Immunotherapy for polyomavirus

Abstract

Provided herein are methods and compositions relating to polyomavirus epitopes that are useful for treating cancer or polyomavirus infection.

Description

[Technical Field]

[0001] Related applications This application claims priority to U.S. Provisional Patent Application No. 62 / 878,105, filed on 24 July 2019, which is incorporated in its entirety by reference. [Background technology]

[0002] Polyomaviruses are ubiquitous viruses that infect a wide range of mammalian species. Currently, more than 12 different human polyomavirus species have been identified, including BK polyomavirus (BKV / human polyomavirus 1), John Cunningham polyomavirus (JCV / human polyomavirus 2), and Merkel cell polyomavirus (MCV / human polyomavirus 5).

[0003] Most such polyomaviruses are typically asymptomatic in humans. However, disease-related human polyomaviruses often infect childhood and / or immunocompromised hosts. For example, early JCV infection may occur via the amygdala or gastrointestinal tract, remaining latent in the gastrointestinal tract, and possibly lymphoid organs, nerve cell tissue, and kidneys, where the virus continues to reproduce and shed viral particles. Subsequently, under conditions of immunodeficiency, immunosuppression, or immune deficiency, both JCV and BKV can reactivate and progress to serious organ disease.

[0004] Of particular note is the JC virus, which can cross the blood-brain barrier and invade the neurotropic central nervous system (CNS), infecting glial cells (e.g., oligodendrocytes and astrocytes) and meningeal cells. Once reactivated in the brain (e.g., in immunocompromised subjects), JCV infection is associated with white matter demyelination and several pathological syndromes, such as JCV granule cell layer neuropathy (JCV GCN), JCV encephalopathy (JCV CPN / JCVE), JCV meningitis (JCVM), and especially progressive multifocal leukoencephalopathy (PML), a demyelinating disease of the central nervous system with a high mortality rate. PML is observed primarily in patients with acquired immunodeficiency syndrome (AIDS), as well as in patients with severe immunodeficiency, such as those receiving immunosuppressive therapy (e.g., steroids, cell division inhibitors and antiproliferative agents, therapeutic antibodies, calcineurin inhibitors, anti-rejection drugs, etc.), including those with organ transplants, Hodgkin lymphoma, multiple sclerosis, psoriasis, and other autoimmune diseases. Currently, there are no drugs that effectively inhibit or cure the viral infection. Treatment mainly relies on reversing or alleviating the patient's immunodeficiency to delay or halt disease progression. Unfortunately, such strategies require the temporary suspension or discontinuation of treatment in immunosuppressed patients, creating a dilemma that leaves these patients vulnerable to one of two conditions. Therefore, new therapies are needed to treat and prevent polyomavirus infection and / or polyomavirus-related diseases. [Overview of the project]

[0005] Compositions and methods relating to polyomavirus epitopes (e.g., epitopes listed in Tables 1, 2, 3, 4, 5, and / or 6) that are recognized by T lymphocytes (e.g., cytotoxic T lymphocytes (CTLs) and / or helper T lymphocytes) and are useful for the prevention and / or treatment of polyomavirus infection (e.g., JCV infection) and / or cancer (e.g., polyomavirus-associated cancer, e.g., JCV-associated cancer). In some embodiments, the compositions and methods relate to JCV epitopes (e.g., epitopes listed in Tables 1, 2, and 3). In some embodiments, the compositions and methods relate to hybrid epitopes (e.g., epitopes listed in Table 4) that incorporate sequence mutations found within and / or across related virus strains.

[0006] In certain embodiments, peptides (e.g., isolated and / or recombinant polypeptides) comprising one or more epitopes derived from one or more JCV antigens (e.g., epitopes derived from LTA, STA, or VP1 virus antigens, e.g., epitopes listed in Tables 1, 2, and / or 3) and / or one or more hybrid epitopes (e.g., epitopes listed in Table 4) are provided herein. In some embodiments, the polypeptide comprises a plurality of such epitopes. In some embodiments, the polypeptide further comprises an intervening amino acid sequence between at least two of the plurality of epitopes. In some embodiments, the peptide can induce an immune response upon administration to a subject (e.g., a mammalian subject, e.g., a human subject).

[0007] In some embodiments, the epitope is selected to provide broad coverage in the human population. In some embodiments, the epitope has HLA class I constraint for HLA-A1, -A2, -A3, -A11, -A23, -A24, -A26, -A29, -A30, -B7, -B8, -B27, -B35, -B38, -B40, -B41, -B44, -B51, -B56, -B57, or -B58. In some embodiments, the epitope has HLA class II constraint for HLA-DP, -DM, -DOA, -DOB, -DQ, or -DR. In some embodiments, the epitope has HLA class II constraint for HLA-DRB or -DQB. In some embodiments, the peptide contains, is essentially, or consists of the epitope amino acid sequence described in SEQ ID NOs: 1-21. In some embodiments, pharmaceutical compositions comprising the peptides provided herein are provided herein.

[0008] In certain embodiments, nucleic acids encoding peptides disclosed herein (e.g., isolated nucleic acids) are provided herein. In some embodiments, expression constructs comprising such nucleic acids are provided herein. In some embodiments, host cells comprising such expression constructs are provided herein. In certain embodiments, methods for producing isolated peptides are provided herein, comprising expressing the isolated peptide in host cells provided herein and purifying the isolated peptide at least partially. In some embodiments, pharmaceutical compositions comprising the nucleic acids provided herein are provided herein.

[0009] In certain embodiments, T lymphocytes (e.g., isolated T lymphocytes, CD4+ T lymphocytes, CD8+ T lymphocytes) containing T cell receptors (TCRs) that specifically bind to epitopes presented herein on HLA (e.g., class I HLA, class II HLA). In certain embodiments, a method for growing BK virus-specific T lymphocytes for adoptive immunotherapy is provided herein, comprising (i) contacting one or more cells isolated from a subject, including T lymphocytes, with antigen-presenting cells that present epitopes provided herein, and (ii) culturing one or more cells under conditions such that BK virus-specific T lymphocytes grow from the one or more cells. In certain embodiments, culturing one or more cells is carried out in the presence of IL-2 and / or IL-21. In some embodiments, cells are cultured in the presence of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 ng / ml of IL-2 and / or IL-21. In some embodiments, cells are cultured in IL-2 and / or IL-21 at concentrations of 30, 35, 40, 45, 50, 60, 70, 80, 90, or 100 ng / ml or less. In some embodiments, cells are cultured in IL-2 and / or IL-21 at concentrations of 10-50, 20-40, 25-35, or about 30 ng / ml. In some embodiments, cells are cultured in IL-2 and / or IL-21 at concentrations of 30 ng / ml. In certain embodiments, proliferation in the presence of IL-2 and / or IL-21 results in an increase in the ratio of the absolute number of polyomavirus-specific CD8 T cells to the absolute number of polyomavirus-specific CD4 T cells in the proliferated T lymphocyte population, compared to proliferation in the absence of IL-2 and / or IL-21.

[0010] In certain embodiments, methods are provided herein for treating or preventing polyomavirus infection (e.g., JCV infection) and / or treating polyomavirus-associated cancer (e.g., JCV-associated cancer) and / or inducing a T lymphocyte immune response in a subject, comprising administering to the subject a peptide, nucleic acid, T cell or pharmaceutical composition provided herein. In some embodiments, the subject is a mammal. In some embodiments, the subject is a human. In some embodiments, the subject is immunocompromised.

[0011] In certain embodiments, a method is provided for detecting JC virus infection in a subject, comprising detecting the presence of JCV-specific T lymphocytes by contacting T lymphocytes isolated from the subject with an isolated peptide provided herein. In some embodiments, the method further comprises treating the JC virus infection in the subject according to the method described herein. In some embodiments, the subject is a mammal. In some embodiments, the subject is a human. In some embodiments, the subject is immunodeficient.

[0012] In certain embodiments, methods for treating cancer (e.g., polyomavirus-associated cancer, e.g., JCV-associated cancer) in a subject are provided herein. In some embodiments, the method involves administering to a subject a pharmaceutical composition comprising cytotoxic T cells (CTLs) containing T cell receptors (TCRs) that recognize one or more epitopes listed in Tables 1, 2, 3 and / or 4 (e.g., at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more). In some embodiments, the subject expresses human leukocyte antigen (HLA) to which one or more epitopes are constrained. In some embodiments, the CTLs are autologous to the subject. In some embodiments, the CTLs are not autologous to the subject. In some embodiments, the CTLs are obtained from a CTL library or bank. In some embodiments, the method involves administering a vaccine composition containing one or more epitopes listed in Tables 1, 2, 3 and / or 4 (for example, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more). In some embodiments, the method involves administering a pharmaceutical composition antigen-presenting cell (APC) that presents one or more epitopes listed in Tables 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more, as a target. In some embodiments, the target expresses a human leukocyte antigen (HLA) to which one or more epitopes are constrained.

[0013] In certain embodiments, methods for treating a polyomavirus infection (e.g., JCV infection) in a subject are provided herein. In some embodiments, the subject is immunocompromised. In some embodiments, the method involves administering to a subject a pharmaceutical composition comprising a CTL containing a TCR that recognizes one or more epitopes listed in Tables 1, 2, 3 and / or 4 (e.g., at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more). In some embodiments, the subject expresses an HLA to which one or more epitopes are constrained. In some embodiments, the CTL is autologous to the subject. In some embodiments, the CTL is not autologous to the subject. In some embodiments, the CTL is obtained from a CTL library or bank. In some embodiments, the method involves administering a vaccine composition containing one or more epitopes listed in Tables 1, 2, 3 and / or 4 (for example, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more). In some embodiments, the method involves administering a pharmaceutical composition antigen-presenting cell (APC) that presents one or more epitopes listed in Tables 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more, as a target. In some embodiments, the target expresses a human leukocyte antigen (HLA) to which one or more epitopes are constrained.

[0014] In some embodiments, a population of CTLs comprising T cell receptors (TCRs) that recognize one or more epitopes listed in Tables 1, 2, 3 and / or 4 (for example, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more) is provided herein.

[0015] In some embodiments, a population of APCs presenting one or more epitopes listed in Tables 1, 2, 3 and / or 4 (e.g., at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more) is provided herein. In some embodiments, the APCs include B cells, antigen-presenting T cells, dendritic cells and / or artificial antigen-presenting cells, such as aK562 cells. In some embodiments, the antigen-presenting cells (e.g., aK562 cells) express CD80, CD83, 41BB-L and / or CD86. In some embodiments, methods for treating or preventing cancer (e.g., polyomavirus-associated cancer, e.g., JCV-associated cancer) and / or polyomavirus (e.g., JCV) infection in a subject are provided herein, comprising administering the APC described herein to the subject.

[0016] In some embodiments, polypeptides comprising one or more epitopes listed in Tables 1, 2, 3 and / or 4 (for example, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more) are provided herein. In certain embodiments, nucleic acid molecules (e.g., DNA molecules or RNA molecules) encoding polypeptides containing one or more epitopes listed in Tables 1, 2, 3 and / or 4 (e.g., at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more) are provided herein. In some embodiments, the nucleic acid molecule is a vector (e.g., an adenovirus vector). In some embodiments, vaccine compositions comprising the polypeptides and / or nucleic acid molecules described herein are provided herein.

[0017] In some embodiments, a method for generating, activating, and / or inducing the proliferation of polyomavirus-specific CTLs (e.g., JCV-specific CTLs) is provided herein, comprising contacting an APC that presents one or more epitopes listed in Tables 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more) with a CTL. In some embodiments, the CTL is contacted with the APC in vitro. In some embodiments, the APC includes B cells, antigen-presenting T cells, dendritic cells, and / or artificial antigen-presenting cells, such as aK562 cells. In some embodiments, antigen-presenting cells (e.g., aK562 cells) express CD80, CD83, 41BB-L, and / or CD86. In some embodiments, CTLs are contacted with APCs in the presence of one or more cytokines.

[0018] In some embodiments, a polypeptide comprising one or more epitopes listed in Table 1, Table 2, Table 3 and / or Table 4 (for example, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more) and / or one or more epitopes listed in Table 1, Table 2, Table 3 and / or Table 4 ( For example, a method for generating an APC that displays the epitopes provided herein is provided, comprising contacting the APC with a nucleic acid encoding a polypeptide containing at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more epitopes. In some embodiments, the APC expresses an HLA to which one or more of its epitopes are constrained.

[0019] In some embodiments, one or more epitopes include epitopes shared by two or more polyomaviruses. In some embodiments, the shared epitopes include regions of sequence homology between at least two polyomaviruses, and the region of sequence homology is at least 3, 4, 5, 6, or 7 amino acids over the full length of the epitope sequence. In some embodiments, the two polyomaviruses are BKV and JCV. In some embodiments, at least three amino acids are LLL.

[0020] In other aspects, provided herein are methods of identifying a subject suitable for a treatment method provided herein (e.g., administration of a CTL, APC, or vaccine composition provided herein), including isolating a sample from the subject (e.g., a blood or tumor sample) and detecting the presence of an epitope provided herein or a nucleic acid encoding an epitope provided herein. In certain embodiments, a subject is identified as suitable for a treatment method provided herein if the subject expresses an HLA to which one or more epitopes described herein are bound. In some embodiments, a subject identified as suitable for a treatment method provided herein is treated using that treatment method. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] [Figure 1] FIG. 1 is a graph showing the T cell response to JCV antigen. Briefly, PBMCs from 17 healthy subjects were stimulated with JCV overlapping peptide pools (OPPs), and the T cells were grown for 14 days in the presence of IL-2. On day 14 after stimulation with each peptide pool, the cells were assayed for IFN-γ expression using flow cytometry. Edited data for all 17 donors are represented in graphs showing the response of (A) BKV-specific CD8+ T cells and (B) CD4+ T cells after stimulation with each BKV OPP. [Figure 2]Figure 2 is a graph showing representative data demonstrating the identification of T cell determinants using a two-dimensional peptide matrix. JCV-specific T cells, grown in vitro using OPP, were further characterized by the identification of specific T cell determinants. Subpools (24 pools; LTA1-LTA24) were created using individual duplicate peptides, and the T cell response for each pool was measured by the intracellular cytokine staining (ICS) IFN-γ assay shown in the bar graph of Panel A. The responses using the subpools were superimposed on a two-dimensional matrix to show individual peptides common to each pool. The FACs plot in Panel B shows the T cell response for each individual peptide (P29, P30, and P32) when used in the IFN-γ ICS assay, thereby identifying the peptides that induce the JCV-specific T cell response. [Figure 3] Figure 3 is a graph showing representative data from HLA class II restrictiveness analysis of epitopes mapped from the JCV-LT antigen. Specifically, it shows the HLA class II restrictiveness of the VDLHAFLSQAVFSNR(LT29), FLSQAVFSNRTVASF(LT30), and TVASFAVYTTKEKAQ(LT32) peptides as HLA DRB1*10:01. Briefly, a panel of lymphoblastoid cells (LCLs) matching a single allele of the donor's HLA type was selected and each peptide was loaded for 1 hour. The loaded LCLs were then used as stimulating cells in an IFN-γ ICS assay. When presented by MHC, the peptides induce an IFN-γ response in JCV-specific T cells. [Figure 4]Figure 4 is a graph showing T cell cross-reactivity between the BKV epitope and the JCV epitope. The ICS FACS plot shows IFN-γ expression in T cells grown with either the BKV epitope (SSGTQQWRGLARYFK) or the JCV epitope (RSGSQQWRGLSRYFK). These primed T cells were stimulated with both BKV and JCV peptides, demonstrating that T cells grown with either of these epitopes recognize both the BKV and JCV peptide sequences. [Figure 5] Figure 5 is a graph illustrating JCV-specific T cell proliferation from healthy subjects. The frequency of IFN-γ-expressing CD4+ T cells was evaluated, and the response of each individual subject is shown in the graph. [Figure 6] Figure 6 is a graph showing the pluripotency of JCV-specific T cells proliferated using pooled peptides (pepdtide). The dot plots of representative FACs show the expression of individual effector molecules in CD4+ T cells when restimulated with the JCV peptide pool (a). The pluripotency of JCV-specific T cells expressing multiple cytokines is shown in panels (b) and (c). [Figure 7] Figure 7 is a graph showing the transcription factor and effector profiles of JCV-specific T cells grown in vitro. [Modes for carrying out the invention]

[0022] general T lymphocytes (e.g., cytotoxic (CD8) + )T lymphocytes (CTLs) and / or helper cells (CD4 +Compositions and methods relating to polyomavirus epitopes (e.g., epitopes listed in Tables 1, 2, 3, and / or 4) that are recognized by T lymphocytes and are useful for the prevention and / or treatment of polyomavirus infection (e.g., JCV infection) and / or cancer (e.g., polyomavirus-associated cancer, e.g., JCV-associated cancer). In some embodiments, the compositions and methods provided herein relate to JCV epitopes (e.g., epitopes listed in Tables 1, 2, and 3). In some embodiments, the compositions and methods relate to hybrid epitopes that encompass mutations found within or across BKV and JCV epitopes (e.g., epitopes listed in Table 4).

[0023] definition For convenience, the specific terms used in this specification, the examples, and the appended claims are set forth herein.

[0024] The articles “a” and “an” are used herein to refer to one or more than one (i.e., at least one) of the grammatical objects of the article. For example, “one element” means one element or more than one element.

[0025] As used herein, the term “administer” means to provide a drug or composition to a subject, and includes, but is not limited to, administration by a healthcare professional and self-administration. Such drugs may include, for example, the peptides described herein, antigen-presenting cells provided herein, and / or CTLs provided herein.

[0026] The term "amino acid" encompasses all natural or synthetic molecules that contain both amino and acid functional groups and can be included in polymers of natural amino acids. Examples of amino acids include natural amino acids, their analogues, derivatives and homologues, amino acid analogues with mutant side chains, and all stereoisomers of any of the above.

[0027] The terms "binding" or "interacting" refer to an association that may be a stable association between two molecules, for example, between a TCR and a peptide / HLA, through electrostatic, hydrophobic, ionic, and / or hydrogen bonding interactions under physiological conditions. A TCR "recognizes" a T cell epitope that it can bind to when presented on appropriate HLA.

[0028] The terms “biological sample,” “tissue sample,” or simply “sample” refer, respectively, to a collection of cells obtained from the tissue of the subject. Sources of tissue samples may include solid tissue such as fresh, frozen, and / or preserved organs, tissue samples, biopsies, or aspirates; blood or any blood component, serum, blood; body fluids, such as cerebrospinal fluid, amniotic fluid, ascites, or interstitial fluid, urine, saliva, feces, tears; or cells from any point in time of the subject’s pregnancy or development.

[0029] As used herein, the term “cancer” includes, but is not limited to, solid tumors and hematological tumors. The term cancer includes diseases of the skin, tissues, organs, bones, cartilage, blood, and blood vessels. The term “cancer” further encompasses primary cancer and metastatic cancer.

[0030] As used herein, the term “homologous” refers to sequence similarity (e.g., nucleic acid or amino acid sequences) between two regions of the same sequence chain or between regions of two different sequence chains. The term “homologous” is also used to refer to sequence similarity between two regions of the same sequence chain or between regions of two different sequence chains. For example, if the positions of amino acid residues in both regions are occupied by the same amino acid residues, then these regions are homologous in that position. A first region is homologous to a second region if at least one nucleotide residue position in each region is occupied by the same residue. The homology between two regions is expressed in terms of the proportion (ratio) of nucleotide or amino acid residue positions in the two regions that are occupied by the same nucleotide or amino acid residue. For example, a region having the nucleotide sequence 5'-ATTGCC-3' and a region having the sequence 5'-TATGGC-3' have 50% homology. Preferably, the first region includes a first portion, and the second region includes a second portion, so that at least about 50%, preferably at least about 75%, at least about 90%, or at least about 95% of the nucleotide residue positions in each of these portions are occupied by the same nucleotide residue. More preferably, all the nucleotide residue positions in each of these portions are occupied by the same nucleotide residue.

[0031] The term "isolated" refers to a material that has been removed from its natural state or otherwise subjected to human manipulation. An isolated material may be manipulated to be substantially or essentially devoid of its naturally associated components, or to be in an artificial state with its naturally associated components.

[0032] The term "peptide" refers to two or more amino acids linked together by a peptide bond or modified peptide bond. The terms "peptide," "polypeptide," and "protein" as used herein may be used interchangeably. In certain embodiments, the peptide is prepared from recombinant DNA or RNA, or DNA or RNA of synthetic origin, or any combination thereof, and (1) does not associate with peptides commonly found in nature, (2) is isolated from cells in which it commonly resides, (3) is isolated without other proteins from the same cellular source, (4) is expressed by cells of different species, or (5) does not occur in nature.

[0033] The term "epitope" refers to a peptide determinant that can specifically bind to an antibody or TCR. Epitopes typically consist of chemically active surface groups of a molecule, such as amino acids or sugar side chains. A specific epitope can be defined by a particular sequence of amino acids to which an antibody can bind.

[0034] As used herein, the term “pharmaceutically acceptable” means a drug, compound, material, composition, and / or dosage form that is suitable for use in contact with human and animal tissues, within the bounds of sound medical judgment, without excessive toxicity, irritation, allergic reactions, or other problems or complications, and in proportion to a reasonable benefit-to-risk ratio.

[0035] As used herein, the term “pharmaceutically acceptable carrier” means a pharmaceutically acceptable material, composition, or vehicle involved in transporting a drug from one organ or part of the body to another, such as a liquid or solid filler, diluent, excipient, or material encapsulating a solvent. Each carrier must be “acceptable” in the sense that it is compatible with the other components of the formulation and is not harmful to the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers include: (1) sugars, e.g., lactose, glucose and sucrose; (2) starches, e.g., corn starch and potato starch; (3) cellulose and its derivatives, e.g., sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, e.g., cocoa butter and suppository wax; (9) oils, e.g., peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (1 (0) Glycols, e.g., propylene glycol; (11) Polyols, e.g., glycerin, sorbitol, mannitol, and polyethylene glycol; (12) Esters, e.g., ethyl oleate and ethyl laurate; (13) Agar; (14) Buffering agents, e.g., magnesium hydroxide and aluminum hydroxide; (15) Alginic acid; (16) Phenothermally hydrated substances; (17) Isotonic saline; (18) Ringer's solution; (19) Ethyl alcohol; (20) pH buffered solutions; (21) Polyesters, polycarbonates, and / or polyanhydrides; and (22) Other non-toxic, suitable substances used in pharmaceutical formulations.

[0036] The terms “polynucleotide” and “nucleic acid” are used interchangeably. They refer to polymeric forms of nucleotides of any length, whether deoxyribonucleotides, ribonucleotides, or their analogues. Polynucleotides may have any three-dimensional structure and may perform any function. The following are non-limiting examples of polynucleotides: coding or non-coding regions of genes or gene fragments, loci(s) determined from linkage analysis, exons, introns, messenger RNA (mRNA), transfer RNA, ribosomal RNA, ribozymes, cDNA, recombinant polynucleotides, branched polynucleotides, plasmids, vectors, isolated DNA of any sequence, isolated RNA of any sequence, nucleic acid probes, and primers. Polynucleotides may include modified nucleotides, such as methylated nucleotides and nucleotide analogues. Modifications to the nucleotide structure, if present, may be made before or after the construction of the polymer. Polynucleotides may be further modified, for example, by conjugation with labeling components. In all nucleic acid sequences provided herein, U nucleotides are interchangeable with T nucleotides.

[0037] As used herein, a therapeutic agent that “prevents” a condition means a compound that, when administered to a statistical sample before the onset of the disorder or condition, reduces the incidence of the disorder or condition in the treated sample compared to an untreated control sample, or delays the onset or reduces the severity of one or more symptoms of the disorder or condition compared to an untreated control sample.

[0038] As used herein, “specific binding” refers to the ability of an antibody to bind to a given antigen or a peptide to bind to a given binding partner. Typically, an antibody or peptide binds to about 10 -7 K below M DIt binds specifically to the given antigen or binding partner with an affinity equivalent to (K), and has an affinity at least 10 times smaller, at least 100 times smaller, or at least 1000 times smaller than its affinity for binding to nonspecific and unrelated antigens / binding partners (e.g., BSA, casein). D It binds to a predetermined antigen / binding partner (such as represented by ).

[0039] As used herein, the term “subject” means a human or non-human animal selected for treatment or therapy.

[0040] As used herein, the terms “therapeutic effective dose” and “effective dose” mean the amount of a drug that, when administered to a subject, elicits an appropriate therapeutic response in the subject and produces a beneficial outcome in the subject with a reasonable benefit-to-risk ratio applicable to any medical treatment.

[0041] To "treat" a disease in a subject, or to "treat" a subject with a disease, means to administer a medical treatment, such as the administration of a drug, to the subject in such a way that at least one symptom of the disease is reduced or prevented from worsening.

[0042] The term "vector" refers to a means by which nucleic acids can be replicated and / or transferred between organisms, cells, or cellular components. Examples of vectors include plasmids, viruses, bacteriophages, proviruses, phagemids, transposons, and artificial chromosomes, which may or may not be able to replicate autonomously, or may be incorporated into the chromosomes of a host cell.

[0043] Epitope In certain embodiments, methods and compositions relating to polyomavirus epitopes, e.g., JCV epitopes, that are recognized by immunoeffector cells (e.g., cytotoxic T cells / CTLs) when presented on HLA are provided herein. In certain embodiments, the epitopes described herein are useful in the prevention and / or treatment of polyomavirus infection (e.g., JCV virus infection) and / or cancer (e.g., JVC-associated cancer expressing the epitopes provided herein), and for the production of pharmaceutical agents and compositions thereof (e.g., sensitized immunoeffector cells and / or APCs) useful in the prevention and / or treatment of polyomavirus infection (e.g., JCV virus infection) and / or cancer (e.g., polyomavirus-associated cancer expressing the epitopes provided herein). In certain embodiments, the epitopes are the JCV epitopes listed in Tables 1, 2 and / or 3. In some embodiments, the epitope is a hybrid epitope comprising amino acids derived from both the BKV epitope and the homologous JCV epitope, and / or amino acid variants found in different BKV or JCV strains. Exemplary hybrid epitopes are listed in Table 4. In some embodiments, the compositions and methods described herein further relate to epitopes from additional viruses, such as EBV, CMV, or ADV. In some embodiments, the epitope is an HLA class I-restricted T cell epitope. In other embodiments, the epitope is an HLA class II-restricted T cell epitope.

[0044] [Table 1]

[0045] [Table 2]

[0046] [Table 3] TIFF0007844327000004.tif142167

[0047] Table 4 TIFF0007844327000006.tif50169

[0048] In some embodiments, peptides (e.g., polypeptides) comprising one or more epitopes from Tables 1, 2, 3, and / or 4 are provided herein. In some embodiments, the peptides disclosed herein are full-length viral proteins (e.g., full-length BKV and / or JCV proteins). In some embodiments, the peptides are not full-length viral proteins (e.g., not full-length BKV and / or JCV proteins). In some embodiments, the peptides disclosed herein comprise BKV and JCV epitopes having sequence homology (e.g., epitopes listed in Tables 2, 3, and 4). In some embodiments, the peptides disclosed herein comprise 100, 90, 80, 70, 60, 50, 40, 30, 25, 20, 15, or fewer than 10 consecutive amino acids of the viral protein. In some embodiments, the peptides disclosed herein comprise two or more epitopes listed in Tables 1, 2, 3, and / or 4. For example, in some embodiments, the peptides disclosed herein include two or more epitopes listed in Tables 1, 2, 3 and / or 4, linked by a polypeptide linker. In some embodiments, the peptides provided herein include at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, or 27 epitopes (for example, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, or 27 peptides listed in Tables 1, 2, 3 and / or 4). In preferred embodiments, the peptides disclosed herein include one or any combination thereof of the JCV epitopes listed in Table 1, i.e., the JCV epitopes listed in SEQ ID NOs: 1 to 21. For example, the peptide may include at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or all 21 epitopes encoded by the amino acid sequences listed in SEQ ID NOs: 1 to 21.

[0049] In certain embodiments, polypeptides (e.g., isolated polypeptides and / or recombinant polypeptides) comprising a plurality of epitopes derived from BKV or JCV antigens (e.g., epitopes derived from large T antigen (LTA), small T antigen (STA), or major capsid protein VP1 viral antigen, e.g., epitopes listed in Tables 1, 2, 3, or 4), preferably those listed in Table 1, are provided herein. More preferably, the polypeptides disclosed herein comprise one or any combination thereof of the JCV epitopes listed in SEQ ID NOs: 1 to 21. In some such embodiments, the polypeptide further comprises an intervening amino acid sequence between at least two of the plurality of epitopes. In some embodiments, the intervening amino acid or amino acid sequence is a proteasome liberation amino acid or amino acid sequence. Non-limiting examples of proteasome liberation amino acids or amino acid sequences are AD, K, or R, or include them. In some embodiments, the intervening amino acid or amino acid sequence is a TAP recognition motif. Typically, the TAP recognition motif is represented by the following formula: (R / N:I / Q:W / Y) n The formula may follow (wherein n is any integer of 1 or more). Non-limiting examples of TAP recognition motifs include RIW, RQW, NIW, and NQY. In some embodiments, the epitopes provided herein are linked or associated at the carboxyl terminus of each epitope by proteasome dissociated amino acid sequences, and optionally by TAP recognition motifs. In some such embodiments, the polypeptide comprises, or is essentially derived from, each epitope encoded by the amino acid sequences described in SEQ ID NOs: 1-21.

[0050] In some embodiments, the polypeptides provided herein further comprise epitopes derived from at least one additional virus (e.g., Epstein-Barr virus (EBV), cytomegalovirus (CMV), and / or adenovirus (ADV)). In some embodiments, the peptide comprises epitopes from two or more viruses. In some embodiments, the peptide comprises epitopes from three or more viruses. In some embodiments, the peptide comprises epitopes from four or more viruses. In some embodiments, the peptide comprises epitopes from five or more viruses. For example, in some embodiments, the peptide comprises sequences derived from at least two, three, four, or five of JCV, BKV, EBV, CMV, and / or ADV.

[0051] In some embodiments, polyepitope polypeptides containing two or more epitopes described herein (i.e., a single chain of amino acid residues containing multiple T cell epitopes that are not naturally linked) are provided herein. In some embodiments, the T cell epitopes in the polypeptide are linked via amino acid linkers. In some embodiments, the T cell epitopes in the polypeptide are linked directly without intervening amino acids. Examples of polyepitope polypeptides, methods for producing polyepitope polypeptides, and vectors encoding polyepitope polypeptides can be found in Dasari et al., Molecular Therapy-Methods & Clinical Development (2016) 3, 16058, which is incorporated herein by reference in its entirety.

[0052] In certain embodiments, a pool of immunogenic peptides is provided, comprising HLA class I and class II-restricted polyomavirus peptide epitopes (e.g., epitopes listed in Tables 1, 2, 3, 4, 5, and / or 6) that can induce the proliferation of peptide-specific T cells. In some embodiments, the immunogenic peptide pool includes at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, or 27 epitopes (e.g., at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, or 27 epitopes listed in Tables 1, 2, 3 and / or 4) or combinations thereof. In preferred embodiments, the peptide pool includes at least one JCV epitope listed in Table 1, i.e., any one of the JCV epitopes listed in Sequence IDs 1 to 21 or any combination thereof. For example, a pool of immunogenic peptides may contain at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or all 21 epitopes encoded by the amino acid sequences described in SEQ ID NOs: 1-21. Most preferably, such a peptide pool contains each of the JCV peptide epitope amino acid sequences described in SEQ ID NOs: 1-21. Immunogenic peptides and their pools are peptide-specific T cells (e.g., peptide-specific cytotoxic T cells and / or CD4). + It can induce the proliferation of T cells.

[0053] In some embodiments, the compositions and methods provided herein include or relate to natural variants of epitopes listed in Tables 1, 2, and / or 3. For example, in some embodiments, polyepitope polypeptides are provided herein that include two or more natural variants (e.g., at least 3, 4, 5, 6, 7, 8, 9, or 10) of epitopes listed in Tables 1, 2, and / or 3.

[0054] In some embodiments, the epitope sequences provided herein have the sequence disclosed herein with one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) conservative sequence modifications. As used herein, the term “conservative sequence modification” is intended to mean an amino acid modification that does not significantly affect or alter the interaction between the TCR and the peptide containing the amino acid sequence presented on the HLA. Such conservative modifications include amino acid substitutions, additions (e.g., addition of an amino acid to the N-terminus or C-terminus of a peptide) and deletions (e.g., deletion of an amino acid from the N-terminus or C-terminus of a peptide). A conservative amino acid substitution is one in which an amino acid residue is replaced by an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains are defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), amino acids with acidic side chains (e.g., aspartic acid, glutamic acid), amino acids with non-charged side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), amino acids with nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), amino acids with beta-branched side chains (e.g., threonine, valine, isoleucine), and amino acids with aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Therefore, one or more amino acid residues of the peptides described herein can be substituted with other amino acid residues from the same side chain family, and the modified peptides can be tested for retention of TCR binding (e.g., antigenicity) using methods known in the art. Modifications can be introduced into antibodies by standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis.

[0055] In some embodiments, the peptides (e.g., polypeptides) described herein are immunogenic and can induce an immune response upon administration to a subject (e.g., a mammalian subject, e.g., a human subject). In further embodiments, the peptides (e.g., polypeptides) described herein can induce an immune response after endogenous or exogenous processing and / or presentation of the peptide by immune cells (e.g., immune cells of the subject and / or immune cells derived from a donor, e.g., allogeneic PBMCs).

[0056] In some embodiments, cells presenting one or more peptides described herein (e.g., peptides comprising at least one epitope listed in Tables 1, 2, 3, and / or 4) are provided herein. In some embodiments, the cells are mammalian cells. In some embodiments, the cells are antigen-presenting cells (APCs) (e.g., antigen-presenting T cells, dendritic cells, B cells, macrophages, or artificial antigen-presenting cells, e.g., aK562 cells). Cells presenting the peptides described herein can be produced by standard techniques known in the art. For example, cells may be pulsed to promote peptide uptake. In some embodiments, cells are transfected with nucleic acids encoding the peptides provided herein. In some embodiments, a method for producing antigen-presenting cells (APCs) is provided herein, comprising the step of pulsing cells with the peptides described herein. An exemplary example of producing antigen-presenting cells can be found in WO2013088114, which is incorporated herein in its entirety.

[0057] The peptides provided herein can be isolated from cell or tissue sources by appropriate purification schemes using standard protein purification techniques, produced by recombinant DNA techniques, and / or chemically synthesized using standard peptide synthesis techniques. The peptides described herein can be produced in prokaryotic or eukaryotic host cells by expression of nucleotides encoding the peptide(s) of the present invention. Alternatively, such peptides can be synthesized by chemical methods. Methods for the expression of heterologous peptides in recombinant hosts, the chemical synthesis of peptides, and in vitro translation are well known in the art. Furthermore, the following are incorporated herein by reference: Maniatis et al., Molecular Cloning: A Laboratory Manual (1989), 2nd edition, Cold Spring Harbor, NY; Berger and Kimmel, Methods in Enzymology, Vol. 152; Guide to Molecular Cloning Techniques (1987), Academic Press, Inc., San Diego, Calif.; Merrifield, J. (1969) J. Am. Chem. Soc. 91:501; Chaiken IM (1981) CRC Crit. Rev. Biochem. 11:255; Kaiser et al. (1989) Science 243:187; Merrifield, B. (1986) Science 232:342; Kent, SBH (1988) Annu. Rev. This information is found in Biochem. 57:957, Offord, RE (1980) Semisynthetic Proteins, Wiley Publishing.

[0058] nucleic acid molecule Nucleic acid molecules encoding epitopes and peptides described herein are provided herein. Nucleic acids may exist, for example, in whole cells, in cell lysates, or in partially purified or substantially pure forms. The nucleic acid molecules described herein can be isolated using standard molecular biological techniques and sequence information provided herein. For example, oligonucleotides corresponding to one or more nucleotide sequences of epitopes listed in Tables 1, 2, 3, or 4 can be prepared by standard synthetic techniques, i.e., using an automated DNA synthesizer.

[0059] In some embodiments, vectors containing nucleic acid molecules described herein (e.g., viral vectors, e.g., adenovirus-based expression vectors) are provided herein. Viral vectors may include additional DNA segments that can be ligated to a viral genome. Certain vectors can autonomously replicate in the host cell into which they are introduced (e.g., bacterial vectors with bacterial origins of replication, episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) can be integrated into the host cell's genome upon introduction into the host cell and thereby replicate together with the host genome. Furthermore, certain vectors can direct gene expression. Such vectors are referred herein as “recombinant expression vectors” (or simply “expression vectors”). In some embodiments, nucleic acids functionally linked to one or more regulatory sequences (e.g., promoters) in an expression vector are provided herein. In some embodiments, cells transcribe the nucleic acids provided herein and thereby express antibodies, their antigen-binding fragments, or peptides described herein. The nucleic acid molecules may be integrated into the cell's genome, or they may be extrachromosomal.

[0060] In some embodiments, the nucleic acid vectors or recombinant adenoviruses provided herein encode one or more epitopes listed in Tables 1, 2, 3, and / or 4. For example, a nucleic acid vector or recombinant adenovirus may consist of one or more epitopes from the same table (e.g., one or more epitopes from Table 1, one or more epitopes from Table 2, one or more epitopes from Table 3, or one or more epitopes from Table 4). Alternatively, a nucleic acid vector or recombinant adenovirus may consist of one or more epitopes from the same table (e.g., Table 1) and one or more epitopes from a different table (e.g., Table 2). In some embodiments, in addition to the epitopes listed in Tables 1, 2, 3, or 4, the nucleic acid vectors or recombinant adenoviruses provided herein encode 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acids.

[0061] In some embodiments, the nucleic acid vector contains a codon-optimized nucleic acid sequence. In such embodiments, the coding sequence is constructed by modifying the codons in each nucleic acid used to assemble the coding sequence. Generally, a method for identifying a nucleotide sequence that optimizes codon usage for peptide production comprises at least the following steps (a) to (e): Step (a) provides an oligomer encoding a portion of a polypeptide, which contains a degenerate codon for the amino acid it encodes, together with an oligomer extended to yield an adjacent coding sequence with a duplicate sequence. Step (b) processes the oligomers so that assembly of the peptide coding sequence occurs. The reassembled peptide is placed in an expression system functionally linked to a control sequence and its expression is performed. Step (c) transfects a culture of compatible host cells with the expression system. Step (d) tests colonies obtained from the transformed host cells for polypeptide production levels. Step (e) obtains at least one colony from the expression system that shows the maximum or satisfactory polypeptide production. The sequence of the portion of the expression system encoding that protein is determined. A further explanation of codon optimization is provided in U.S. Patent Application Publication US2010 / 035768, which is incorporated herein by reference in its entirety.

[0062] antigen presenting cells In some embodiments, an APC is provided herein that presents one or more T cell epitopes (e.g., one or more T cell epitopes listed in Tables 1, 2, 3 and / or 4) (e.g., on HLA). In some embodiments, the HLA is a class I HLA. In some embodiments, the HLA is a class II HLA. In some embodiments, the class I HLA has an α-chain polypeptide which is HLA-A, HLA-B, HLA-C, HLA-E, HLA-F, HLA-g, HLA-K or HLA-L. In some embodiments, the class II HLA has an α-chain polypeptide which is HLA-DMA, HLA-DOA, HLA-DPA, HLA-DQA or HLA-DRA. In some embodiments, the class II HLA has a β-chain polypeptide which is HLA-DMB, HLA-DOB, HLA-DPB, HLA-DQB or HLA-DRB. In some embodiments, the APC has at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, or 38 T cell epitopes (e.g., Table 1, Table 2, Present at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, or 39 T cell epitopes listed in Table 3 and / or Table 4.

[0063] In some embodiments, the APC is a B cell, antigen-presenting T cell, dendritic cell, or artificial antigen-presenting cell (e.g., aK562 cell). Dendritic cells for use in this process can be prepared by collecting PBMCs from a patient sample and attaching them to plastic. Generally, the monocyte population remains, and all other cells can be washed away. The attached cell population is then differentiated with IL-4 and GM-CSF to produce monocyte-derived dendritic cells. These cells may be matured by adding IL-1β, IL-6, PGE-1 and TNF-α (which upregulate key costimulatory molecules on the surface of dendritic cells) and then contacted with the recombinant adenovirus described herein.

[0064] In some embodiments, the APC is an artificial antigen-presenting cell, such as aK562 cells. In some embodiments, the artificial antigen-presenting cell is engineered to express CD80, CD83, 41BB-L, and / or CD86. An example of an artificial antigen-presenting cell, such as aK562 cells, is described in U.S. Patent Application Publication 2003 / 0147869, incorporated herein by reference.

[0065] In certain embodiments, a method for producing an APC that presents two or more T cell epitopes as described herein is provided, comprising contacting the APC with a nucleic acid vector and / or recombinant adenovirus encoding a T cell epitope as described herein and / or a polyepitope produced by the nucleic acid vector or recombinant adenovirus as described herein. In some embodiments, the APC is irradiated.

[0066] T cells In certain embodiments, T cells and T cell populations (e.g., CD4 T cells and / or CD8 T cells) that express a TCR (e.g., an αβ TCR or a γδ TCR) that recognizes a peptide described herein presented on HLA (e.g., an epitope listed in Table 1, Table 2, Table 3, and / or Table 4) are provided herein. In some embodiments, the T cell is a CD8 T cell (CTL) that expresses a TCR that recognizes a peptide described herein presented on class I HLA. In some embodiments, the T cell is a CD4 T cell (helper T cell) that recognizes a peptide described herein presented on class II HLA. Most preferably, the present disclosure relates to the stimulation and expansion of multifunctional T cells that provide a more effective immune response against an epitope (e.g., an epitope listed in Table 1, Table 2, Table 3, and / or Table 4) than, for example, cells that produce only a single immune effector (e.g., a single biomarker, e.g., a cytokine or CD107a), i.e., T cells that can induce multiple immune effector functions. Multifunctionally impaired, single-functional, or even "exhausted" T cells can dominate the immune response during chronic infection or disease states (e.g., cancer) and thus may have an adverse effect on the treatment or prevention of viral complications. The functional competence and activity of such T cells may be further evaluated by determining the expression patterns (e.g., expression profiles by ICS assay) of transcription factors, such as T-bet and Eomes, and / or cytotoxic effector molecules, such as perforin and granzyme B. In some embodiments, the expression of each of T-bet, Eomes, perforin, and granzyme B is determined for the T cells disclosed herein. Such expression levels may be determined and evaluated as relative measurements, e.g., ratios. In preferred embodiments, the expression profiles of T-bet / Eomes and / or granzyme B / perforin are determined. In preferred embodiments, the T cells disclosed herein (e.g., JCV-specific T cells) have high expression of T-bet and low expression of Eomes (i.e., T-bet hi / Eomes low) shows, and similarly, the T cells disclosed herein exhibit high expression of granzyme B and low expression of perforin (i.e., granzyme hi / Perforin low ) may be shown. In some such embodiments, T-bet hi / Eomes low and / or Granzyme hi / Perforin low T cells exhibiting an expression profile (e.g., JCV-specific T cells) are identified as functionally responsive and active. Such T cells may be selected for use and / or proliferation in adoptive T cell immunotherapy. Most preferably, the T cells disclosed herein (e.g., JCV-specific T cells) are multifunctional (i.e., producing two or more cytokines as described herein) and T-bet hi / Eomes low and / or Granzyme hi / Perforin low The expression profile is shown.

[0067] In some embodiments, methods are provided herein for generating, activating, and / or inducing proliferation of T cells (e.g., CTLs) that recognize one or more of the epitopes described herein. In some embodiments, a sample containing CTLs (i.e., a PBMC sample) is incubated in a culture with an APC provided herein (e.g., an APC that presents a peptide containing the BKV and / or JCV epitopes described herein on a class I HLA complex). In some embodiments, a sample containing T cells is incubated two or more times with the APC provided herein. In some embodiments, T cells are incubated with the APC in the presence of at least one cytokine. In some embodiments, the cytokine is IL-4, IL-7, and / or IL-15. An exemplary method for inducing T cell proliferation using an APC is provided, for example, in U.S. Patent Application Publication 2015 / 0017723, which is incorporated herein by reference.

[0068] In some embodiments, a population of CTLs comprising collectively T cell receptors that recognize one or more T cell epitopes (e.g., one or more T cell epitopes listed in Table 1, Table 2, Table 3, and / or Table 4) is provided herein. In some embodiments, the CTLs recognize two or more T cell epitopes from Table 1, Table 2, Table 3, and / or Table 4. In some embodiments, the population of CTLs comprises collectively T cell receptors that recognize T cell epitopes derived from any combination of JCV, BKV, EBV, CMV, ADV, and / or other viruses. In some embodiments, a population of CTLs has at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, or 38 T cell epitopes (for example, at least T cell epitopes from Table 1). It collectively comprises T cell receptors that recognize 1, 2, 3, 4, 5, 6, or 7 epitopes and / or at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 epitopes listed in Table 1, Table 2, Table 3, and / or Table 4.

[0069] In some embodiments, methods are provided herein for preventing or treating polyomavirus infection (e.g., JCV infection) or cancer (e.g., polyomavirus-associated cancer, e.g., JVC-associated cancer) in a subject, comprising administering to the subject a composition (e.g., a therapeutic composition) comprising a nucleic acid vector as described herein, a peptide produced by the nucleic acid vector as described herein, CTLs and / or APCs provided herein (e.g., including the nucleic acid vector as described herein) and a pharmaceutically acceptable carrier. In some embodiments, the CTLs and / or APCs are not autologous to the subject (i.e., the CTLs and / or APCs are allogeneic to the subject). In some embodiments, the T cells and / or APCs are autologous to the subject. In some embodiments, the T cells and / or APCs are stored in a cell bank before being administered to the subject.

[0070] Pharmaceutical composition In some embodiments, compositions (e.g., pharmaceutical compositions such as vaccine compositions) containing peptides described herein (e.g., peptides containing epitopes from Table 1), nucleic acids, nucleic acid vectors, recombinant adenoviruses, antibodies, CTLs, or APCs, formulated with a pharmaceutically acceptable carrier, are provided herein, as well as methods for treating cancer (e.g., polyomavirus-associated cancer, e.g., JVC-associated cancer) or polyomavirus infection (e.g., JCV, CMV, EBV, or ADV infection) using such pharmaceutical compositions. In some embodiments, the composition comprises a combination of several (e.g., two or more) agents provided herein.

[0071] In some embodiments, the pharmaceutical composition further comprises an adjuvant. As used herein, the term “adjuvant” broadly refers to any agent that influences an immunological or physiological response in a patient or subject. For example, an adjuvant may increase the presence of an antigen over time or in a region of interest such as a tumor, help antigen-presenting cells absorb antigens, activate macrophages and lymphocytes, and support cytokine production. By altering the immune response, an adjuvant may allow a lower dose of an immunoassayant to increase the efficacy or safety of a particular dose of an immunoassayant. For example, an adjuvant may prevent T cell depletion and thus increase the efficacy or safety of a particular immunoassayant. Examples of adjuvants include, but are not limited to, immunomodulatory proteins, adjuvant 65, α-GalCer, aluminum phosphate, aluminum hydroxide, calcium phosphate, β-glucan peptides, CpG oligodeoxynucleotides, non-CpG oligodeoxynucleotides, GPI-0100, lipid A and its modified forms (e.g., monophosphorylated lipid A), lipopolysaccharides, lipovant, montanide, N-acetyl-muramyl-L-alanyl-D-isoglutamine, Pam3CSK4, quil A, TLR9 agonists, ODN1a, cationic antimicrobial peptides (CAMP), such as KLK, IC31, and trehalose dimycolate.

[0072] Methods for preparing these formulations or compositions involve combining the agents described herein with a carrier and optionally one or more auxiliary components. Generally, formulations are prepared by homogeneously and closely associating the agents described herein with a liquid carrier, a fine solid carrier, or both, and then shaping the product as necessary.

[0073] The pharmaceutical compositions of the present invention, suitable for parenteral administration, comprise one or more of the agents described herein in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders that can be reconstituted into sterile injection solutions or sterile injection dispersions immediately before use, and may also comprise sugars, alcohols, antioxidants, buffers, bacteriostatic agents, solutes that are isotonic with the blood of the recipient to whom the formulation is intended, or suspending agents or thickeners. Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions of the present invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils (such as olive oil), and organic esters for injection (such as ethyl oleate). Appropriate fluidity can be maintained, for example, by the use of coating materials such as lecithin, by maintaining the required particle size in the case of dispersions, and by the use of surfactants.

[0074] Regardless of the selected route of administration, the agents and / or pharmaceutical compositions of the present invention, which can be used in an appropriate hydrated form, are formulated into pharmaceutically acceptable dosage forms by conventional methods known to those skilled in the art.

[0075] Treatment method JCV sequence and / or protein expression can be observed in several malignancies and is frequently reported in immunocompromised (e.g., immunodeficiency, immunodysfunction, and / or immunosuppression) patients with or without PMLs. Therefore, in certain embodiments, methods for treating and / or preventing cancer (e.g., polyomavirus-associated cancer, e.g., JCV-associated cancer) or polyomavirus infection (e.g., JCV infection) are provided herein. In some embodiments, the method involves administering a pharmaceutical composition comprising the CTLs, APCs, polypeptides, and / or nucleic acid molecules described herein to the target.

[0076] In some embodiments, the subject being treated is immunocompromised. For example, in some embodiments, the subject has T-cell deficiency. In some embodiments, the subject has leukemia, lymphoma (e.g., Hodgkin lymphoma), or multiple myeloma. In some embodiments, the subject has multiple sclerosis, psoriasis, and / or other autoimmune diseases. In some embodiments, the subject is infected with HIV and / or has AIDS. In some embodiments, the subject has received a tissue, organ, and / or bone marrow transplant. In some embodiments, the subject has received immunosuppressive therapy, such as steroids, cell division inhibitors and antiproliferative agents, therapeutic antibodies, calcineurin inhibitors, anti-rejection drugs, etc., or a combination thereof. In some embodiments, the subject has received and / or is receiving chemotherapy. In some embodiments, the subject has received and / or is receiving radiotherapy.

[0077] In preferred embodiments, the subject suffers from a JCV infection in the central nervous system (e.g., reactivation of JCV infection or seeding of newly reactivated virus). In some such embodiments, the JCV infection is associated with the destruction of oligodendrocytes and / or white matter demyelination. In further embodiments, the subject suffers from JCV granule cell layer neuropathy (JCV GCN), JCV encephalopathy (JCV CPN / JCVE), JCV meningitis (JCVM), and / or progressive multifocal leukoencephalopathy (PML), preferably PML. In some such embodiments, the pathogen (e.g., JCV) is detectable in the subject's cerebrospinal fluid.

[0078] In some embodiments, the subject has cancer. In some embodiments, the methods described herein may be used to treat any cancerous or precancerous tumor. In some embodiments, the cancer expresses one or more polyomavirus epitopes provided herein (e.g., BKV / JCV epitopes listed in Tables 1, 2, 3 and / or 4). In some embodiments, the cancer is JVC-associated cancer. In some embodiments, the cancer includes solid tumors. Preferably, the cancer is a gastrointestinal malignancy, e.g., colon cancer, gastric cancer, gastrointestinal tumors, etc. Most preferably, the cancer is a CNS malignancy, e.g., glioma and all its subtypes (e.g., ependymoma, astrocytoma, brainstem glioma, oligodendroglioma, optic glioma, mixed glioma, etc.), medulloblastoma, primary neuroectodermal tumor, and neuroblastoma. Cancers that can be treated by methods and compositions provided herein, but not limited to, include cancer cells originating from the bladder, blood, bone, bone marrow, brain, breast, colon, esophagus, gastrointestinal tract, gums, head, kidney, liver, lung, nasopharynx, neck, ovary, prostate, skin, stomach, testis, tongue, or uterus. In addition, cancer can be of the following histological types, though not limited to: neoplasms, malignant; carcinoma; undifferentiated carcinoma; giant cell carcinoma and spindle cell carcinoma; small cell carcinoma; papillary carcinoma; squamous cell carcinoma; lymphoepithelial carcinoma; basal cell carcinoma; piloma carcinoma; transitional cell carcinoma; papillary transitional cell carcinoma; adenocarcinoma; gastrin-producing tumors, malignant; cholangiocarcinoma; hepatocellular carcinoma; combination of hepatocellular carcinoma and cholangiocarcinoma; cord-like adenocarcinoma; adenoid cystic carcinoma; adenocarcinoma in adenomatous polyps; adenocarcinoma; familial colonic polyposis; solid tumors; carcinoid tumors, malignant; bronchioloalveolar adenocarcinoma; papillary adenocarcinoma; pigmentaphobic carcinoma; eosinophil carcinoma; eosinophilic adenocarcinoma Adenocarcinoma, basophilic carcinoma, clear cell adenocarcinoma, granular cell carcinoma, follicular adenocarcinoma, papillary adenocarcinoma and follicular adenocarcinoma, non-encapsulated sclerosing carcinoma, adrenocortical carcinoma, endometrioid carcinoma, cutaneous adnexal carcinoma, apocrine gland carcinoma, sebaceous gland carcinoma, ceruminous gland carcinoma, mucosal epidermal carcinoma, cystadenocarcinoma, papillary cystadenocarcinoma, papillary serous cystadenocarcinoma, mucinous cystadenocarcinoma, mucinous adenocarcinoma, signet ring cell carcinoma, invasive ductal carcinoma, medullary carcinoma, lobular carcinoma, inflammatory carcinoma, Paget's disease of the breast, acinar cell carcinoma, adenosquamous cell carcinoma, adenocarcinoma with squamous metaplasia, malignant thymoma, malignant ovarian stromal carcinoma, malignant theca cell carcinoma, malignant granulosa cell carcinoma, malignant androgen-producing cell carcinoma,Sertoli cell tumor, malignant Leydig cell tumor, malignant lipid cell tumor, malignant paraganglioma, malignant extramammary paraganglioma, pheochromocytoma, angioglobulosarcoma, malignant melanoma, achromatic melanoma, superficial spreading melanoma, malignant melanoma in giant pigmented nevi, epithelioid cell melanoma, malignant blue nevus, sarcoma, fibrosarcoma, malignant fibrous histiocytoma, myxosarcoma, liposarcoma, leiomyosarcoma, rhabdomyosarcoma, embryonal rhabdomyosarcoma, alveolar rhabdomyosarcoma, stromal sarcoma, malignant mixed tumor, Müllerian mixed tumor, kidney Hepatoblastoma, carcinosarcoma, malignant mesenchymal tumor, malignant Brenner tumor, malignant phyllodes tumor, synovial sarcoma, malignant mesothelioma, undifferentiated germ cell tumor, fetal carcinoma, malignant teratoma, malignant ovarian goiter, choriocarcinoma, malignant mesonephroma, angiosarcoma, malignant hemangioendothelioma, Kaposi's sarcoma, malignant hemangioectocytoma, lymphangiosarcoma, osteosarcoma, paraosteal osteosarcoma, chondrosarcoma, malignant chondroblastoma, mesenchymal chondrosarcoma, giant cell tumor of bone, Ewing's sarcoma, malignant odontogenic tumor, ameloblastic odontosarcoma, malignant ameloblastoma, ameloblastoma Physofibrosarcoma, malignant pineal tumor, chordoma, malignant glioma, ependymoma, astrocytoma, protoplasmic astrocytoma, fibrous astrocytoma, astroblastoma, glioblastoma, glioblastoma pleomorphic, glioblastoma (pineal tumor), oligodendroglioblastoma, oligodendroglioblastoma, primitive neuroectodermal, cerebellar sarcoma, ganglioblastoma, neuroblastoma, retinoblastoma, olfactory neurogenic tumor, malignant meningioma, neurofibrosarcoma, malignant schwannoma, malignant granulocyte, malignant lymphoma, Hodgkin's disease, Hodgkin's lymphoma, flank tissue Lymphoma, small lymphocytic lymphoma, diffuse large cell lymphoma, follicular lymphoma, mycosis fungoides, other designated non-Hodgkin lymphomas, malignant histiocytosis, multiple myeloma, mast cell sarcoma, immunoproliferative bowel disease, leukemia, lymphocytic leukemia, plasma cell leukemia, erythroleukemia, lymphosarcoma, myeloid leukemia, basophilic leukemia, eosinophilic leukemia, monocytic leukemia, mast cell leukemia, megakaryoblastic leukemia, myelosarcoma, and hairy cell leukemia.

[0079] In some embodiments, subjects are also administered antiviral drugs that inhibit the replication of polyomaviruses. For example, in some embodiments, subjects are administered ganciclovir, valganciclovir, foscarnet, cidofovir, acyclovir, holmivirsen, maribavir, BAY 38-4766, or GW275175X.

[0080] In some embodiments, the subjects are also administered immune checkpoint inhibitors. Immune checkpoint inhibition broadly refers to inhibiting checkpoints that cancer cells can produce to prevent or downregulate the immune response. Examples of immune checkpoint proteins include, but are not limited to, CTLA4, PD-1, PD-L1, PD-L2, A2AR, B7-H3, B7-H4, BTLA, KIR, LAG3, TIM-3, or VISTA. Immune checkpoint inhibitors may be antibodies or antigen-binding fragments thereof that bind to and inhibit immune checkpoint proteins. Examples of immune checkpoint inhibitors include atezolizumab, avelumab, camrelizumab, semiprimab, cetrerimab, durvalumab (MEDI-4736), genolimuzumab, ipilimumab, nivolumab, pembrolizumab, pizilizumab, cintilimab, spartalizumab, tislerizumab, tripalimab, AMP-224, AMP-514, AK-104, and ASP-8374. Examples include, but are not limited to, AUR-012, BCD-135, BGB-A333, BMS-936559, CBT-502, MCLA-145, KN-046, MGD-019, MK-4830, MSB-0020718C, RG-7446, SL-279252, STI-A1010, STI-A1110, TSR-042, XmAb20717, and XmAb23104.

[0081] In some embodiments, the compositions provided herein are administered prophylactically to prevent cancer and / or polyomavirus infection (e.g., JCV infection). In some embodiments, the compositions may be administered before or after the detection of cancer cells or polyomavirus-infected cells in a subject. Accordingly, in some such embodiments, the compositions provided herein are administered before or after the administration of immunosuppressive therapy (e.g., steroids, cell division inhibitors and antiproliferative agents, therapeutic antibodies, calcineurin inhibitors, anti-rejection agents, etc., or combinations thereof). In some such embodiments, the compositions are administered before or after chemotherapy. Similarly, in some embodiments, the compositions are administered before or after radiotherapy. In some embodiments, a pro-inflammatory response is induced after administration of a composition comprising the peptides, nucleic acids, CTLs and / or APCs described herein. The pro-inflammatory immune response includes the production of pro-inflammatory cytokines and / or chemokines, such as interferon-gamma (IFN-γ) and / or interleukin-2 (IL-2).

[0082] Conjunctive therapy involves sequential, simultaneous, separate, and / or co-administration of active compounds in such a manner that the therapeutic effect of the administered first agent is not completely lost when subsequent treatments are administered. In some embodiments, the second agent may be co-formulated with the first agent or formulated in separate pharmaceutical compositions.

[0083] In some embodiments, methods for identifying subjects suitable for the treatments provided herein (methods for treating polyomavirus infection, JCV infection and / or cancer in a subject, including, for example, administering pharmaceutical compositions provided herein to the subject) are provided herein. In some embodiments, the method includes isolating a sample (e.g., a blood sample, a tissue sample, a tumor sample) from the subject and detecting the presence of epitopes listed in Tables 1, 2, or 3 in the sample. In some embodiments, the epitopes are detected using ELISA assays, Western blot assays, FACS assays, fluorescence microscopy assays, Edman degradation assays and / or mass spectrometry assays (e.g., protein sequencing). In some such embodiments, for example, the presence of a JCV epitope is detected by detecting the nucleic acid encoding the JCV epitope. In some embodiments, the nucleic acid encoding the JCV epitope is detected using nucleic acid probes, nucleic acid amplification assays and / or sequencing assays. Notably, the JC virus genome consists of two conserved coding regions separated by a highly variable non-coding regulatory region (NCCR) that contains both the sequence necessary for replication (the ORI, which is the origin of viral replication) and the sequence necessary for transcription (several promoters and cis regulatory elements). The highly conserved region containing the ORI is followed by sections a, b, c, d, e, and f. JC viruses found in the CNS of PML patients are often found to have rearranged NCCRs (e.g., the absence of sections b and d and duplication of the ace sequence). Differences in NCCR sequences can be a factor in the compatibility of the virus in the CNS and, therefore, in the development of PML. Accordingly, a method for identifying subjects suitable for the treatment provided herein is provided, which includes isolating a sample (e.g., blood, urine, tissue, cerebrospinal fluid, tumor) from the subject and detecting the presence of PML-associated JCV sequence rearrangements (e.g., using nucleic acid amplification techniques such as nested PCR). Such sequences and detection methods are known in the art and are described, for example, in L' Honner et al., PLoS ONE, 13(6), 2018, which is incorporated in whole by reference.

[0084] In some embodiments, the method includes determining the HLA type of the subject. In some embodiments, the subject is identified as suitable for treatment by the method provided herein if the subject expresses HLA bound to the epitope provided herein. In some embodiments, the method provided herein further includes treating the identified subject using the therapeutic method provided herein (for example, by administering the pharmaceutical composition provided herein to the subject). In some embodiments, the subject is administered a composition comprising a CTL described herein, the CTL comprising a TCR that recognizes the HLA-bound epitope provided herein to the HLA expressed by the subject. In some embodiments, the subject is administered a composition comprising a polypeptide comprising the HLA-bound epitope provided herein to the HLA expressed by the subject. In some embodiments, the subject is administered a composition comprising an APC that displays the polypeptide comprising the HLA-bound epitope provided herein to the HLA expressed by the subject. In some embodiments, the subject is administered a composition comprising a nucleic acid encoding a polypeptide comprising the HLA-bound epitope provided herein to the HLA expressed by the subject. [Section 1] A peptide containing one or more of the epitopes listed in Tables 1-4. [Section 2] The peptide described in item 1, wherein one or more epitopes contain the JC virus (JCV) epitopes listed in Table 1. [Section 3] The peptide according to item 1 or 2, wherein one or more epitopes contain the JCV epitopes described in SEQ ID NOs: 1 to 21. [Section 4] The peptide described in item 1, wherein one or more epitopes contain JC virus (JCV) epitopes listed in Table 2 and / or Table 3. [Section 5] The peptide described in item 1, wherein one or more epitopes include hybrid epitopes as shown in Table 4. [Section 6] A peptide described in any one of items 1 to 5, wherein the peptide contains multiple epitopes listed in Tables 1 to 4. [Section 7] The peptide described in item 6, comprising multiple epitopes, including multiple JCV epitopes listed in Table 1. [Section 8] The peptide according to item 7, wherein multiple epitopes include multiple JCV epitopes described in SEQ ID NOs: 1 to 21. [Section 9] The peptide described in item 5, wherein the multiple epitopes include multiple JCV epitopes listed in Table 2 and / or Table 3. [Section 10] The peptide described in item 6, wherein the multiple epitopes include the JVC epitopes listed in Table 1 and the JCV epitopes listed in Table 2 and / or Table 3. [Section 11] A peptide according to any one of items 6 to 10, further comprising an intervening amino acid sequence between at least two of multiple epitopes. [Section 12] A peptide according to any one of items 1 to 11, wherein the peptide can induce an immune response when administered to a target. [Section 13] A peptide as described in any one of sections 1 to 12, wherein the epitope is selected to provide broad coverage in the human population. [Section 14] The peptide according to claim 13, wherein the epitope has HLA class II restriction to HLA-DP, -DM, -DOA, -DOB, -DQ, or -DR. [Section 15] The peptide according to item 14, wherein the epitope has HLA class II restriction to HLA-DRB or -DQB. [Section 16] A peptide according to any one of items 1 to 15, containing each of the epitope amino acid sequences described in SEQ ID NOs: 1 to 21. [Section 17] A peptide described in any one of items 1 to 16, which essentially consists of the epitope amino acid sequences described in SEQ ID NOs: 1 to 21. [Section 18] The peptide is one of the peptides described in any one of the items 1 to 17, wherein the peptide consists of the epitope amino acid sequences described in SEQ ID NOs: 1 to 21. [Section 19] The peptide is the peptide according to any one of items 1 to 18, further comprising one or more epitopes derived from Merkel cell virus (MCV). [Section 20] A peptide according to any one of items 1 to 19, further comprising one or more epitopes derived from a non-polyomavirus. [Section 21] The peptide according to item 20, wherein one or more epitopes derived from a non-polyomavirus contain one or more epitopes derived from adenovirus (ADV), Epstein-Barr virus (EBV), or cytomegalovirus (CMV). [Section 22] An isolated nucleic acid encoding a peptide as described in any one of items 1 to 21. [Section 23] An expression construct comprising the isolated nucleic acid described in item 22. [Section 24] A host cell containing the expression construct described in item 23. [Section 25] A method for producing a peptide, comprising expressing the peptide in a host cell as described in item 24, and purifying the peptide at least partially. [Section 26] A pharmaceutical composition comprising a peptide as described in any one of items 1 to 21, and a pharmaceutically acceptable carrier. [Section 27] A pharmaceutical composition comprising the isolated nucleic acid described in item 22. [Section 28] A vaccine composition comprising a peptide as described in any one of items 1 to 21, and a pharmaceutically acceptable carrier. [Section 29] The vaccine composition according to item 28, further comprising an adjuvant. [Section 30] A method for treating or preventing polyomavirus infection in a subject, comprising administering to the subject a pharmaceutical composition described in item 26 or 27 or a vaccine composition described in item 28 or 29. [Section 31] The method described in paragraph 29, wherein the polyomavirus infection is a JC virus (JCV) infection. [Section 32] The method according to item 30 or 31, wherein the subject has JCV granule cell layer neuropathy (JCV GCN), JCV encephalopathy (JCVE), JCV meningitis (JCVM), and / or progressive multifocal leukoencephalopathy (PML). [Section 33] A method for treating or preventing polyomavirus-related cancer in a subject, comprising administering to the subject a pharmaceutical composition described in item 26 or 27 or a vaccine composition described in item 28 or 29. [Section 34] The method according to item 33, wherein polyomavirus-associated cancer is JCV-associated cancer. [Section 35] The method according to item 33 or 34, wherein the polyomavirus-associated cancer is a gastrointestinal malignancy, such as colon cancer, gastric cancer, and / or gastrointestinal tumor. [Section 36] The method according to item 33 or 34, wherein the polyomavirus-associated cancer is a central nervous system (CNS) malignancy, such as glioma, medulloblastoma, primary neuroectodermal tumor and / or neuroblastoma. [Section 37] A method for inducing a T-cell immune response in a subject, comprising administering to the subject a pharmaceutical composition described in item 26 or 27 or a vaccine composition described in item 28 or 29. [Section 38] A pool of immunogenic peptides comprising HLA class I and / or class II-restricted JCV peptide epitopes capable of inducing peptide-specific T cell proliferation, and comprising at least one or a combination thereof of the epitope amino acid sequences described in SEQ ID NOs: 1-21. [Section 39] A pool of immunogenic peptides as described in item 38, further comprising at least one or a combination thereof of the JCV peptide epitope amino acid sequences listed in Table 1. [Section 40] A pool of immunogenic peptides as described in section 38 or 39, comprising each of the JCV peptide epitope amino acid sequences described in SEQ ID NOs: 1 to 21. [Section 41] A pool of immunogenic peptides according to any one of claims 38 to 40, wherein the epitope has HLA class II restriction to HLA-DP, -DM, -DOA, -DOB, -DQ, or -DR. [Section 42] A pool of immunogenic peptides as described in item 41, wherein the epitope has HLA class II restriction to HLA-DRB or -DQB. [Section 43] A pool of immunogenic peptides described in any one of sections 38 to 42, wherein each epitope is constrained by one of the HLA specificities selected from DRB1*01:01, DRB1*03:01, DRB1*04:01, DRB1*10:01, DRB1*11:01, DRB1*13:01, DRB1*14:04, DRB1*15:01, DRB1*16:01, DQB1*02:02, or DQB1*05:03. [Section 44] A pool of immunogenic peptides described in any one of sections 38-43, wherein peptide-specific T cells exhibit a multifunctional immune effector profile. [Section 45] A pool of immunogenic peptides according to any one of items 38 to 44, wherein the immunogenic peptide can induce proliferation of peptide-specific cytotoxic T cells (CTLs). [Section 46] A pool of immunogenic peptides described in Section 45, in which peptide-specific cytotoxic T cells (CTLs) exhibit a multifunctional immune effector profile. [Section 47] A method for increasing JC virus-specific T cells for adoptive immunotherapy, (i) Contacting one or more cells isolated from the subject with a pool of peptides described in any one of items 1 to 21 or immunogenic peptides described in any one of items 38 to 46, and (ii) Culturing the one or more cells under conditions such that JC virus-specific T cells proliferate from the one or more cells. A method that includes this. [Section 48] The method according to item 47, wherein the pool of peptides or immunogenic peptides essentially consists of each of the JCV peptide epitope amino acid sequences described in SEQ ID NOs: 1 to 21. [Section 49] The method according to item 47 or 46, wherein one or more cells isolated from the subject include peripheral blood mononuclear cells (PBMCs) from a healthy donor. [Section 50] The method according to paragraph 47 or 46, wherein one or more cells isolated from the subject include PBMCs from an immunodeficient donor. [Section 51] The donor is receiving immunosuppressive therapy, as described in item 50. [Section 52] The method according to either paragraph 50 or 51, wherein the donor is an organ transplant recipient. [Section 53] The method described in any one of paragraphs 50 to 52, wherein the donor is a donor receiving antiviral therapy. [Section 54] The method according to any one of items 47 to 53, wherein JC virus-specific T cells exhibit a multifunctional immune effector profile. [Section 55] The method according to any one of claims 47 to 54, further comprising administering JC virus-specific T cells to a subject suffering from JCV infection. [Section 56] A method for treating or preventing JCV infection in a subject, comprising administering JC virus-specific T cells as described in any one of paragraphs 47 to 54. [Section 57] CTLs prepared by the method described in any one of items 47 to 54. [Section 58] A method for treating or preventing JCV infection in a subject, including administering the CTLs described in item 57. [Section 59] The method according to item 58, wherein exposure to an immunogenic peptide or a pool of immunogenic peptides induces stimulation and proliferation of JCV peptide-specific T cells. [Section 60] The method according to item 58, wherein the CTL administered to the subject is autologous. [Section 61] The method described in paragraph 58, wherein the CTL administered to the subject is not autologous. [Section 62] The method according to any one of items 56 to 61, wherein the infection is a recurrent JCV infection. [Section 63] The method according to any one of items 56 to 62, wherein the JCV infection is drug-resistant. [Section 64] The method described in any one of paragraphs 56 to 63, wherein the subject is an organ transplant recipient. [Section 65] The method according to any one of paragraphs 56-64, wherein the subject has JCV granule cell layer neuropathy (JCV GCN), JCV encephalopathy (JCVE), JCV meningitis (JCVM), and / or progressive multifocal leukoencephalopathy (PML). [Section 66] A method for treating or preventing polyomavirus-associated cancer in a subject, comprising administering proliferated JC virus-specific T lymphocytes described in any one of paragraphs 45 to 54. [Section 67] The method according to item 66, wherein polyomavirus-associated cancer is JCV-associated cancer. [Section 68] A method for detecting JC virus infection in a subject, comprising detecting the presence of JCV-specific T lymphocytes by contacting T lymphocytes isolated from the subject with a peptide described in any one of items 1 to 21. [Section 69] The method according to claim 68, further comprising detecting JCV-specific DNA in a cerebrospinal fluid sample isolated from a subject. [Section 70] The method according to claim 69, wherein the JCV-specific DNA includes the absence of sequences b and d in the non-coding control region (NCCR) and the duplication of sequence ace. [Section 71] The method according to any one of claims 68 to 70, further comprising administering to an adoptive immunotherapy composition comprising JC virus-specific T cells that target any one or a combination thereof of the epitope amino acid sequences described in SEQ ID NOs: 1 to 21. [Section 72] The method described in any one of paragraphs 30-37 or 47-71, wherein the subject is a mammal. [Section 73] The method described in paragraph 72, wherein the subject is a human. [Section 74] The method according to paragraph 72 or 73, wherein the subject is immunocompromised. [Section 75] A method for treating or preventing cancer in a subject, comprising administering a pharmaceutical composition containing cytotoxic T cells (CTLs) that include a T cell receptor (TCR) that recognizes one or more epitopes listed in Tables 1-4 to the subject. [Section 76] The method according to item 75, wherein one or more epitopes include JC virus (JCV) epitopes listed in Table 1. [Section 77] The method according to item 75, wherein one or more epitopes include JC virus (JCV) epitopes described in SEQ ID NOs: 1 to 21. [Section 78] The method according to item 75 or 76, wherein one or more epitopes include JC virus (JCV) epitopes listed in Table 2 and / or Table 3. [Section 79] The method according to any one of items 76 to 78, wherein one or more epitopes include hybrid epitopes listed in Table 4. [Section 80] The method according to any one of paragraphs 76 to 79, wherein the cancer is a polyomavirus-associated cancer. [Section 81] The method according to item 80, wherein the polyomavirus is JC virus (JCV). [Section 82] The method according to item 80 or 81, wherein the polyomavirus-associated cancer is a gastrointestinal malignancy, such as colon cancer, gastric cancer, and / or gastrointestinal tumor. [Section 83] The method according to item 80 or 81, wherein the polyomavirus-associated cancer is a central nervous system (CNS) malignancy, such as glioma, medulloblastoma, primary neuroectodermal tumor and / or neuroblastoma. [Section 84] A method for treating or preventing polyomavirus infection in a subject, comprising administering a pharmaceutical composition containing cytotoxic T cells (CTLs) that include one or more T cell receptors (TCRs) that recognize one or more epitopes listed in Tables 1-4 to the subject. [Section 85] The method according to item 84, wherein one or more epitopes include JC virus (JCV) epitopes listed in Table 1. [Section 86] The method according to item 85, wherein one or more epitopes include JC virus (JCV) epitopes described in SEQ ID NOs: 1 to 21. [Section 87] The method according to any one of items 84 to 86, wherein one or more epitopes include JC virus (JCV) epitopes listed in Table 2 and / or Table 3. [Section 88] The method according to any one of items 84 to 87, wherein one or more epitopes include hybrid epitopes listed in Table 4. [Section 89] The method according to any one of items 84 to 88, wherein the polyomavirus is JC virus (JCV). [Section 90] The method according to any one of items 75-89, wherein at least one TCR recognizes a VP1 epitope derived from JCV. [Section 91] The method according to any one of items 75-90, wherein at least one TCR recognizes an LTA epitope derived from JCV. [Section 92] The method according to any one of items 75-91, wherein at least one TCR recognizes an STA epitope derived from JCV. [Section 93] The method according to any one of paragraphs 75 to 92, wherein the CTLs collectively comprise TCRs that recognize at least two of the epitopes described in sequence numbers 1 to 21. [Section 94] The method according to paragraph 93, wherein the CTLs collectively comprise TCRs that recognize at least five of the epitopes described in sequence numbers 1 to 21. [Section 95] The method according to paragraph 94, wherein the CTL collectively comprises a TCR that recognizes at least 10 of the epitopes described in sequence numbers 1 to 21. [Section 96] The method according to paragraph 95, wherein the CTL collectively comprises TCRs that recognize each of the epitopes described in sequence numbers 1 to 21. [Section 97] The method according to any one of paragraphs 75 to 96, wherein the TCR collectively recognizes epitopes derived from at least two different viruses. [Section 98] The method according to paragraph 97, wherein the TCR collectively recognizes epitopes derived from at least three different viruses. [Section 99] The method according to paragraph 98, wherein the TCR collectively recognizes at least four different virus-derived epitopes. [Section 100] The method according to paragraph 99, wherein the TCR collectively recognizes at least five different virus-derived epitopes. [Section 101] The method according to any one of items 75 to 100, wherein the TCR collectively recognizes one or more epitopes derived from non-polyomaviruses. [Section 102] The method according to item 101, wherein one or more epitopes derived from a non-polyomavirus include one or more epitopes derived from adenovirus (ADV), Epstein-Barr virus (EBV), or cytomegalovirus (CMV). [Section 103] The method according to any one of items 75 to 102, wherein the subject expresses a human leukocyte antigen (HLA) in which one or more epitopes are constrained. [Section 104] The method according to any one of paragraphs 75 to 103, wherein the CTL is autologous to the subject. [Section 105] The method according to any one of paragraphs 75 to 103, wherein the CTL is not autologous to the subject. [Section 106] The method according to paragraph 105, wherein CTLs are obtained from a CTL library or bank. [Section 107] The method described in any one of items 75 to 106, wherein the subject is immunocompromised. [Section 108] The method according to any one of sections 75-107, wherein the CTL exhibits a multifunctional immune effector profile. [Section 109] A method for inducing the proliferation of polyomavirus-specific cytotoxic T cells (CTLs), comprising contacting CTLs with antigen-presenting cells (APCs) that present one or more polyomavirus peptides containing one of the epitopes listed in Tables 1-4. [Section 110] The method according to item 109, wherein one or more peptides contain the JC virus (JCV) epitopes listed in Table 1. [Section 111] The method according to item 110, wherein one or more peptides contain a JC virus (JCV) epitope selected from any one of SEQ ID NOs. 1 to 21. [Section 112] The method according to any one of items 109 to 111, wherein one or more peptides contain a JC virus (JCV) epitope listed in Table 2 and / or Table 3. [Section 113] The method according to any one of items 109 to 112, wherein one or more peptides contain a hybrid epitope listed in Table 4. [Section 114] The method according to any one of paragraphs 109 to 113, wherein a CTL is brought into contact with an APC in vitro. [Section 115] The method according to any one of claims 109 to 114, wherein one or more peptides contain one or more epitopes derived from non-polyomaviruses. [Section 116] The method according to item 115, wherein one or more peptides derived from a non-polyomavirus contain one or more epitopes derived from adenovirus (ADV), Epstein-Barr virus (EBV), or cytomegalovirus (CMV). [Section 117] The method according to any one of items 109 to 116, wherein the CTL is contacted with the APC in the presence of one or more cytokines. [Section 118] The method according to any one of items 109 to 117, wherein the APC contains B cells. [Section 119] The method according to any one of items 109 to 118, wherein the APC includes antigen-presenting T cells. [Section 120] The method according to any one of items 109 to 119, wherein the APC includes dendritic cells. [Section 121] The method according to any one of items 109 to 120, wherein the APC contains aK562 cells. [Section 122] The method according to any one of items 109 to 121, wherein the CTLs are derived from a sample of peripheral blood mononuclear cells (PBMCs). [Section 123] The method according to any one of items 109 to 122, wherein polyomavirus-specific cytotoxic T cells are stored in a CTL library or bank. [Section 124] The method according to any one of items 109 to 123, wherein polyomavirus-specific cytotoxic T cells exhibit a multifunctional immune effector profile. [Section 125] A method for treating or preventing cancer in a subject, comprising administering a vaccine composition containing one or more epitopes listed in Tables 1-4 to the subject. [Section 126] The method according to item 125, wherein one or more epitopes include JC virus (JVC) epitopes listed in Table 1. [Section 127] The method according to item 126, wherein one or more epitopes include JC virus (JVC) epitopes described in SEQ ID NOs: 1 to 21. [Section 128] The method according to either item 125 or 127, wherein one or more epitopes include JC virus (JCV) epitopes listed in Table 2 and / or Table 3. [Section 129] The method according to any one of items 125 to 128, wherein one or more epitopes include hybrid epitopes listed in Table 4. [Section 130] The method according to any one of paragraphs 125 to 129, wherein the cancer is a polyomavirus-associated cancer. [Section 131] The method according to paragraph 130, wherein the polyomavirus-associated cancer is a gastrointestinal malignancy, such as colon cancer, gastric cancer, and / or gastrointestinal tumor. [Section 132] The method according to paragraph 130, wherein the polyomavirus-associated cancer is a central nervous system (CNS) malignancy, such as glioma, medulloblastoma, primary neuroectodermal tumor and / or neuroblastoma. [Section 133] The method according to any one of items 130 to 132, wherein the polyomavirus is JC virus (JCV). [Section 134] A method for treating or preventing polyomavirus infection in a subject, comprising administering a vaccine composition containing one or more epitopes listed in Tables 1-4 to the subject. [Section 135] The method according to item 134, wherein one or more epitopes include JC virus (JCV) epitopes listed in Table 1. [Section 136] The method according to item 135, wherein one or more epitopes contain the JC virus (JCV) epitopes described in SEQ ID NOs: 1 to 21. [Section 137] The method according to any one of items 134 to 136, wherein one or more epitopes include JC virus (JCV) epitopes listed in Table 2 and / or Table 3. [Section 138] The method according to any one of items 134 to 137, wherein one or more epitopes include hybrid epitopes listed in Table 4. [Section 139] The method according to any one of items 134 to 138, wherein the polyomavirus is JC virus (JCV). [Section 140] The method according to any one of claims 125 to 139, wherein the vaccine composition further comprises one or more epitopes derived from a non-polyomavirus. [Section 141] The method according to item 140, wherein one or more epitopes derived from a non-polyomavirus include one or more epitopes derived from adenovirus (ADV), Epstein-Barr virus (EBV), or cytomegalovirus (CMV). [Section 142] The method according to any one of claims 125 to 141, wherein one or more epitopes include at least two of the epitopes described in SEQ ID NOs. 1 to 21. [Section 143] The method according to claim 142, wherein one or more epitopes include at least five of the epitopes described in SEQ ID NOs: 1 to 21. [Section 144] The method according to paragraph 143, wherein one or more epitopes include at least 10 of the epitopes described in SEQ ID NOs: 1 to 21. [Section 145] The method according to paragraph 144, wherein one or more epitopes include at least each of the epitopes described in Sequence IDs 1 to 21. [Section 146] The method according to any one of items 125 to 145, wherein the subject expresses a human leukocyte antigen (HLA) in which one or more epitopes are constrained. [Section 147] The method according to any one of claims 125 to 146, wherein the vaccine composition further comprises an adjuvant. [Section 148] The method according to any one of paragraphs 125 to 147, wherein the subject is immune deficiency, immune dysfunction, or immunodeficiency. [Section 149] The method described in any one of items 75 to 148, wherein the subject is a human. [Section 150] ⏎ The method according to item 47, wherein cells are cultured in the presence of IL-2. [Section 151] The method according to item 150, wherein IL-2 is present at a concentration of approximately 120 IU / ml. [Section 152] The method according to any one of items 109 to 123, further comprising culturing CTLs in the presence of IL-2. [Section 153] The method described in item 152, wherein IL-2 is present at a concentration of approximately 120 IU / ml.

[0085] <s [Example] [Example 1] CD8 + and CD4 + T cell responses PBMC from 17 healthy volunteers were incubated with JVC overlapping peptide pools (OPP), and these cells were cultured for 14 days in the presence of IL-2. The peptide matrices for each of the large T antigen (LTA), small T antigen (STA), and viral protein 1 (VP1), and the composition of the peptide pools for each matrix were arranged as follows.

[0086] TIFF0007844327000007.tif104168

[0087] TIFF0007844327000008.tif65155

[0088] TIFF0007844327000009.tif53149

[0089] On the 14th day, these T cell cultures were evaluated for JVC specificity using an intracellular cytokine (ICS) assay. Notably, 14 days of in vitro culture of T cells with JVC peptide resulted in the proliferation of virus-specific T cells. These initial analyses clearly showed that the T cell response was directed towards LTA, VP1, and STA (see Figure 1).

[0090] [Example 2] JCV epitope HLA restriction To accurately map HLA class I and class II restricted T cell responses, individual overlapping peptides (15 amino acids long with a 10 amino acid overlap) regarding LTA, STA, and VP1 proteins were procured for T cell epitope mapping. Using a two-dimensional peptide matrix, all individual peptides were distributed into small overlapping peptide pools. For example, in the case of the large T antigen, the matrix was set such that each peptide of the pool (LTA1 - LTA24) appears once on the ordinate. The T cell response for each pool was measured by an intracellular cytokine staining (ICS) IFN-γ assay (see Figure 2A), and the data was overlaid on a two-dimensional matrix as follows.

[0091] TIFF0007844327000010.tif99159

[0092] Thus, individual peptides common to the pools that induced T cell responses were identified: peptide 32 (P32) at the intersection of row LTA4 and column LTA16, and peptides P29 and P30 at the intersections of row LTA3 and columns LTA23 and LTA24. Fluorescence-activated cell sorting (FACS) confirmed that the individual peptides P29, P30, and P32 induced JCV-specific T cell responses (see Figure 2B).

[0093] These individual peptides were further evaluated for T cell proliferation and ICS analysis to identify potential JCV antigens. The resulting peptides provide high HLA allele coverage against JCV (see Figure 3 and Table 5).

[0094] [Table 5]

[0095] [Example 3] JCV-specific T cell proliferation and characterization JCV-specific T cells were stimulated with a pooled JCV epitope and then proliferated in vitro. Specifically, PBMCs from healthy volunteers were stimulated with synthetic JCV peptide (Table 1) for 1 hour, and then cultured for 12–14 days in the presence of different cytokine combinations including IL-2 (10 ng / ml), IL-7 (10 ng / ml), IL-12 (10 ng / ml), and / or IL-15 (10 ng / ml). The JCV specificity of the proliferated T cells was evaluated using a standard intracellular cytokine assay (Table 6).

[0096] [Table 6]

[0097] [Example 4] T cell cross-reactivity between JCV epitopes and BKV epitopes Peptide-specific T cells were stimulated with a JCV epitope or a BKV epitope, grown in vitro, and then restimulated with the corresponding homologous peptide epitope (see Table 2). Any cross-reactivity response between the JCV and BKV epitopes was observed. Specifically, PBMCs from healthy volunteers were cultured with either the synthetic JCV peptide epitope RSGSQQWRGLSRYFK or the synthetic BKV peptide epitope SSGTQQWRGLARYFK. After initial growth, each sample was restimulated (recalled) with either the JCV epitope RSGSQQWRGLSRYFK or the BKV epitope SSGTQQWRGLARYFK (samples recalled with the same epitope served as internal controls). The responsiveness of the grown T cells was evaluated using a standard intracellular cytokine assay (see Figure 4). T cells proliferated on either homologous epitope recognize both BKV and JCV peptide sequences.

[0098] [Example 5] Profiling the functional and phenotypic characteristics of JCV-specific T cells in healthy individuals and transplant recipients. In recent years, T-box transcription factors (T-bet) and Eomesodermin (Eomes) have been found to be involved in the infection of CD8 +These transcription factors have been shown to play a crucial role in T cell fate determination. High levels of T-bet are associated with the differentiation of cytotoxic T cells as well as the upregulation of perforin and granzyme B in antigen-specific cells. High levels of Eomes are associated with long-term memory formation. Various studies have shown that their coordinated expression is important for infection control. Mouse studies have also shown that deletion of either transcription factor results in failure to suppress infection. Therefore, it is important to study the expression of these transcription factors, which can help in understanding the phenotypic characterization and T cell differentiation of T cells during both acute and chronic viral infections. The expression patterns of T-bet and Eomes in JCV-specific T cells are still unknown, and analysis of these transcription factors on such T cells may enable a deeper understanding of JCV-specific T cell differentiation. Detailed studies of the functional characteristics of T cells may also lead to the development of effective immunotherapies for JCV-related diseases. An initial series of experiments are studying the transcription factors that regulate their differentiation on T cells. The expression of T-bet, Eomes, perforin, and granzyme B is assayed in JCV-specific and CMV-specific T cells using ICS. Initial analysis shows moderate to low levels of T-bet expression in JCV-specific T cells, while high levels of T-bet are observed in CMV-specific T cells. Furthermore, very low levels of Eomes expression are found in JCV-specific T cells compared to CMV-specific T cells. Similarly, low levels of perforin and granzyme B are also observed in JCV-specific T cells. This suggests that JCV-specific T cells are functionally less effective in effector function compared to CMV-specific T cells. Therefore, driving the effector function of JCV-specific CTLs will be a focus of research contributing to the development of effective adoptive T cell immunotherapy.

[0099] [Example 6] Feasibility of T cell proliferation using the proposed peptide pool PBMCs from 15 healthy donors were randomly selected regardless of their HLA type and stimulated with a peptide pool containing the peptides disclosed herein (i.e., peptides containing the amino acid sequences described in SEQ ID NOs: 1-21), resulting in proliferation of JCV-specific T cells. The T cells were grown for 17 days, and the JCV response was evaluated using an intracellular cytokine staining assay. T cells from 13 of the 15 donors exhibited a JCV-specific T cell response, as evidenced by IFN-γ production upon restimulation with the peptide pool (see Figure 5).

[0100] [Example 7] Functional characterization of JCV-specific T cell products To determine the multifunctionality of JCV-specific T cells grown using a peptide pool, T cells were analyzed for the expression of IL-2, TNF, IFN-γ, and CD107 by intracellular staining (see Figure 6a). JCV-specific T cells showed higher expression of TNF, along with other cytokines.

[0101] Boolean analysis of the expression patterns of different cytokine combinations revealed that JCV-specific T cell products are multifunctional and produce two or more cytokines (see Figure 6, b and c). Further characterization of JCV-specific T cell products was investigated by the expression of transcription factors (T-bet and Eomes) and effector molecules (perforin and granzyme B). Most cells produced T-bet hi / Eomes low and Granzyme hi / Perforin low The profile revealed that JCV-specific T cells are functionally active and can exhibit cytotoxic effects against JCV-infected cells. Such T cells can be proliferated in vitro and used to treat JCV-related diseases (see Figure 7).

Claims

1. A pharmaceutical composition comprising the peptide of SEQ ID NO: 1 and a pharmaceutically acceptable carrier.

2. A pool of immunogenic peptides, including the peptide of SEQ ID NO:

1.

3. The pharmaceutical composition according to claim 1, or the pool of immunogenic peptides according to claim 2, further comprising one or more peptides listed in Table 1.

4. A pool of immunogenic peptides according to claim 2 or 3, wherein the immunogenic peptides can induce the proliferation of peptide-specific cytotoxic T cells (CTLs).

5. A method for increasing JC virus-specific T cells for adoptive immunotherapy, (i) Contacting one or more cells isolated from the subject with the pool of immunogenic peptides described in claim 4, and (ii) Culturing the one or more cells under conditions such that JC virus-specific T cells proliferate from the one or more cells. A method that includes this.

6. The method according to claim 5, wherein one or more cells isolated from the subject include peripheral blood mononuclear cells (PBMCs) from a healthy donor.

7. The method according to claim 6, wherein one or more cells isolated from the subject include PBMCs from an immunodeficient donor.

8. A composition comprising T cells prepared by the method of any one of claims 5 to 7, for use in the treatment or prevention of JCV infection in a subject, wherein the T cells recognize the epitope of SEQ ID NO:

1.

9. A population of proliferated cytotoxic T cells (CTLs) prepared by the method according to any one of claims 5 to 7, wherein the CTLs recognize the epitope of SEQ ID NO:

1.

10. A group of CTLs according to claim 9 for use in the treatment or prevention of JCV infection in a subject.

11. A population of CTLs according to claim 10, wherein exposure to an immunogenic peptide or a pool of immunogenic peptides induces stimulation and proliferation of JCV peptide-specific T cells.

12. The population of CTLs according to claim 11, wherein the CTLs administered to the subject are autologous.

13. The population of CTLs according to claim 11, wherein the CTLs administered to the subject are not autologous.

14. The target patients are JCV granule cell layer neuropathy (JCV GCN), JCV encephalopathy (JCVE), JCV meningitis (JCVM), and / or progressive multifocal white blood cell disease. A group of CTLs according to any one of claims 10 to 13, who are suffering from polyencephalopathy (PML).

15. A population of proliferated JC virus-specific CTLs according to claim 9, for use in the treatment or prevention of JCV polyomavirus-associated cancer in a subject.

16. A group of CTLs according to claim 9 for use in the treatment or prevention of cancer in a subject.

17. A population of CTLs according to claim 16, wherein the cancer is JCV polyomavirus-associated cancer.

Images