID NO: 329. In some embodiments, the T cells and / or natural killer cells may additionally express CD8 encoded by the nucleic acid(s) and / or vector(s). Finally, compositions comprising said nucleic acids, vectors and / or said transduced T cells and / or natural killer cells are also provided. Said compositions are envisaged for medical purposes and in particular in the treatment of cancer.

[0008] In some embodiments, a membrane-bound IL- 15 polypeptide (membrane-bound IL- 15 or mbIL-15) may be provided. In some embodiments, nucleic acids described herein may comprise and / or encode a membrane-bound IL- 15 polypeptide. In some embodiments,vectors described herein may comprise and / or encode a membrane-bound IL- 15 polypeptide. In some embodiments, cells described herein may comprise and / or express a membranebound IL- 15 polypeptide. In some embodiments, compositions described herein may comprise a membrane-bound IL- 15 polypeptide or may comprise cells comprising and / or expressing a membrane-bound IL-15 polypeptide. In some embodiments, IL-15 may be rendered membrane-bound by expressing an IL-15 polypeptide and an IL-15Ra polypeptide in an IL-15 / IL-15Ra fusion polypeptide (IL-15 / IL-15Ra). IL-15 / IL-15Ra fusion polypeptides and other membrane-bound forms of IL- 15 may be referred to as membrane-bound IL- 15 (mbIL-15).

[0009] In some embodiments, isolated membrane-bound IL- 15 polypeptides may be provided. Isolated nucleic acid sequences comprising one or more nucleic acid sequences encoding one or more membrane-bound IL- 15 polypeptides may be provided. In some embodiments, isolated vectors comprising one or more nucleic acid sequences comprising one or more nucleic acid sequences encoding one or more membrane-bound IL- 15 polypeptides may be provided. In some embodiments, cells comprising and / or expressing one or more membrane-bound IL-15 polypeptides may be provided. In some embodiments, cells comprising or expressing one or more nucleic acid sequences comprising one or more nucleic acid sequences encoding one or more membrane-bound IL- 15 polypeptides may be provided. In some embodiments, cells comprising or expressing one or more vectors comprising one or more nucleic acid sequences comprising one or more nucleic acid sequences encoding one or more membrane-bound IL- 15 polypeptides may be provided. In some embodiments, compositions comprising such polypeptides, nucleic acids, vectors, and / or cells may be provided.

[0010] In some embodiments, an IL-15 polypeptide may be located N-terminal to an IL- 15Ra polypeptide in a membrane-bound IL- 15 polypeptide. (FIG. 67A). In some embodiments, an IL- 15 polypeptide may be located C-terminal to an IL-15Ra polypeptide in a membrane-bound IL- 15 polypeptide. (FIG. 67B). The IL- 15 polypeptide in FIGS. 67 A and 67B may be immature wild type (“wt”), immature mutated, mature wild type, or mature mutated. The IL-15Ra polypeptide in FIGS. 67A and 67B may be immature wild type, immature mutated, mature wild type, or mature mutated. In some embodiments, the IL- 15 polypeptide in 67A and 67B is mature and may or may not be mutated, and the IL-15Ra polypeptide in 67A and 67B is mature and may or may not be mutated. In some embodiments, the IL-15 polypeptide in 67A and 67B is mature and may or may not be mutated, and the IL-15Ra polypeptide in in 67A and 67B is mature and mutated. Although alinker is depicted in FIGS: 67 A and 67B, a mbIL-15 polypeptide may or may not comprise a linker.

[0011] In some embodiments, an IL- 15 polypeptide and an IL-15Ra polypeptide may be linked by one or more linker. In some embodiments, a membrane-bound IL- 15 may comprise and / or be encoded by a structure as shown in FIG. 67A or FIG. 67B. In FIGs. 67A and 67B, the lines connecting the IL- 15 to the one or more linker (L) and the one or more linker (L) to the IL-15Ra may represent direct linkages, with no intervening sequences, or may represent intervening sequences, such as, but not limited to, a linker, an untranslated sequence (in the case of a nucleic acid sequence), a translated sequence, a sequence comprising one or more restriction endonuclease sites (in the case of a nucleic acid sequence), or a combination thereof.

[0012] In some embodiments, an IL-15 / IL-15Ra polypeptide may comprise one or more signal peptide. In some embodiments, a membrane-bound IL- 15 comprising one or more signal peptide and, optionally, one or more linker may comprise and / or be encoded by a structure as shown in FIG. 68 A or FIG. 68B. The IL- 15 polypeptide in FIGS. 68 A and 68B may be immature wild type, immature mutated, mature wild type, or mature mutated. The IL- 15Ra polypeptide in FIGS. 68A and 68B may be immature wild type, immature mutated, mature wild type, or mature mutated. In some embodiments, the IL- 15 polypeptide in FIG. 68 A and FIG. 68B is mature and may or may not be mutated, and the IL-15Ra polypeptide in FIG. 68A and FIG. 68B is mature and may or may not be mutated. In some embodiments, the IL- 15 polypeptide in FIG. 68 A and FIG. 68B is mature and may or may not be mutated, and the IL-15Ra polypeptide in FIG. 68 A and FIG. 68B is mature and mutated. Although a linker is depicted in FIGS: 68A and 68B, a mbIL-15 polypeptide comprising a signal peptide may or may not comprise a linker. In FIGS. 68A and 68B, the lines connecting (a) the one or more signal peptide (SP) to the IL-15, the IL-15 to the one or more linker (L), and the one or more linker to the IL-15Ra (as in FIG. 68A) or (b) the one or more signal peptide (SP) to the IL- 15a, the IL- 15a to the one or more linker (L), and the one or more linker to the IL- 15 (as in FIG. 68B) may represent direct linkages, with no intervening sequences, or may represent intervening sequences, such as, but not limited to, a linker, an untranslated sequence (in the case of a nucleic acid sequence), a translated sequence, a sequence comprising one or more restriction endonuclease sites (in the case of a nucleic acid sequence), or a combination thereof.

[0013] In some embodiments, CD8 polypeptides described herein may comprise a CD8a immunoglobulin (Ig)-like domain, a CD8P region, a CD8a transmembrane domain, and aCD8a cytoplasmic domain. In some embodiments, a CD8P region may be a CD8P stalk region or domain.

[0014] In some embodiments, CD8 polypeptides described herein may 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 CD8P 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% identity sequence identity to the amino acid sequence of SEQ ID NO: 2, (c) a transmembrane 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: 3, and (d) a cytoplasmic 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: 4.

[0015] In some embodiments, CD8 polypeptides described herein have at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% sequence identity to the amino acid sequence of SEQ ID NO: 5.

[0016] In some embodiments, CD8 polypeptides described herein have at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% sequence identity to the amino acid sequence of SEQ ID NO: 7.

[0017] In some embodiments, CD8 polypeptides described herein may comprise one or more signal peptide with at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 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 directly or indirectly fused to the N-terminus or to the C-terminus of CD8 polypeptides described herein.

[0018] In some embodiments, 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 some embodiments, amino acid substitutions may be conservative or non-conservative. In some embodiments, amino acid substitution(s) may be conservative amino acid substitution(s).

[0019] In some embodiments, CD8 polypeptides described herein may be CD8a or modified CD8a polypeptides.

[0020] In some embodiments, CD8 polypeptides described herein may be CD8aP or modified CD8a polypeptides.

[0021] In some embodiments, a CD8P polypeptide may comprise the amino acid sequence of any one of SEQ ID NO: 8, 9, 10, 11, 12, 13, or 14.

[0022] In some embodiments, a TCR a chain and a TCR P chain may 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.

[0023] In some embodiments, a TCR a chain variable domain and a TCR P chain variable domain may be selected from SEQ ID NO: 455 and 456.

[0024] In some embodiments, an isolated nucleic acid may comprise a nucleic acid sequence encoding a T-cell receptor comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain and a P chain. In some embodiments, a CD8 polypeptide may comprise a CD8a chain and / or a CD8P chain, and the CD8a chain and / or CD8P chain may independently be modified or unmodified. An isolated nucleic acid may comprise a nucleic acid at least about 80% identical to the nucleic acid sequence of SEQ ID NO: 267, 269, 271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291, 295, 297, 299, or 301. An isolated nucleic acid may 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 an aspect, polypeptide sequences and / or nucleic acid sequences described herein may be isolated and / or recombinant sequences.

[0026] In an aspect, cells described herein may be isolated and / or recombinant cells.

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

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

[0029] In some embodiments, isolated polypeptide(s) may be encoded by nucleic acids described herein or, due, for example, to codon degeneration, by nucleic acids encoding the same polypeptide.

[0030] In some embodiments, an isolated polypeptide may comprise an amino acid sequence 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. An 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 comprise 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 comprise at most 1, 2, 3, 4, 5, 10, 15, or 20 amino acid substitutions or deletions.

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

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

[0033] In some embodiments, a nucleic acid encoding a fusion polypeptide of Formula I: N-terminus-P6-PL-P7-C-terminus [I] wherein P6 and P7 are each independently a first and second polypeptides and PL is a linker, wherein PL comprises SEQ ID NO: 387 or 389 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: 387 or 389 may be provided.

[0034] In some embodiments, a nucleic acid comprising formula II: 5’-N6-NL-N7-3’ [II] wherein N6 and N7 each independently encode a first and second polypeptides and NL encodes a linker, wherein NL comprises SEQ ID NO: 388 or 390 or a sequence at least about95%, 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 may be provided.

[0035] In some embodiments, a nucleic acid encoding a polypeptide comprising SEQ ID NO: 309, 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: 309, 311, 313, or 315 may be provided.

[0036] In some embodiments, 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 may be provided.

[0037] In some embodiments, a nucleic acid comprising SEQ ID NO: 310, 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: 310, 312, 314, or 316 may be provided. In some embodiments, a nucleic acid comprising 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 may be provided.

[0038] In some embodiments, the nucleic acid comprises the sequence SEQ ID NO: 314, 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: 314.

[0039] In some embodiments, a nucleic acid encoding (i) a polypeptide comprising SEQ ID NO: 307 fused directly or indirectly to an N terminus of a polypeptide comprising any of SEQ ID NO: 309, 311, 313, or 315 or (ii) 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: 307 fused directly or indirectly to 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 any of SEQ ID NO: 309, 311, 313, or 315 may be provided.

[0040] In some embodiments, a nucleic acid encoding (i) a polypeptide comprising SEQ ID NO: 307 fused directly or indirectly to an N terminus of a polypeptide comprising any of SEQ ID NO: 311, 313, or 315 or (ii) 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: 307 fused directly or indirectly to 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 any of SEQ ID NO: 311, 313, or 315 may be provided.

[0041] In some embodiments, a nucleic acid encoding (i) a polypeptide comprising SEQ ID NO: 307 fused directly or indirectly to an N terminus of a polypeptide comprising any of SEQ ID NO: 313 or (ii) 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: 307 fused directly or indirectly to 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 any of SEQ ID NO: 313 may be provided.

[0042] In some embodiments, a nucleic acid may further comprise a nucleic acid encoding a signal peptide directly or indirectly fused to an N terminus of SEQ ID NO: 307 or to an 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 some embodiments, the signal peptide may be derived from an IgE polypeptide. In some embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367 or a sequence 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.

[0043] In some embodiments, a nucleic acid comprising (i) SEQ ID NO: 308 fused directly or indirectly to a 5’ end of any of SEQ ID NO: 310, 312, 314, or 316 or (ii) 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 fused directly or indirectly to the 5’ end of any 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 any of SEQ ID NO: 310, 312, 314, or 316 may be provided.

[0044] In some embodiments, a nucleic acid comprising (i) SEQ ID NO: 308 fused directly or indirectly to a 5’ end of any of SEQ ID NO: 312, 314, or 316 or (ii) 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 fused directly or indirectly to the 5’ end of any 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 any of SEQ ID NO: 312, 314, or 316 may be provided.

[0045] In some embodiments, a nucleic acid comprising (i) SEQ ID NO: 308 fused directly or indirectly to a 5’ end of any of SEQ ID NO: 314 or (ii) 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 fused directly or indirectly to the 5’ end of any 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 any of SEQ ID NO: 314 may be provided.

[0046] In some embodiments, a nucleic acid may further comprise a nucleic acid encoding a signal peptide directly or indirectly fused to the 5’ end of SEQ ID NO: 308 or to 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 some embodiments, the signal peptide may be derived from an IgE polypeptide. In some embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 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.

[0047] In some embodiments, a nucleic acid encoding a polypeptide comprising SEQ ID NO: 317, 321, 325, 327, 329, 331, 333, or 335 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, 333, or 335 may be provided.

[0048] In some embodiments, a nucleic acid 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 may be provided.

[0049] In some embodiments, a nucleic acid encoding a polypeptide comprising SEQ ID NO: 329 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: 329 may be provided.

[0050] In some embodiments, a nucleic acid may further comprise a nucleic acid encoding a signal peptide directly or indirectly fused to an N terminus of SEQ ID NO: 317, 321, 325, 327, 329, 331, or 333 or to an 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 some embodiments, thesignal peptide may be derived from an IgE polypeptide. In some embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367 or a sequence 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.

[0051] In some embodiments, a nucleic acid may further comprise a nucleic acid encoding a signal peptide directly or indirectly fused to an N terminus of SEQ ID NO.: 329 or to an 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: 329. In some embodiments, the signal peptide may be derived from an IgE polypeptide. In some embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367 or a sequence 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 some embodiments, a nucleic acid comprising SEQ ID NO: 318, 322, 326, 328, 330, 332, 334, or 336 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: 318, 322, 326, 328, 330, 332, 334, or 336 may be provided.

[0053] In some embodiments, a nucleic acid comprising SEQ ID NO: 318, 322, 326, 328, 330, 332, or 334 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: 318, 322, 326, 328, 330, 332, or 334 may be provided.

[0054] In some embodiments, a nucleic acid comprising SEQ ID NO: 330 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: 330 may be provided.

[0055]

[0056] In some embodiments, a nucleic acid may further comprise a nucleic acid encoding a signal peptide directly or indirectly fused to the 5’ end of SEQ ID NO: 318, 322, 326, 328, 330, 332, or 334 or to 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: 318, 322, 326, 328, 330, 332, or 334. In some embodiments, the signal peptide may be derived from an IgEpolypeptide. In some embodiments, the signal peptide derived from an IgE polypeptide may be 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.

[0057] In some embodiments, a nucleic acid may further comprise a nucleic acid encoding a signal peptide directly or indirectly fused to the 5’ end of SEQ ID NO: 330, or to 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: 330. In some embodiments, the signal peptide may be derived from an IgE polypeptide. In some embodiments, the signal peptide derived from an IgE polypeptide may be 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.

[0058] In some embodiments, a nucleic acid encoding a polypeptide comprising SEQ ID NO: 337, 341, 345, 347, 349, 351, 353, or 355 or a polypeptide at least about 95%, 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 may be provided.

[0059] In some embodiments, a nucleic acid encoding 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 may be provided.

[0060] In some embodiments, a nucleic acid encoding a polypeptide comprising SEQ ID NO: 349 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: 349 may be provided.

[0061] In some embodiments, a nucleic acid comprising SEQ ID NO: 338, 342, 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 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 338, 342, 346, 348, 350, 352, 354, or 356 may be provided.

[0062] In some embodiments, a nucleic acid comprising 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 about99%, or about 100% identical to SEQ ID NO: 338, 342, 346, 348, 350, 352, or 354 may be provided.

[0063] In some embodiments, a nucleic acid comprising SEQ ID NO: 350 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: 350 may be provided.

[0064] In some embodiments, a nucleic acid described herein may further comprise a nucleic acid encoding (a) at least one TCR polypeptide comprising an a chain and a P chain, (b) at least one CD8 polypeptide comprising (i) an a chain, (ii) a P chain, or (iii) both an a chain and a P chain, or (c) at least one TCR polypeptide comprising an a chain and a P chain and at least one CD8 polypeptide comprising (i) an a chain, (ii) a P chain, or (iii) both an a chain and a P chain.

[0065] In some embodiments, a polypeptide, polypeptides, or fusion polypeptide encoded by a nucleic acid described herein may be provided.

[0066] In some embodiments, a polypeptide or fusion polypeptide comprising an amino acid sequence 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 may be provided.

[0067] In some embodiments, a polypeptide or fusion polypeptide comprising an amino acid sequence 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 may be provided.

[0068] In some embodiments, the polypeptide or fusion polypeptide comprises an amino acid sequence at least about 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 329.

[0069] In some embodiments, a fusion polypeptide comprising a polypeptide 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 an N terminus of any of 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 any of SEQ ID NO: 309, 311, 313, or 315 may be provided.

[0070] In some embodiments, a fusion polypeptide comprising a polypeptide 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 an N terminus of any of apolypeptide at 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 may be provided.

[0071] In some embodiments, a fusion polypeptide may further comprise a signal peptide directly or indirectly fused to an N terminus of 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: 307. In some embodiments, the signal peptide may be derived from an IgE polypeptide. In some embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367 or a sequence 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.

[0072] In some embodiments, a nucleic acid comprising: (a) a nucleic acid encoding (i) a T-cell receptor (TCR) comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain and a P chain, or (ii) a TCR comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain without a P chain, and (b) a nucleic acid encoding an IL-15 / IL-15Ra fusion polypeptide, wherein the TCR a chain and the TCR P 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 and60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, 71 and 303, 304 and 74, 75 and76, 77 and 78, 79 and 80, 81 and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90, and 91 and92; wherein the CD8 a chain is SEQ ID NO: 7, 258, 259, 262, or a variant thereof; wherein, if present, the CD8 P chain is SEQ ID NO: 8, 9, 10, 11, 12, 13, or 14; and wherein the IL- 15 / IL-15Ra 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, directly or indirectly fused to an 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, directly or indirectly fused to an 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; (iii) SEQ ID NO: 307 or a sequence at least about 95%, at least about 96%, at least about 97%, at least about 98%, atleast about 99%, or about 100% identical thereto, directly or indirectly fused to an 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, directly or indirectly fused to an 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 may be provided.

[0073] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may further comprise a signal peptide directly or indirectly fused to the N terminus of SEQ ID NO: 307 of any of (i), (ii), (iii), or (iv) 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 of any of (i), (ii), (iii), or (iv).

[0074] In some embodiments, a nucleic acid comprising: (a) a nucleic acid encoding (i) a T-cell receptor (TCR) comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain and a P chain, or (ii) a TCR comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain without a P chain, and (b) a nucleic acid encoding an IL-15 / IL-15Ra fusion polypeptide, wherein the TCR a chain and the TCR P 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 a chain is SEQ ID NO: 7, 258, 259, 262, or a variant thereof; wherein, if present, the CD8 P chain is SEQ ID NO: 8, 9, 10, 11, 12, 13, or 14; and wherein the IL- 15 / IL-15Ra 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, directly or indirectly fused to an 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, directly or indirectly fused to an N terminus of SEQ ID NO: 313 or a sequence at least about95%, 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 (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 an 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 may be provided.

[0075] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may further comprise a signal peptide directly or indirectly fused to the N terminus of SEQ ID NO: 307 of any of (i), (ii), or (iii) or to the N terminus of a sequence at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 307 of any of (i), (ii), or (iii).

[0076] In some embodiments, the signal peptide may be derived from an IgE polypeptide. In some embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367 or a sequence 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.

[0077] In some embodiments, a nucleic acid comprising: (a) a nucleic acid encoding (i) a T-cell receptor (TCR) comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain and a P chain, or (ii) a TCR comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain without a P chain, and (b) a nucleic acid encoding an IL-15 / IL-15Ra fusion polypeptide, wherein the TCR a chain and the TCR P 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 a chain is SEQ ID NO: 7, 258, 259, 262, or a variant thereof; wherein, if present, the CD8 P chain is SEQ ID NO: 8, 9, 10, 11, 12, 13, or 14; and wherein the IL- 15 / IL-15Ra fusion polypeptide is selected from SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, 333, or 335, 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: 317, 319, 321, 323, 325, 327, 329, 331, 333, or 335 may be provided.

[0078] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may further comprise a signal peptide directly or indirectly fused to an N terminus of SEQ ID NO: SEQID NO: 317, 319, 321, 323, 325, 327, 329, 331, 333, or 335, or directly or indirectly fused to an 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, 319, 321, 323, 325, 327, 329, 331, 333, or 335.

[0079] In some embodiments, a nucleic acid comprising: (a) a nucleic acid encoding (i) a T-cell receptor (TCR) comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain and a P chain, or (ii) a TCR comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain without a P chain, and (b) a nucleic acid encoding an IL-15 / IL-15Ra fusion polypeptide, wherein the TCR a chain and the TCR P 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 a chain is SEQ ID NO: 7, 258, 259, 262, or a variant thereof; wherein, if present, the CD8 P chain is SEQ ID NO: 8, 9, 10, 11, 12, 13, or 14; and wherein the IL- 15 / IL-15Ra fusion polypeptide is selected from SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, or 333, 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: 317, 319, 321, 323, 325, 327, 329, 331, or 333 may be provided.

[0080] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may further comprise a signal peptide directly or indirectly fused to an 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 an 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, 319, 321, 323, 325, 327, 329, 331, or 333.

[0081] In some embodiments, the signal peptide may be derived from an IgE polypeptide. In some embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367 or a sequence 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.

[0082] In some embodiments, a nucleic acid comprising: (a) a nucleic acid encoding (i) a T-cell receptor (TCR) comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain and a P chain, or (ii) a TCR comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain without a P chain, and (b) a nucleic acid encoding afusion polypeptide comprising (i) 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 to SEQ ID NO: 367 fused to (ii) an N terminus of an IL-15 / IL-15Ra fusion polypeptide, wherein the TCR a chain and the TCR P 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 a chain is SEQ ID NO: 7, 258, 259, 262, or a variant thereof; wherein, if present, the CD8 P chain 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 at least about 95%, 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 may be provided.

[0083] In some embodiments, a nucleic acid comprising: (a) a nucleic acid encoding (i) a T-cell receptor (TCR) comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain and a P chain, or (ii) a TCR comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain without a P chain, and (b) a nucleic acid encoding a fusion polypeptide comprising (i) 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 to SEQ ID NO: 367 fused to (ii) an N terminus of an IL-15 / IL-15Ra fusion polypeptide, wherein the TCR a chain and the TCR P 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 a chain is SEQ ID NO: 7, 258, 259, 262, or a variant thereof; wherein, if present, the CD8 P chain 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 at least about 95%, 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 may be provided.

[0084] In some embodiments, a nucleic acid comprising: (a) a nucleic acid encoding (i) a T-cell receptor (TCR) comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain and a P chain, or (ii) a TCR comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain without a P chain, and (b) a nucleic acid encoding an IL-15 / IL-15Ra fusion polypeptide, wherein the TCR a chain and the TCR P 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 a chain is SEQ ID NO: 7, 258, 259, 262, or a variant thereof; wherein, if present, the CD8 P chain is SEQ ID NO: 8, 9, 10, 11, 12, 13, or 14; and wherein the IL- 15 / IL-15Ra fusion polypeptide is selected from SEQ ID NO: 317, 321, 325, 327, 329, 331, 333, or 335 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: 317, 321, 325, 327, 329, 331, 333, or 335 may be provided.

[0085] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may further comprise a signal peptide directly or indirectly fused to an N terminus of SEQ ID NO: 317, 321, 325, 327, 329, 331, 333, or 335, or to an 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, 333, or 335.

[0086] In some embodiments, a nucleic acid comprising: (a) a nucleic acid encoding (i) a T-cell receptor (TCR) comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain and a P chain, or (ii) a TCR comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain without a P chain, and (b) a nucleic acid encoding an IL-15 / IL-15Ra fusion polypeptide, wherein the TCR a chain and the TCR P 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 a chain is SEQ ID NO: 7, 258, 259, 262, or a variant thereof; wherein, if present, the CD8 P chain is SEQ ID NO: 8, 9, 10, 11, 12, 13, or 14; and wherein the IL- 15 / IL-15Ra fusion polypeptide is selected from SEQ ID NO: 317, 321, 325, 327, 329, 331, or333 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: 317, 321, 325, 327, 329, 331, or 333 may be provided.

[0087] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may further comprise a signal peptide directly or indirectly fused to an N terminus of SEQ ID NO: 317, 321, 325, 327, 329, 331, or 333, or to an 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.

[0088] In some embodiments, the signal peptide may be derived from an IgE polypeptide. In some embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367 or a sequence 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 some embodiments, a nucleic acid comprising: (a) a nucleic acid encoding (i) a T-cell receptor (TCR) comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain and a P chain, or (ii) a TCR comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain without a P chain, and (b) a nucleic acid encoding a fusion polypeptide comprising (i) 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 to SEQ ID NO: 367 fused to (ii) an N terminus of an IL-15 / IL-15Ra fusion polypeptide, wherein the TCR a chain and the TCR P 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 a chain is SEQ ID NO: 7, 258, 259, 262, or a variant thereof; wherein, if present, the CD8 P chain 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 at least about 95%, 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 may be provided.

[0090] In some embodiments, a nucleic acid comprising: (a) a nucleic acid encoding (i) a T-cell receptor (TCR) comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain and a P chain, or (ii) a TCR comprising an a chain and a P chain and aCD8 polypeptide comprising an a chain without a P chain, and (b) a nucleic acid encoding a fusion polypeptide comprising (i) 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 to SEQ ID NO: 367 fused to (ii) an N terminus of an IL-15 / IL-15Ra fusion polypeptide, wherein the TCR a chain and the TCR P 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 a chain is SEQ ID NO: 7, 258, 259, 262, or a variant thereof; wherein, if present, the CD8 P chain 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 at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 337, 341, 345, 347, 349, 351, or 353 may be provided.

[0091] In some embodiments, a nucleic acid comprising: (a) a nucleic acid encoding (i) a T-cell receptor (TCR) comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain and a P chain, or (ii) a TCR comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain without a P chain, and (b) a nucleic acid encoding an IL-15 / IL-15Ra fusion polypeptide, wherein the TCR a chain and the TCR P chain are selected from SEQ ID 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 a chain is SEQ ID NO: 7, 258, 259, 262, or a variant thereof; and wherein, if present, the CD8 P chain is SEQ ID NO: 8, 9, 10, 11, 12, 13, or 14 may be provided.

[0092] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may comprise SEQ ID NO: 317 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.

[0093] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may comprise SEQ ID NO: 319 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.

[0094] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may comprise SEQ ID NO: 321 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.

[0095] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may comprise SEQ ID NO: 323 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.

[0096] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may comprise SEQ ID NO: 325 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 some embodiments, the IL-15 / IL-15Ra fusion polypeptide may comprise SEQ ID NO: 327 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.

[0098] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may comprise SEQ ID NO: 329 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.

[0099] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may comprise SEQ ID NO: 331 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.

[0100] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may comprise SEQ ID NO: 333 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.

[0101] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may comprise SEQ ID NO: 335 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.

[0102] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may further comprise a signal peptide directly or indirectly fused to an N terminus of the IL-15 / IL-15Ra fusion polypeptide. In some embodiments, the signal peptide may be derived from an IgE polypeptide. In some embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367 or a sequence 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.

[0103] In some embodiments, a nucleic acid comprising: (a) a nucleic acid encoding (i) a T-cell receptor (TCR) comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain and a P chain, or (ii) a TCR comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain without a P chain, and (b) a nucleic acid encoding a fusion polypeptide comprising (i) 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 to SEQ ID NO: 367 fused to (ii) an N terminus of an IL-15 / IL-15Ra fusionpolypeptide, wherein the TCR a chain and the TCR P chain are selected from SEQ ID 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 a chain is SEQ ID NO: 7, 258, 259, 262, or a variant thereof; and wherein, if present, the CD8 P chain is SEQ ID NO: 8, 9, 10, 11, 12, 13, or 14 may be provided.

[0104] In some embodiments, the nucleic acid of (b) may encode SEQ ID NO: 337, 339, 341, 343, 345, 347, 349, 351, 353 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 any of the aforementioned SEQ ID NOs. 337, 339, 341, 343, 345, 347, 349, 351, 353.

[0105] In some embodiments, a nucleic acid comprising: (a) a nucleic acid 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-15Ra fusion polypeptide may be provided.

[0106] In some embodiments, a nucleic acid comprising: (a) a nucleic acid 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-15Ra fusion polypeptide may be provided.

[0107] In some embodiments, the nucleic acid encoding an IL-15 / IL-15Ra fusion polypeptide may 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, directly or indirectly fused to a 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, directly or indirectly fused to a 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; (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, directly or indirectly fused to a 5’ end of SEQ ID NO: 314 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; or (iv) SEQ ID NO: 308 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 a 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.

[0108] In some embodiments, a 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 of (i), (ii), (iii), or (iv) or to the 5’ end of 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: 308 of any of (i), (ii), (iii), or (iv).

[0109] In some embodiments, the nucleic acid encoding an IL-15 / IL-15Ra fusion polypeptide may 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, directly or indirectly fused to a 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, directly or indirectly fused to a 5’ end of SEQ ID NO: 314 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; 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, directly or indirectly fused to a 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.

[0110] In some embodiments, a 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 of (i), (ii), or (iii) or to the 5’ end of 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: 308 of any of (i), (ii), or (iii).

[0111] In some embodiments, the signal peptide may be derived from an IgE polypeptide. In some embodiments, the nucleic acid encoding the signal peptide may 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.

[0112] In some embodiments, the nucleic acid encoding an IL-15 / IL-15Ra fusion polypeptide may be selected from SEQ ID NO: 318, 320, 322, 324, 326, 328, 330, 332, 334, or 336 or a sequence 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.

[0113] In some embodiments, the nucleic acid encoding an IL-15 / IL-15Ra fusion polypeptide may further comprise a nucleic acid encoding a signal peptide, wherein the nucleic acid encoding the signal peptide is directly or indirectly fused to a 5’ end of SEQ ID NO: 318, 320, 322, 324, 326, 328, 330, 332, 334, or 336 or a sequence 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.

[0114] In some embodiments, the nucleic acid encoding an IL-15 / IL-15Ra fusion polypeptide may be selected from SEQ ID NO: 318, 320, 322, 324, 326, 328, 330, 332, or 334 or a sequence 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.

[0115] In some embodiments, the nucleic acid encoding an IL-15 / IL-15Ra fusion polypeptide may further comprise a nucleic acid encoding a signal peptide, wherein the nucleic acid encoding the signal peptide is directly or indirectly fused to a 5’ end of SEQ ID NO: 318, 320, 322, 324, 326, 328, 330, 332, or 334 or a sequence 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.

[0116] In some embodiments, the signal peptide may be derived from an IgE polypeptide. In some embodiments, the nucleic acid encoding the signal peptide may comprise SEQ ID NO: 368 or a sequence 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.

[0117] In some embodiments, the nucleic acid encoding an IL-15 / IL-15Ra 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 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.

[0118] In some embodiments, the nucleic acid encoding an IL-15 / IL-15Ra 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 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.

[0119] In some embodiments, the nucleic acid encoding an IL-15 / IL-15Ra fusion polypeptide may be selected from SEQ ID NO: 318, 322, 326, 328, 330, 332, 334, or 336 or a sequence 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.

[0120] In some embodiments, the nucleic acid encoding an IL-15 / IL-15Ra fusion polypeptide may further comprise an nucleic acid encoding a signal peptide, wherein the nucleic acid encoding the signal peptide is directly or indirectly fused to a 5’ end of SEQ ID NO: 318, 322, 326, 328, 330, 332, 334, or 336 or a sequence 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.

[0121] In some embodiments, the nucleic acid encoding an IL-15 / IL-15Ra fusion polypeptide may be selected from SEQ ID NO: 318, 322, 326, 328, 330, 332, or 334 or a sequence 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.

[0122] In some embodiments, the nucleic acid encoding an IL-15 / IL-15Ra fusion polypeptide may further comprise an nucleic acid encoding a signal peptide, wherein the nucleic acid encoding the signal peptide is directly or indirectly fused to a 5’ end of SEQ ID NO: 318, 322, 326, 328, 330, 332, or 334 or a sequence 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.

[0123] In some embodiments, the signal peptide may be derived from an IgE polypeptide. In some embodiments, the nucleic acid encoding the signal peptide may comprise SEQ ID NO: 368 or a sequence 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.

[0124] In some embodiments, the nucleic acid encoding an IL-15 / IL-15Ra 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 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.

[0125] In some embodiments, the nucleic acid encoding an IL-15 / IL-15Ra fusion polypeptide (i) may further comprise a nucleic acid encoding a signal peptide derived from an- 11 -IgE polypeptide and (ii) may be selected from SEQ ID NO: 338, 342, 346, 348, 350, 352, or 354 or a sequence 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.

[0126] In some embodiments, a vector comprising a nucleic acid encoding at least one CD8a chain, at least one TCRa chain, at least one TCRP chain, at least one IL-15 / IL-15Ra fusion polypeptide, and optionally at least one CD8P chain may be provided.

[0127] In some embodiments, a vector comprising Nl, N2, N3, N4, N5, LI, L2, L3, and L4, in any order, wherein Nl comprises a nucleic acid encoding a CD8P chain and is present or absent, N2 comprises a nucleic acid encoding a CD8a chain, N3 comprises a nucleic acid encoding a TCRP chain, N4 comprises a nucleic acid encoding a TCRa chain, and N5 comprises a nucleic acid encoding an IL-15 / IL-15Ra 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 may be provided.

[0128] In some embodiments, a vector may comprise Formula III or Formula IV:5 ’ -Nl -L 1 -N2-L2-N3 -L3 -N4-L4-N5-3 ’ [III] 5’-N5-Ll-Nl-L2-N2-L3-N3-L4-N4 -3’ [IV],

[0129] In some embodiments, Nl may comprise a nucleic acid encoding SEQ ID NO: 8, 9, 10, 11, 12, 13, or 14.

[0130] In some embodiments, N2 comprises a nucleic acid encoding a SEQ ID NO: 7, 258, 259, 262, or a variant thereof.

[0131] In some embodiments, N4 and N3 may comprise nucleic acids encoding SEQ ID NO: 15 and 16, 17 and 18, 19 and 20, 21 and 22, 23 and 24, 25 and 26, 27 and 28, 29 and 30, 31 and 32, 33 and 34, 35 and 36, 37 and 38, 39 and 40, 41 and 42, 43 and 44, 45 and 46, 47 and 48, 49 and 50, 51 and 52, 53 and 54, 55 and 56, 57 and 58, 59 and 60, 61 and 62, 63 and 64, 65 and 66, 67 and 68, 69 and 70, 71 and 303, 304 and 74, 75 and 76, 77 and 78, 79 and 80, 81 and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90, or 91 and 92.

[0132] In some embodiments, 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, directly or indirectly fused to an 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 about99%, or about 100% identical to SEQ ID NO: 307, directly or indirectly fused to an 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, directly or indirectly fused to an 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, directly or indirectly fused to an 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 therebetween.

[0133] In some embodiments, N5 may further comprise a nucleic acid encoding a signal peptide directly or indirectly fused to the 5’ end of the nucleic acid encoding SEQ ID NO: 307 of any of (i), (ii), (iii), or (iv) or to the 5’ end of the nucleic acid encoding the 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 of any of (i), (ii), (iii), or (iv).

[0134] In some embodiments, 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, directly or indirectly fused to an 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, directly or indirectly fused to an 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 at least about 95%, 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 an N terminus of SEQ ID NO: 315 or a sequence at least about 95%, at least about 96%, at least about 97%, atleast about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 315, with or without a linker therebetween.

[0135] In some embodiments, N5 may further comprise a nucleic acid encoding a signal peptide directly or indirectly fused to the 5’ end of the nucleic acid encoding SEQ ID NO: 307 of any of (i), (ii), or (iii) or to the 5’ end of the nucleic acid encoding the 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 of any of (i), (ii), or (iii).

[0136] In some embodiments, the signal peptide may be derived from an IgE polypeptide. In some embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367 or a sequence 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.

[0137] In some embodiments, N5 may comprise a nucleic acid encoding SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, 333, or 335 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: 317, 319, 321, 323, 325, 327, 329, 331, 333, or 335.

[0138] In some embodiments, N5 may further comprise a nucleic acid encoding a signal peptide directly or indirectly fused to the 5’ end of the nucleic acid encoding SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, 333, or 335 or to the 5’ end of the nucleic acid encoding the 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, 319, 321, 323, 325, 327, 329, 331, 333, or 335.

[0139] In some embodiments, N5 may comprise a nucleic acid encoding SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, or 333 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: 317, 319, 321, 323, 325, 327, 329, 331, or 333.

[0140] In some embodiments, N5 may further comprise a nucleic acid encoding a signal peptide directly or indirectly fused to the 5’ end of the nucleic acid encoding SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, or 333 or to the 5’ end of the nucleic acid encoding the 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, 319, 321, 323, 325, 327, 329, 331, or 333.

[0141] In some embodiments, the signal peptide may be derived from an IgE polypeptide. In some embodiments, the signal peptide derived from an IgE polypeptide may compriseSEQ 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.

[0142] In some embodiments, (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 at least about 95%, 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.

[0143] In some embodiments, (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 at least about 95%, 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.

[0144] In some embodiments, N5 may comprise a nucleic acid encoding SEQ ID NO: 317, 321, 325, 327, 329, 331, 333, or 335 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: 317, 321, 325, 327, 329, 331, 333, or 335.

[0145] In some embodiments, N5 may further comprise a nucleic acid encoding a signal peptide directly or indirectly fused to the 5’ end of the nucleic acid encoding SEQ ID NO: 317, 321, 325, 327, 329, 331, 333, or 335 or to the 5’ end of the nucleic acid encoding the 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, 333, or 335.

[0146] In some embodiments, N5 may comprise a nucleic acid encoding SEQ ID NO: 317, 321, 325, 327, 329, 331, or 333 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: 317, 321, 325, 327, 329, 331, or 333.

[0147] In some embodiments, N5 may further comprise a nucleic acid encoding a signal peptide directly or indirectly fused to the 5’ end of the nucleic acid encoding SEQ ID NO: 317, 321, 325, 327, 329, 331, or 333 or to the 5’ end of the nucleic acid encoding the 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.

[0148] In some embodiments, the signal peptide may be derived from an IgE polypeptide. In some embodiments, the signal peptide derived from an IgE polypeptide may compriseSEQ 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.

[0149] In some embodiments, (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 at least about 95%, 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.

[0150] In some embodiments, (i) the vector may further encode a 2A peptide or an internal ribosome entry site (IRES) positioned between N1 and LI, between LI 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) positioned between N5 and LI, between LI and Nl, 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.

[0151] In some embodiments, (i) the vector may further encode a furin positioned between Nl and LI, between LI 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 positioned between N5 and LI, between LI and Nl, between Nl 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.

[0152] In some embodiments, the 2A peptide may be P2A (SEQ ID NO: 93), T2A (SEQ ID NO: 94), E2A (SEQ ID NO: 95), or F2A (SEQ ID NO: 96).

[0153] In some embodiments, the IRES may be selected from the group consisting of IRES from picomavirus, IRES from flavivirus, IRES from pestivirus, IRES from retrovirus, IRES from lentivirus, IRES from insect RNA virus, and IRES from cellular mRNA.

[0154] In some embodiments, a T cell and / or natural killer cell comprising: (a) (i) a T- cell receptor (TCR) comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain and a P chain, or (ii) a TCR comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain without a P chain, and (b) an IL-15 / IL-15Ra fusion polypeptide, wherein the TCR a chain and the TCR P 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, 81and 82, 83 and 84, 85 and 86, 87 and 88, 89 and 90, and 91 and 92; wherein the CD8 a chain is SEQ ID NO: 7, 258, 259, 262, or a variant thereof; wherein, if present, the CD8 P chain is SEQ ID NO: 8, 9, 10, 11, 12, 13, or 14; and wherein the IL-15 / IL-15Ra 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, directly or indirectly fused to an 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, directly or indirectly fused to an 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, directly or indirectly fused to an 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, directly or indirectly fused to an 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 therebetween may be provided.

[0155] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may further comprise a signal peptide directly or indirectly fused to the N terminus of SEQ ID NO: 307 of any of (i), (ii), (iii), or (iv) 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 of any of (i), (ii), (iii), or (iv).

[0156] In some embodiments, a T cell and / or natural killer cell comprising: (a) (i) a T- cell receptor (TCR) comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain and a P chain, or (ii) a TCR comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain without a P chain, and (b) an IL-15 / IL-15Ra fusion polypeptide, wherein the TCR a chain and the TCR P 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, 31and 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 a chain is SEQ ID NO: 7, 258, 259, 262, or a variant thereof; wherein, if present, the CD8 P chain is SEQ ID NO: 8, 9, 10, 11, 12, 13, or 14; and wherein the IL-15 / IL-15Ra 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, directly or indirectly fused to an 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, directly or indirectly fused to an 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 at least about 95%, 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 an 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 therebetween may be provided.

[0157] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may further comprise a signal peptide directly or indirectly fused to the N terminus of SEQ ID NO: 307 of any of (i), (ii), or (iii) or to the N terminus of a sequence at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 307 of any of (i), (ii), or (iii).

[0158] In some embodiments, the signal peptide may be derived from an IgE polypeptide. In some embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367 or a sequence 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.

[0159] In some embodiments, a T cell and / or natural killer cell comprising: (a) (i) a T- cell receptor (TCR) comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain and a P chain, or (ii) a TCR comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain without a P chain, and (b) an IL-15 / IL-15Ra fusionpolypeptide, wherein the TCR a chain and the TCR P 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 a chain is SEQ ID NO: 7, 258, 259, 262, or a variant thereof; wherein, if present, the CD8 P chain is SEQ ID NO: 8, 9, 10, 11, 12, 13, or 14; and wherein the IL-15 / IL-15Ra fusion polypeptide is selected from SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, 333, or 335, 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: 317, 319, 321, 323, 325, 327, 329, 331, 333, or 335 may be provided.

[0160] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may further comprise a signal peptide directly or indirectly fused to an N terminus of SEQ ID NO: SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, 333, or 335, or directly or indirectly fused to an 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, 319, 321, 323, 325, 327, 329, 331, 333, or 335.

[0161] In some embodiments, a T cell and / or natural killer cell comprising: (a) (i) a T- cell receptor (TCR) comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain and a P chain, or (ii) a TCR comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain without a P chain, and (b) an IL-15 / IL-15Ra fusion polypeptide, wherein the TCR a chain and the TCR P 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 a chain is SEQ ID NO: 7, 258, 259, 262, or a variant thereof; wherein, if present, the CD8 P chain is SEQ ID NO: 8, 9, 10, 11, 12, 13, or 14; and wherein the IL-15 / IL-15Ra fusion polypeptide is selected from SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, or 333, 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: 317, 319, 321, 323, 325, 327, 329, 331, or 333 may be provided.

[0162] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may further comprise a signal peptide directly or indirectly fused to an 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 an 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, 319, 321, 323, 325, 327, 329, 331, or 333.

[0163] In some embodiments, the signal peptide may be derived from an IgE polypeptide. In some embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367 or a sequence 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.

[0164] In some embodiments, a T cell and / or natural killer cell comprising: (a) (i) a T- cell receptor (TCR) comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain and a P chain, or (ii) a TCR comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain without a P chain, and (b) a fusion polypeptide comprising (i) a signal peptide comprising 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 to SEQ ID NO: 367 fused to (ii) an IL-15 / IL-15Ra fusion polypeptide, wherein the TCR a chain and the TCR P 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 a chain is SEQ ID NO: 7, 258, 259, 262, or a variant thereof; wherein, if present, the CD8 P chain 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 at least about 95%, 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 may be provided.

[0165] In some embodiments, a T cell and / or natural killer cell comprising: (a) (i) a T- cell receptor (TCR) comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain and a P chain, or (ii) a TCR comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain without a P chain, and (b) a fusion polypeptide comprising (i) a signal peptide comprising SEQ ID NO: 367 or a sequence at least about 95%, at leastabout 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 367 fused to (ii) an IL-15 / IL-15Ra fusion polypeptide, wherein the TCR a chain and the TCR P 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 a chain is SEQ ID NO: 7, 258, 259, 262, or a variant thereof; wherein, if present, the CD8 P chain 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 at least about 95%, 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 may be provided.

[0166] In some embodiments, a T cell and / or natural killer cell transduced to express (a) (i) a T-cell receptor (TCR) comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain and a P chain, or (ii) a TCR comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain without a P chain, and (b) an IL-15 / IL-15Ra fusion polypeptide, wherein the TCR a chain and the TCR P 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 a chain is SEQ ID NO: 7, 258, 259, 262, or a variant thereof; wherein, if present, the CD8 P chain is SEQ ID NO: 8, 9, 10, 11, 12, 13, or 14; and wherein the IL-15 / IL-15Ra fusion polypeptide is selected from SEQ ID NO: 317, 321, 325, 327, 329, 331, 333, or 335 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: 317, 321, 325, 327, 329, 331, 333, or 335 may be provided.

[0167] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may further comprise a signal peptide directly or indirectly fused to an N terminus of SEQ ID NO: SEQ ID NO: 317, 321, 325, 327, 329, 331, 333, or 335, or directly or indirectly fused to an N terminus of a sequence at least about 95%, at least about 96%, at least about 97%, at leastabout 98%, at least about 99%, or about 100% identical to SEQ ID NO: 317, 321, 325, 327, 329, 331, 333, or 335.

[0168] In some embodiments, a T cell and / or natural killer cell transduced to express (a) (i) a T-cell receptor (TCR) comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain and a P chain, or (ii) a TCR comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain without a P chain, and (b) an IL-15 / IL-15Ra fusion polypeptide, wherein the TCR a chain and the TCR P 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 a chain is SEQ ID NO: 7, 258, 259, 262, or a variant thereof; wherein, if present, the CD8 P chain is SEQ ID NO: 8, 9, 10, 11, 12, 13, or 14; and wherein the IL-15 / IL-15Ra fusion polypeptide is selected from SEQ ID NO: 317, 321, 325, 327, 329, 331, or 333 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: 317, 321, 325, 327, 329, 331, or 333 may be provided.

[0169] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may further comprise a signal peptide directly or indirectly fused to an N terminus of SEQ ID NO: SEQ ID NO: 317, 321, 325, 327, 329, 331, or 333, or directly or indirectly fused to an 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.

[0170] In some embodiments, the signal peptide may be derived from an IgE polypeptide. In some embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367 or a sequence 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 some embodiments, a T cell and / or natural killer cell comprising: (a) (i) a T- cell receptor (TCR) comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain and a P chain, or (ii) a TCR comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain without a P chain, and (b) a fusion polypeptide comprising (i) a signal peptide comprising 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 to SEQ ID NO: 367 directly or indirectly fused to (ii) an IL-15 / IL-15Ra fusionpolypeptide, wherein the TCR a chain and the TCR P 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 a chain is SEQ ID NO: 7, 258, 259, 262, or a variant thereof; wherein, if present, the CD8 P chain 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 at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 337, 341, 345, 347, 349, 351, or 353 may be provided.

[0172] In some embodiments, a T cell and / or natural killer cell comprising: (a) (i) a T- cell receptor (TCR) comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain and a P chain, or (ii) a TCR comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain without a P chain, and (b) an IL-15 / IL-15Ra fusion polypeptide, wherein the TCR a chain and the TCR P chain are selected from SEQ ID 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 a chain is SEQ ID NO: 7, 258, 259, 262, or a variant thereof; and wherein, if present, the CD8 P chain is SEQ ID NO: 8, 9, 10, 11, 12, 13, or 14.

[0173] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may comprise SEQ ID NO: 317 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.

[0174] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may comprise SEQ ID NO: 319 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.

[0175] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may comprise SEQ ID NO: 321 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.

[0176] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may comprise SEQ ID NO: 323 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.

[0177] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may comprise SEQ ID NO: 325 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.

[0178] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may comprise SEQ ID NO: 327 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.

[0179] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may comprise SEQ ID NO: 329 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 some embodiments, the IL-15 / IL-15Ra fusion polypeptide may comprise SEQ ID NO: 331 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.

[0181] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may comprise SEQ ID NO: 333 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.

[0182] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may comprise SEQ ID NO: 335 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.

[0183] In some embodiments, the IL-15 / IL-15Ra fusion polypeptide may further comprise a signal peptide directly or indirectly fused to an N terminus of the IL-15 / IL-15Ra fusion polypeptide. In some embodiments, the signal peptide may be derived from an IgE polypeptide. In some embodiments, the signal peptide derived from an IgE polypeptide may comprise SEQ ID NO: 367 or a sequence 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 some embodiments, T cell and / or natural killer cell comprising: (a) (i) a T-cell receptor (TCR) comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain and a P chain, or (ii) a TCR comprising an a chain and a P chain and a CD8 polypeptide comprising an a chain without a P chain, and (b) a fusion polypeptide comprising(i) 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 to SEQ ID NO: 367 fused to(ii) an N terminus of an IL-15 / IL-15Ra fusion polypeptide, wherein the TCR a chain and the TCR P chain are selected from SEQ ID 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 a chain is SEQ ID NO: 7, 258, 259, 262, or a variant thereof; and wherein, if present, the CD8 P chain is SEQ ID NO: 8, 9, 10, 11, 12, 13, or 14.

[0185] In some embodiments, the fusion polypeptide of (b) may comprise SEQ ID NO: 337 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.

[0186] In some embodiments, the fusion polypeptide of (b) may comprise SEQ ID NO: 339 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.

[0187] In some embodiments, the fusion polypeptide of (b) may comprise SEQ ID NO: 341 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.

[0188] In some embodiments, the fusion polypeptide of (b) may comprise SEQ ID NO: 343 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.

[0189] In some embodiments, the fusion polypeptide of (b) may comprise SEQ ID NO: 345 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.

[0190] In some embodiments, the fusion polypeptide of (b) may comprise SEQ ID NO: 347 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.

[0191] In some embodiments, the fusion polypeptide of (b) may comprise SEQ ID NO: 349 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.

[0192] In some embodiments, the fusion polypeptide of (b) may comprise SEQ ID NO: 351 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.

[0193] In some embodiments, the fusion polypeptide of (b) may comprise SEQ ID NO: 353 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.

[0194] In some embodiments, the fusion polypeptide of (b) may comprise SEQ ID NO: 355 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.

[0195] In some embodiments, the T cell may be an aP T cell, a y5 T cell, a natural killer T cell, or any combination thereof. In some embodiments, the aP T cell may be a CD4+ T cell. In some embodiments, the aP T cell may be a CD8+ T cell. In some embodiments, the y5 T cell may be a Vy9V52+ T cell.

[0196] In some embodiments, a composition comprising a T cell and / or natural killer cell described herein may be provided. In some embodiments, the composition may be a pharmaceutical composition. In some embodiments, the composition may further comprise an adjuvant, excipient, carrier, diluent, buffer, stabilizer, or a combination thereof.

[0197] In some embodiments, the adjuvant may be an anti-CD40 antibody, imiquimod, resiquimod, GM-CSF, cyclophosphamide, sunitinib, bevacizumab, atezolizumab, interferonalpha, interferon-beta, CpG oligonucleotides and derivatives, poly(I:C) and derivatives, RNA, sildenafil, particulate formulations with 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 some embodiments, the adjuvant may be IL-2, IL-7, IL-12, IL-15, IL-21, or any combination thereof.

[0198] In some embodiments, a method of preparing T cells and / or natural killer cells for immunotherapy may be provided, the method comprising: isolating or enriching 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 expanding the transduced T cells and / or natural killer cells. In some embodiments, the method may further comprise isolating or enriching CD4+CD8+ T cells from the transduced T cells and / or natural killer cells and expanding the isolated CD4+CD8+ transduced T cells. In some embodiments, the blood sample may comprise peripheral blood mononuclear cells (PMBC). In some embodiments, the activating may comprise contacting the T cells and / or natural killer cells with an anti-CD3 and an anti-CD28 antibody. In some embodiments, the T cell may be a CD4+ T cell. In some embodiments, the T cell may be a CD8+ T cell. In some embodiments, the T cell may be a y5 T cell or an aP T cell. In some embodiments, the activation, the expanding, or both may be in the presence of a combination of IL-2 and IL- 15 and optionally with zoledronate.

[0199] In some embodiments, a method of increasing persistence, functionality, naivety, longevity, capacity to kill antigen-presenting cells, or a combination thereof, of T cells and / or natural killer cells may be provided, the method comprising: isolating or enriching 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 the transduced T cellsand / or natural killer cells, wherein the persistence, longevity, naivety, capacity to kill antigen- presenting cells, or a combination thereof of the transduced T cells and / or natural killer cells is increased as compared with that of control cells. In some embodiments, the method may further comprise expanding the transduced T cells and / or natural killer cells.

[0200] In some embodiments, the control cells may comprise non-transduced T cells and / or natural killer cells, T cells and / or natural killer cells transduced with TCR only, or a combination thereof. In some embodiments, the control may cells comprise non-transduced T cells and / or natural killer cells, T cells and / or natural killer cells transduced with TCR only, T cells and / or natural killer cells transduced with TCR and CD8 only, or a combination thereof. In some embodiments, the persistence, longevity, functionality, naivety, capacity 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 one challenge with antigen-presenting cells, two challenges with antigen-presenting cells, three challenges with antigen-presenting cells, four challenges with antigen-presenting cells, five challenges with antigen-presenting cells, six challenges with antigen-presenting cells, seven challenges with antigen-presenting cells, or more challenges with antigen-presenting cells, the persistence, longevity, functionality, naivety, capacity 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 five or more challenges with antigen-presenting cells or more challenges with antigen- presenting cells.

[0201] In some embodiments, a method of increasing interferon y (IFNy) secretion by T cells and / or natural killer cells may be provided, the method comprising: isolating or enriching 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 the transduced T cells and / or natural killer cells, wherein the IFNy secretion of the transduced T cells and / or natural killer cells is increased as compared with that of control cells. In some embodiments, the method may further comprise expanding the transduced T cells and / or natural killer cells. In some embodiments, the control cells may comprise non-transduced T cells and / or natural killer cells, T cells and / or natural killer cells transduced with TCR only, or a combination thereof.

[0202] In some embodiments, the control cells may comprise non-transduced T cells and / or natural killer cells, T cells and / or natural killer cells transduced with TCR only, T cells and / or natural killer cells transduced with TCR and CD8 only, or a combination thereof. In some embodiments, the IFNy secretion by the transduced T cells and / or natural killer cells and control cells may be determined after one challenge with antigen-presenting cells, two challenges with antigen-presenting cells, three challenges with antigen-presenting cells, four challenges with antigen-presenting cells, five challenges with antigen-presenting cells, six challenges with antigen-presenting cells, seven challenges with antigen-presenting cells, or more challenges with antigen-presenting cells. In some embodiments, the IFNy secretion by the transduced T cells and / or natural killer cells and control cells may be determined after five or more challenges with antigen-presenting cells or more challenges with antigen- presenting cells.

[0203] In some embodiments, the antigen presenting cells may present an antigen on a cell surface, and the transduced T cells and / or natural killer cells and control cells may be capable of killing the antigen presenting cells. In some embodiments, the antigen may comprise a peptide. In some embodiments, the peptide may be in a complex with an MHC molecule on the cell surface.

[0204] In some embodiments, a polypeptide, polypeptides, or fusion polypeptide encoded by a nucleic acid described herein may be provided.

[0205] In some embodiments, a nucleic acid described herein may be isolated, recombinant, or both isolated and recombinant.

[0206] In some embodiments, a vector described herein may be isolated, recombinant, or both isolated and recombinant.

[0207] In some embodiments, a T cell and / or natural killer cell described herein may be isolated, recombinant, engineered, or any combination thereof.

[0208] In some embodiments, a polypeptide, polypeptides, or fusion polypeptide described herein may be isolated, recombinant, or both isolated and recombinant.

[0209] In some embodiments, a vector comprising a nucleic acid described herein may be provided. In some embodiments, a vector described herein may further comprise a nucleic acid encoding a 2A peptide or an internal ribosome entry site (IRES) positioned between a nucleic acid encoding a CD8 a chain and a nucleic acid encoding a CD8 p chain. In some embodiments, the vector may further comprise a nucleic acid encoding a 2A peptide or an IRES positioned between a nucleic acid encoding a TCR a chain and a nucleic acid encodinga TCR P chain. In some embodiments, the vector may further comprise a nucleic acid encoding a 2A peptide or an IRES positioned 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- 15Ra fusion polypeptide. In some embodiments, the 2A peptide may be P2A (SEQ ID NO: 93), T2A (SEQ ID NO: 94), E2A (SEQ ID NO: 95), or F2A (SEQ ID NO: 96). In some embodiments, the IRES may be selected from the group consisting of IRES from picornavirus, IRES from flavivirus, IRES from pestivirus, IRES from retrovirus, IRES from lentivirus, IRES from insect RNA virus, and IRES from cellular mRNA. In some embodiments, the vector may further comprise a post-transcriptional regulatory element (PRE) sequence selected from a 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 some embodiments, the post-transcriptional regulatory element (PRE) sequence may be a Woodchuck PRE (WPRE) mutant 1 comprising the nucleic acid sequence of SEQ ID NO: 256. In some embodiments, the post-transcriptional regulatory element (PRE) sequence may be a Woodchuck PRE (WPRE) mutant 2 comprising the nucleic acid sequence of SEQ ID NO: 257. In some embodiments, the vector may further comprise a promoter selected from cytomegalovirus (CMV) promoter, phosphoglycerate kinase (PGK) promoter, myelin basic protein (MBP) promoter, glial fibrillary acidic protein (GFAP) promoter, modified MoMuLV LTR comprising myeloproliferative sarcoma virus enhancer (MNDU3), Ubiqitin C promoter, EF-1 alpha promoter, or Murine Stem Cell Virus (MSCV) promoter. In some embodiments, the promoter may be a Murine Stem Cell Virus (MSCV) promoter. In some embodiments, vector may be a viral vector or a non-viral vector. In some embodiments, the vector may be a viral vector. In some embodiments, the viral vector may be selected from adenoviruses, poxviruses, alphaviruses, arenaviruses, flaviviruses, rhabdoviruses, retroviruses, lentiviruses, herpesviruses, paramyxoviruses, picornaviruses, and any combination thereof. In some embodiments, the viral vector may be pseudotyped with an envelope protein of a virus selected from the native feline endogenous virus (RD114), a version of RD 114 (RD114TR), gibbon ape leukemia virus (GALV), a 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 retroviral envelope glycoprotein (BaEV), and lymphocytic choriomeningitis virus (LCMV). In some embodiments, the vector may be a lentiviral vector. In some embodiments, the vector may further comprise a nucleic acid encoding a chimeric antigen receptor (CAR).

[0210] In some embodiments, a T cell and / or natural killer cell expressing a polypeptide as described herein and / or comprising a vector described herein and / or produced by a method described herein may be provided. In some embodiments, the T cell may be an aP T cell, a y5 T cell, a natural killer T cell, or any combination thereof. In some embodiments, the aP T cell may be a CD4+ T cell. In some embodiments, the aP T cell may be a CD8+ T cell. In some embodiments, the y5 T cell may be a Vy9V52+ T cell.

[0211] In some embodiments, a composition comprising a T cell and / or natural killer cell described herein may be provided. In some embodiments, the composition may be a pharmaceutical composition. In some embodiments, the composition may further comprise an adjuvant, excipient, carrier, diluent, buffer, stabilizer, or a combination thereof. In some embodiments, the adjuvant may be an anti-CD40 antibody, imiquimod, resiquimod, GM- CSF, cyclophosphamide, sunitinib, bevacizumab, atezolizumab, interferon-alpha, interferonbeta, CpG oligonucleotides and derivatives, poly(I:C) and derivatives, RNA, sildenafil, particulate formulations with 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 some embodiments, the adjuvant may be IL-2, IL-7, IL-12, IL- 15, IL-21, or any combination thereof.

[0212] In some embodiments, a method of treating a patient who has 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, urinary bladder cancer, kidney cancer, leukemia, ovarian cancer, esophageal cancer, brain cancer, gastric cancer, and prostate cancer. In some embodiments, a method of eliciting an immune response in a patient who has 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, urinary bladder cancer, kidney cancer, leukemia, ovarian cancer, esophageal cancer, brain cancer, gastric cancer, and prostate cancer. In some embodiments, the T cell and / or natural killer cell may kill cancer cells that present a peptide in a complex with an MHC molecule on a cell surface.

[0213] In some embodiments, nucleic acid sequences disclosed herein may be mutated such that the amino acids encoded remain the same, but the nucleic acid codons are changed to maintain improved expression in a target cell and / or by a target vector. In some embodiments, nucleic acids disclosed herein may be codon optimized. In some embodiments, nucleic acid sequences set forth herein are codon optimized. In some embodiments, nucleic acid sequences set forth herein may be codon optimized, and nucleic acid sequences encoding polypeptides set forth herein may be codon optimized. In some embodiments, mutation of nucleic acid sequences set forth herein may encompass codon optimization.

[0214] In some embodiments, expression of membrane-bound IL-15 may improve immune cell, such as but not limited to, T cell and / or natural killer cell, persistence, functionality, growth, viability, expansion, or any combination thereof, as compared to cells not expressing membrane-bound IL-15. In some embodiments, expression of membranebound IL- 15 may improve immune cell, such as but not limited to, T cell and / or natural killer cell, persistence, functionality, growth, viability, expansion, or any combination thereof, in a tumor microenvironment, as compared to cells not expressing membrane-bound IL-15. In some embodiments, expression of membrane-bound IL- 15 may increase efficacy of immune cells, such as, but not limited to, T cells and / or natural killer cells, in killing tumor cells, as compared to cells not expressing membrane-bound IL-15. In some embodiments, expression of membrane-bound IL-15 may increase 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, as compared to cells not expressing membranebound IL- 15. In some embodiments, expression of membrane-bound IL- 15 may increase ability of immune cells, such as, but not limited to, T cells and / or natural killer cells, to maintain a naive phenotype.

[0215] Persistence may be assessed, as a non-limiting example, by the length of time cells are detectable in an individual (e.g., patient) after infusion. As non-limiting examples, persistence may be measured at days, weeks, months, or years after infusion, as non-limiting examples, at about 1 week, about 2 weeks, about 4 weeks, about 1 month, about 2 months, about 3 months, about 6 months, about 9 months, about 12 months, about 18 months, about 24 months, and / or about 30 months after infusion. Persistence may be assessed, as nonlimiting examples, by PCR of peripheral blood sample(s), by flow cytometry of peripheral blood samples(s), and / or by analysis of tumor biopsy sample(s). Persistence of cells expressing membrane-bound IL- 15 may be compared, as non-limiting examples, to typicalpersistence of infused ACT cells or persistence of similar cells not expressing membranebound IL- 15.

[0216] Continued ability to kill tumor cells may be measured, as non-limiting examples, via (i) serial killing assays using an IncuCyte (wherein ability to kill / impair tumor growth as measured by fold growth during repeated tumor stimulations over a duration of time is assessed), and / or (ii) via cytokine / effector molecule production (IFNy via ELISAs and other pro-inflammatory cytokines via Luminex (cytokines measured may include, as non-limiting examples, IFNY, TNFa, Granzyme B, perforin, IL-2, IL-6, MIP-ip, MIP-la, GM-CSF, RANTES, IL-18, IL-4, IL-10, and IP10)). Continued ability of cells expressing membranebound IL- 15 to kill tumor cells may be compared, as non-limiting examples, to continued ability of similar cells not expressing membrane-bound IL- 15 to kill tumor cells or continued ability of other control cells to kill tumor cells.

[0217] Naivety of phenotype may be assessed, as a non-limiting example, via Tmem panel assay via flow cytometry. Typically, flow cytometer gating is off of CD8+TCR+ cells. Typically, a more naive phenotype may be indicated by higher frequencies of the T memory subsets Tnaive / scm (CD45RA+CCR7+), and Tcm (CD45RA-CCR7+) and an increase or retention of the CD39-CD69- and CD27+CD28+ populations. Low CD57 expression may also be desirable.

[0218] When assessing the persistence, functionality, growth, viability, expansion, tumor killing efficacy, naivety, or other characteristics of cells expressing dnTGFRpRII, cells such as non-transduced cells, cells transduced with TCR only, cells transduced with CD8 and TCR, or a combination thereof, may serve as control cells, as non-limiting examples.

[0219] In some embodiments, membrane-bound IL- 15 may act in a cis manner (e.g., affecting cells in which it is expressed), in a trans manner (e.g., affecting cells in which it is not expressed), or any combination thereof. In some embodiments in which membrane-bound IL- 15 acts in trans, cells adjacent to or near (e.g., within the tumor microenvironment) cells expressing membrane-bound IL- 15 may exhibit any or combination of improvements the same or similar to those described for cells expressing membrane-bound IL- 15, as compared to cells not adjacent to or near cells expressing membrane-bound IL-15.

[0220] In some embodiments, the disclosure provides for nucleic acid(s) encoding polypeptide(s) described herein. In some embodiments, the disclosure provides for vectors comprising nucleic acids encoding polypeptide(s) described herein. In some embodiments, one or more vector may comprise a nucleic acid encoding an IL-15 / IL-15Ra fusionpolypeptide. In some embodiments, one or more vector may comprise a nucleic acid encoding a CD8 polypeptide. In some embodiments, one or more vector may comprise a nucleic acid encoding a CD8a polypeptide. In some embodiments, one or more vector may comprise a nucleic acid encoding a CD8P polypeptide.

[0221] In some embodiments, one or more vector may comprise one or more nucleic acid encoding a T cell receptor (TCR) comprising an a chain and a P chain. In some embodiments, one or more vector may comprise one or more nucleic acid encoding a T cell receptor (TCR) comprising an y chain and a 5 chain. In some embodiments, one or more vector may comprise one or more nucleic acid encoding a chimeric antigen receptor (CAR).

[0222] In some embodiments, a vector or vectors comprising one or more nucleic acid(s) encoding one or any combination of a TCR comprising an a chain and a P chain, a TCR comprising a y chain and a 5 chain, a CAR, an IL-15 polypeptide, an IL-15Ra polypeptide, an IL-15 / IL-15Ra fusion polypeptide, and / or a CD8 polypeptide may be provided. In some embodiments, a CD8 polypeptide may comprise a CD8a chain and / or a CD8P chain, and the CD8a chain and / or CD8P chain may independently be modified or unmodified.

[0223] In some embodiments, a vector or vectors comprising one or more nucleic acid(s) encoding one or any combination of a TCR comprising an a chain and a P chain, an IL-15 / IL- 15Ra fusion polypeptide, and / or a CD8 polypeptide may be provided. In some embodiments, a vector or vectors comprising one or more nucleic acid(s) encoding one or any combination of a TCR comprising a y chain and a 5 chain, an IL-15 / IL-15Ra fusion polypeptide, and / or a CD8 polypeptide may be provided. In some embodiments, a vector or vectors comprising one or more nucleic acid(s) encoding one or any combination of a CAR, an IL-15 / IL-15Ra fusion polypeptide, and / or a CD8 polypeptide may be provided. In some embodiments, a CD8 polypeptide may comprise a CD8a chain and / or a CD8P chain, and the CD8a chain and / or CD8P chain may independently be modified or unmodified.

[0224] In some embodiments, a vector or vectors comprising one or more nucleic acid(s) encoding a TCR comprising an a chain and a P chain, an IL-15 / IL-15Ra fusion polypeptide, and a CD8 polypeptide may be provided. In some embodiments, a vector or vectors comprising one or more nucleic acid(s) encoding a TCR comprising a y chain and a 5 chain, an IL-15 / IL-15Ra fusion polypeptide, and a CD8 polypeptide may be provided. In some embodiments, a vector or vectors comprising one or more nucleic acid(s) encoding a CAR, an IL-15 / IL-15Ra fusion polypeptide, and a CD8 polypeptide may be provided. In someembodiments, a CD8 polypeptide may comprise a CD8a chain and / or a CD8P chain, and the CD8a chain and / or CD8P chain may independently be modified or unmodified.

[0225] In some embodiments, a vector or vectors comprising one or more nucleic acid(s) encoding a TCR comprising an a chain and a P chain and an IL-15 / IL-15Ra fusion polypeptide may be provided. In some embodiments, a vector or vectors comprising one or more nucleic acid(s) encoding a TCR comprising a y chain and a 5 chain and an IL-15 / IL- 15Ra fusion polypeptide may be provided. In some embodiments, a vector or vectors comprising one or more nucleic acid(s) encoding a CAR and an IL-15 / IL-15Ra fusion polypeptide may be provided.

[0226] In some embodiments, a vector or vectors comprising one or more nucleic acid(s) encoding a TCR comprising an a chain and a P chain and a CD8 polypeptide may be provided. In some embodiments, a vector or vectors comprising one or more nucleic acid(s) encoding a TCR comprising a y chain and a 5 chain and a CD8 polypeptide may be provided. In some embodiments, a cell or cells comprising one or more nucleic acid(s) encoding a CAR and a CD8 polypeptide may be provided. In some embodiments, a CD8 polypeptide may comprise a CD8a chain and / or a CD8P chain, and the CD8a chain and / or CD8P chain may independently be modified or unmodified.

[0227] In some 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 some embodiments, a cell or cells may comprise an aP T cell, a y5 T cell, a natural killer (NK) cell, a natural killer T cell, a CD4+ T cell, CD8+ T cell, a CD4+ / CD8+ cell, or any combination thereof. Specifically, the cell or cells may be a y51 T cell or a y52 T cell.

[0228] In some embodiments, more than one vector may comprise a nucleic acid or nucleic acids encoding one or any combination of an IL- 15 polypeptide, an IL-15Ra polypeptide, an IL-15 / IL-15Ra fusion polypeptide, a CD8 polypeptide, a TCR comprising an a chain and a P chain, a TCR comprising an y chain and a 5 chain, and / or a CAR. In some embodiments, a CD8 polypeptide may comprise a CD8a chain and / or a CD8P chain, and the CD8a chain and / or CD8P chain may independently be modified or unmodified.

[0229] In some embodiments, a single vector may comprise a nucleic acid or nucleic acids encoding one or any combination of an IL- 15 polypeptide, an IL-15Ra polypeptide, an IL-15 / IL-15Ra fusion polypeptide, a CD8 polypeptide, a TCR comprising an a chain and a P chain, a TCR comprising an y chain and a 5 chain, and / or a CAR. In some embodiments, aCD8 polypeptide may comprise a CD8a chain and / or a CD8P chain, and the CD8a chain and / or CD8P chain may independently be modified or unmodified.

[0230] In some embodiments, nucleic acids may be polycistronic, and one or more polycistronic nucleic acids may be utilized. Expression of multiple (e.g., 2, 3, 4, 5, or more) polypeptides from polycistronic nucleic acid may be achieved by any suitable method, such as i) pre-mRNA splicing; ii) proteolytic cleavage sites; iii) fusion proteins; iv) inclusion of one or more 2A peptide-encoding nucleic acid(s) (such as, but not limited to P2A, T2A, E2A, and F2A), v) inclusion of one or more internal ribosome entry site (IRES), or other mechanisms, as well. Each of these approaches has some advantages and disadvantages to provide multiple transcription units. Among the five approaches, the most widely used are the self-cleaving 2A peptides and IRESs. In some embodiments, nucleic acids may be monocistronic, and one or more monocistronic nucleic acid(s) may be utilized.

[0231] In some embodiments, a 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).

[0232] In some embodiments, an IRES may be selected from the group consisting of IRES from picomavirus, IRES from flavivirus, IRES from pestivirus, IRES from retrovirus, IRES from lentivirus, IRES from insect RNA virus, and IRES from cellular mRNA.

[0233] In some embodiments, a vector may comprise nucleic acid encoding a 2A peptide or an internal ribosome entry site (IRES) positioned between a nucleic acid encoding a modified CD8a polypeptide and a nucleic acid encoding a CD8P polypeptide.

[0234] In some embodiments, a vector may comprise nucleic acid encoding a 2A peptide positioned between a nucleic acid encoding a TCR a chain and a nucleic acid encoding a TCR P chain.

[0235] In some embodiments, a vector may comprise nucleic acid encoding a 2A peptide or an internal ribosome entry site (IRES) positioned between a nucleic acid encoding a modified CD8a polypeptide, a nucleic acid encoding a CD8P polypeptide, a nucleic acid encoding a TCR a chain, or a nucleic acid encoding a TCR P chain and a nucleic acid encoding a membrane-bound IL-15.

[0236] In some embodiments, a single vector may comprise a nucleic acid or nucleic acids encoding one or any combination of an IL- 15 polypeptide, an IL-15Ra polypeptide, an IL-15 / IL-15Ra fusion polypeptide, a CD8 polypeptide, a TCR comprising an a chain and a P chain, a TCR comprising an y chain and a 5 chain, and / or a CAR, and a vector may comprise a nucleic acid encoding a 2A peptide or an internal ribosome entry site (IRES) positionedbetween any or each of nucleic acids encoding polypeptides or fusion polypeptides. In some embodiments, a CD8 polypeptide may comprise a CD8a chain and / or a CD8P chain, and the CD8a chain and / or CD8P chain may independently be modified or unmodified.

[0237] In some embodiments, a vector may further comprise a post-transcriptional regulatory element (PRE) sequence. In some embodiments, the post-transcriptional regulatory element (PRE) sequence may be selected from a 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 a mutated WPRE, such as but not limited to WPREmutl (SEQ ID NO: 256) or WPREmut2 (SEQ ID NO: 257)) or a hepatitis B virus (HBV) PRE (HPRE) (SEQ ID NO: 437), variant(s) thereof, or any combination thereof.

[0238] In some embodiments, a vector may further comprise one or more promoter. In some embodiments, promoter(s) may be selected from cytomegalovirus (CMV) promoter, phosphoglycerate kinase (PGK) promoter, myelin basic protein (MBP) promoter, glial fibrillary acidic protein (GFAP) promoter, modified MoMuLV LTR comprising myeloproliferative sarcoma virus enhancer (MNDU3), Ubiqitin C promoter, EF-1 alpha promoter, Murine Stem Cell Virus (MSCV) promoter, the promoter from CD69, nuclear factor of activated T-cells (NF AT) promoter, IL-2 promoter, minimal IL-2 promoter, or a combination thereof.

[0239] In some embodiments, a vector may be a viral vector or a non-viral vector.

[0240] In some embodiments, a vector may be selected from adenoviruses, poxviruses, alphaviruses, arenaviruses, flaviviruses, rhabdoviruses, retroviruses, lentiviruses, herpesviruses, paramyxoviruses, picornaviruses, or a combination thereof.

[0241] In some embodiments, a vector may be pseudotyped with an envelope protein of a virus selected from the native feline endogenous virus (RD114), a chimeric version of RD 114 (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 retroviral envelope glycoprotein (BaEV), lymphocytic choriomeningitis virus (LCMV), or a combination thereof.

[0242] In some embodiments, a vector may comprise one or more Kozak sequence. In some embodiments, a Kozak sequence may initiate, increase, or facilitate translation, or a combination thereof. In some embodiments, the Kozak sequence may be GCCACC. In some embodiments, the Kozak sequence may be ACCATGG. In some embodiments, the Kozak sequence may be GCCNCCATGG. where N is a purine (A or G) (SEQ ID NO:382).

[0243] In some embodiments, a vector may comprise one or more Factor Xa sites.

[0244] In some embodiments, a vector may comprise one or more enhancer. In some embodiments, an enhancer may comprise Conserved Non-Coding Sequence (CNS) 0, CNS 1, CNS2, CNS 3, CNS 4, or portions or any combination thereof.

[0245] In some embodiments, the disclosure provides for one or more cells transduced with and / or expressing one or more vectors comprising nucleic acids encoding polypeptide(s).

[0246] In some embodiments, a cell may comprise an aP T cell, a y5 T cell, a natural killer cell, a natural killer T cell, a CD4+ T cell, CD8+ T cell, a CD4+ / CD8+ cell, or any combination thereof.

[0247] In some embodiments, a T cell may be a CD4+ T cell. In some embodiments, a T cell may be a CD8+ T cell. In some embodiments, a T cell may be a CD4+ / CD8+ T cell. In some embodiments, a T cell may be a aP T cell. In some embodiments, a T cell may be a y5 T cell.

[0248] In some embodiments, a T cell may be an aP T cell and may express a CD8 polypeptide described herein. In some embodiments, a CD8 polypeptide may comprise a CD8a chain and / or a CD8P chain, and the CD8a chain and / or CD8P chain may independently be modified or unmodified. In some embodiments, a T cell may be an aP T cell and may express a modified CD8 polypeptide described herein, for example, a modified CD8a polypeptide or a modified CD8a polypeptide with a CD8P stalk region, e.g., mlCD8a in Constructs #11 and #12 (FIG. 4) and CD8a* (FIG. 55B). In some embodiments, a T cell may be an aP T cell and may express one or any combination of an IL- 15 polypeptide, an IL- 15Ra polypeptide, an IL-15 / IL-15Ra fusion polypeptide, a modified CD8 polypeptide, and / or a CAR.

[0249] In some embodiments, a T cell may be a y5 T cell and may express a CD8 polypeptide described herein. In some embodiments, a CD8 polypeptide may comprise a CD8a chain and / or a CD8P chain, and the CD8a chain and / or CD8P chain may independently be modified or unmodified. In some embodiments, a T cell may be a y5 T cell and may express a modified CD8 polypeptide described herein, for example, a modified CD8a polypeptide or a modified CD8a polypeptide with a CD8P stalk region, e.g., mlCD8a in Constructs #11 and #12 (FIG. 4) and CD8a* (FIG. 55B). In some embodiments, a T cell may be a y5 T cell and may express one or any combination of an IL- 15 polypeptide, an IL- 15Ra polypeptide, an IL-15 / IL-15Ra fusion polypeptide, a CD8 polypeptide, and / or a CAR. In some embodiments, a CD8 polypeptide may comprise a CD8a chain and / or a CD8P chain, and the CD8a chain and / or CD8P chain may independently be modified or unmodified.

[0250] In some embodiments, a cell or cells comprising, or comprising one or more nucleic acid(s) encoding, one or any combination of a TCR comprising an a chain and a P chain, a TCR comprising a y chain and a 5 chain, a CAR, an IL- 15 polypeptide, an IL-15Ra polypeptide, an IL-15 / IL-15Ra fusion polypeptide, and / or a CD8 polypeptide may be provided. In some embodiments, a CD8 polypeptide may comprise a CD8a chain and / or a CD8P chain, and the CD8a chain and / or CD8P chain may independently be modified or unmodified.

[0251] In some embodiments, a cell or cells comprising, or comprising one or more nucleic acid(s) encoding, one or any combination of a TCR comprising an a chain and a P chain, an IL-15 / IL-15Ra fusion polypeptide, and / or a CD8 polypeptide may be provided. In some embodiments, a cell or cells comprising, or comprising one or more nucleic acid(s) encoding, one or any combination of a TCR comprising a y chain and a 5 chain, an IL-15 / IL- 15Ra fusion polypeptide, and / or a CD8 polypeptide may be provided. In some embodiments, a cell or cells comprising, or comprising one or more nucleic acid(s) encoding, one or any combination of a CAR, an IL-15 / IL-15Ra fusion polypeptide, and / or a CD8 polypeptide may be provided. In some embodiments, a CD8 polypeptide may comprise a CD8a chain and / or a CD8P chain, and the CD8a chain and / or CD8P chain may independently be modified or unmodified.

[0252] In some embodiments, a cell or cells comprising, or comprising one or more nucleic acid(s) encoding, a TCR comprising an a chain and a P chain, an IL-15 / IL-15Ra fusion polypeptide, and a CD8 polypeptide may be provided. In some embodiments, a cell or cells comprising, or comprising one or more nucleic acid(s) encoding, a TCR comprising a y chain and a 5 chain, an IL-15 / IL-15Ra fusion polypeptide, and a CD8 polypeptide may be provided. In some embodiments, a cell or cells comprising, or comprising one or more nucleic acid(s) encoding, a CAR, an IL-15 / IL-15Ra fusion polypeptide, and a CD8 polypeptide may be provided. In some embodiments, a CD8 polypeptide may comprise a CD8a chain and / or a CD8P chain, and the CD8a chain and / or CD8P chain may independently be modified or unmodified.

[0253] In some embodiments, a cell or cells comprising, or comprising one or more nucleic acid(s) encoding, a TCR comprising an a chain and a P chain and an IL-15 / IL-15Ra fusion polypeptide may be provided. In some embodiments, a cell or cells comprising, or comprising one or more nucleic acid(s) encoding, a TCR comprising a y chain and a 5 chain and an IL-15 / IL-15Ra fusion polypeptide may be provided. In some embodiments, a cell orcells comprising, or comprising one or more nucleic acid(s) encoding, a CAR and an IL- 15 / IL-15Ra fusion polypeptide may be provided.

[0254] In some embodiments, a cell or cells comprising, or comprising one or more nucleic acid(s) encoding, a TCR comprising an a chain and a P chain and a CD8 polypeptide may be provided. In some embodiments, a cell or cells comprising, or comprising one or more nucleic acid(s) encoding, a TCR comprising a y chain and a 5 chain and a CD8 polypeptide may be provided. In some embodiments, a cell or cells comprising, or comprising one or more nucleic acid(s) encoding, a CAR and a CD8 polypeptide may be provided. In some embodiments, a CD8 polypeptide may comprise a CD8a chain and / or a CD8P chain, and the CD8a chain and / or CD8P chain may independently be modified or unmodified.

[0255] In some embodiments, one or more nucleic acid(s) may be comprised in and / or expressed from a vector or vectors.

[0256] In some embodiments, a cell or cells may comprise an aP T cell, a y5 T cell, a natural killer cell, a CD4+ T cell, CD8+ T cell, a CD4+ / CD8+ cell, or any combination thereof.

[0257] In some embodiments, populations of cells as described herein may be provided. As a non-limiting example, the disclosure provides for a population of modified cells comprising, or comprising one or more nucleic acid(s) encoding one or any combination of an exogenous CD8 co-receptor comprising a polypeptide described herein, for example, amino acid sequences at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 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-15Ra fusion polypeptide), as described herein; and / or a T cell receptor. In some embodiments, populations of cells may comprise aP T cells, y5 T cells, natural killer cells, a natural killer T cell, CD4+ T cells, CD8+ T cells, CD4+ / CD8+ cells, or any combination thereof.

[0258] In an aspect, polypeptide sequences and / or nucleic acid sequences described herein may be isolated and / or recombinant sequences.

[0259] In an aspect, cells described herein may be isolated and / or recombinant cells.

[0260] In some embodiments, a method of preparing cells for immunotherapy may comprise isolating or enriching cells from a blood sample of a human subject, activating theisolated cells, transducing the activated cells with one or more vector, and expanding the transduced cells. In some embodiments, a cell may comprise an aP T cell, a y5 T cell, a natural killer cell, a natural killer T cell, a CD4+ T cell, CD8+ T cell, a CD4+ / CD8+ cell, or any combination thereof.

[0261] In some embodiments, a method of treating a patient who has cancer may comprise administering to the patient a composition comprising the population of expanded cells, wherein the cells kill cancer cells that present a peptide in a complex 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, urinary bladder cancer, kidney cancer, leukemia, ovarian cancer, esophageal cancer, brain cancer, gastric cancer, prostate cancer, or a combination thereof. In some embodiments, a cell may comprise an aP T cell, a y5 T cell, a natural killer cell, a natural killer T cell, a CD4+ T cell, CD8+ T cell, a CD4+ / CD8+ cell, or any combination thereof.

[0262] In some embodiments, the composition may further comprise an adjuvant. In some embodiments, the adjuvant may be selected from anti-CD40 antibody, imiquimod, resiquimod, GM-CSF, cyclophosphamide, sunitinib, bevacizumab, atezolizumab, interferonalpha, interferon-beta, CpG oligonucleotides and derivatives, poly(I:C) and derivatives, RNA, sildenafil, particulate formulations with poly(lactide co-glycolide) (PLG), virosomes, interleukin (IL)-l, IL-2, IL-4, IL-7, IL-12, IL-13, IL-15, IL-21, IL-23, or any combination thereof.

[0263] In some embodiments, a method of eliciting an immune response in a patient who has cancer may comprise administering to the patient a composition comprising the population of expanded cells, wherein the cells kill cancer cells that present a peptide in a complex 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, urinary bladder cancer, kidney cancer, leukemia, ovarian cancer, esophageal cancer, brain cancer, gastric cancer, prostate cancer, or a combination thereof. In some embodiments, a cell may comprise an aP T cell, a y5 T cell, a natural killer cell, a natural killer T cell, a CD4+ T cell, CD8+ T cell, a CD4+ / CD8+ cell, or any combination thereof. Specifically, the cell may comprise a y51 T cell and / or a y52 T cell.BRIEF DESCRIPTION OF THE DRAWINGS

[0264] FIG. 1 shows a representative CD8a subunit, e.g., SEQ ID NO: 258 (CD8al) . CD8al includes five domains: (1) signal peptide, (2) Ig-like domain-1, (3) a stalk region, (4) transmembrane (TM) domain, and (5) a cytoplasmic tail (Cyto) comprising a / c#-binding motif.

[0265] FIG. 2 shows a sequence alignment between CD8al (SEQ ID NO: 258) and mlCD8a (SEQ ID NO: 7).

[0266] FIG. 3 shows a sequence alignment between CD8a2 (SEQ ID NO: 259) and m2CD8a (SEQ ID NO: 262), in which the cysteine substitution at position 112 is indicated by an arrow.

[0267] FIG. 4 shows exemplary vectors according to an aspect of the disclosure. In some embodiments, vectors may also comprise additional elements, such as those described herein, such as, but not limited to one or more promoter or one or more post-transcriptional regulatory element. In FIG. 4, the lines may represent direct linkages, with no intervening sequences, or may represent intervening sequences, such as, but not limited to, a linker, a furin, a sequence encoding a 2A polypeptide, a factor Xa site, an untranslated sequence, a translated sequence, a sequence comprising one or more restriction endonuclease sites, or a combination thereof.

[0268] FIG. 5 A shows titers of viral vectors shown in FIG. 4.

[0269] FIG. 5B shows titers of further viral vectors in accordance with the present disclosure. Construct #13; Construct #14; Construct #15; Construct #16; Construct #17; Construct #18; Construct #19; Construct #21; Construct #10n; Construct #1 In; and TCR: R1 IKEA (SEQ ID NO: 15 and SEQ ID NO: 16, which may be encoded by SEQ ID NO: 72 and SEQ ID NO: 73, respectively) (Construct #8), which binds PRAME-004 (SLLQHLIGL) (SEQ ID NO: 147). Note that Constructs #10 and #10n are different batches of the same construct (SEQ ID NO: 291 and 292) and Constructs #11 and #1 In are different batches of the same construct (SEQ ID NO: 285 and 286).

[0270] FIG. 6 shows T cell manufacturing.

[0271] FIG. 7A shows expression of activation markers before and after activation in CD3+CD8+ cells.

[0272] FIG. 7B shows expression of activation markers before and after activation in CD3+CD4+ cells.

[0273] FIG. 8A shows fold expansion of cells transduced with various constructs from Donor #1. The constructs are as follows: Construct #9b; Construct #10; Construct #11; Construct #12; Construct #1; Construct #2; TCR = R1 IKEA. WPRE"1(TCR with wild type WPRE); NT = Non-transduced T cells (as a negative control). Note that Constructs #9 and #9b are different batches of the same construct (SEQ ID NO: 287 and 288).

[0274] FIG. 8B shows fold expansion of cells transduced with various constructs from Donor #2. The constructs are as follows: Construct #9b; Construct #10; Construct #11; Construct #12; Construct #1; Construct #2; TCR = R1 IKEA. WPRE"1(TCR with wild type WPRE) (Construct #8); NT = Non-transduced T cells (as a negative control).

[0275] FIG. 9A shows flow plots of cells transduced with Construct #9 .

[0276] FIG. 9B shows flow plots of cells transduced with Construct #10 in accordance with one embodiment of the present disclosure.

[0277] FIG. 9C shows flow plots of cells transduced with Construct #11.

[0278] FIG. 9D shows flow plots of cells transduced with Construct #12.

[0279] FIG. 10 shows % CD8+CD4+ of cells transduced with various constructs for Donor #1 and Donor #2. The constructs are as follows: Construct #9b; Construct #10; Construct #11; Construct #12; Construct #1; Construct #2; TCR = R11KEA.WPRE"1TCR with wild type WPRE); NT = Non-transduced T cells (as a negative control).

[0280] FIG. 11 shows % Tet of CD8+CD4+ of cells transduced with various constructs. The constructs are as follows: Construct #9b; Construct #10; Construct #11; Construct #12; Construct #1; Construct #2; TCR = R11KEA.WPRE"1(TCR with wild type WPRE); NT = Non-transduced T cells (as a negative control).

[0281] FIG. 12 shows Tet MFI (CD8+CD4+Tet+) of cells transduced with various constructs. The constructs are as follows: Construct #9b; Construct #10; Construct #11; Construct #12; Construct #1; Construct #2; TCR = R1 IKEA. WPRE"1(TCR with wild type WPRE); NT = Non-transduced T cells (as a negative control).

[0282] FIG. 13 shows CD8a MFI (CD8+CD4+Tet+) of cells transduced with various constructs. The constructs are as follows: Construct #9b; Construct #10; Construct #11; Construct #12; Construct #1; Construct #2; TCR = R1 IKEA. WPRE"1(TCR with wild type WPRE); NT = Non-transduced T cells (as a negative control).

[0283] FIG. 14 shows % CD8+CD4 (of CD3+) of cells transduced with various constructs. The constructs are as follows: Construct #9b; Construct #10; Construct #11; Construct #12; Construct #1; Construct #2; TCR = R1 IKEA. WPRE"1(TCR with wild type WPRE); NT = Non-transduced T cells (as a negative control).

[0284] FIG. 15 shows % CD8+Tet+ (of CD3+) of cells transduced with various constructs. The constructs are as follows: Construct #9b; Construct #10; Construct #11; Construct #12; Construct #1; Construct #2; TCR = R1 IKEA. WPRE"1TCR with wild type WPRE); NT = Non-transduced T cells (as a negative control).

[0285] FIG. 16 shows Tet MFI (CD8+Tet+) of cells transduced with various constructs. The constructs are as follows: Construct #9b; Construct #10; Construct #11; Construct #12; Construct #1; Construct #2; TCR = R1 I KEA.WPRE"1(TCR with wild type WPRE); NT = Non-transduced T cells (as a negative control).

[0286] FIG. 17 shows CD8a MFI (CD8+Tet+) of cells transduced with various constructs. The constructs are as follows: Construct #9b; Construct #10; Construct #11; Construct #12; Construct #1; Construct #2; TCR = R1 IKEA. WPRE"1(TCR with wild type WPRE); NT = Non-transduced T cells (as a negative control).

[0287] FIG. 18 shows % Tet+ (of CD3+) of cells transduced with various constructs. The constructs are as follows: Construct #9b; Construct #10; Construct #11; Construct #12; Construct #1; Construct #2; TCR = R11KEA.WPRE"1(TCR with wild type WPRE); NT = Non-transduced T cells (as a negative control).

[0288] FIG. 19 shows VCN (upper panel) and CD3+Tet+ / VCN (lower panel) of cells transduced with various constructs. The constructs are as follows: Construct #9b; Construct #10; Construct #11; Construct #12; Construct #1; Construct #2; TCR = R11KEA.WPRE"1(TCR with wild type WPRE); NT = Non-transduced T cells (as a negative control).

[0289] FIG. 20A-20C depicts data showing that constructs (#10, #11, & #12) are comparable to TCR-only in mediating cytotoxicity against target positive cells lines expressing antigen at different levels (UACC257 at 1081 copies per cell and A375 at 50 copies per cell).

[0290] FIG. 21 A-21B depict data showing that IFNy secretion in response to UACC257 is comparable among constructs, however with A375, #10 expressing is the highest among all constructs. However, comparing #9 with #11 expressing wild type and modified CD8 coreceptor sequences respectively, T cells transduced with #11 induced stronger cytokine response measured as IFNY quantified in the supernatants from Incucyte plates. Construct #9; Construct #10; Construct #11; Construct #12; Construct #1; Construct #2; Construct #8 = R1 IKEA TCR only.

[0291] FIG. 22 depicts an exemplary experiment design to assess DC maturation and cytokine secretion by PBMC-derived product in response to UACC257 and A375 targets. N=2.

[0292] FIG. 23A-23B depicts data showing that the IFNy secretion in response to A375 increases in the presence of iDCs. In the tri-cocultures with iDCs, IFNY secretion is higher in Construct #10 compared to the other constructs. However, comparing Construct #9 with Construct #11 expressing wild type and modified CD8 coreceptor sequences respectively, T cells transduced with #11 induced stronger cytokine response measured as IFNy quantified in the culture supernatants of three-way cocultures using donor D600115, E:T:iDC:: l : l / 10: l / 4. Construct #9; Construct #10; Construct #11; Construct #12; Construct #1; Construct #2; Construct #8 = R1 IKEA TCR only.

[0293] FIG. 24A-24B depicts data showing that IFNY secretion in response to A375 increases in the presence of iDCs. In the tri-cocultures with iDCs, IFNY secretion was higher in Construct #10 compared to the other constructs. IFNy quantified in the culture supernatants of three-way cocultures using donor D150081, E:T:iDC:: l : l / 10: l / 4. Construct #9; Construct #10; Construct #11; Construct #12; Construct #1; Construct #2; Construct #8 = R1 IKEA TCR only.

[0294] FIG. 25A-25B depicts data showing that IFNY secretion in response to UACC257 increases in the presence of iDCs. In the tri-cocultures with iDCs, IFNY secretion is higher in Construct #10 compared to the other constructs. However, comparing Construct #9 with Construct #11 expressing wild type and modified CD8 coreceptor sequences respectively, T cells transduced with Construct #11 induced stronger cytokine response measured as IFNy quantified in the culture supernatants of three-way cocultures using donor D600115, E:T:iDC:: l : l / 10: l / 4. Construct #9; Construct #10; Construct #l l; Construct #12; Construct #1; Construct #2; Construct #8 = R1 IKEA TCR only.

[0295] FIG. 26 shows T cell manufacturing in accordance with one embodiment of the present disclosure.

[0296] FIG. 27A shows expression of activation markers before and after activation in CD3+CD8+ cells.

[0297] FIG. 27B shows expression of activation markers before and after activation in CD3+CD4+ cells in accordance with one embodiment of the present disclosure.

[0298] FIG. 28 shows fold expansion of cells transduced with various constructs.

[0299] FIG. 29A & 29B show % CD8+CD4+ of cells transduced with various constructs in accordance with one embodiment of the present disclosure.

[0300] FIG. 30A & 30B show % Tet of CD8+CD4+ of cells transduced with various constructs in accordance with one embodiment of the present disclosure.

[0301] FIG. 31 A & 3 IB show Tet MFI (CD8+CD4+Tet+) of cells transduced with various constructs in accordance with one embodiment of the present disclosure.

[0302] FIG. 32A & 32B show % CD8+CD4- (of CD3+) of cells transduced with various constructs in accordance with one embodiment of the present disclosure.

[0303] FIG. 33A & 33B show % CD8+Tet+ (of CD3+) of cells transduced with various constructs in accordance with one embodiment of the present disclosure.

[0304] FIG. 34A & 34B show Tet MFI (CD8+Tet+) of cells transduced with various constructs in accordance with one embodiment of the present disclosure.

[0305] FIG. 35A & 35B show % Tet+ (of CD3+) of cells transduced with various constructs in accordance with one embodiment of the present disclosure.

[0306] FIG. 36A & 36B show VCN of cells transduced with various constructs in accordance with one embodiment of the present disclosure.

[0307] FIG. 37 shows T cell manufacturing in accordance with one embodiment of the present disclosure.

[0308] FIG. 38 shows % Tet of CD8+CD4+ of cells transduced with various constructs.

[0309] FIG. 39 shows Tet MFI of CD8+CD4+Tet+ of cells transduced with various constructs.

[0310] FIG. 40 shows Tet MFI of CD8+Tet+ of cells transduced with various constructs.

[0311] FIG. 41 shows % Tet+ of CD3+ cells transduced with various constructs.

[0312] FIG. 42 shows vector copy number (VCN) of cells transduced with various constructs.

[0313] FIG. 43 shows the % T cell subsets in cells transduced with various constructs. FACS analysis was gated on CD3+TCR+.

[0314] FIG. 44A and FIG. 44B shows % T cell subsets in cells transduced with various constructs. FACS analysis was gated on CD4+CD8+ for FIG. 44A and on CD4-CD8+TCR+ f or FIG. 44B.

[0315] FIG. 45 A and 45B depicts data showing that Constructs #13 and #10 are comparable to TCR-only in mediating cytotoxicity against UACC257 target positive cells lines expressing high levels of antigen (1081 copies per cell). Construct # 15 was also effective but slower in killing compared to Constructs #13 and #10. The effectortarget ratio used to generate these results was 4: 1.

[0316] FIG. 46 shows IFNy secretion in response in UACC257 cell line was higher with Construct #13 compared to Construct #10. IFNy quantified in the supernatants from Incucyte plates. The effectortarget ratio used to generate these results was 4: 1.

[0101] FIG. 47 shows ICI marker frequency (2B4, 41BB, LAG3, PD-1, TIGIT, TIM3, CD39+CD69+, and CD39-CD69-).

[0317] FIG. 48 A - 48G show increased expression of IFNy, IL-2, and TNFa with CD4+CD8+ cells transduced with Construct #10 (WT signal peptide, CD8pi) compared to other constructs. FACS analysis was gated on CD3+CD4+CD8+ cells against UACC257, 4: 1 E:T.

[0318] FIG. 49A-49G show increased expression of IFNy, IL-2, MIP-ip, and TNFa with CD4-CD8+ cells transduced with Construct #10 (WT signal peptide, CD8pi) compared to other constructs. FACS analysis was gated on CD3+CD4-CD8+ cells against UACC257, 4: 1 E:T.

[0319] FIG. 50A-50G show increased expression of IL-2 and TNFa with CD3+TCR+ cells transduced with Construct #10 (WT signal peptide, CD8pi) compared to other constructs. FACS analysis was gated on CD3+TCR+ cells against UACC257, 4: 1 E:T.

[0320] FIG. 51A-51C show results from FACS analysis gated on CD4+CD8+ cells against A375, 4: 1 E:T.

[0321] FIG. 52A-52C show results from FACS analysis gated on CD4-CD8+ cells against A375, 4: 1 E:T.

[0322] FIG. 53A-53C show results from FACS analysis gated on CD3+TCR+ cells against A375, 4: 1 E:T.

[0323] FIG. 54 shows T cell manufacturing in accordance with one embodiment of the present disclosure.

[0324] FIG. 55A-55C show interaction between peptide / MHC complex of antigen- presenting cell (APC) with T cell by binding a complex of TCR and CD8aP heterodimer (FIG. 55A, e.g., produced by transducing T cells with Constructs #2, #3, #4, #10, #13, #14, #15, #16, #17, #18, or #21), a complex of TCR and homodimer CD8a having its stalk region replaced with CD8P stalk region (CD8aa*) (FIG. 55B, e.g., produced by transducing T cells with Construct #11, #12, or #19), and a complex of TCR and CD8a homodimer (FIG. 55C, e.g., produced by transducing T cells with Constructs #1, #5, #6, #7, or #9).

[0325] FIG. 56 shows the levels of IL-12 secretion by dendritic cells (DC) in the presence of CD4+ T cells transduced with Construct #10 or #11 and immature dendritic cells (iDCs) in accordance with one embodiment of the present disclosure.

[0326] FIG. 57 shows the levels of TNF-a secretion by dendritic cells (DC) in the presence of CD4+ T cells transduced with Construct #10 or #11 and immature dendritic cells (iDCs) in accordance with one embodiment of the present disclosure.

[0327] FIG. 58 shows the levels of IL-6 secretion by dendritic cells (DC) in the presence of CD4+ T cells transduced with Construct #10 or #11 and immature dendritic cells (iDCs) in accordance with one embodiment of the present disclosure.

[0328] FIG. 59 shows a scheme of determining the levels of cytokine secretion by dendritic cells (DC) in the presence of PBMCs transduced with various constructs and target cells in accordance with one embodiment of the present disclosure.

[0329] FIG. 60 shows the levels of IL- 12 secretion by dendritic cells (DC) in the presence of PBMCs transduced with various constructs and target cells in accordance with one embodiment of the present disclosure.

[0330] FIG. 61 shows the levels of TNF-a secretion by dendritic cells (DC) in the presence of PBMCs transduced with various constructs and target cells in accordance with one embodiment of the present disclosure

[0331] FIG. 62 shows the levels of IL-6 secretion by dendritic cells (DC) in the presence of PBMCs transduced with various constructs and target cells in accordance with one embodiment of the present disclosure.

[0332] FIG. 63A-63C show IFNy production from the transduced CD4+ selected T cells obtained from Donor #1 (FIG. 63 A), Donor #2 (FIG. 63B), and Donor #3 (FIG. 63C) in accordance to one embodiment of the present disclosure.

[0333] FIG. 63D shows EC50 values (ng / ml) in FIG. 63A-63C.

[0334] FIG. 64A-64C show IFNy production from the transduced PBMC obtained from Donor #4 (FIG. 64 A), Donor #1 (FIG. 64B), and Donor #3 (FIG. 64C) and their respective EC50 values (ng / ml) in accordance to one embodiment of the present disclosure.

[0335] FIG. 64D shows comparison of EC50 values (ng / ml) among different donors in FIG. 64A-64C.

[0336] FIG. 65A-65C show IFNy production from the transduced PBMC (FIG. 65A), CD8+ selected T cells (FIG. 65B), and CD4+ selected T cells (FIG. 65C) and their respective EC50 values (ng / ml) from a single donor in accordance to one embodiment of the present disclosure.

[0337] FIG. 66 schematically depicts an exemplary membrane-bound IL- 15 bound to the membrane of a T cell, which may be provided In some embodiments. In some embodiments membrane-bound IL-15 may signal via an intermediate IL-2 / IL-15 receptor, as a non-limiting example.

[0338] FIG. 67 A depicts an exemplary membrane-bound IL- 15 polypeptide in which an IL-15 polypeptide is located N-terminal to an IL-15Ra polypeptide, which may be providedIn some embodiments. FIG. 67B depicts an exemplary membrane-bound IL-15 polypeptide in which an IL-15 polypeptide is located C-terminal to an IL-15Ra polypeptide, which may be provided In some embodiments. Nucleic acids encoding such constructs are also provided, in embodiment. In FIG. 67A and 67B, L represents one or more optional linker, and the lines may represent direct linkages, with no intervening sequences, or may represent intervening sequences, such as, but not limited to, a linker, an untranslated sequence (in the case of a nucleic acid sequence), a translated sequence, a sequence comprising one or more restriction endonuclease sites (in the case of a nucleic acid sequence), or a combination thereof.

[0339] FIG. 68 A depicts an exemplary membrane-bound IL- 15 polypeptide in which an IL-15 polypeptide is located N-terminal to an IL-15Ra polypeptide, and a signal peptide (SP) is located N terminal to the IL- 15 polypeptide, which may be provided In some embodiments. FIG. 68B depicts an exemplary membrane-bound IL- 15 polypeptide in which an IL- 15 polypeptide is located C-terminal to an IL-15Ra polypeptide, and a signal peptide (SP) is located N terminal to the IL-15Ra polypeptide, which may be provided In some embodiments. Nucleic acids encoding such constructs are also provided, In some embodiments. In FIG. 68A and 68B, L represents one or more optional linker, and the lines may represent direct linkages, with no intervening sequences, or may represent intervening sequences, such as, but not limited to, a linker, an untranslated sequence (in the case of a nucleic acid sequence), a translated sequence, a sequence comprising one or more restriction endonuclease sites (in the case of a nucleic acid sequence), or a combination thereof.

[0340] FIG. 69A depicts exemplary polypeptide constructs, which may be provided In some embodiments. FIG. 69B depicts exemplary nucleic acid constructs, which may be provided In some embodiments. In FIG. 69A and 69B, the lines may represent direct linkages, with no intervening sequences, or may represent intervening sequences, such as, but not limited to, a linker, an untranslated sequence (in the case of a nucleic acid sequence), a translated sequence, a sequence comprising one or more restriction endonuclease sites (in the case of a nucleic acid sequence), or a combination thereof.

[0341] FIG. 70 depicts exemplary vector constructs, which may be provided In some embodiments. In some embodiments, vectors may also comprise additional elements, such as those described herein, such as, but not limited to one or more promoter or one or more post- transcriptional regulatory element. In FIG. 70, the lines may represent direct linkages, with no intervening sequences, or may represent intervening sequences, such as, but not limited to, a linker, a furin, a sequence encoding a 2A polypeptide, a factor Xa site, an untranslatedsequence, a translated sequence, a sequence comprising one or more restriction endonuclease sites, or a combination thereof.

[0342] FIG. 71A-D shows %TCR+ (A), % IL15Ra+TCR+(B), fold expansion (C) and cell viability (D) of different vector constructs co-expressing TCR and mbIL15 (Alt vl-v4, v7) compared to TCR only (“TCR”) and non-transduced cells (“NT”) as control in an exemplary assay. The flow cytometer was gated on CD8+. Data are grouped (n=2) and represented as mean.

[0343] FIG. 72A-D shows %TCR+ (A), % IL15Ra+TCR+(B), fold expansion (C) and total TCR+ cells (D) of different vector constructs co-expressing TCR and mbIL15 (Alt vl- v4, v7) compared to TCR only (“TCR”) and non-transduced cells (“NT”) as control in an exemplary assay. The flow cytometer was gated off CD8+. Data are grouped (n=4) and represented as mean.

[0344] FIG. 73 A-C shows exemplary kinetic killing (A), tumor growth indices (B) and IFNy release (C) in a co-culture assay of UACC257 tumor cells expressing red fluorescent protein (RFP) (“UACC257-RFP”) with non-transduced (“NT”) cells, cells transduced with TCR only (“TCR”) or cells transduced with various constructs co-expressing TCR and mbIL15 (Alt vl-v4, v7). Cells were challenged with UACC257 cells four times over 9-10 days at about 0 hours, about 70 hours, about 120 hours, and about 170 hours, at an effectortarget ratio of 1 : 1. UACC257-RFP cells alone were assayed as a control. The data are grouped (n=4), represented as mean, and TCR+ normalized. Data were gathered using IncuCyte and ELISA.

[0345] FIG. 74A-C shows exemplary kinetic killing (A), tumor growth indices (B) and IFNy release (C) in a co-culture assay of hs695T tumor cells expressing red fluorescent protein (RFP) (“hs695T-RFP”) with non-transduced (“NT”) cells, cells transduced with TCR only (“TCR”) or cells transduced with various constructs co-expressing TCR and mbIL15 (Alt vl-v4, v7). Cells were challenged with hs695T cells four times over 9-10 days at about 0 hours, about 70 hours, about 120 hours, and about 170 hours at an effectortarget ratio of 2: 1. hs695T-RFP cells alone were assayed as a control. The data are grouped (n=4), represented as mean, and TCR+ normalized. Data were gathered using IncuCyte and ELISA.

[0346] FIG. 75 A-C shows exemplary kinetic killing (A), tumor growth indices (B) and IFNy release (C) in a co-culture assay of A375 tumor cells expressing red fluorescent protein (RFP) (“A375 -RFP”) with non-transduced (“NT”) cells, cells transduced with TCR only (“TCR”) or cells transduced with various constructs. Cells were challenged with A375 cells four times over 9-10 days at about 0 hours, about 70 hours, about 120 hours, and about 170hours at an effectortarget ratio of 4: 1. A375-RFP cells alone were assayed as a control. The data are grouped (n=4), represented as mean, and TCR+ normalized. Data were gathered using IncuCyte and ELISA.

[0347] FIG. 76A-D show the percentage of TemRA, Tern, T naive / scm, and Tcm cells, of cells transduced with TCR only (“TCR”) and various constructs in an example. Nontransduced cells (“NT”) were assayed as a control. The panel in FIG. 76 A was performed on cells that were not exposed to antigen-presenting tumor cells (pre-Ag). The panels in FIG. 76B-D were performed after four tumor stimulations with UACC257 cells (B), hs695T cells (C) and A375 cells (D) over 9-10 days. The flow cytometer was gated on CD3+CD8+ cells. Data are represented as mean.

[0348] FIG. 77A-D show the percentage of CD8+ cells that were positive for each of LAG-3, PD-1, TIGIT, TIM-3, CD39, and CD69 prior to (A) or after exposure of the cells to antigen-bearing UACC257 cells (B), hs695T cells (C) or A375 cells (D) in an example. Cells were challenged four times with tumor cells over 9-10 days. Expression percentages are shown by each of non-transduced (“NT”) cells and cells transduced with TCR only (“TCR”), and cells transduced with various constructs. Data are grouped (n=4) and are represented as mean.

[0349] FIG. 78A-F shows flow plots of cells transduced with TCR only (“TCR”) and various constructs in an example. Non-transduced cells (“NT”) were assayed as a control. X- axis shows staining for cell viability markers (Helix NP), Y axis shows staining for apoptosis markers (ApoTracker™). Flow plots were performed on cells after four antigen challenges over 9-10 days with antigen-presenting UACC257 tumor cells. FIG. 78G-H show frequencies of live and dead apoptotic cells, respectively. Cells with similar results were obtained after challenges with hs695T and A375 tumor cells (data not shown). The flow cytometer was gated on CD8+ cells. Data are grouped (n=4) and represented as mean.

[0350] FIG. 79A-C shows proliferation indices of cells transduced with TCR only (“TCR”) and various constructs. Cells were challenged twice with UACC257 tumor cells (A), hs695T tumor cells (B) or A375 tumor cells (C) over 6 days. The flow cytometer was gated on CD8+ cells. Data are grouped (n=4) and represented as mean.

[0351] FIG. 80A-C shows total cell count (A), fold expansion (B) and cell viability (C) of cells transduced with TCR only (“TCR”) and various constructs in the absence or presence of exogenous Interleukin-7 and Interleukin- 15 (“IL7 / 15”) and in the absence of antigen stimulation. Conditions with exogenous Interleukin-7 and Interleukin- 15 (“IL7 / 15”)wereterminated on day 17 while conditions without exogenous Interleukin-7 and Interleukin- 15 (“IL7 / 15”) were in culture up to 31 days. Data are grouped (n=4) and represented as mean.

[0352] FIG. 81A-B shows exemplary kinetic killing of cells transduced with various constructs after 31 days in culture without exogenous cytokine addition or antigen stimulation. To assess the killing capacities, transduced cells were co-cultured with UACC257 tumor cells at an effectortarget ratio of 1 : 1 (A) or hs695T tumor cells at an effectortarget ratio of 2: 1 (B). Data are grouped (n=4), represented as mean, and TCR+ normalized.

[0353] FIG. 82A shows exemplary kinetic killing of T cells transduced with various constructs after 44 days in culture. To assess the killing capacities, transduced cells were cocultured with UACC257 tumor cells at an effectortarget ratio of 3: 1. FIG. 82B shows AUC for TCR & mbIL15 constructs up to 668-hours at which point viable effectors were harvested from the original co-culture plate and transferred to a new plate containing the next tumor stimulation. FIG: 82C shows AUC for mbIL15 constructs for the entire duration of the assay (excludes TCR). Data are grouped (n=3) and represented as mean. FIG. 82D-F shows IFN-y production for various timepoints after stimulation rounds #1, #8 (last stimulation before wells were harvested and transferred to a new plate), and #13, respectively. Data are grouped (n=3) and represented as mean.

[0354] Figure 83 A-C shows pooled tumor growth indices from two studies, with a combined n=9 (6 healthy donors & 3 patients); each symbol corresponds to a different donor / patient. Cells were challenged with 3-4 tumor stimulations over the course of 9-10 days (3 tumor cell lines in total; (A) UACC257, (B) hs695T, & (C) A375).DETAILED DESCRIPTIONMembrane-Bound IL-15

[0355] In some embodiments a membrane-bound IL- 15 polypeptide (membrane-bound IL- 15 or mbIL-15) is provided. In some embodiments nucleic acids described herein comprise and / or encode a membrane-bound IL- 15 polypeptide. In some embodiments vectors described herein comprise and / or encode a membrane-bound IL-15 polypeptide. In some embodiments cells described herein comprise and / or express a membrane-bound IL- 15 polypeptide. In some embodiments compositions described herein comprise a membranebound IL- 15 polypeptide or comprise cells comprising and / or expressing a membrane-bound IL- 15 polypeptide. In some embodiments IL- 15 is rendered membrane-bound by expressing an IL- 15 polypeptide and an IL-15Ra polypeptide in an IL-15 / IL-15Ra fusion polypeptide (IL-15 / IL-15Ra).

[0356] Membrane-bound IL- 15 polypeptides are provided. Isolated nucleic acid sequences comprising one or more nucleic acid sequences encoding one or more membranebound IL-15 polypeptides are provided. Vectors comprising one or more nucleic acid sequences comprising one or more nucleic acid sequences encoding one or more membranebound IL-15 polypeptides are provided. Cells comprising and / or expressing one or more membrane-bound IL-15 polypeptides are provided. Cells comprising or expressing one or more nucleic acid sequences comprising one or more nucleic acid sequences encoding one or more membrane-bound IL- 15 polypeptides are provided. Cells comprising or expressing one or more vectors comprising one or more nucleic acid sequences comprising one or more nucleic acid sequences encoding one or more membrane-bound IL- 15 polypeptides are provided. In some embodiments, cells described herein may comprise a membrane-bound IL- 15 polypeptide, a CD8 polypeptide, a cell receptor (TCR) comprising an a chain and a P chain, a TCR comprising an y chain and a 5 chain, a chimeric antigen receptor (CAR), or any combination thereof. In some embodiments a cell may comprise an aP T cell, an y5 T cell, a natural killer cell, a natural killer T cell, a CD4+ cell, a CD8+ cell, a CD4+ / CD8+ cell, or any combination thereof. In some embodiments 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.

[0357] In an aspect, polypeptide sequences and / or nucleic acid sequences described herein may be isolated and / or recombinant sequences.

[0358] In an aspect, cells described herein may be isolated and / or recombinant cells.

[0359] Membrane-bound IL- 15 may comprise, for example, an IL-15 / IL-15Ra fusion polypeptide and / or an IL-15Ra / IL-15 fusion polypeptide. One or more linkers may be disposed between IL-15 and IL-15Ra or between IL-15Ra and IL-15. In some embodiments an IL-15 polypeptide is located N-terminal to an IL-15Ra polypeptide in a membrane-bound IL- 15 polypeptide. (FIG. 67A). In some embodiments, an IL- 15 polypeptide is located C- terminal to an IL-15Ra polypeptide in a membrane-bound IL- 15 polypeptide. (FIG. 67B). The IL- 15 polypeptide in FIGS. 67 A and 67B may be immature wild type, immature mutated, mature wild type, or mature mutated. The IL-15Ra polypeptide in FIGS. 67 A and 67B may be immature wild type, immature mutated, mature wild type, or mature mutated. In some embodiments the IL-15 polypeptide in FIG. 67A and FIG. 67B is mature and may or may not be mutated, and the IL-15Ra polypeptide in FIG. 67A and FIG. 67B is mature and may or may not be mutated. In some embodiments the IL- 15 polypeptide in FIG. 67 A andFIG. 67B is mature and may or may not be mutated, and the IL-15Ra polypeptide in FIG. 67A and FIG. 67B is mature and mutated. Although a linker is depicted in FIGS. 67A and 67B, a mbIL-15 may or may not comprise a linker.

[0360] In some embodiments an IL- 15 polypeptide and an IL-15Ra polypeptide is linked by one or more linker. An IL-15 / IL-15Ra fusion polypeptide and / or an IL-15Ra / IL-15 fusion polypeptide may also comprise one or more linker. In some embodiments a membrane-bound IL- 15 comprises and / or is encoded by a structure as shown in FIG. 67A or FIG. 67B. In FIGS. 67A and 67B, the lines connecting the IL-15 to the one or more linker (L) and the one or more linker (L) to the IL-15Ra may represent direct linkages, with no intervening sequences, or may represent intervening sequences, such as, but not limited to, a linker, an untranslated sequence (in the case of a nucleic acid sequence), a translated sequence, a sequence comprising one or more restriction endonuclease sites (in the case of a nucleic acid sequence), or a combination thereof.

[0361] In some embodiments IL-15 / IL-15Ra fusion polypeptide and / or an IL-15Ra / IL-15 fusion polypeptide may comprise one or more signal peptide. In some embodiments a membrane-bound IL-15 comprising one or more signal peptide and, optionally, one or more linkers may comprise and / or be encoded by a structure as shown in FIG. 68A or FIG. 68B. An exemplary IL-15 / IL-15Ra fusion polypeptide comprising, optionally, at least one linker and at least one signal peptide is depicted in FIG. 68 A. An exemplary 15Ra / IL-15 fusion polypeptide comprising, optionally, at least one linker and at least one signal peptide is depicted in FIG. 68B. The IL- 15 polypeptide in FIGS. 68 A and 68B may be immature wild type, immature mutated, mature wild type, or mature mutated. The IL-15Ra polypeptide in FIGS. 68 A and 68B may be immature wild type, immature mutated, mature wild type, or mature mutated. In some embodiments the IL- 15 polypeptide in FIG. 68 A and FIG. 68B is mature and may or may not be mutated, and the IL-15Ra polypeptide in FIG. 68 A and FIG. 68B is mature and may or may not be mutated. In some embodiments the IL- 15 polypeptide in FIG. 68 A and 68B is mature and may or may not be mutated, and the IL-15Ra polypeptide in FIG. 68A and 68B is mature and mutated. In FIGs. 68A and 68B, the lines connecting (a) the one or more signal peptide (SP) to the IL- 15, the IL- 15 to the one or more linker (L), and the one or more linker to the IL-15Ra (as in FIG. 68 A) or (b) the one or more signal peptide (SP) to the IL-15a, the IL-15a to the one or more linker (L), and the one or more linker to the IL- 15 (as in FIG. 68B) may represent direct linkages, with no intervening sequences, or may represent intervening sequences, such as, but not limited to, a linker, an untranslated sequence (in the case of a nucleic acid sequence), a translated sequence, a sequencecomprising one or more restriction endonuclease sites (in the case of a nucleic acid sequence), or a combination thereof.

[0362] In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprises an entire IL- 15 polypeptide, an entire IL-15Ra polypeptide, or both. In some embodiments an entire, or full, wild type IL-15 polypeptide may comprise SEQ ID NO: 305. In some embodiments an entire, or full, wild type IL-15Ra polypeptide may comprise SEQ ID NO: 306.

[0363] In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprises a mature IL-15 polypeptide (e.g., SEQ ID NO: 307), a mature IL-15Ra polypeptide (e.g., SEQ ID NO: 309), which may be mutated (e.g., SEQ ID NO: 311, 313, 315), or both. In some embodiments a 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 some embodiments a mature wild type IL-15Ra 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 some embodiments a mature wild type IL- 15 polypeptide is encoded by a nucleic acid comprising or consisting of the nucleic acid set forth in SEQ ID NO: 308. In some embodiments a mature wild type IL-15Ra polypeptide is encoded by a nucleic acid comprising or consisting of the nucleic acid set forth in SEQ ID NO: 310. However, In some embodiments an IL-15 / IL- 15Ra fusion polypeptide does not comprise a mature wild type IL-15Ra as in SEQ ID NO: 309 or sequences having about 95% or more sequence identity thereto. In some embodiments an IL-15 / IL-15Ra fusion polypeptide does not comprise a mature wild type IL-15Ra encoded by SEQ ID NO: 310 or sequences having about 80%, about 85%, about 90%, or about 95% or more sequence identity thereto.

[0364] In some embodiments an IL- 15 polypeptide is mutated and / or truncated, an IL- 15Ra polypeptide is mutated and / or truncated, or both are mutated and / or truncated.

[0365] In some embodiments an 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.

[0366] In some embodiments an IL-15Ra polypeptide may comprise or may lack a native signal sequence (which may have a sequence comprising SEQ ID NO: 370).

[0367] In some embodiments an IL-15Ra polypeptide, which may be a mature IL-15Ra polypeptide (e.g., SEQ ID NO: 309), may be mutated. In some embodiments an IL-15Ra polypeptide IL-15Ra polypeptide, may comprise a mutated transmembrane domain. In some embodiments the transmembrane domain of an IL-15Ra polypeptide may comprise or consistof SEQ ID NO: 376 or SEQ ID NO: 378. In some embodiments the transmembrane domain of an IL-15Ra polypeptide may be encoded by a nucleic acid comprising or consisting of the sequence set forth in SEQ ID NO: 377 or SEQ ID NO: 379. In some embodiments a mutant IL-15Ra may comprise a heterologous transmembrane domain. In some embodiments, a heterologous transmembrane domain may be derived from CD25. In some embodiments a 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 thereto. In some embodiments a transmembrane domain derived from CD25 is encoded by a nucleic acid comprising or consisting of the nucleic acid seq 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 thereto. In some embodiments, an IL-15Ra 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 thereto. In some embodiments, an IL-15Ra 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 thereto. In some embodiments, a heterologous transmembrane domain may be derived from CD28. In some embodiments a 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 thereto. In some embodiments a transmembrane domain derived from CD28 is encoded by a nucleic acid comprising or consisting of the nucleic acid seq 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 about97%, at least about 98%, at least about 99%, or about 100% identity thereto. In some embodiments, an IL-15Ra 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 thereto. In some embodiments, an IL-15Ra 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 thereto. In some embodiments an IL-15Ra may be mutated by deleting exon 3 of human IL-15Ra genomic DNA. In some embodiments, an IL-15Ra polypeptide comprising a deletion of exon 3 comprises or consists of the sequence set forth in SEQ ID NO: 315 or a sequence having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity thereto. In some embodiments, an IL-15Ra polypeptide comprising a deletion of exon 3 is encoded by a nucleic acid comprising or consisting of the sequence set forth in SEQ ID NO: 316 or a sequence having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity thereto. In some embodiments, function(s) of IL-15Ra, such as, but not limited to, the ability of IL-15Ra be membrane-bound and one or more signaling function(s) of IL-15a are preserved and / or enhanced in an IL-15Ra polypeptide having a heterologous transmembrane domain or deleted exon 3.

[0368] In some embodiments the disclosure provides for nucleic acids encoding polypeptide(s) described herein.

[0369] In an aspect, polypeptide sequences and / or nucleic acid sequences described herein may be isolated and / or recombinant sequences.

[0370] In an aspect, cells described herein may be isolated and / or recombinant cells.

[0371] In some embodiments an 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 some embodiments, function(s) of IL-15, such as, but not limited to, one or more signaling function(s) of IL- 15, are preserved and / or enhanced in a mutated IL- 15 polypeptide.

[0372] In some embodiments an 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 some embodiments, function(s) of IL-15, such as, but not limited to, one or more signaling function(s) of IL- 15, are preserved and / or enhanced in a mutated IL- 15 polypeptide.

[0373] In some embodiments an 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 some embodiments, amnio acid substitutions are conservative or non-conservative. In some embodiments amino acid substitution(s) are conservative amino acid substitution(s). In some embodiments, function(s) of IL-15, such as, but not limited to, one or more signaling function(s) of IL-15, are preserved and / or enhanced in a mutated IL- 15 polypeptide.

[0374] In some embodiments an 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 some embodiments, function(s) of IL-15, such as, but not limited to, one or more signaling function(s) of IL- 15, are preserved and / or enhanced in an IL- 15 polypeptide encoded by a mutated nucleic acid sequence.

[0375] In some embodiments an 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 some embodiments, one or more nucleic acid substitution in a codon may result in a codon encoding the same amino acid or may result in a codon encoding a different amino acid. In some embodiments, one or more nucleic acid substitution in a codon may result in a codon encoding a conservative amino acid substitution. In some embodiments, one or more nucleic acid substitution in a codon may result in a codon encoding the same amino acid. In some embodiments, function(s) of IL- 15, such as, but not limited to, one or moresignaling function(s) of IL- 15, are preserved and / or enhanced in an IL- 15 polypeptide encoded by a mutated nucleic acid sequence.

[0376] In some embodiments, a nucleic acid encoding an IL- 15 polypeptide may comprise a stop codon (such as TAA, TAG, or TGA), positioned at, as a non-limiting example, at the 3’ end of a nucleotide encoding an IL- 15 polypeptide.

[0377] In some embodiments an IL-15Ra 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 some embodiments an IL-15Ra 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 some embodiments an IL-15Ra 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 some embodiments an IL-15Ra 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: 311. In some embodiments an IL- 15Ra 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 some embodiments an IL-15Ra 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 some embodiments, function(s) of IL-15Ra, such as, but not limited to, the ability of IL-15Ra be membranebound and one or more signaling function(s) of IL- 15a are preserved and / or enhanced in a mutated IL-15Ra polypeptide.

[0378] In some embodiments an IL-15Ra polypeptide may comprise (a) SEQ ID NO: 306 comprising one, two, three, four, or five amino acid substitutions; (b) SEQ ID NO: 309comprising 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 (e) SEQ ID NO: 315 comprising one, two, three, four, or five amino acid substitutions. However, In some embodiments an IL-15Ra 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 some embodiments, amnio acid substitutions are conservative or non-conservative. In some embodiments amino acid substitution(s) are conservative amino acid substitution(s). In some embodiments, function(s) of IL-15Ra, such as, but not limited to, the ability of IL-15Ra be membrane-bound and one or more signaling function(s) of IL- 15a are preserved and / or enhanced in a mutated IL-15Ra polypeptide.

[0379] In some embodiments an IL-15Ra 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 some embodiments an IL-15Ra polypeptide is not encoded by a nucleic acid comprising SEQ ID NO: 310 or a having about 85%, about 90%, about 95% or more sequence identity thereto. In some embodiments an IL- 15Ra 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 some embodiments an IL-15Ra 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 some embodiments an IL-15Ra 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 some embodiments, function(s) of IL-15Ra, such as, but not limited to, the ability of IL-15Ra be membrane-bound and one or more signaling function(s) of IL-15a are preserved and / or enhanced in an IL-15a polypeptide encoded by a mutated nucleic acid sequence.

[0380] In some embodiments an IL-15Ra 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 some embodiments an IL-15Ra polypeptide is not encoded by a nucleic acid comprising SEQ ID NO: 310 or a having about 85%, about 90%, about 95% or more sequence identity thereto. In some embodiments, one or more nucleic acid substitution in a codon may result in a codon encoding the same amino acid or may result in a codon encoding a different amino acid. In some embodiments, one or more nucleic acid substitution in a codon may result in a codon encoding a conservative amino acid substitution. In some embodiments, one or more nucleic acid substitution in a codon may result in a codon encoding the same amino acid. In some embodiments, function(s) of IL-15Ra, such as, but not limited to, the ability of IL-15Ra be membrane-bound and one or more signaling function(s) of IL-15a are preserved and / or enhanced in an IL-15a polypeptide encoded by a mutated nucleic acid sequence.

[0381] In some embodiments, a nucleic acid encoding an IL-15Ra polypeptide may comprise a stop codon (such as TAA, TAG, or TGA), positioned at, as a non-limiting example, at the 3’ end of a nucleotide encoding an IL-15Ra polypeptide.

[0382] In some embodiments an IL- 15 polypeptide and an IL-15Ra polypeptide is linked by one or more linker. In some embodiments, a linker is a peptide linker. In some embodiments, a peptide linker is rigid or flexible. In some embodiments, a linker is cleavable. In some embodiments, a linker may promote stability or proper folding of a fusion polypeptide, may increase expression of a fusion polypeptide, may improve biological activity of a fusion polypeptide, may facilitate targeting of a fusion polypeptide, may alter the PK of a fusion polypeptide, or any combination thereof.

[0383] In some embodiments a linker comprises about 2-40 amnio 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.

[0384] In some embodiments one or more linker of an IL-15 / IL-15Ra fusion polypeptide independently comprises or consists 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 some embodiments, one or more linker of an IL-15 / IL- 15Ra fusion polypeptide is not SEQ ID NO: 391 and / or SEQ ID NO: 395. In some embodiments one or more linker of an IL-15 / IL-15Ra fusion polypeptide independently comprises or consists of any 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 some embodiments one or more linker of an IL-15 / IL-15Ra fusion polypeptide independently comprises or consists 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.

[0385] In some embodiments one or more linker of an IL-15 / IL-15Ra fusion polypeptide is independently encoded by one or more nucleic acid comprising or consisting of any of SEQ ID NO: 384, 386, 388, 390, or 392, by a sequence having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity to any of SEQ ID NO: 384, 386, 388, 390, or 392, by one or more nucleic acid encoding any linker comprising or consisting of GSG, LE, or one or more linker set forth in SEQ ID NO: 266 or 393-432, or by one or more nucleic acid encoding any linker having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity to any of SEQ ID NO: 266 or 393-432. However, In some embodiments, one or more linker of an IL-15 / IL-15Ra fusion polypeptide is not encoded by SEQ ID NO: 392 and is not encoded by a nucleic acid encoding SEQ ID NO: 391 or 395.-

[0386] In some embodiments one or more linker of an IL-15 / IL-15Ra fusion polypeptide is independently encoded by one or more nucleic acid comprising or consisting of any of SEQ ID NO: 384, 386, 388, or 390, by a sequence having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity to any of SEQ ID NO: 384, 386, 388, or 390, by one or more nucleic acid encoding any linker comprising or consisting of GSG or one or more linker set forth in SEQ ID NO: 266, 383, 385, 387, 389, 393, or 396-432, or by one or more nucleic acid encoding any linker having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity to any of SEQ ID NO: 266, 383, 385, 387, 389, 393, or 396-432.

[0387] In some embodiments one or more linker of an IL-15 / IL-15Ra fusion polypeptide is independently encoded by any of SEQ ID NO: 384, 386, 388, or 390 or by a sequence having at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity to any of SEQ ID NO: 384, 386, 388, or 390.

[0388] In some embodiments a 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 some embodiments a 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 some embodiments a 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 some embodiments a 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% sequenceidentity to the amino acid sequence of SEQ ID NO: 389. In some embodiments, one or more function(s) of a linker, such as, but not limited to, one or more of flexibility, rigidity, cleavability, ability to promote stability or proper folding of a fusion polypeptide, ability to increase expression of a fusion polypeptide, ability improve biological activity of a fusion polypeptide, ability facilitate targeting of a fusion polypeptide, ability to alter the PK of a fusion polypeptide, or a combination thereof, of the linker, are preserved and / or enhanced in a mutated linker.

[0389] In some embodiments a 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 (e) SEQ ID NO: 391 comprising one, two, three, four, or five amino acid substitutions. In some embodiments, amnio acid substitutions may be conservative or non-conservative. In some embodiments amino acid substitution(s) may be conservative amino acid substitution(s). In some embodiments, one or more function(s) of a linker, such as, but not limited to, one or more of flexibility, rigidity, cleavability, ability to promote stability or proper folding of a fusion polypeptide, ability to increase expression of a fusion polypeptide, ability improve biological activity of a fusion polypeptide, ability facilitate targeting of a fusion polypeptide, ability to alter the PK of a fusion polypeptide, or a combination thereof, of the linker, are preserved and / or enhanced in a mutated linker.

[0390] In some embodiments a 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 some embodiments a 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 some embodiments a 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 some embodiments a linker is encoded by a nucleic acid comprising at least about 80%, at least about 85%, at least about90%, at least about 91%, at least about 92%, at least about 93%, at least about 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 some embodiments, one or more function(s) of a linker, such as, but not limited to, one or more of flexibility, rigidity, cleavability, ability to promote stability or proper folding of a fusion polypeptide, ability to increase expression of a fusion polypeptide, ability improve biological activity of a fusion polypeptide, ability facilitate targeting of a fusion polypeptide, ability to alter the PK of a fusion polypeptide, or a combination thereof, of the linker, are preserved and / or enhanced in a mutated linker.

[0391] In some embodiments a 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 (e) SEQ ID NO: 392 comprising one, two, three, four, or five nucleic acid substitutions. In some embodiments, one or more nucleic acid substitution in a codon may result in a codon encoding the same amino acid or may result in a codon encoding a different amino acid. In some embodiments, one or more nucleic acid substitution in a codon may result in a codon encoding a conservative amino acid substitution. In some embodiments, one or more nucleic acid substitution in a codon may result in a codon encoding the same amino acid. In some embodiments, one or more function(s) of a linker, such as, but not limited to, one or more of flexibility, rigidity, cleavability, ability to promote stability or proper folding of a fusion polypeptide, ability to increase expression of a fusion polypeptide, ability improve biological activity of a fusion polypeptide, ability facilitate targeting of a fusion polypeptide, ability to alter the PK of a fusion polypeptide, or a combination thereof, of the linker, are preserved and / or enhanced in a mutated linker.

[0392] In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker comprises SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, 333, or 335. In some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker does not comprise or consist of SEQ ID NO: 335 or sequences having about 95% or more sequence identity thereto. In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker is not encoded by a nucleic acid comprising or consisting of SEQ ID NO:336 or by sequences having about 80%, about 85%, about 90%, or about 95% or more sequence identity thereto. In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker comprises SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, or 333. In some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker comprises SEQ ID NO: 317, 321, 325, 327, 329, 331, or 333. In some embodiments an IL- 15 / IL-15Ra fusion polypeptide comprising a linker is encoded by a nucleic acid comprising SEQ ID NO: 318, 322, 326, 328, 330, 332, or 334.

[0393] In some embodiments an IL-15 / IL-15Ra 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: 333 comprising one, two, three, four, or five amino acid substitutions; or (j) SEQ ID NO: 335 comprising one, two, three, four, or five amino acid substitutions. However, In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker does not comprise or consist of SEQ ID NO: 335 or sequences having about 95% or more sequence identity thereto. In some embodiments, amnio acid substitutions may be conservative or non-conservative. In some embodiments amino acid substitution(s) may be conservative amino acid substitution(s). In some embodiments, (i) function(s) of IL- 15, such as, but not limited to, one or more signaling function(s) of IL- 15, (ii) function(s) of IL-15Ra, such as, but not limited to, the ability of IL-15Ra be membrane-bound, and signaling function(s) of IL-15Ra or (iii) both (i) and (ii), are preserved and / or enhanced in a mutated IL-15 / IL-15Ra fusion polypeptide.

[0394] In some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra fusion polypeptidecomprising 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 some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL- 15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL- 15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker may comprise at leastabout 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 some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker does not comprise or consist of SEQ ID NO: 335 or sequences having about 95% or more sequence identity thereto. In some embodiments, (i) function(s) of IL-15, such as, but not limited to, one or more signaling function(s) of IL- 15, (ii) function(s) of IL-15Ra, such as, but not limited to, the ability of IL-15Ra be membrane-bound, and signaling function(s) of IL-15Ra or (iii) both (i) and (ii), are preserved and / or enhanced in an IL-15 / IL-15Ra fusion polypeptide encoded by a mutated nucleic acid sequence.

[0395] In some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL-15 / IL- 15Ra 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 some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker may be encoded by a nucleic acidcomprising at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 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 some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra 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 some embodiments, (i) function(s) of IL- 15, such as, but not limited to, one or more signaling function(s) of IL- 15, (ii) function(s) of IL-15Ra, such as, but not limited to, the ability of IL-15Ra be membrane-bound and one or more signaling function(s) of IL-15Ra or (iii) both (i) and (ii), are preserved and / or enhanced in an IL-15 / IL-15Ra fusion polypeptide encoded by a mutated nucleic acid sequence.

[0396] In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker may 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 some embodiments, one or more nucleic acid substitution in a codon may result in a codon encoding the same amino acid or may result in a codon encoding a different amino acid. In some embodiments, one or more nucleic acid substitution in a codon may result in a codon encoding a conservative amino acid substitution. In some embodiments, one or more nucleic acid substitution in a codon may result in a codon encoding the same amino acid. In some embodiments, (i) function(s) of IL- 15, such as, but not limited to, one or more signaling function(s) of IL-15, (ii) function(s) of IL-15Ra, such as, but not limited to, the ability of IL-15Ra be membrane-bound and one or more signaling function(s) of IL-15Ra or (iii) both (i) and (ii), are preserved and / or enhanced in an IL-15 / IL- 15Ra fusion polypeptide encoded by a mutated nucleic acid sequence.

[0397] In some embodiments an IL-15 / IL-15Ra fusion polypeptide, optionally comprising one or more linker, 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) directly or indirectly fused to an 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; 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) directly or indirectly fused to an 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: 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 leastabout 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to an 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) directly or indirectly fused to an 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, 333, or 335 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 to SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, 333, or 335. However, In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker does not comprise or consist of (i) SEQ ID NO: 307 directly or indirectly fused to an N terminus of SEQ ID NO: 309 with a linker therebetween; (ii) SEQ ID NO: 335, (iii) sequences having about 95% or more sequence identity to SEQ ID NO: 307 directly or indirectly fused to an N terminus of SEQ ID NO: 309 with a linker therebetween; or (iv) sequences having about 95% or more sequence identity to SEQ ID NO: 335. In some embodiments one or more linkers comprises or consists of a linker sequence set forth herein.

[0398] In some embodiments an IL-15 / IL-15Ra fusion polypeptide, optionally comprising one or more linker, 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) directly or indirectly fused to an 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 orwithout a linker therebetween; 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) directly or indirectly fused to an 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) directly or indirectly fused to an 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 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 to SEQ ID NO: 317, 319, 321, 323, 325, 327, 329, 331, or 333. In some embodiments one or more linkers comprises or consists of a linker sequence set forth herein.

[0399] In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising one or more linker comprises or consists of, e.g., SEQ ID NO: 317, 321, 325, 327, 329, 331, or 333 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 to SEQ ID NO: 317, 321, 325, 327, 329, 331, or 333.

[0400] In some embodiments an IL-15 / IL-15Ra fusion polypeptide, optionally comprising one or more linker, 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) directly or indirectly fused 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%, atleast about 92%, at least about 93%, at least about 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: 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) directly or indirectly fused 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: 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) directly or indirectly fused 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) directly or indirectly fused 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, 334, or 336 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 to SEQ ID NO: 318, 320, 322, 324, 326, 328, 330, 332, 334, or 336. However, In some embodiments an IL-15 / IL-15Ra 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) sequences having about 80%, about 85%, about 90%, or about 95% or more sequenceidentity to SEQ ID NO: 308 fused to the 5’ end of SEQ ID NO: 310 with a linker therebetween; or (iv) sequences having about 80%, about 85%, about 90%, or about 95% or more sequence identity to SEQ ID NO: 336. In some embodiments one or more linkers is encoded by one or more nucleic acid comprising or consisting of a nucleic acid encoding a linker set forth herein.

[0401] In some embodiments an IL-15 / IL-15Ra fusion polypeptide, optionally comprising one or more linker, 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) directly or indirectly fused 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: 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) directly or indirectly fused 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) directly or indirectly fused 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 at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 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 some embodiments one or more linkers is encoded by one or more nucleic acid comprising or consisting of a nucleic acid encoding a linker set forth herein.

[0402] In some embodiments an IL-15 / IL-15Ra fusion polypeptide is encoded by a nucleic acid comprising or consisting of, e.g., SEQ ID NO: 318, 322, 326, 328, 330, 332, or 334 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 to SEQ ID NO: 318, 322, 326, 328, 330, 332, or 334.

[0403] In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker (L) comprises or consists of any construct A-J as set forth in FIG. 69A. In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker (L) comprises or consists of any construct A-I as set forth in FIG. 69 A. In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker comprises or consists of any construct A, C, or E-I as set forth in FIG. 69A. In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker is encoded by one or more nucleic acid comprising or consisting of any construct A’ -J’ as set forth in FIG. 69B. In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker is encoded by one or more nucleic acid comprising or consisting of any construct A’-F as set forth in FIG. 69B. In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker is encoded by one or more nucleic acid comprising or consisting of any construct A’, C’, or E’-I’ as set forth in FIG. 69B. In some embodiments, sequences 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% identity to any of the sequences set forth in FIGs. 69A and 69B are also provided. In FIGs. 69 A and 69B, the lines connecting the IL- 15 to the linker and the linker to the IL-15Ra may represent direct linkages, with no intervening sequences, or may represent intervening sequences, such as, but not limited to, a linker, an untranslated sequence (in the case of FIG. 69B), a translated sequence, a sequence comprising one or more restriction endonuclease sites (in the case of FIG. 69B), or a combination thereof.

[0404] In some embodiments, a nucleic acid encoding an IL-15 / IL-15Ra fusion polypeptide may comprise a stop codon (such as TAA, TAG, or TGA), positioned at, as nonlimiting examples, at the 3’ end of a nucleotide encoding an IL-15Ra polypeptide, such as where the encoded fusion polypeptide is in an orientation shown in FIG. 67A or FIG. 668Aor at the 3’ end of the IL- 15 polypeptide, such as where the encoded fusion polypeptide is in an orientation shown in FIG. 67B or FIG. 668B.

[0405] In some embodiments IL-15 / IL-15Ra fusion polypeptide and / or an IL-15Ra / IL-15 fusion polypeptide may comprise one or more signal peptide. In some embodiments a fusion polypeptide may comprise the entirety or a portion(s) of the short or the long signal peptide of IL- 15 or the entirety or a portion(s) of the signal peptide of IL-15Ra. In some embodiments the entire signal peptide or part of the signal peptide of IL-15, IL-15Ra, or both, may be mutated or deleted. In some embodiments a fusion polypeptide may comprise one or more heterologous signal peptide, i.e., the entirety or a portion of the signal peptide from a molecule other than IL- 15 and IL-15Ra. In some embodiments, a heterologous signal peptide may be derived from IL-2, CD33, IgVK, or IgE. In some embodiments, a signal peptide may be a signal peptide derived from IgE. In some embodiments a signal peptide derived from IgE may comprise or consist of SEQ ID NO: 367. In some embodiments a signal peptide derived from IgE may be encoded by a nucleic acid comprising or consisting of the sequence set forth in SEQ ID NO: 368.

[0406] In some embodiments a signal peptide may be cleaved or otherwise removed from an IL-15 / IL-15Ra fusion polypeptide.

[0407] In some embodiments, a signal peptide may increase or facilitate transcription, translation, translocation, or a combination thereof, of a fusion polypeptide, as compared to a native IL-15Ra signal peptide, a native IL-15 signal peptide, or both. In some embodiments, the signal peptide may be directly or indirectly fused to the N-terminus or to the C-terminus of an IL-15 / IL-15Ra fusion polypeptide.

[0408] In some embodiments a 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 some embodiments, function(s) of a signal peptide, such as, but not limited to, one or more signaling function(s) of the signal peptide, are preserved and / or enhanced in a mutated signal peptide.

[0409] In some embodiments a signal peptide may comprise SEQ ID NO: 367 comprising one, two, three, four, or five amino acid substitutions. In some embodiments, function(s) of a signal peptide, such as, but not limited to, one or more signaling function(s) of the signal peptide, are preserved and / or enhanced in a mutated signal peptide.

[0410] In some embodiments a 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 some embodiments, function(s) of a signal peptide, such as, but not limited to, one or more signaling function(s) of the signal peptide, are preserved and / or enhanced in a signal peptide that is encoded by a mutated nucleic acid sequence.

[0411] In some embodiments a signal peptide may be encoded by a nucleic acid comprising SEQ ID NO: 368 comprising one, two, three, four, or five nucleic acid substitutions. In some embodiments, one or more nucleic acid substitution in a codon may result in a codon encoding the same amino acid or may result in a codon encoding a different amino acid. In some embodiments, one or more nucleic acid substitution in a codon may result in a codon encoding a conservative amino acid substitution. In some embodiments, one or more nucleic acid substitution in a codon may result in a codon encoding the same amino acid. In some embodiments, function(s) of a signal peptide, such as, but not limited to, one or more signaling function(s) of the signal peptide, are preserved and / or enhanced in a signal peptide that is encoded by a mutated nucleic acid sequence.

[0412] In some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker and a signal peptide derived from IgE may comprise at least about 80%, at least about 85%, at least about90%, at least about 91%, at least about 92%, at least about 93%, at least about 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 some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker and a signal peptide derived from IgE does not comprise or consist of SEQ ID NO: 355 or sequences having about 95% or more sequence identity thereto. In some embodiments, (i) function(s) of IL-15, such as, but not limited to, one or more signalingfunction(s) of IL-15, (ii) function(s) of IL-15Ra, such as, but not limited to, the ability of IL- 15Ra be membrane-bound and one or more signaling function(s) of IL-15Ra (iii) function(s) of a signal peptide derived from IgE, such as, but not limited to, one or more signaling function(s) of the signal peptide, or (iv) all of (i), (ii), and (iii), are preserved and / or enhanced in a mutated IL-15 / IL-15Ra fusion polypeptide comprising a signal peptide derived from IgE.

[0413] In some embodiments an IL-15 / IL-15Ra 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: 353 comprising one, two, three, four, or five amino acid substitutions; or (j) SEQ ID NO: 355 comprising one, two, three, four, or five amino acid substitutions. However, In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker and a signal peptide derived from IgE does not comprise or consist of SEQ ID NO: 355 or sequences having about 95% or more sequence identity thereto. In some embodiments, amnio acid substitutions may be conservative or non-conservative. In some embodiments amino acid substitution(s) may be conservative amino acid substitution(s). In some embodiments, (i) function(s) of IL-15, such as, but not limited to, one or more signaling function(s) of IL-15, (ii) function(s) of IL-15Ra, such as, but not limited to, the ability of IL- 15Ra be membrane-bound and one or more signaling function(s) of IL-15Ra, (iii) function(s) of a signal peptide derived from IgE, such as, but not limited to, one or more signaling function(s) of the signal peptide, or (iv) all of (i), (ii), and (iii), are preserved and / or enhanced in a mutated IL-15 / IL-15Ra fusion polypeptide comprising a signal peptide derived from IgE.

[0414] In some embodiments an IL-15 / IL-15Ra 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: 338. In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker and a signal peptide derived from IgE may be encoded by a nucleic acidcomprising at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 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 some embodiments an IL-15 / IL-15Ra 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: 342. In some embodiments an IL- 15 / IL-15Ra 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: 344. In some embodiments an IL-15 / IL-15Ra 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: 346. In some embodiments an IL-15 / IL-15Ra 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: 348. In some embodiments an IL-15 / IL-15Ra 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: 350. In some embodiments an IL-15 / IL-15Ra 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% sequenceidentity to the nucleic acid of SEQ ID NO: 352. In some embodiments an IL-15 / IL-15Ra 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: 354. In some embodiments an IL- 15 / IL-15Ra 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 some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker and a signal peptide derived from IgE is not encoded by SEQ ID NO: 356 or sequences having about 80%, about 85%, about 90%, or about 95% or more sequence identity thereto. In some embodiments, (i) function(s) of IL-15, such as, but not limited to, one or more signaling function(s) of IL-15, (ii) function(s) of IL-15Ra, such as, but not limited to, the ability of IL- 15Ra be membrane-bound and one or more signaling function(s) of IL-15Ra, (iii) function(s) of a signal peptide derived from IgE, such as, but not limited to, one or more signaling function(s) of the signal peptide, or (iv) all of (i), (ii), and (iii), are preserved and / or enhanced in an IL-15 / IL-15Ra fusion polypeptide comprising a signal peptide derived from Ige, that is encoded by a mutated nucleic acid sequence.

[0415] In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker and a signal peptide derived from IgE may 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: 354 comprising one, two, three, four, or five nucleic acid substitutions; or (j) SEQ ID NO: 356 comprising one, two, three, four, or five nucleic acid substitutions. However, In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker and a signal peptidederived from IgE is not encoded by SEQ ID NO: 356 or sequences having about 80%, about 85%, about 90%, or about 95% or more sequence identity thereto. In some embodiments, one or more nucleic acid substitution in a codon may result in a codon encoding the same amino acid or may result in a codon encoding a different amino acid. In some embodiments, one or more nucleic acid substitution in a codon may result in a codon encoding a conservative amino acid substitution. In some embodiments, (i) function(s) of IL- 15, such as, but not limited to, one or more signaling function(s) of IL- 15, (ii) function(s) of IL-15Ra, such as, but not limited to, the ability of IL-15Ra be membrane-bound and one or more signaling function(s) of IL-15Ra, (iii) function(s) of a signal peptide derived from IgE, such as, but not limited to, one or more signaling function(s) of the signal peptide, or (iv) all of (i), (ii), and (iii), are preserved and / or enhanced in an IL-15 / IL-15Ra fusion polypeptide comprising a signal peptide derived from IgE, that is encoded by a mutated nucleic acid sequence.

[0416] In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a signal peptide derived from IgE, and optionally comprising one or more linker, 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) directly or indirectly fused 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) directly or indirectly fused 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; 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) directly or indirectly fused 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) directly or indirectly fused to the N terminus of SEQ ID NO: 311 (or a sequence at least about 80%, at least about 85%, at leastabout 90%, at least about 91%, at least about 92%, at least about 93%, at least about 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) directly or indirectly fused 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) directly or indirectly fused 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) directly or indirectly fused 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) directly or indirectly fused 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 at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 337, 339, 341, 343, 345, 347, 349, 351, 353, or 355. However, In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a signal peptide derived from IgE, and optionally comprising one or more linker does not comprise or consist of (i) SEQ ID NO: 307 directly or indirectly fused to the N terminus of SEQ ID NO: 309 with a linker therebetween; (ii) SEQ ID NO: 335 or SEQ ID NO: 355; (iii)sequences having about 95% or more sequence identity to SEQ ID NO: 307 directly or indirectly fused to the N terminus of SEQ ID NO: 309 with a linker therebetween; or (iv) sequences having about 95% or more sequence identity to SEQ ID NO: 335 or SEQ ID NO: 355. In some embodiments one or more linkers comprises or consists of a linker sequence set forth herein.

[0417] In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a signal peptide derived from IgE, and optionally comprising one or more linker, 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) directly or indirectly fused 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) directly or indirectly fused 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) directly or indirectly fused 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) directly or indirectly fused 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) directly or indirectly fused to the N terminus of SEQ ID NO: 307 (or asequence at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the N terminus of SEQ ID NO: 315 (or a sequence at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 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, or 353 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 to SEQ ID NO: 337, 339, 341, 343, 345, 347, 349, 351, or 353. In some embodiments one or more linkers comprises or consists of a linker sequence set forth herein.

[0418] In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a signal peptide derived from IgE, and comprising a linker, comprises or consists of, e.g., SEQ ID NO: 337, 341, 345, 347, 349, 351, or 353 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 to SEQ ID NO: 337, 341, 345, 347, 349, 351, or 353.

[0419] In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a signal peptide derived from IgE, and optionally comprising one or more linker, 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) directly or indirectly fused 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) directly or indirectly fused 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; 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) directly or indirectly fused 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) directly or indirectly fused 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: 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) directly or indirectly fused 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) directly or indirectly fused 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: 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) directly or indirectly fused 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) directly or indirectly fused 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%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 338, 340, 342, 344, 346, 348, 350, 352, 354, or 356.

[0420] However, In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a linker and a signal peptide derived from IgE is not encoded by a nucleic acid comprising or consisting of (i) SEQ ID NO: 356 or by sequences having about 80%, about 85%, about 90%, or about 95% or more sequence identity thereto or (ii) SEQ ID NO: 368 directly or indirectly fused to the 5’ end of SEQ ID NO: 308 directly or indirectly fused to the 5’ end of SEQ ID NO: 310 or by sequences having about 80%, about 85%, about 90%, or about 95% or more sequence identity thereto. In some embodiments one or more linkers is encoded by one or more nucleic acid comprising or consisting of a nucleic acid encoding a linker set forth herein.

[0421] In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a signal peptide derived from IgE, and optionally comprising one or more linker, 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) directly or indirectly fused 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) directly or indirectly fused 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: 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) directly or indirectly fused 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 leastabout 97%, at least about 98%, at least about 99%, or about 100% identical thereto) directly or indirectly fused to the 5’ end of SEQ ID NO: 314 (or a sequence at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 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: 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) directly or indirectly fused 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) directly or indirectly fused 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, or 354 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 to SEQ ID NO: 338, 340, 342, 344, 346, 348, 350, 352, or 354. In some embodiments one or more linkers is encoded by one or more nucleic acid comprising or consisting of a nucleic acid encoding a linker set forth herein.

[0422] In some embodiments an IL-15 / IL-15Ra fusion polypeptide comprising a signal peptide derived from IgE, and comprising one or more linker is encoded by a nucleic acid comprising or consisting of, e.g., 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 91%, at least about 92%, at least about 93%, at least about 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.

[0423] In some embodiments a vector may further comprise a post-transcriptional regulatory element (PRE) sequence. In some embodiments the post-transcriptional regulatory element (PRE) sequence may be selected from a 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 amutated WPRE, such as but not limited to WPREmutl (SEQ ID NO: 256) or WPREmut2 (SEQ ID NO: 257)) or a hepatitis B virus (HBV) PRE (HPRE) (SEQ ID NO: 437), variant(s) thereof, or any combination thereof.

[0424] In some embodiments (i) an IL-15 / IL-15Ra fusion polypeptide comprising a linker and a signal peptide derived from IgE (ii) followed by WPREmut2 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: 357. In some embodiments (i) an IL- 15 / IL-15Ra fusion polypeptide comprising a linker and a signal peptide derived from IgE (ii) followed by WPREmut2 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: 358. In some embodiments (i) an IL-15 / IL-15Ra fusion polypeptide comprising a linker and a signal peptide derived from IgE (ii) followed by WPREmut2 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: 359. In some embodiments (i) an IL- 15 / IL-15Ra fusion polypeptide comprising a linker and a signal peptide derived from IgE (ii) followed by WPREmut2 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: 360. In some embodiments (i) an IL-15 / IL-15Ra fusion polypeptide comprising a linker and a signal peptide derived from IgE (ii) followed by WPREmut2 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: 361. In some embodiments (i) an IL- 15 / IL-15Ra fusion polypeptide comprising a linker and a signal peptide derived from IgE (ii) followed by WPREmut2 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 about93%, at least about 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 some embodiments (i) an IL-15 / IL-15Ra fusion polypeptide comprising a linker and a signal peptide derived from IgE (ii) followed by WPREmut2 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: 363. In some embodiments (i) an IL- 15 / IL-15Ra fusion polypeptide comprising a linker and a signal peptide derived from IgE (ii) followed by WPREmut2 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: 364. In some embodiments (i) an IL-15 / IL-15Ra fusion polypeptide comprising a linker and a signal peptide derived from IgE (ii) followed by WPREmut2 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: 365. In some embodiments (i) an IL- 15 / IL-15Ra fusion polypeptide comprising a linker and a signal peptide derived from IgE (ii) followed by WPRE 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: 366. However, In some embodiments (i) an IL-15 / IL-15Ra 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 sequences having about 80%, about 85%, about 90%, or about 95% or more sequence identity thereto. In some embodiments, (i) function(s) of IL-15, such as, but not limited to, one or more signaling function(s) of IL- 15, (ii) function(s) of IL- 15Ra, such as, but not limited to, the ability of IL-15Ra be membrane-bound and signaling function(s) of IL-15Ra, (iii) function(s) of a signal peptide derived from IgE, such as, but not limited to, one or more signaling function(s) of the signal peptide, (iv) post-transcriptional regulatory function(s) of wild type or mutant WPRE, or (v) all of (i), (ii), (iii), and (iv) arepreserved and / or enhanced in an IL-15 / IL-15Ra fusion polypeptide comprising a signal peptide derived from IgE that is encoded by a mutated nucleic acid sequence.

[0425] In some embodiments (i) an IL-15 / IL-15Ra fusion polypeptide comprising a linker and a signal peptide derived from IgE (ii) followed by WPREmut2 or wild type WPRE (wt where indicated) may 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; (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 some embodiments (i) an IL-15 / IL-15Ra 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 sequences having about 80%, about 85%, about 90%, or about 95% or more sequence identity thereto. In some embodiments, one or more nucleic acid substitution in a codon may result in a codon encoding the same amino acid or may result in a codon encoding a different amino acid. In some embodiments, one or more nucleic acid substitution in a codon may result in a codon encoding a conservative amino acid substitution. In some embodiments, one or more nucleic acid substitution in a codon may result in a codon encoding the same amino acid. In some embodiments, (i) function(s) of IL- 15, such as, but not limited to, one or more signaling function(s) of IL-15, (ii) function(s) of IL-15Ra, such as, but not limited to, the ability of IL-15Ra be membrane-bound and signaling function(s) of IL-15Ra, (iii) function(s) of a signal peptide derived from IgE, such as, but not limited to, one or more signaling function(s) of the signal peptide, (iv) post-transcriptional regulatory function(s) of mutant or wild type WPRE, or (v) all of (i), (ii), (iii), and (iv) are preserved and / or enhanced in an IL-15 / IL-15Ra fusion polypeptide comprising a signal peptide derived from IgE that is encoded by a mutated nucleic acid sequence.

[0426] In some embodiments nucleic acid sequences encoding a mb IL-15 polypeptide operatively coupled to a promoter are provided. In some embodiments nucleic acid sequences encoding a mbIL-15 polypeptide operatively coupled to a post-transcriptional regulatoryelement are provided. In some embodiments the promoter is an MSCV promoter and / or the post-transcriptional regulatory element is a WPRE, optionally a mutated WPRE, optionally WPREmut2. In some embodiments the promoter is MSCV promoter. In some embodiments the WPRE is WPREmut2.

[0427] In some embodiments one or more vectors comprising one or more nucleic acids encoding SEQ ID NO: 305, 306, 307, 309, 311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331, 333, 335, 337, 339, 341, 343, 345, 347, 349, 351, 353, 355, or combinations thereof are provided. In some embodiments one or more vectors comprising one or more nucleic acids encoding SEQ ID NO: 307, 311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331, 333, 337, 339, 341, 343, 345, 347, 349, 351, 353, or combinations thereof are provided. In some embodiments one or more vectors comprising one or more nucleic acids encoding SEQ ID NO: 311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331, 333, 337, 339, 341, 343, 345,347, 349, 351, 353, or combinations thereof are provided. In some embodiments one or more vectors comprising one or more nucleic acids encoding SEQ ID NO: 311, 313, 315, 317, 321, 325, 327, 329, 331, 333, 337, 341, 345, 347, 349, 351, 353, or combinations thereof are provided. Such vectors may also comprise one or more nucleic acids encoding one or more TCRa, one or more TCRP, one or more CD8a, one or more CD8P, or combinations thereof. Each of TCRa, TCRP, CD8a, and CD8P may indep...

Claims

CLAIMS1. A nucleic acid comprising a nucleotide sequence of SEQ ID NOs: 454, 451, 448, 449, 450, 452 or 453 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 NOs: 454, 451, 448, 449, 450, 452 or 453.

2. The nucleic acid of claim 1 comprising a nucleotide sequence of SEQ ID NOs: 454, 451 or 448, 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 NOs: 454, 451 or 448.

3. The nucleic acid of claim 2 comprising a nucleotide sequence of SEQ ID NO: 454 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: 454.

4. A vector comprising the nucleic acid of claim 1.

5. The vector of claim 4, wherein the vector is a viral vector or a non-viral vector.

6. The vector of claim 5, wherein the viral vector is selected from adenoviruses, poxviruses, alphaviruses, arenaviruses, flaviviruses, rhabdoviruses, retroviruses, lentiviruses, herpesviruses, paramyxoviruses, picomaviruses, and any combination thereof.

7. A method of preparing T cells and / or natural killer cells for immunotherapy comprising: isolating or enriching 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 the vector of claim 4, expanding the transduced T cells and / or natural killer cells.

8. The method of claim 7, wherein the blood sample comprises peripheral blood mononuclear cells (PMBC).

9. The method of claim 7, wherein (i) the activating comprises contacting the T cells and / or natural killer cells with an anti-CD3 and an anti-CD28 antibody; and / or (ii) the activation, the expanding, or both are in the presence of a combination of IL-2 and IL- 15 and optionally with zoledronate.

10. A T cell and / or natural killer cell transduced with the nucleic acid of claim 1.

11. The T cell and / or natural killer cell of claim 10, expressing a) a TCR alpha variable domain having an amino acid sequence as comprised in SEQ ID NO: 15, and a TCR beta variable domain having an amino acid sequence as comprised in SEQ ID NO: 16, and b) an IL-15 / IL-15 Receptor alpha fusion polypeptide having an amino acid sequence as comprised in any one of SEQ ID NOs: 329, 327, 325, 323 or 321.

12. The T cell and / or natural killer cell of claim 1 1, wherein the TCR alpha variable domain has an amino acid sequence comprising the sequence of SEQ ID NO: 455, the TCR beta variable domain has an amino acid sequence comprising SEQ ID NO: 456, and the IL-15 / IL-15 Receptor alpha fusion polypeptide has an amino acid sequence as comprised in SEQ ID NO: 329.

13. The T cell and / or natural killer cell of claim 10, wherein the T cell is an aP T cell, a yb T cell, a natural killer T cell, or any combination thereof.

14. The T cell and / or natural killer cell of claim 11, wherein the a T cell is a CD4+ or CD8+ T cell.

15. A composition comprising the vector of claim 4.

16. The composition of claim 15, wherein the composition is a pharmaceutical composition.

17. The composition of claim 16, wherein the composition further comprises an adjuvant, excipient, carrier, diluent, buffer, stabilizer, or a combination thereof.

18. A method of treating cancer in a subject in need thereof, comprising administering the T cell and / or natural killer cell of claim 10 to said subject.

19. The nucleic acid of claim 2 comprising a nucleotide sequence of SEQ ID NO: 451 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: 451.

20. The nucleic acid of claim 2 comprising a nucleotide sequence of SEQ ID NO: 448 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: 448.

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