Antigenic peptides for the prevention and treatment of b-cell malignancies

Abstract

This invention relates to antigen-based immunotherapy, specifically cancer immunotherapy. Specifically, the invention provides antigenic peptides that have epitopes different from human tumor antigens but share amino acid similarities, particularly the same core sequence. The invention further provides immunogenic compounds, nanoparticles, cells, and pharmaceutical compositions comprising such antigenic peptides, as well as nucleic acids encoding such antigenic peptides.

Description

[0001] This invention relates to the field of cancer treatment, and more specifically to immunotherapy. Specifically, this invention provides various peptides that can be used for cancer immunotherapy, particularly for the prevention and treatment of B-cell malignancies.

[0002] Among all B-cell malignancies, such as B-cell lymphoma and non-Hodgkin lymphoma (NHL), are the seventh leading cause of new cancer cases, accounting for approximately 3% of cancer-related deaths in the United States. Of all NHL cases, diffuse large B-cell lymphoma (DLBCL) is the most common lymphoma subtype, accounting for 32.5% of all new diagnoses, followed by follicular lymphoma (FL) at 17.1%, and mantle cell lymphoma (MCL) at 3-5%. More than 25,000 new cases of DLBCL are diagnosed annually in the United States, with an incidence rate of 6.9 per 100,000 people. Adding an anti-CD20 monoclonal antibody, rituximab, to standard chemotherapy R-CHOP has resulted in significant improvements in complete response (CR) rates, event-free survival (EFS), and overall survival (OS) in DLBCL. Unfortunately, approximately 30-40% of cases relapse or progress after R-CHOP. Certain patient subgroups have poor responses and outcomes to standard R-CHOP, such as MYC rearranged DLBCL, high-grade B-cell lymphomas with MYC, BCL2, or BCL rearrangements, and activated B-cell (ABC) DLBCL, who may benefit from the new approach (Chavez et al., CAR T-cell therapy for B-cell lymphomas: clinical trial results of available products; Ther AdvHematol. 2019).

[0003] In this new approach, chimeric antigen receptor (CAR) T cells, such as CD19-targeted CAR T cells, represent a new standard of care for patients with refractory DLBCL who have undergone at least two prior lines of therapy. Two CAR T-cell products, axicabtagene ciloleucel (axi-cel) (KTE-019) and tisagenlecleucel (CTL019), have been approved by the U.S. Food and Drug Administration for the treatment of refractory DLBCL after two lines of therapy. While this represents a significant addition to the treatment options for DLBCL, approximately 50% of cases will continue to die from the disease. Therefore, future research must focus on identifying disease-, treatment-, or patient-related factors that contribute to successful prediction of treatment outcomes.

[0004] Therefore, tumor antigen-based vaccination represents a unique cancer treatment approach that has attracted considerable interest because it can utilize the patient's own immune system to recognize, attack, and destroy tumors in a specific and persistent manner. Tumor cells are indeed known to express large amounts of peptide antigens that are readily recognized by the immune system. Therefore, vaccines based on such antigens offer significant opportunities not only to improve overall patient survival but also, due to the low toxicity and low molecular weight of tumor antigens, to monitor immune responses and prepare GMP-grade products. Examples of tumor antigens include byproducts of proteins transcribed from normally silenced or overexpressed genes and proteins expressed by oncoviruses (Kvistborg et al., Human cancerregression antigens. Curr Opin Immunol. 2013 Apr; 25(2):284-90) and neoantigens resulting from point mutations in cellular proteins. However, most tumor-associated antigens (TAAs) and tumor-specific antigens (TSAs) are (existing) human proteins and are therefore considered autoantigens. During thymic selection, T cells with sufficient affinity to recognize the peptide / autoMHC complex are clonally depleted. By providing protection against autoimmune diseases, this T-cell repertoire selection mechanism also reduces the likelihood of developing immunity to TAAs and TSAs. This can be illustrated by the fact that cancer-reactive TCRs generally have weak affinity. Furthermore, to date, most vaccine trials using TAAs and TSAs selected for their high binding affinity to MHC have failed to elicit strong immunity, which may reflect the consequences of thymic selection. Therefore, an effective antitumor response will depend on the presentation of immunoreactive peptides and the presence of sufficient reactive cells “trained” to recognize these antigens. Consequently, there is a need in the art to identify alternative antigenic peptides that can overcome the limitations encountered in this field.

[0005] The object of this invention is to satisfy the aforementioned needs. This object is achieved by the subject matter set forth below, specifically the invention and the items provided in the appended claims.

[0006] The invention is described in more detail below.

[0007] definition

[0008] Unless otherwise defined herein, the scientific and technical terms used in this application will have the meanings commonly understood by one of ordinary skill in the art. Furthermore, unless the context requires otherwise, the nomenclature and cell and tissue culture techniques used herein are well-known and commonly used in the art.

[0009] Such technology is used in fields such as Owen et al. (Kuby Immunology, 7) thIt is well described in the literature of WH Freeman (edition, 2013) and Sambrook et al. (Molecular cloning: A laboratory manual 4th edition, Cold Spring Harbor Laboratory Press-Cold Spring Harbor, NY, USA, 2012).

[0010] However, the following definitions are particularly applicable regarding the use of different terms throughout this specification.

[0011] The terms “peptide,” “polypeptide,” “protein,” and variations thereof refer to a peptide, oligopeptide, polypeptide, or protein comprising at least two amino acids linked together—preferably by ordinary peptide bonds or optionally by modified peptide bonds, such as, for example, in the case of isotopes. The term “(poly)peptide” refers to peptides and / or polypeptides. Specifically, the terms “peptide,” “polypeptide,” and “protein” refer to a sequential chain of amino acids of any length linked together by peptide bonds (-NHCO-). Peptides, polypeptides, and proteins can perform structural and / or functional roles in vitro and / or in vivo within cells. The terms “peptide,” “polypeptide,” and “protein” preferably comprise amino acid chains ranging in size from 2 to at least about 1000 amino acid residues. The term “peptide” herein preferably comprises an amino acid chain of less than about 30 amino acids, while the terms “polypeptide” and “protein” preferably comprise an amino acid chain of at least 30 amino acids. The terms “polypeptide” and “protein” are used interchangeably herein. In a preferred embodiment, the terms "peptide," "polypeptide," and "protein" also include "peptide mimics," which are defined as peptide analogs containing non-peptide structural elements that mimic or antagonize the biological effects (one or more) of the natural parent peptide. Peptide mimics lack typical peptide characteristics, such as enzymatically cleaved peptide bonds. Specifically, peptides, polypeptides, or proteins may contain amino acids other than the 20 amino acids defined by the genetic code, or may be composed of amino acids other than the 20 amino acids defined by the genetic code. Specifically, in the context of this invention, peptides, polypeptides, or proteins may be equivalent to amino acids modified by natural processes known to those skilled in the art, such as post-translational maturation or chemical processes. Such modifications are described in detail in the literature. These modifications can occur at any position in the polypeptide: in the peptide backbone, in the amino acid chain, or even at the carboxyl terminus or amino terminus. Specifically, peptides or polypeptides may be branched after ubiquitination or may be cyclic with or without branches. This type of modification may be the result of natural or synthetic post-translational processes known to those skilled in the art. In the context of this invention, the terms "peptide," "polypeptide," and "protein" specifically also include modified peptides, polypeptides, and proteins. For example, modifications to peptides, polypeptides, or proteins may include acetylation, acylation, ADP-ribosylation, amidation, covalent fixation of nucleotides or nucleotide derivatives, covalent fixation of lipids or lipid derivatives, covalent fixation of phosphatidylinositol, covalent or non-covalent crosslinking, cyclization, disulfide bond formation, demethylation, glycosylation—including PEGylation, hydroxylation, iodination, methylation, myristylation, oxidation, proteolytic processes, phosphorylation, isopreneation, racemization, seneloylation, sulfation, and amino acid additions such as arginination or ubiquitination.Such modifications are described in detail in the literature (Proteins Structure and Molecular Properties (1993) 2nd Ed., TECreighton, New York; Post-translational Covalent Modifications of Proteins (1983) BC Johnson, Ed., Academic Press, New York; Seifter et al. (1990) Analysis for protein modifications and nonprotein cofactors, Meth. Enzymol. 182: 626-646; and Rattan et al. (1992) Protein Synthesis: Post-translational Modifications and Aging, Ann NY Acad Sci, 663: 48-62). Therefore, the terms "peptide," "polypeptide," and "protein" preferably include, for example, lipopeptides, lipoproteins, glycopeptides, and glycoproteins.

[0012] In a preferred embodiment, the (poly)peptide or protein is a "typical" (poly)peptide or protein, wherein the "typical" (poly)peptide or protein is typically composed of amino acids selected from 20 amino acids defined by the genetic code, which are linked together by ordinary peptide bonds.

[0013] As is known in the art, peptides, polypeptides, and proteins can be encoded by nucleic acids. The terms “nucleic acid,” “nucleic acid molecule,” “nucleic acid sequence,” “polynucleotide,” and “nucleotide sequence” are used interchangeably herein and refer to precisely contiguous natural nucleotides (e.g., A, T, G, C, and U) or synthetic nucleotides, i.e., a chain of at least two nucleotides. Specifically, the terms “nucleic acid,” “nucleic acid molecule,” “nucleic acid sequence,” “polynucleotide,” and “nucleotide sequence” refer to DNA or RNA. Nucleic acids preferably comprise single-stranded, double-stranded, or partially double-stranded DNA or RNA, preferably selected from genomic DNA (gDNA), complementary DNA (cDNA), ribosomal DNA (rDNA), and transcripts of said DNA, such as RNA. Preferred examples of nucleic acids include ribosomal RNA (rRNA), messenger RNA (mRNA); antisense DNA, antisense RNA; complementary RNA and / or DNA sequences, ribozymes, (complementary) RNA / DNA sequences with or without expression elements, vectors; minigenes, gene fragments, regulatory elements, promoters, and combinations thereof. Further preferred examples of nucleic acids (molecules) and / or polynucleotides include, for example, recombinant polynucleotides, vectors, oligonucleotides, RNA molecules such as rRNA, mRNA, or transfer RNA (tRNA), or the aforementioned DNA molecules. Therefore, it is preferred that the nucleic acid (molecule) is a DNA molecule or an RNA molecule; preferably selected from gDNA; cDNA; rRNA; mRNA; antisense DNA; antisense RNA; complementary RNA and / or DNA sequences; RNA and / or DNA sequences with or without expression elements, regulatory elements, and / or promoters; vectors; and combinations thereof. Determining a nucleotide sequence that can encode a specific amino acid sequence is within the capabilities of those skilled in the art.

[0014] The (poly)peptides and / or nucleic acids according to the invention can be prepared by any method known in the art, including but not limited to any synthetic method, any recombinant method, any in vitro generation method, and any combination thereof. Such techniques are fully described in the foregoing literature.

[0015] As used herein, the term "antigenic peptide" refers to a peptide that readily induces / triggers, enhances, prolongs, or maintains an immune response in its administered object. Specifically, an antigenic peptide is a sequence variant of a (human) tumor antigen (fragment / epitope). In other words, the antigenic peptide is preferably distinct from the (human) tumor antigen (fragment / epitope), but preferably has amino acid similarity to it. Importantly, the antigenic peptide shares the same core sequence as its corresponding (fragment / epitope) of the (human) tumor antigen. Preferably, the immune response induced / triggered, enhanced, prolonged, or maintained by the antigenic peptide is also directed against the corresponding (human) tumor antigen (fragment / epitope).

[0016] As used herein, the term “tumor antigen” includes tumor-specific antigens (TSA) and tumor-associated antigens (TAA). Generally, the term “tumor antigen” or “tumor protein” refers herein to antigenic substances produced in tumor cells and sometimes in normal cells that can trigger an immune response when administered to a subject. In humans, these (tumor antigens) have been classified according to their expression patterns, function, or genetic origin, and include, without limitation, overexpressed autoantigens (e.g., BIRC5); testicular cancer (CT) antigens (e.g., MAGE-1); mutant antigens, also known as neoantigens (e.g., mutants of p53); tissue-specific differentiation antigens (e.g., melanoma antigen Melan A / MART-1); viral antigens expressed by oncoviruses (e.g., HPV, EBV); fetal antigens (e.g., alpha-fetoprotein AFP and carcinoembryonic antigen CEA); and universal antigens (terminal enzymes).

[0017] As used herein, the term "B-cell tumor antigen" refers to an antigen that is etiologically related to and / or involved in B-cell malignancies, such as those involved in or expressed in B-cell malignancies. In other words, the antigen is expressed by or on B cells (including human B cells), such as any of a variety of known B-cell markers. Preferably, the B-cell tumor antigen is highly expressed (overexpressed) in B-cell lymphomas such as CD19, CD20, CD22, CD37, or TNFRSF13C. Antigenic peptides "derived" from B-cell tumor antigens generally share the same core sequence as the epitope ("reference epitope") of said B-cell tumor antigen.

[0018] As used herein, the term "core sequence" refers to the middle amino acid of a sequence, such as the middle amino acid in an antigenic peptide and / or (reference) epitope (also referred to as the "central amino acid" of the sequence). Thus, a core sequence consists of all amino acids except for the two most N-terminal and two most C-terminal amino acids. For example, in a nine-amino acid peptide (e.g., the corresponding (fragment / epitope) of an antigenic peptide or (human) tumor antigen according to the invention), five middle amino acids represent the core sequence, and changes may occur only at either of the two N-terminal and two C-terminal amino acid positions. Therefore, a "shared core sequence" (or a "maintained" core sequence) generally means that mutations / differences are only permitted at the two most N-terminal and two most C-terminal amino acids of the (reference) epitope / sequence.

[0019] As used herein, the term "prevalence" refers to the cumulative frequency of each protein in the human microbiota where a shared core sequence (fragment / epitope) corresponding to a (human) tumor antigen is found and present in the antigenic peptide. In reality, the target core sequence may be present in one or more different antigenic peptides, each of which may be present in one or more different proteins expressed in the human microbiota. Therefore, the global prevalence of the core sequence stems from the frequency of each protein in the human microbiota where a shared core sequence (fragment / epitope) corresponding to a (human) tumor antigen is found, and similar peptides (i.e., different antigenic peptides) sharing the same core sequence are identified by considering the frequency of each protein in the human microbiota.

[0020] As used herein, the term "microbial community" refers to the symbiotic microorganisms found in and on all multicellular organisms, from plants to animals, studied to date. Specifically, microbial communities have been found to be essential for the immune system, hormones, and metabolic homeostasis of their hosts. Microbial communities include bacteria, archaea, protozoa, fungi, and viruses. Thus, a "microbial community sequence variant" (or "microbial community variant") is a sequence variant of a (human) reference sequence (specifically, an epitope / fragment of a human tumor antigen) present in a microbial community such as bacteria (e.g., it may be contained in a microbial community protein, such as a bacterial protein). Preferably, the antigenic peptide of the present invention is a microbial community sequence variant of the (reference epitope / fragment of a human B-cell tumor antigen). Therefore, the antigenic peptide is preferably present (e.g., contained in) at least one protein expressed by a human microbial community.

[0021] The “sequence variant” generally shares at least 50% sequence identity with the reference sequence (i.e., a fragment / epitope of the (reference) tumor antigen), particularly along the full length of the sequence. Preferably, the sequence variant shares at least 60%, more preferably at least 70%, more preferably at least 75%, more preferably at least 80%, even more preferably at least 85%, even more preferably at least 90%, particularly preferably at least 95%, and most preferably at least 99% sequence identity with the reference sequence (i.e., a fragment / epitope of the (reference) tumor antigen). Sequence identity can be calculated as is known in the art, specifically as described below. Preferably, the sequence variant retains a specific function of the reference sequence, such as its function as a tumor epitope and / or its ability to elicit or maintain an immune response. Microbial cluster sequence variants are preferably selected from bacterial sequence variants, archaea sequence variants, protist sequence variants, fungal sequence variants, and viral sequence variants. More preferably, the microbial cluster sequence variant is a bacterial sequence variant.

[0022] Anatomically, microbial communities reside on or within a variety of tissues and biological fluids, including the skin, conjunctiva, mammary glands, vagina, placenta, semen, uterus, follicles, lungs, saliva, oral cavity (specifically, oral mucosa), and gastrointestinal tract, specifically the intestine. In the context of this invention, the microbial community sequence variant is preferably a sequence variant of the gastrointestinal microbial community (microorganisms residing in the gastrointestinal tract), more preferably a sequence variant of the intestinal microbial community (microorganisms residing in the intestine). Therefore, the most preferred microbial community sequence variant is a (human) intestinal bacterial sequence variant (i.e., a sequence variant of bacteria residing in the (human) intestine).

[0023] While microbial communities can be found in and on many multicellular organisms (all multicellular organisms from plants to animals studied to date), those found in and on the human body are preferred. Such microbial communities are referred to herein as “human microbial communities” (wherein the term human specifically refers to the location / habitat of the microbial community). In the context of this invention, the microbial community sequence variant is the human microbial community sequence variant.

[0024] The term "immunogenic compound" refers to a compound containing an antigenic peptide according to the invention. An "immunogenic compound" is capable of inducing / initiating, increasing, prolonging, or maintaining an immune response against said antigenic peptide in the subject to which it is administered. In some embodiments, the immunogenic compound contains at least one antigenic peptide or optionally at least one compound containing such an antigenic peptide, which is linked to a protein such as a carrier protein.

[0025] "Carrier protein" is generally a protein capable of transporting cargo, such as the antigenic peptide according to the invention. For example, a carrier protein can transport its cargo across a membrane. In the context of the invention, carrier proteins specifically (also) include peptides or polypeptides capable of eliciting an immune response against the antigenic peptide to which they are attached. Carrier proteins are known in the art.

[0026] Alternatively, this carrier peptide or polypeptide can be co-administered as an immune adjuvant.

[0027] Preferably, the antigenic peptides described herein can be co-administered or, for example, covalently or non-covalently linked to proteins / peptides with adjuvant properties, such as providing stimulation of CD4+Th1 cells. Although the antigenic peptides described herein are preferably bound to MHC class I, CD4+ helper epitopes can be additionally used to provide an effective immune response. Th1 helper cells are able to maintain effective dendritic cell (DC) activation and specific CTL activation—by secreting interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), and interleukin-2 (IL-2) and enhancing the expression of co-stimulatory signals on DCs and T cells (Galaine et al., Interest of Tumor-Specific CD4 T Helper Cells for Therapeutic Antiticancer Vaccine. Vaccines (Basel). 2015 Jun 30; 3(3):490-502).

[0028] For example, the adjuvant peptide / protein may preferably be different from the antigen peptide according to the invention. Preferably, the adjuvant peptide / protein is capable of restoring immune memory or providing nonspecific help, or may be a specific helper peptide. Several helper peptides for providing nonspecific T cell help have been described in the literature, such as tetanus helper peptide, scutellarin peptide, or PADRE peptide (Adotévi et al., Targeting antitumor CD4 helper T cells with universal tumor-reactive helper peptides derived from telomerase for cancer vaccine. HumVaccin Immunother. May 2013; 9(5):1073-7, Slingluff CL, The present and future of peptide vaccines for cancer: single or multiple, long or short, alone or incombination? Cancer J. Sep-October 2011; 17(5):343-50). Therefore, tetanus helper peptide, scutellarin peptide, and PADRE peptide are preferred examples of such adjuvant peptides / proteins. The HHD-DR3 peptide with the sequence MAKTIAYDEEARRGLERGLN (SEQ ID NO: 473). This peptide represents another example of an accessory peptide (with immune adjuvant properties), which is preferred in the context of this invention. Another preferred example is h-pAg T13L (sequence: TPPAYRPPNAPIL; SEQ ID NO: 474; Bhasin M, Singh H, Raghava GP (2003) MHCBN: a comprehensive database of MHC binding and non-binding peptides. Bioinformatics 19:665–666).Other examples of preferred accessory peptides include UCP2 peptides (e.g., WO2013 / 135553A1 or Dosset M, Godet Y, Vauchy C, Beziaud L, Lone YC, Sedlik C, Liard C, Levionnois E, Clerc B, Sandoval F, Daguindau E, Wain-Hobson S, Tartou E, Langlade-Demoyen P, Borg C, Adotévi O: Universal cancer peptide-based therapeutic vaccine breaks out of olerance against telomerase and eradicates established tumor. Clin Cancer Res. 2012 Nov 15; 18(22):6284-95. doi:10.1158 / 1078-0432.CCR-12-0896.Epub (as described on October 2, 2012) and BIRC5 peptides (e.g., EP2119726A1 or Wildenmeyer M, Griesemann H,). S,Feyerabend S,Klein R,Attig S,Hennenlotter J,Wernet D,Kuprash DV,Sazykin AY,Pascolo S,Stenzl A,Gouttefangeas C,Rammensee HG:Promiscuous survivin peptide induces robust CD4+ T-cell responses in the majority of vaccinated cancer patients. Int J Cancer. 2012 Jul 1; 131(1):140-9. doi:10.1002 / ijc.26365.Epub (described on 2011 Sep 14). The most preferred accessory peptide is the UCP2 peptide (amino acid sequence: KSVWSKLQSIGIRQH; SEQ ID NO: 475, for example, as described in WO2013 / 135553 A1 or Dosset et al., Clin Cancer Res. 15 Nov 2012; 18(22):6284-95). Specifically, the antigenic peptide described herein or a polypeptide containing the antigenic peptide may, for example, be linked to the accessory peptide by covalent or non-covalent bonds.

[0029] As used herein, the term "immunogenic composition" refers to a composition capable of inducing, enhancing, prolonging, or maintaining an immune response, particularly when administered to mammals, and especially when administered to human individuals. Preferably, the immunogenic composition further comprises one or more immune adjuvant substances.

[0030] "Pharmaceutically acceptable excipients or carriers" herein means pharmaceutical-grade compounds that improve the delivery, stability, or bioavailability of an active agent and can be metabolized by the subject to which they are given and are non-toxic to the subject. Preferred excipients and carriers according to the invention include any excipients or carriers commonly used in pharmaceutical products, such as, for example, water, saline, phosphate-buffered saline, dextran, glycerol, ethanol, and combinations thereof. In many cases, it is preferred to include isotonic agents in the composition, such as sugars, polyols such as mannitol, sorbitol, or sodium chloride. Pharmaceutically acceptable excipients or carriers may further contain small amounts of excipients, such as wetting agents, emulsifiers, or preservatives.

[0031] In this context, "vaccine" is defined as a composition capable of stimulating the immune system of a living organism to provide protection against harmful antigens through prevention or treatment. Prophylactic vaccines are preferred. Preferably, the vaccine or vaccine composition further comprises one or more immune adjuvant substances.

[0032] According to the various aspects and embodiments of the invention described herein, "object" or "host" preferably refers to a mammal, and most preferably to a human. The object may have B-cell malignancy, be suspected of having B-cell malignancy, or be at risk of developing B-cell malignancy.

[0033] The term "B-cell malignancy" refers to diseases associated with B-cell transformation. These include B-cell lymphoma, acute lymphoblastic (or lymphoblastic) leukemia (ALL), chronic lymphocytic leukemia (CLL, Richter's disease), etc. In the context of this invention, B-cell lymphoma, such as non-Hodgkin's lymphoma (NHL), is preferred. For example, NHL is selected from indolent (slow-growing) NHL, aggressive NHL, diffuse large B-cell lymphoma (DLBCL), NOS (derived from de novo and indolent transformation), primary mediastinal large B-cell lymphoma (PMBCL), T-cell / histiocytic rich large B-cell lymphoma (TCHRBCL), Burkitt lymphoma, mantle cell lymphoma (MCL), and / or follicular lymphoma (FL), optionally, follicular lymphoma grade 3B (FL3B).

[0034] As used herein, the term “preventing” (“prevention”, “prophylaxis”, or “prevent”) generally means avoiding or minimizing the onset or development of a disease or condition before it occurs, while the term “treating” (“treatment”, or “treat”) includes reducing, improving, or curing a disease or condition (or its symptoms) after it has occurred. The term “prevention” includes “reducing the likelihood of occurrence” or “reducing the likelihood of recurrence”.

[0035] As used herein, "effective amount" or "effective dose" refers to the amount that provides the desired effect. For therapeutic purposes, an effective amount is an amount sufficient to provide a beneficial or desired clinical outcome. A preferred effective amount for a given application can be readily determined by a person skilled in the art by considering factors such as the size, age, weight of the subject, the type of disease / disorder to be prevented or treated, and the time elapsed since the onset of the disease / disorder. In the context of this invention, for prevention or treatment, an effective amount of the composition is an amount sufficient to induce a humoral and / or cell-mediated immune response against the disease / disorder.

[0036] Throughout the specification and appended claims, unless the context otherwise requires, the term "comprising" and variations such as "including" and "containing" will be understood to mean including the stated members, integers, or steps, but not excluding any other unstated members, integers, or steps. The term "consisting of" is one specific embodiment of the term "comprising," in which any other unstated members, integers, or steps are excluded. In the context of this invention, the term "comprising" encompasses the term "consisting of." Therefore, the term "comprising" covers both "including" and "consisting of," for example, a composition "comprising" X may consist of only X, or may include other elements, such as X+Y.

[0037] The terms “a,” “an,” and “the,” as well as similar references, used in the context of the description of this invention (particularly in the context of the claims) should be interpreted to cover both the singular and plural, unless otherwise stated herein or clearly contradicted by the context. The descriptions of numerical ranges herein are intended only as a concise way of referring to each individual numerical value falling within that range. Unless otherwise stated herein, each individual numerical value is incorporated into the specification as if it were referred to separately herein. No language in the specification should be construed as indicating that any unclaimed element is necessary for the practice of this invention.

[0038] The word "substantially" does not exclude "completely," for example, a composition that is "substantially free of" Y can be completely free of Y. The word "substantially" may be omitted from the definition of this invention when necessary.

[0039] The term "about" related to the numerical value x refers to x ± 10%.

[0040] Other definitions are provided throughout the specification.

[0041] The invention can be more readily understood by referring to the following detailed description, including preferred embodiments and examples included herein.

[0042] Detailed description

[0043] Although the invention is described in detail below, it should be understood that the invention is not limited to the specific methods, schemes, and reagents described herein, as these can be modified. It should also be understood that the terminology used herein is not intended to limit the scope of the invention, which is limited only by the appended claims. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

[0044] The elements of the invention will be described below. These elements are set forth along with the specific embodiments; however, it should be understood that they can be combined in any manner and in any number to create other embodiments. The various examples and preferred embodiments described should not be construed as limiting the invention to the explicitly described embodiments. This description should be understood to support and cover embodiments that combine the explicitly described embodiments with any number of disclosed and / or preferred elements. Furthermore, unless the context otherwise requires, the description of this application should be considered to disclose any permutation and combination of all the elements described in this application.

[0045] The antigenic peptide according to the present invention

[0046] In a first aspect, the present invention provides antigenic peptides derived from tumor antigens, particularly B-cell tumor antigens, wherein the antigenic peptides share the same core sequence as the reference epitope of the tumor antigen, and wherein the shared core sequence is highly prevalent in the human microbiome.

[0047] The present invention also provides an antigenic peptide comprising or consisting of the amino acid sequence shown in any one of SEQ ID NO 316, 304-315, 317-472 and 501-509. Preferably, the antigenic peptide may comprise or consist of the amino acid sequence shown in any one of SEQ ID NO 316, 304-315 and 317-326.

[0048] Furthermore, the present invention provides an antigenic peptide comprising or consisting of the amino acid sequence shown in any of SEQ ID NO 1-257 and 476-500, wherein, optionally, one or two amino acid residues may be substituted, deleted, or added. Preferably, if one or two amino acid residues are substituted, deleted, or added, the core sequence (of SEQ ID NO 1-257 and 476-500) is retained. More preferably, the antigenic peptide comprises (without mutation) or consists of the amino acid sequence shown in any of SEQ ID NO 1-257 and 476-500. The amino acid sequences according to SEQ ID NO 1-257 and 476-500 are human tumor epitopes, specifically microbial cluster sequence variants (microbial cluster variants, specifically bacterial sequence variants) of the reference epitope / fragment of human B-cell tumor antigens. In other words, the amino acid sequences of SEQ ID NO 1-257 and 476-500 can be found in bacterial proteins and show sequence similarity to reference epitopes / fragments of human B-cell tumor antigens, as detailed in Table 1A.

[0049] The inventors have identified a set of antigenic peptides that can be used to induce specific immune responses against tumor cells. These antigenic peptides are distinct from fragments of human tumor antigens, particularly tumor antigens highly expressed in B-cell lymphomas, such as CD19, CD20, CD22, CD37, or TNFRSF13C, but share amino acid similarities, as shown in Tables 1A, 1B, and 1C. Importantly, the antigenic peptides according to the invention have a core sequence identical to the core sequence of the epitope (fragment) of the reference tumor antigen. Furthermore, the core sequence exhibits high universality based on the frequency of proteins present in the human microbiome where it is found.

[0050] Specifically, the antigenic peptides according to the invention are contained in polypeptides and proteins produced by symbiotic bacteria from the human gut. Therefore, the antigenic peptides according to the invention are not human sequences, but bacterial sequences. Not wishing to be bound by any specific theory, the inventors believe that the human immune repertoire contains T-cell clones reactive to bacterial peptides (contained in proteins produced by symbiotic bacteria from the gut) that have amino acid similarity to fragments of human tumor antigens. Specifically, the antigenic peptides according to the invention can elicit a stronger immune response than the corresponding human peptides because T cells capable of strictly recognizing human peptides are depleted during maturation when recognizing their own antigens, a situation not present with the antigenic peptides according to the invention. This explains why the antigenic peptides described herein can induce an immune response, and especially a T-cell response, when these peptides are administered to (human) individuals.

[0051] Therefore, without being bound by any theoretical constraints, the inventors hypothesize that proteins produced by symbiotic bacteria from the gut can "mimic" tumor antigens and can be used to trigger a specific immune response against tumor cells. These findings provide further evidence that symbiotic bacteria can contribute to tumor cell elimination.

[0052] The antigenic peptides disclosed herein can be prepared using known techniques. For example, peptides can be prepared synthetically using recombinant DNA technology or chemical synthesis. The peptides disclosed herein can be synthesized individually or as longer polypeptides comprising two or more peptides (e.g., two or more peptides, or peptides and non-peptides). The antigenic peptides can be isolated, i.e., purified to be substantially free of other naturally occurring host cells.

[0053] Cellular proteins and their fragments, for example, purified to at least about 70%, 80%, or 90%. Preferably, the antigenic peptide according to the invention is an isolated antigenic peptide.

[0054] Therefore, the core sequence represents the principal characteristics of the antigenic peptide according to the invention. The inventors have thus identified a highly targeted core sequence with high universality, as it is present at high frequency in several sequence variants of (reference) tumor antigens and / or several human microbiome protein fragments in a significant portion of the general population. The inventors have selected the antigenic peptide of the invention that readily triggers an optimal cross-reactive tumor-specific cytotoxic T-cell immune response for the prevention and treatment of B-cell malignancies.

[0055] In general, when the universality is greater than 30%, preferably greater than 40%, preferably greater than 50%, more preferably greater than 60%, even more preferably greater than 70%, even more preferably greater than 80%, particularly preferably greater than 90%, and most preferably greater than 95%, the shared core sequence has high universality in the human microbiome.

[0056] The universality of a core sequence is derived from the frequency of each protein in the human microbiome, where a core sequence shared by corresponding (fragments / epitopes) of (human) tumor antigens is found, and similar peptides (i.e., different antigenic peptides) sharing the same core sequence are identified by considering the frequency of each protein in the human microbiome. For example, to evaluate whether a core sequence has high universality, the frequency of each protein in the human microbiome where the core sequence is found is calculated from a microbiome sequence database. Universality is then derived from the cumulative frequency calculated from each protein in the human microbiome where the target core sequence is found and present in the antigenic peptide. Such a database may preferably include microbiome (sequence) data from multiple individuals (objects). An example of such a database is the "Integrated reference catalog of the human gut microbiome" (version 1.0, March 2014; Li et al. MetaHIT Consortium. An integrated catalog of reference genes in the human gut microbiome. Nat Biotechnol. August 2014; 32(8):834-41 URL: http: / / meta.genomics.cn / meta / home), which includes data from major human microbiome analysis efforts, the National Institutes of Health Human Microbiome Project (NIH-HMP), and the European Metagenomics (MetaHIT) initiative of the Human Gut Initiative.

[0057] Therefore, the present invention relates to antigenic peptides that have amino acid similarity to tumor antigens because they are derived from such tumor antigens (or tumor epitopes). As used herein, the expression "having amino acid similarity to a tumor antigen" specifically refers to a sequence variant of a fragment of a (reference) human tumor antigen such as CD22 or other exemplary human tumor antigens described in Tables 1A, 1B, and 1C below. A "sequence variant" generally shares (specifically, over the full length of the sequence) at least 50% sequence identity with a reference sequence, i.e., a fragment of the (reference) tumor antigen. Preferably, the sequence variant shares at least 55%, preferably at least 60%, preferably 66%, preferably at least 70%, preferably at least 77%, more preferably at least 80%, even more preferably at least 88%, and even more preferably at least 90% sequence identity with a reference sequence, i.e., a fragment of the (reference) tumor antigen. Sequence identity can be calculated as is known in the art, specifically as described below. Preferably, the sequence variant retains a specific function of the reference sequence, such as its function as a tumor epitope and / or its ability to initiate or maintain an immune response. Specifically, an amino acid sequence variant has an altered sequence in which one or more amino acids in the reference sequence are mutated, for example, deleted or substituted, or one or more amino acids are inserted into the reference amino acid sequence. For example, a variant sequence having at least 90% identity has no more than 10 changes per 100 amino acids in the reference sequence, i.e., any combination of deletions, insertions, or substitutions.

[0058] Methods for comparing the identity (similarity) of two or more sequences are well known in the art. The percentage of identity between two sequences can be determined, for example, using mathematical algorithms. A preferred, but not limiting, example of an available mathematical algorithm is the one by Karlin et al. (1993), PNAS USA, 90:5873-5877. Such algorithms are integrated into the BLAST family of programs, such as BLAST or NBLAST (see also Altschul et al., 1990, J.Mol.Biol. 215, 403-410 or Altschul et al. (1997), Nucleic Acids Res, 25: 3389-3402) – accessible via the NCBI webpage ncbi.nlm.nih.gov, and FASTA (Pearson (1990), Methods Enzymol. 183, 63-98; Pearson and Lipman (1988), Proc. Natl. Acad. Sci. USA 85, 2444-2448). These programs can identify sequences that share a certain degree of identity with other sequences. In addition, programs available in Wisconsin sequence analysis software package version 9.1 (Devereux et al., 1984, Nucleic Acids Res., 387-395), such as the programs BESTFIT and GAP, can also be used to determine the percentage of identity between two polynucleotides and between two (poly)peptide sequences. BESTFIT utilizes the "local homology" algorithm of Smith and Waterman (1981), J. Mol. Biol. 147, 195-197, and finds the best single similar region between two sequences.

[0059] In general, the antigenic peptide according to the present invention binds to MHC class I (major histocompatibility complex class I, MHC I) molecules.

[0060] MHC class I molecules present epitopes to cytotoxic T cells, also known as cytotoxic T lymphocytes (CTLs). In addition to the TCR (T cell receptor), CTLs also express the CD8 receptor. When the CTL's CD8 receptor docks with an MHC class I molecule, if the CTL's TCR matches an epitope within the MHC class I molecule, the CTL triggers programmed cell death via apoptosis. This pathway is particularly useful in cancer prevention and / or treatment, as cancer cells are directly attacked. In humans, MHC class I includes HLA-A, HLA-B, and HLA-C molecules. Typically, peptides (epitopes) of 8-10 amino acids in length are presented by MHC class I.

[0061] In general, the antigenic peptide according to the invention can have any length. Preferably, the length of the antigenic peptide according to the invention does not exceed 350 amino acids. For example, the maximum length of the antigenic peptide according to the invention can be 300 or 250 amino acids. More preferably, the maximum length of the antigenic peptide according to the invention does not exceed 200 amino acids, for example, not exceeding 190, 180, 170, 160, 150, 140, 130, 120, 110, 100, 95, 90, 85, 80, 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, or 13 amino acids. Specifically, the length of the antigenic peptide according to the invention is preferably at most 30 or 25 amino acids, more preferably at most 20 or 15 amino acids, and even more preferably at most 10 amino acids. Particularly preferably, the antigenic peptide according to the invention comprises at least 8 or 9 amino acids, such as 10 amino acids. Even more preferably, the antigenic peptide has a length of 9 or 10 amino acids. Specifically, the antigenic peptide is not a full-length protein produced by the human microbiome (the antigenic peptide may be derived from this). In other words, the antigenic peptide of the invention is preferably a fragment of a full-length protein (produced by the human microbiome).

[0062] Similarly, the "fragment / epitope" of a (reference) tumor antigen, which generally serves as a reference sequence, preferably comprises nine consecutive amino acids of the tumor antigen, ultimately containing ten amino acids. It should be understood that the "fragment / epitope" of the (reference) tumor antigen is not the full-length tumor antigen (protein).

[0063] As used herein, the maximum length of the "fragment" (of a protein or nucleic acid (sequence)) is preferably 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% of the full-length (reference) protein / nucleic acid / sequence. In some embodiments, the length of the fragment does not exceed 50% of the length of the (full-length) (reference) protein / nucleic acid. In other embodiments, the length of the fragment of the (reference) protein / nucleic acid does not exceed 20% or 10% of the length of the (full-length) (reference) protein / nucleic acid.

[0064] More generally, the present invention provides antigenic peptides comprising or composed of: sequence variants of fragments / epitopes of human (reference) tumor antigens, particularly microbial cluster sequence variants of fragments / epitopes of human tumor antigens. The human tumor antigen may be selected from CD19, CD20, CD22, CD37, and TNFRSF13C. The fragments / epitopes of the human (reference) tumor antigen may be selected from any of SEQ ID 258-280.

[0065] Preferably, the antigenic peptide comprises or consists of a microbial cluster variant of or is composed of a human reference peptide according to any of SEQ ID NO 258-280. Specifically, SEQ ID NO 258-280 refers to a human tumor epitope, specifically a reference epitope / fragment of a human B-cell tumor antigen. Examples of microbial cluster variants of the human reference peptide according to any of SEQ ID NO 258-280 are peptides according to SEQ ID NO 1-257 and 476-500 (examples are shown in Table 1A below). Preferred microbial cluster sequence variants (microbial cluster variants) are bacterial sequence variants. In other words, the amino acid sequences according to SEQ ID NO 1-257 and 476-500 can be found in bacterial proteins and show sequence similarity to the reference epitope / fragment of the human B-cell tumor antigen, as detailed in Table 1A. More preferably, the antigenic peptide comprises or is composed of a microbial cluster variant of a human reference peptide according to any one of SEQ ID NO 258, 260-266, 270, 271, 279 and 280. Even more preferably, the antigenic peptide comprises or is composed of a microbial cluster variant of a human reference peptide according to any one of SEQ ID NO 260, 264, 270, 271, 279 and 280. Even more preferably, the antigenic peptide comprises or is composed of a microbial cluster variant of a human reference peptide according to any one of SEQ ID NO 264, 270, 271 and 279.

[0066] Therefore, the present invention also provides antigenic peptides comprising, or consisting of, any of SEQ ID NO 258-280; preferably any of SEQ ID NO 258, 260-266, 270, 271, 279 and 280; more preferably any of SEQ ID NO 260, 264, 270, 271, 279 and 280; and even more preferably, microbial cluster variants of or composed of any of SEQ ID NO 264, 270, 271 and 279 of a human reference peptide.

[0067] In specific embodiments, the present invention provides antigenic peptides derived from CD19, CD20, CD22, CD37, or TNFRSF13C, wherein the antigenic peptide shares the same core sequence as a fragment / epitope of a (reference) tumor antigen; and wherein the shared core sequence is highly prevalent in the human microbiome. This prevalence stems from the frequency of at least one protein present in the human microbiome in which said core sequence is found. Preferably, the antigenic peptide according to the invention comprises or consists of any of SEQ ID Nos. 304-326 (MHC I common sequences).

[0068] In one embodiment, the antigenic peptide binds moderately, strongly, or very strongly to MHC class I (major histocompatibility complex class I, MHC I) molecules.

[0069] The binding of at least one antigenic peptide to MHC class I (major histocompatibility complex class I) molecules can be tested by a computer-aided or in vitro binding assay as described herein. Therefore, strong and very strong binding agents can be selected as described above. Preferably, the binding of at least one antigenic peptide to an MHC I molecule and another (corresponding reference) tumor antigen (fraction / epitope of the (reference) tumor antigen) to an MHC I molecule (as described herein, computer-aided and / or in vitro) is tested for MHC I binding, and preferably, the binding affinity of both (tumor antigen and fragment / epitope of antigenic peptide) is obtained.

[0070] Following the binding assay, it is preferable to select only antigenic peptides that bind moderately, strongly, or very strongly to MHC I. More preferably, only strong and very strong binders are selected, and most preferably, only antigenic peptides that bind very strongly to MHC I are selected. More preferably, only antigenic peptides that bind strongly or very strongly to the MHC I molecule are selected, wherein the (reference) tumor antigen epitope / fragment ("corresponding" tumor antigen epitope sequence) binds weakly (e.g., weakly or moderately) to the MHC I molecule. Even more preferably, only microbial cluster sequence variants that bind very strongly to MHC I are selected, wherein the (reference) tumor antigen epitope / fragment binds weakly to MHC I.

[0071] Therefore, the binding affinity of the antigenic peptide of the present invention to the MHC I molecule is preferably stronger (i.e., higher) than that of the corresponding human reference epitope of the B-cell tumor antigen (which shares the same core sequence). In other words, the binding affinity of the reference epitope of the human B-cell tumor antigen (which shares the same core sequence) to the MHC I molecule is preferably weaker (i.e., lower) than that of the corresponding antigenic peptide.

[0072] Publicly available tools, such as NetMHCpan, including NetMHCpan 3.0Server or NetMHCpan 4.0Server (Center for biological sequence analysis, Technical University of Denmark DTU; URL: http: / / www.cbs.dtu.dk / services / NetMHCpan / ), can be used to predict MHC class I binding (MHC computer binding test). The NetMHCpan method, specifically NetMHCpan 3.0 or later, is trained on over 180,000 quantitative binding data points, covering 172 MHC molecules from humans (HLA-A, B, C, E) and other species. Generally, affinity can be predicted by preserving default thresholds for strong and weak binders. For example, for HLA-A*0201, a calculated affinity below 50 nM can indicate a "strong binder," while an affinity between 50 and 300 nM can indicate a "moderate binder." In NetMHCpan, such as NetMHCpan 3.0 or NetMHCpan 4.0, the predicted affinity rank can be compared to a set of 400,000 random natural peptides, which can be used as a measure of percentage-rank binding affinity. This value is not affected by the inherent bias of some molecules towards higher or lower average predicted affinity. For example (e.g., for HLA-A*0201), a very strong binder can be defined as having a % rank < 0.5, a strong binder as having a % rank < 1.0, a moderate binder as having a % rank between 1.0 and 2.0, and a weak binder as having a % rank > 2.0. The methods used for in vitro testing are well known to those skilled in the art. For example, technicians can utilize experimental protocols for validation of peptides represented by HLA-A*0201, as described in Tourdot et al., A general strategy to enhance immunogenicity of low-affinity HLA-A2.1-associated peptides: implication in the identification of cryptic tumor epitopes. Eur J Immunol. 2000 Dec; 30(12):341 1-21. In this context, reference peptides, such as HIV pol589-597, can be additionally used for testing.This allows for the calculation of the in vitro affinity of the observed binding relative to a reference peptide, for example, by the following equation: Relative affinity = Concentration of each peptide inducing 20% ​​expression of HLA-A*0201 / Concentration of a reference peptide inducing 20% ​​expression of HLA-A*0201 (where 100% is the level of HLA-A*0201 expression detected using a reference peptide, such as HIV pol 589-597, for example, at a concentration of 100 μM). For example, peptides showing a relative affinity below 1 can be considered “strong binders,” peptides showing a relative affinity between 1 and 2 can be considered “moderate binders,” and peptides showing a relative affinity greater than 3 can be considered “weak binders.”

[0073] In humans, there are three distinct loci encoding MHC class I molecules (human MHC molecules are also known as human leukocyte antigens (HLA)): HLA-A, HLA-B, and HLA-C. HLA-A*01, HLA-A*02, HLA-A*24, and HLA-B*07 are examples of different MHC class I alleles that can be expressed by these loci. For example, the antigenic peptide according to the invention can bind to HLA-A*01, HLA-A*02, HLA-A*24, and HLA-B*07 molecules. Specifically, the antigenic peptide according to the invention binds to HLA-A*02. In one embodiment, the antigenic peptide according to the invention is a microbial cluster sequence variant of an epitope of a human tumor antigen.

[0074] Microbial cluster sequence variants having epitopes of human tumor antigens were identified in at least one protein expressed in the human microbial cluster. Preferably, at least one protein present in the human microbial cluster is secreted or includes a transmembrane domain.

[0075] Cellular localization, specifically whether a protein is secreted or includes a transmembrane domain, can be performed computer-based or in vitro using methods known to those skilled in the art. For example, SignalP 4.1Server (Center for biological sequence analysis, Technical University of Denmark DTU; URL: www.cbs.dtu.dk / services / SignalP) and / or Phobius (a combined transmembrane topology and signal peptide predictor, Stockholm Bioinformatics Centre; URL: phobius.sbc.su.se) can be used. Preferably, both predictive tools (e.g., SignalP 4.1Server and Phobius) can be combined.

[0076] For example, to test whether a protein is secreted, the presence of a signal peptide can be evaluated. Signal peptides are ubiquitous protein sorting signals that target their passenger (cargo) proteins for translocation across the cytoplasmic membrane in prokaryotes. To test for the presence of a signal peptide, for example, SignalP 4.1Server (Center for biological sequence analysis, Technical University of Denmark DTU; URL: www.cbs.dtu.dk / services / SignalP) and / or Phobius (a combined transmembrane topology and signal peptide predictor, Stockholm Bioinformatics Centre; URL: phobius.sbc.su.se) can be used. Preferably, both predictive tools (e.g., SignalP 4.1Server and Phobius) can be combined.

[0077] Furthermore, it can be determined whether a protein contains a transmembrane domain. Both signal peptides and transmembrane domains are hydrophobic, but transmembrane helices typically have longer hydrophobic regions. For example, SignalP 4.1Server and Phobius have the ability to distinguish between signal peptides and transmembrane domains. Preferably, a minimum number of two predicted transmembrane helices are set to distinguish between membrane proteins and cytoplasmic proteins to provide a final common list.

[0078] In one implementation, a microbial cluster sequence variant of a fragment / epitope of a human tumor antigen is presented intact after cleavage of a bacterial protein expressed in the human microbiome. In this context, the term "cleavage" refers to the processing of a peptide for MHC presentation, specifically MHC-I processing. For example, cleavage prediction scores and / or affinity can be used to predict antigen cleavage for appropriate MHC binding (for presentation to CD8 T cells). This "cleavage probability score" can be calculated using software (preferably, such a score is above 70%, more preferably above 80%, and more preferably above 90%). For example, "IEDB" (Immune Epitope Database and Analysis Resource, IEDB Analysis Resource, supported by the National Institutes of Health in the Department of Health and Human Services, National Institute of Allergy and Infectious Diseases) can be used, which provides, for example, MHC-I processing prediction (URL: http: / / tools.iedb.org / mhcnp / ). In another instance, NetChop3 (The role of the proteasome ingenerating cytotoxic T cell epitopes: Insights obtained from improved predictions of proteasomal cleavage. M. Nielsen, C. Lundegaard, O. Lund, and C. Kesmir. Immunogenetics., 57(1-2):33-41, 2005; Prediction of proteasome cleavage motifs by neural networks. C. Kesmir, A. Nussbaum, Hansjorg Schild, Vincent Detours, and S. Brunak, Prot. Eng., 15(4):287-296, 2002; URL: http: / / www.cbs.dtu.dk / services / NetChop / ) can be used for cleavage prediction. This algorithm is based on a neural network trained on experimental data of proteasome cleavage and known HLA ligands in vitro. The network output is a score from 0.0 to 1.0. The random predictor score is 0.5, and the perfect predictor score is 1.Therefore, a high score for a peptide (e.g., 0.7 or higher, preferably 0.8 or higher, more preferably 0.9 or higher, as described above) indicates that it is efficiently cleaved at its N and / or C-terminus (rather than its center), while a low score is associated with peptides cleaved at their center. In this way, information on proteasome cleavage, TAP transport, and MHC class 1 analytical tools can be combined to predict peptide presentation.

[0079] In one implementation, the antigenic peptide induces T-cell cross-reactivity against a human epitope of a (reference) tumor antigen. T-cell cross-reactivity is an immune system phenomenon defined as the recognition of two or more peptide-MHC complexes (pMHC) by the T-cell receptor (TCR).

[0080] Epitope mimicry involves the concept of sequence and structural similarity between exogenous antigens and self-antigens as a triggering mechanism for cross-reactive immune responses against self-antigens. Interestingly, due to the clonal exhaustion of T cells that recognize self-antigens, this epitope mimicry offers a possible way to circumvent the limitations of the human T cell pool.

[0081] Specifically, antigens that are distinct from self-antigens (e.g., human epitopes of tumor antigens) but share sequence similarity with self-antigens (i.e., antigenic peptides according to the invention) (i) can still be recognized due to cross-reactivity with T cell receptors and (ii) are expected to be recognized by T cells / TCRs and have not been depleted during T cell culture. Therefore, such antigens can elicit a strong immune response, leading to clonal expansion of T cells with potential cross-reactivity with self-antigens.

[0082] Cross-reactivity of T-cell receptors with human (reference) tumor antigen epitopes can be measured by EL1SPOT-IFNγ assay as shown in the Examples section. In short, HLA-A2 transgenic mice (e.g., HHD DR1 mice expressing human HLA-A2 and HLA-DR1 MHC and lacking mouse H-2 class I and II MHC and / or HHD DR3 mice expressing human HLA-A2 and HLA-DR3 MHC) were immunized with a prime injection on day 0 (d0) and boosted with an epitope of the antigen peptide of the present invention on day 14. Seven days after the booster injection (i.e., day 21), the mice were euthanized and their ability to secrete IFN-γ was evaluated in vitro by stimulating spleen cells with the antigen peptide of the present invention, as assessed by ELISPOT.

[0083] Specifically, the present invention provides antigenic peptides that are microbial cluster sequence variants of fragments / epitopes of human tumor antigens, wherein the epitopes of human tumor antigens may include or consist of any of SEQ ID Nos. 1-257 and 476-500.

[0084] Table 1A below provides an overview of the antigenic peptides according to the invention, their amino acid sequences and SEQ ID NOs, and the corresponding fragments / epitopes of human tumor antigens (also referred to herein as "human reference peptides"). Table 1A also provides information related to tumor antigens for each antigenic peptide according to the invention. SEQ ID NOs 1 to 257 and 476-500 refer to the HLA-A*02 antigenic peptides according to the invention.

[0085] Table 1A. HLA-A*02 antigenic peptide according to the present invention.

[0086]

[0087]

[0088]

[0089]

[0090]

[0091]

[0092]

[0093]

[0094] Table 1B below provides an overview of the common sequences (MHC Class I common sequences) of the antigenic peptides according to the present invention, their amino acid sequences and SEQ ID NO, and the corresponding fragments / epitopes of human tumor antigens (also referred to herein as "human reference peptides") and their core sequences.

[0095] Table 1B. Common MHC Class I sequences of the antigenic peptides according to the present invention.

[0096]

[0097]

[0098] Table 1C below provides an overview of the HLA-A*02 common sequences, amino acid sequences, and SEQ ID NOs of the antigenic peptides according to the present invention, as well as the corresponding core sequences of human tumor antigens. Table 1C also provides information on the core sequences and HLA-A*02 common sequences involved in each antigenic peptide according to the present invention.

[0099] Therefore, the antigenic peptide may include or consist of the amino acid sequence shown in any of SEQ ID NO 309–326. In some embodiments, the antigenic peptide includes or consists of any of SEQ ID NO 316, 304-315, 317-472, and 501-509, preferably the amino acid sequence shown in or consisting of any of SEQ ID NO 316, 304-315, and 317-326.

[0100] Table 1C. HLA-A*02 common sequences of the antigenic peptides according to the present invention.

[0101]

[0102]

[0103]

[0104]

[0105]

[0106] The antigenic peptides according to the present invention can be classified based on the corresponding human tumor antigen, the corresponding core sequence, and the corresponding MHC class I and / or HLA-A*02 common sequence, as can be retrieved from Tables 1A-1C.

[0107] Therefore, the antigenic peptide may comprise the (core) sequence of any one of SEQ ID NO 281-303. Preferably, the antigenic peptide comprises the (core) sequence of any one of SEQ ID NO 281, 283-289, 293, 294, 302, and 303. More preferably, the antigenic peptide comprises the (core) sequence of any one of SEQ ID NO 283, 287, 293, 294, 302, and 303. Even more preferably, the antigenic peptide comprises the (core) sequence of any one of SEQ ID NO 287, 293, 294, and 302.

[0108] In one embodiment, the antigenic peptide according to the invention is a sequence variant of a fragment of the tumor antigen CD19 (human reference peptide), such as “FLLFLTPME” (SEQ ID NO:258), “KLSLGLPGL” (SEQ ID NO:259), “SLVGILHLQ” (SEQ ID NO:260), “TLAYLIFCL” (SEQ ID NO:261), or “QQMGGFYLC” (SEQ ID NO:262). In a preferred embodiment, the antigenic peptide according to the invention is a sequence variant of a fragment of the tumor antigen CD19, such as an antigenic peptide having a core sequence consisting of the amino acid sequences shown in any of SEQ ID NOs 281-285. Therefore, the antigenic peptide according to the invention is a sequence variant of a fragment of the tumor antigen CD19, comprising or consisting of the following: the MHC class I common sequence shown in any of SEQ ID NOs 304-308. In specific embodiments, the antigenic peptide according to the invention is a sequence variant of a fragment of the tumor antigen CD19 that binds to a specific HLA molecule (HLA-A2*02), such as an antigenic peptide comprising or consisting of the following: the HLA-A2*02 common sequence shown in any of SEQ ID Nos. 327-346. More preferably, the antigenic peptide according to the invention is a microbial cluster sequence variant of a fragment of the tumor antigen CD19, such as an antigenic peptide comprising or consisting of the following: the amino acid sequence shown in any of SEQ ID Nos. 1-40. More specifically, the antigenic peptide according to the invention is a sequence variant of the CD19 fragment (human reference peptide) "FLLFLTPME" (SEQ ID No: 258), such as an antigenic peptide comprising or consisting of the following: the amino acid sequences shown in SEQ ID Nos. 1-12, 304, and 327-333, for example, an antigenic peptide comprising or consisting of the following: the amino acid sequence shown in SEQ ID No: 10. More preferably, the antigenic peptide according to the invention is a sequence variant of the CD19 fragment (human reference peptide) "KLSLGLPGL" (SEQ ID NO:259), such as an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO:13-33, 305, and 334-343. Even more preferably, the antigenic peptide according to the invention is a sequence variant of the CD19 fragment (human reference peptide) "SLVGILHLQ" (SEQ ID NO:260), such as an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO:34-35, 306, and 344, for example, an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO:34 or 35.More preferably, the antigenic peptide according to the invention is a sequence variant of the CD19 fragment (human reference peptide) "TLAYLIFCL" (SEQ ID NO:261), such as an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO:36-39, 307, and 345-346, for example, an antigenic peptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:39. Even more preferably, the antigenic peptide according to the invention is a sequence variant of the CD19 fragment (human reference peptide) "QQMGGFYLC" (SEQ ID NO:262), such as an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO:40 and 308, for example, an antigenic peptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:40.

[0109] In one embodiment, the antigenic peptide according to the invention is a sequence variant of a fragment of the tumor antigen CD20 (MS4A1; human reference peptide), such as “IALGGLLMI” (SEQ ID NO: 263), “IMNSLSLFA” (SEQ ID NO: 264), “LMIPAGIYA” (SEQ ID NO: 265), or “SLFLGILSV” (SEQ ID NO: 266). In a preferred embodiment, the antigenic peptide according to the invention is a sequence variant of a fragment of the tumor antigen CD20, such as an antigenic peptide having a core sequence consisting of the amino acid sequences shown in any of SEQ ID NOs 286-289. Therefore, the antigenic peptide according to the invention is a sequence variant of a fragment of the tumor antigen CD20, comprising or consisting of the following: the MHC class I common sequence shown in any of SEQ ID NOs 309-312. In specific embodiments, the antigenic peptide according to the present invention is a sequence variant of a fragment of the tumor antigen CD20 that binds to a specific HLA molecule (HLA-A2*02), such as an antigenic peptide comprising or consisting of the following: the HLA-A2*02 common sequence shown in any of SEQ ID Nos. 347-374. More preferably, the antigenic peptide according to the present invention is a microbial cluster sequence variant of a fragment of the tumor antigen CD20, such as an antigenic peptide comprising or consisting of the following: the amino acid sequence shown in any of SEQ ID Nos. 41-87. More preferably, the antigenic peptide according to the present invention is a sequence variant of the CD20 fragment (human reference peptide) "IALGGLLMI" (SEQ ID No: 263), such as an antigenic peptide comprising or consisting of the following: the amino acid sequences shown in SEQ ID Nos. 41-64, 309, and 347-360, for example, an antigenic peptide comprising or consisting of the following: the amino acid sequence shown in SEQ ID No: 61. More preferably, the antigenic peptide according to the invention is a sequence variant of the CD20 fragment (human reference peptide) "IMNSLSLFA" (SEQ ID NO:264), such as comprising or consisting of the amino acid sequences shown in SEQ ID NO:65-70, 310, 361-364, and 476-484, for example comprising or consisting of the amino acid sequences shown in any one of SEQ ID NO:65, 68, 70, and 477. More preferably, the antigenic peptide according to the invention is a sequence variant of the CD20 fragment (human reference peptide) "LMIPAGIYA" (SEQ ID NO:265), such as comprising or consisting of the amino acid sequences shown in SEQ ID NO:71-80, 311, and 365-369, for example comprising or consisting of the amino acid sequence shown in SEQ ID NO:72.More preferably, the antigenic peptide according to the invention is a sequence variant of the CD20 fragment (human reference peptide) "SLFLGILSV" (SEQ ID NO:266), such as an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO:81-87, 312 and 370-374, for example, an antigenic peptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:86.

[0110] In one embodiment, the antigenic peptide according to the invention is a sequence variant of a fragment of the tumor antigen CD22 (human reference peptide), such as “FLSNDTVQL” (SEQ ID NO:267), “HLLGPWLLL” (SEQ ID NO:268), “ILILAICGL” (SEQ ID NO:269), or “WVFEHPETL” (SEQ ID NO:270). In a preferred embodiment, the antigenic peptide according to the invention is a sequence variant of a fragment of the tumor antigen CD22, such as an antigenic peptide having a core sequence consisting of the amino acid sequences shown in any of SEQ ID NOs 290-293. Therefore, the antigenic peptide according to the invention is a sequence variant of a fragment of the tumor antigen CD22, comprising or consisting of the following: the MHC class I common sequence shown in any of SEQ ID NOs 313-316. In specific embodiments, the antigenic peptide according to the present invention is a sequence variant of a fragment of the tumor antigen CD22 that binds to a specific HLA molecule (HLA-A2*02), such as an antigenic peptide comprising or consisting of the following: the HLA-A2*02 common sequence shown in any of SEQ ID Nos. 375-390. More preferably, the antigenic peptide according to the present invention is a microbial cluster sequence variant of a fragment of the tumor antigen CD22, such as an antigenic peptide comprising or consisting of the following: the amino acid sequence shown in any of SEQ ID Nos. 88-110. More preferably, the antigenic peptide according to the invention is a sequence variant of the CD22 fragment (human reference peptide) "FLSNDTVQL" (SEQ ID NO: 267), such as an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO: 88-96, 313, and 375-380. Even more preferably, the antigenic peptide according to the invention is a sequence variant of the CD22 fragment (human reference peptide) "HLLGPWLLL" (SEQ ID NO: 268), such as an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO: 97-101, 314, and 381-383. Even more preferably, the antigenic peptide according to the invention is a sequence variant of the CD22 fragment (human reference peptide) "ILILAICGL" (SEQ ID NO: 269), such as an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO: 102-105, 315, and 384-386.More preferably, the antigenic peptide according to the invention is a sequence variant of the CD22 fragment (human reference peptide) "WVFEHPETL" (SEQ ID NO:270), such as an antigenic peptide comprising or consisting of the following amino acid sequences: SEQ ID NO:106-110, 316 and 387-390 and 485-488, for example, an antigenic peptide comprising or consisting of the following amino acid sequences: any one of SEQ ID NO:107, 108, 109 and 110.

[0111] In one embodiment, the antigenic peptide according to the invention is a sequence variant of a fragment of the tumor antigen CD37 (human reference peptide), such as “GLAFVPLQI” (SEQ ID NO:271), “GLYFGMLLL” (SEQ ID NO:272), “ILILAICGL” (SEQ ID NO:273), “LLLLFATQI” (SEQ ID NO:274), “SIVGICLGV” (SEQ ID NO:275), or “SLIKYFLFV” (SEQ ID NO:276). In a preferred embodiment, the antigenic peptide according to the invention is a sequence variant of a fragment of the tumor antigen CD37, such as an antigenic peptide having a core sequence consisting of the amino acid sequences shown in any of SEQ ID NOs 294-299. Therefore, the antigenic peptide according to the invention is a sequence variant of a fragment of the tumor antigen CD37, comprising or consisting of the following: the MHC class I common sequences shown in any of SEQ ID NOs 317-322. In specific embodiments, the antigenic peptide according to the present invention is a sequence variant of a fragment of the tumor antigen CD37 that binds to a specific HLA molecule (HLA-A2*02), such as an antigenic peptide comprising or consisting of the following: the HLA-A2*02 common sequence shown in any of SEQ ID Nos. 391-425. More preferably, the antigenic peptide according to the present invention is a microbial cluster sequence variant of a fragment of the tumor antigen CD37, such as an antigenic peptide comprising or consisting of the following: the amino acid sequence shown in any of SEQ ID Nos. 111-162. More preferably, the antigenic peptide according to the invention is a sequence variant of the CD37 fragment (human reference peptide) "GLAFVPLQI" (SEQ ID NO:271), such as an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO:111-130, 317 and 391-402 and 489-493, for example, an antigenic peptide comprising or consisting of the amino acid sequences shown in any one of SEQ ID NO:114, 117, 119, 120, 491 and 493. Even more preferably, the antigenic peptide according to the invention is a sequence variant of the CD37 fragment (human reference peptide) "GLYFGMLLL" (SEQ ID NO:272), such as an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO:131-136, 318 and 403-406. More preferably, the antigenic peptide according to the invention is a sequence variant of the CD37 fragment (human reference peptide) “ILILAICGL” (SEQ ID NO:273), such as an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO:137-147, 319 and 407-414.More preferably, the antigenic peptide according to the invention is a sequence variant of the CD37 fragment (human reference peptide) "LLLLFATQI" (SEQ ID NO:274), such as an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO:148-155, 320, and 415-420. More preferably, the antigenic peptide according to the invention is a sequence variant of the CD37 fragment (human reference peptide) "SIVGICLGV" (SEQ ID NO:275), such as an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO:156-157, 321, and 421. More preferably, the antigenic peptide according to the invention is a sequence variant of the CD37 fragment (human reference peptide) "SLIKYFLFV" (SEQ ID NO:276), such as an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO:158-162, 322, and 422-425.

[0112] In one embodiment, the antigenic peptide according to the invention is a sequence variant of a fragment of the tumor antigen TNFRSF13C (human reference peptide), such as “GLAFVPLQI” (SEQ ID NO:277), “GLALVLALV” (SEQ ID NO:278), “LLFGAPALL” (SEQ ID NO:279), “PLPGLLFGA”, or (SEQ ID NO:280). In a preferred embodiment, the antigenic peptide according to the invention is a sequence variant of a fragment of the tumor antigen TNFRSF13C, such as an antigenic peptide having a core sequence consisting of the amino acid sequences shown in any of SEQ ID NOs 300-303. Therefore, the antigenic peptide according to the invention is a sequence variant of a fragment of the tumor antigen TNFRSF13C, comprising or consisting of the following: the MHC class I common sequence shown in any of SEQ ID NOs 323-326. In specific embodiments, the antigenic peptide according to the present invention is a sequence variant of a fragment of the tumor antigen TNFRSF13C that binds to a specific HLA molecule (HLA-A2*02), such as an antigenic peptide comprising or consisting of the following: the HLA-A2*02 common sequence shown in any of SEQ ID Nos. 426-472. More specifically, the antigenic peptide according to the present invention is a microbial cluster sequence variant of a fragment of the tumor antigen TNFRSF13C, such as an antigenic peptide comprising or consisting of the following: the amino acid sequence shown in any of SEQ ID Nos. 163-257. More specifically, the antigenic peptide according to the present invention is a sequence variant of the TNFRSF13C fragment (human reference peptide) "GLAFVPLQI" (SEQ ID No: 277), such as an antigenic peptide comprising or consisting of the following: the amino acid sequences shown in SEQ ID Nos. 163-170, 323, and 426-428. More preferably, the antigenic peptide according to the invention is a sequence variant of the TNFRSF13C fragment (human reference peptide) "GLALVLALV" (SEQ ID NO:278), such as an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO:171-209, 324, and 429-446. Even more preferably, the antigenic peptide according to the invention is a sequence variant of the TNFRSF13C fragment (human reference peptide) "LLFGAPALL" (SEQ ID NO:279), such as an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO:210-225, 325, and 447-455, and 494-500, for example, an antigenic peptide comprising or consisting of the amino acid sequences shown in any one of SEQ ID NO:212, 217, 220, and 224.More preferably, the antigenic peptide according to the invention is a sequence variant of the TNFRSF13C fragment (human reference peptide) “LLFGAPALL” (SEQ ID NO:279), such as an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO:226-257, 326 and 456-472, for example, an antigenic peptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:227.

[0113] Preferably, the antigenic peptide according to the present invention comprises or consists of the following: the MHC class I common sequence shown in any one of SEQ ID Nos. 304-312, 315-316 and 325. More preferably, the antigenic peptide according to the present invention comprises or consists of the following: the HLA-A*02 common sequence shown in any one of SEQ ID Nos. 330, 332-333, 336, 341-344, 346, 354-355, 360, 363-364, 367, 371-372, 374, 387, 390, 392-393, 402 and 450.

[0114] In some embodiments, the antigenic peptide according to the invention comprises or consists of the following amino acid sequences shown in any one of SEQ ID NO 1-12, 34-35, 36-39, 40, 41-64, 65-70, 476-484, 71-80, 81-87, 106-110, 485-488, 111-130, 489-493, 210-225, 494-500, and 226-257; preferably, the antigenic peptide comprises or consists of the following amino acid sequences shown in any one of SEQ ID NO 34-35, 65-70, 476-484, 106-110, 485-488, 111-130, 489-493, 210-225, 494-500, and 226-257; more preferably, the antigenic peptide comprises or consists of the following: SEQ ID NO The amino acid sequence represented by any one of 65-70, 476-484, 106-110, 485-488, 111-130, 489-493, 210-225, and 494-500.

[0115] In some embodiments, the antigenic peptide comprises or consists of the following amino acid sequences shown in any one of SEQ ID NO 10, 34, 35, 39, 40, 61, 68, 70, 72, 86, 107-110, 114, 117, 119, 120, 212, 217, 220, 224, 227, 231, 477, 491, and 493. More specifically, the antigenic peptide according to the invention comprises or consists of the following amino acid sequences shown in any one of SEQ ID NO 10, 21, 33, 35, 39, 40, 61, 65, 68, 72, 86, 110, 114, and 220. Even more preferably, the antigenic peptide according to the invention comprises or consists of the following amino acid sequences shown in any one of SEQ ID NO 10, 21, 33, 35, 39, 40, 110, 114, and 220. Most preferably, the antigenic peptide according to the invention comprises or consists of the amino acid sequence shown in any one of SEQ ID NO 10, 110, 114, and 220. More preferably, the antigenic peptide according to the invention comprises or consists of the amino acid sequence shown in any one of SEQ ID NO 65, 110, 114, and 220. In some embodiments, the antigenic peptide comprises or consists of the amino acid sequence shown in SEQ ID NO: 65. In some embodiments, the antigenic peptide comprises or consists of the amino acid sequence shown in SEQ ID NO: 110. In some embodiments, the antigenic peptide comprises or consists of the amino acid sequence shown in SEQ ID NO: 114. In some embodiments, the antigenic peptide comprises or consists of the amino acid sequence shown in SEQ ID NO: 220.

[0116] As illustrated in the examples herein, the specific antigenic peptides according to the invention allow for a strong immune response against themselves, and, most importantly, allow for a strong immune response against peptides contained in tumor antigens that have amino acid similarity to them, even if the human reference peptides contained in the tumor antigens may be tolerogenic.

[0117] Advantageously, the antigenic peptide according to the invention can be in the form of an immunogenic compound, specifically for the prevention or treatment of B-cell malignancies.

[0118] Immunogenic compounds comprising the antigenic peptide according to the invention

[0119] In a further aspect, the present invention also provides immunogenic compounds comprising the antigenic peptides according to the present invention as described above. Specifically, preferred embodiments of the antigenic peptides described above are also applicable to immunogenic compounds according to the present invention. For example, the antigenic peptides included in the immunogenic compounds preferably comprise or consist of the following: MHC class I common sequences shown in any of SEQ ID Nos. 304-326, such as antigenic peptides comprising or consisting of the following: amino acid sequences shown in any of SEQ ID Nos. 1-247 and 476-500. For example, it is even more preferred to include the following or consist of the following antigenic peptides according to the present invention: amino acid sequences shown in any of SEQ ID Nos. 10, 21, 33, 35, 39, 40, 61, 65, 68, 72, 86, 110, 114, and 220. For example, it is even more preferred to include the following or consist of the following antigenic peptides according to the present invention: amino acid sequences shown in any of SEQ ID Nos. 10, 21, 33, 35, 39, 40, 110, 114, and 220. For example, it is even more preferred to include the antigenic peptides according to the invention comprising or consisting of the following: the amino acid sequences shown in any one of SEQ ID NO 10, 110, 114 and 220. For example, it is even more preferred to include the antigenic peptides according to the invention comprising or consisting of the following: the amino acid sequences shown in any one of SEQ ID NO 65, 110, 114 and 220. Moreover, combinations thereof are preferred, i.e., the immunogenic compound comprises different antigenic peptides according to the invention.

[0120] As used herein, the term "immunogenic compound" refers to a compound that can induce, increase, prolong or maintain an immune response, specifically when administered to mammals and, in particular, to human individuals.

[0121] In general, the term "immunogenic compound" includes all kinds of compounds containing the antigenic peptide according to the invention. For example, the antigenic peptide according to the invention can be linked to a carrier molecule, or the antigenic peptide according to the invention can be contained in a polypeptide or protein (which can exist "alone," i.e., not linked to any other compound, or the polypeptide or protein containing the antigenic peptide can be linked to a carrier molecule).

[0122] Preferably, the immunogenic compound according to the invention comprises an antigenic peptide and a carrier molecule, specifically wherein the antigenic peptide (or a polypeptide or protein containing the antigenic peptide) is linked to the carrier molecule. Preferred carrier molecules are carrier proteins or carrier peptides. According to a preferred embodiment, the antigenic peptide as defined above, or the polypeptide / protein containing said antigenic peptide, is linked to the carrier protein or carrier peptide, for example, via covalent or non-covalent bonds. Optionally, such carrier proteins or carrier peptides described herein may be co-administered (alone) as an immunoadjuvant (i.e., not as an "immunogenic compound," but as a co-administration / combination therapy, as described below).

[0123] The carrier molecule can also be a lipid or lipid-like moiety. In this case, the immunogenic compound can be a lipopeptide. As used herein, the term "lipopeptide" refers to a molecule containing a lipid or lipid-like moiety covalently linked to a peptide moiety. Generally, "lipids" are soluble in nonpolar solvents, but typically "lipids" are insoluble (or poorly soluble) in water. Examples of lipids or lipid-like moiety include, but are not limited to, fatty acids, waxes, sterols, monoglycerides, diglycerides, triglycerides, and phospholipids. Lipids can be fatty acids, glycerolipids, glycerophospholipids, sphingolipids, sterol lipids, prenol lipids, glycolipids, or polyacetylated lipids. Preferably, lipids are fatty acids or derivatives thereof (including monoglycerides, diglycerides, triglycerides, and phospholipids). Fatty acids typically contain a hydrocarbon chain terminated by a carboxyl group. Fatty acids can be saturated or unsaturated. Fatty acids can be attached to functional groups, such as those containing oxygen, halogen, nitrogen, or sulfur. The preferred fatty acids are saturated or unsaturated long-chain fatty acids, such as myristic acid (CH3(CH2)) with 14 carbon atoms. 12 COOH) or palmitic acid (CH3(CH2) with 16 carbon atoms) 14 COOH), and phospholipids, such as phosphatidylglycerol (PG).

[0124] Preferably, the antigenic peptide described herein or a polypeptide / protein containing the antigenic peptide may be co-administered or, for example, covalently or non-covalently linked to a protein / peptide with adjuvant properties, such as providing stimulation of CD4+Th1 cells. Although the antigenic peptide described herein is preferably bound to MHC class I, CD4+ helper epitopes may also be used to provide an effective immune response. Th1 helper cells are able to maintain effective dendritic cell (DC) activation and specific CTL activation by secreting interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), and interleukin-2 (IL-2) and enhancing the expression of co-stimulatory signals on DCs and T cells (Galaine et al., Interest of Tumor-Specific CD4 T Helper Cells for Therapeutic Anticancer Vaccine. Vaccines (Basel). 2015 June 30; 3(3):490-502).

[0125] For example, adjuvant peptides / proteins can preferably be non-tumor antigens that restore immune memory or provide non-specific assistance, or they can be specific tumor-derived helper peptides. Several helper peptides for providing non-specific T cell assistance have been described in the literature, such as tetanus helper peptide, scutellarin peptide, or PADRE peptide (Adotévi et al., Targeting antitumor CD4 helper T cells with universal tumor-reactive helper peptides derived from telomerase for cancer vaccine. Hum Vaccin Immunother. May 2013; 9(5):1073-7, Slingluff CL, The present and future of peptide vaccines for cancer: single or multiple, long or short, alone or in combination? Cancer J. Sep-October 2011; 17(5):343-50). Therefore, tetanus helper peptide, scutellarin peptide, and PADRE peptide are preferred examples of such adjuvant peptides / proteins. In addition, specific tumor-derived helper peptides are preferred. Specific tumor-derived helper peptides are typically presented by MHC class II, specifically by HLA-DR, HLA-DP, or HLA-DQ. These specific tumor-derived helper peptides can be fragments of sequences of shared overexpressed tumor antigens such as HER2, NY-ESO-1, hTERT, or IL13RA2. The length of such fragments is preferably at least 10 amino acids, more preferably at least 11 amino acids, even more preferably at least 12 amino acids, and most preferably at least 13 amino acids. Specifically, fragments of shared overexpressed tumor antigens such as HER2, NY-ESO-1, and hTERT with a length of 13 to 24 amino acids are preferred. Preferred fragments bind to MHC class II and can therefore be determined using IEBB's MHC class II binding prediction tool (immune epitope database and analysis resource; supported by the National Institutes of Health in the Department of Health and Human Services, National Institute of Allergy and Infectious Diseases; URL: http: / / www.iedb.org / ; http: / / tools.iedb.org / mhcii / ).Preferably, the adjuvant peptide / protein can be the HHD-DR3 peptide with the sequence MAKTIAYDEEARRGLERGLN (SEQ ID NO: 473). Another preferred example is h-pAg T13L (sequence: TPPAYRPPNAPIL; SEQ ID NO: 474; Bhasin M, Singh H, Raghava GP (2003) MHCBN: a comprehensive database of MHC binding and non-binding peptides. Bioinformatics 19:665–666). Further examples of preferred adjuvant peptides / proteins (specifically, accessory peptides) include the UCP2 peptide (e.g., as described in WO 2013 / 135553 A1 or Dosset et al. Clin Cancer Res. 2012 Nov 15; 18(22):6284-9) and the BIRC5 peptide (e.g., as described in EP2119726 A1 or Wildenmeyer et al. Int J Cancer. 2012 July 1; 131(1):140-9). The most preferred accessory peptide is the UCP2 peptide (amino acid sequence: KSVWSKLQSIGIRQH; SEQ ID NO: 475, e.g., as described in WO 2013 / 135553 A1 or Dosset et al., Clin Cancer Res. 2012 Nov 15; 18(22):6284-95).

[0126] Preferably, the immunogenic compound according to the invention is a polypeptide or protein comprising the antigenic peptide according to the invention. Preferably, such protein or polypeptide is a recombinant protein or polypeptide, such as a fusion protein. The term "recombinant" means that it does not exist in nature.

[0127] In a preferred embodiment, the immunogenic compound according to the invention comprises or consists of a polypeptide of formula (I).

[0128] PepNt-CORE-PepCt(I)

[0129] in:

[0130] -"PepNt" consists of a polypeptide of varying length between 0 and 500 amino acid residues and is located at the N-terminus of the polypeptide of formula (I).

[0131] -"CORE" is composed of the antigenic peptide according to the invention as defined above; and

[0132] -"PepCt" consists of polypeptides of varying lengths between 0 and 500 amino acid residues and is located in formula (I).

[0133] The C-terminus of the polypeptide.

[0134] For example, immunogenic compounds may include or consist of polypeptides of formula (Ia) or (Ib):

[0135] PepNt-CORE (Ia); or

[0136] CORE-PepCt (Ib)

[0137] The terms “PepNt”, “PepCt”, and “CORE” are defined as above.

[0138] Preferably, the polypeptide of formula (I), (Ia) or (Ib) is a fusion peptide or fusion protein, specifically a recombinant fusion peptide or protein.

[0139] Preferably, the polypeptide or immunogenic compound as defined above comprises 9 to 1000 amino acids; including 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, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 6 4, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, and 1000 amino acids. Therefore, the lengths of “PepNt” and “PepCt”, if applicable, can be limited accordingly.

[0140] Therefore, "PepNt" and "PepCt", as defined above, may comprise 0 to 500 amino acid residues; including 0, 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, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 250, 300, 350, 400, 450, and 500 amino acid residues.

[0141] The types of carrier molecules used to generate the immunogenic compounds of the present invention (such as immunogenic compounds comprising or composed of a polypeptide of formula (I) linked to a carrier molecule) are entirely within the scope of knowledge of those skilled in the art. Specifically, the function of the carrier molecule is to provide cytokine assistance (or T cell assistance) to enhance the immune response against tumor antigens.

[0142] Preferably, the antigenic peptide is linked to a carrier molecule, specifically to a carrier protein, preferably via a covalent or non-covalent bond. The carrier molecule to which the peptide is optionally linked can be selected from a variety of known carriers. Examples of carrier molecules for vaccine use include proteins such as human or bovine serum albumin and keratocyanin peptide (KLH) and fatty acids. Other embodiments of the carrier molecule covalently linked to the antigenic peptide of formula (I) include bacterial toxins or toxins such as diphtheria, cholera, heat-labile or tetanus toxins of E. coli, outer membrane protein of Neisseria meningitidis (European Patent Application No. EP0372501), synthetic peptides (European Patent Application Nos. EP0378881 and EP0427347), heat shock protein (PCT application No. WO93 / 17712), pertussis protein (PCT application No. WO98 / 58668), protein D of Haemophilus influenzae (PCT application No. WO00 / 56360), and toxin A or B of Clostridium difficile (International Patent Application No. WO00 / 61761).

[0143] More preferably, the carrier protein or carrier peptide is a protein / peptide with immune adjuvant properties, such as providing stimulation to CD4+ Th1 cells, as described herein. Preferred examples are non-tumor antigens that restore immune memory or provide non-specific assistance, or can be specific tumor-derived helper peptides, such as tetanus helper peptides, pincer hemocyanin peptides, or PADRE peptides. Another preferred example is a specific tumor-derived helper peptide that can be presented by MHC class II, specifically by HLA-DR, HLA-DP, or HLA-DQ, such as fragments of shared overexpressed tumor antigens, such as HER2, NY-ESO-1, or hTERT. In a preferred embodiment, the carrier protein or carrier peptide is a protein / peptide with immune adjuvant properties, such as the HHD-DR3 carrier peptide MAKXIAYDEEARRGLERGLN (SEQ ID NO: 473). Specifically, “PepNt” and / or “PepCt” can correspond to a carrier protein or carrier peptide, such as the HHD-DR3 carrier peptide MAKTIAYDEEARRGLERGLN (SEQ ID NO: 473). Another preferred example is h-pAg T13L (sequence: TPPAYRPPNAPIL; SEQ ID NO: 474; Bhasin M, Singh H, Raghava GP (2003) MHCBN: a comprehensive database of MHC binding and non-binding peptides. Bioinformatics 19:665–666). Further examples of preferred carrier proteins / peptides (specifically, accessory peptides) include the UCP2 peptide (e.g., as described in WO 2013 / 135553A1 or Dosset et al., Clin Cancer Res. 2012 Nov 15; 18(22):6284-95) and the BIRC5 peptide (e.g., as described in EP2119726 A1 or Wildenmeyer et al., Int J Cancer. 2012 July 1; 131(1):140-9). The most preferred accessory peptide is the UCP2 peptide (amino acid sequence: KSVWSKLQSIGIRQH; SEQ ID NO: 475).

[0144] Furthermore, in the polypeptides according to formula (I), (Ia) or (Ib), “PepNt” and / or “PepCt” may preferably correspond to such proteins / peptides having immune adjuvant properties, such as providing stimulation of CD4+Th1 cells, as described herein.

[0145] Furthermore, the immunogenic compound may comprise or consist of such a protein / peptide having immunoadjuvant properties, covalently linked to the N-terminus of the antigenic peptide according to the invention or to the N-terminus of a polypeptide / protein containing said antigenic peptide, such as providing stimulation of CD4+Th1 cells as described herein.

[0146] Preferably, the antigenic peptide (or polypeptide / protein comprising the antigenic peptide) according to the invention is covalently bound to the carrier molecule via a linker portion.

[0147] Preferred linker agents include linker agents named GMBS, sulfon-GMBS, SMPB, and sulfon-SMPB.

[0148] In some embodiments of the immunogenic compound as defined above, the linker reagent is selected from GMBS (N-[γ-maleiminobutyryl-oxy]succinimide), sulfon-GMBS (N-[γ-maleiminobutyryl-oxy]sulfosuccinimide), SMSB (4-[p-maleiminophenyl]butyrate succinimide), and sulfon-SMPB (4-[p-maleiminophenyl]butyrate sulfosuccinimide).

[0149] Methods for conjugating two proteins using linker reagents, and more specifically using linker reagents selected from GMBS, sulfonyl-GMBS, SMSB, and sulfonyl-SMPB, are well known to those skilled in the art. Exemplarily, such methods are disclosed in publicly available leaflets from Pierce Corporation (Illinois, USA). GMBS, sulfonyl-GMBS, SMSB, and sulfonyl-SMPB consist of a bifunctional linker reagent comprising an N-hydroxysuccinimide (NHS) ester group and a maleimide group. Conjugation using GMBS, sulfonyl-GMBS, SMSB, or sulfonyl-SMPB is typically performed via a two-step procedure. In the first step, the amine-containing protein is reacted with a several molar excess of the linker reagent at pH 7–9 to form an amide bond, and then the excess unreacted linker reagent is removed—usually by desalting or dialysis. In the second step, a thiol-containing molecule (e.g., a peptide of formula (I)) is added to react with the maleimide group already attached to the first protein at pH 6.5-7.5 to form a stable thioether bond.

[0150] Using SMPB or sulfonyl-SMPB as linker reagents, an antigenic peptide (or a polypeptide / protein comprising said antigenic peptide, such as a polypeptide of formula (I)) according to the invention is covalently linked to an amine-containing carrier protein to produce a conjugate of formula (II):

[0151]

[0152] in:

[0153] -R1 consists of a reactive group of an amine-containing transporter protein, wherein the attached NH group originates from (i) the α-amino group at the N-terminus of the amine-containing transporter protein or (ii) the side-chain amino group of the lysine (K) amino acid residue of the amine-containing transporter protein; and

[0154] -R2 is composed of the antigenic peptide according to the invention (or a polypeptide / protein containing said antigenic peptide, such as a polypeptide of formula (I)), wherein the sulfur (S) atom to which it is attached is derived from a thiol (SH) group of a cysteine ​​residue located at the N-terminus or C-terminus of the peptide of formula (I). In some embodiments, the thiol moiety may be part of a non-natural amino acid or any other molecule present at the end of the peptide of formula (I).

[0155] Using GMBS or sulfonyl-GMBS as linker reagents, the antigenic peptide according to the invention (or a polypeptide / protein including said antigenic peptide, such as a polypeptide of formula (I)) is covalently linked to an amine-containing carrier protein (specifically CRM197 carrier protein) to produce a conjugate of formula (III):

[0156]

[0157] in:

[0158] -R1 consists of a reactive group of an amine-containing transporter protein, wherein the attached NH group originates from (i) the α-amino group at the N-terminus of the amine-containing transporter protein or (ii) the side-chain amino group of the lysine (K) amino acid residue of the amine-containing transporter protein; and

[0159] -R2 is composed of the antigenic peptide according to the invention (or a polypeptide / protein containing said antigenic peptide, such as a polypeptide of formula (I)), wherein the sulfur (S) atom to which it is attached is derived from a thiol (SH) group of a cysteine ​​residue located at the N-terminus or C-terminus of the peptide of formula (I). In some embodiments, the thiol moiety may be part of a non-natural amino acid or any other molecule present at the end of the peptide of formula (I).

[0160] Peptide-MHC (pMHC) multimers including antigenic peptides

[0161] In a further aspect, the present invention also provides peptide-MHC (pMHC) polymers comprising the antigenic peptide according to the present invention.

[0162] As used herein, the term "peptide-MHC multimer" (pMHC) refers to a stable multimeric complex composed of major histocompatibility complex (MHC) protein subunits loaded with the antigenic peptide of the present invention. Generally, "MHC multimer" is an oligomeric form of the MHC molecule. The primary function of the MHC molecule is to bind to an antigen. According to the present invention, the antigen is an antigenic peptide according to the present invention. Therefore, an MHC protein complex loaded with the antigenic peptide of the present invention generally means that the antigenic peptide of the present invention binds to one or more MHC proteins. The "peptide-MHC multimer" (pMHC) of the present invention includes, but is not limited to, peptide-MHC dimers, trimers, tetramers, pentamers, hexamers, heptamers, or octamers. MHC tetramers and pentamers are preferred. The term "major histocompatibility complex" (MHC) is a superordinate noun intended to encompass histocompatibility antigen systems described in various species, including human leukocyte antigen (HLA). In humans, there are three genetic loci encoding MHC class I molecules: HLA-A, HLA-B, and HLA-C. HLA-A*01, HLA-A*02, and HLA-A*11 are examples of different MHC class I alleles that can be expressed by these loci.

[0163] In one embodiment of the present invention, the pMHC polymer is a peptide / MHC class I polymer. In another specific embodiment, the pMHC polymer is an HLA corresponding to MHC class I / peptide polymers. Therefore, the pMHC polymer can be an HLA-peptide polymer selected from HLA-A-peptide polymers, HLA-B-peptide polymers, HLA-C-peptide polymers, HLA-E-peptide polymers, MICA-peptide polymers, and MICB-peptide polymers.

[0164] Methods for obtaining pHMC polymers are known in the art and are described, for example, in WO96 / 26962 and WO01 / 18053, which are incorporated herein by reference.

[0165] In addition to the MHC molecule and the antigenic peptide of the present invention, the pMHC may also contain other components, such as polymerizing agents and / or labels (e.g., for visualization). Examples of labels include, but are not limited to, fluorescent labels, such as fluorescently labeled proteins, like streptavidin. Fluorescent labels include allophycocyanin (APC), phycoerythrin (PE), R-phycoerythrin (R-PE), and fluorescein isothiocyanate (FITC). A preferred label is biotin.

[0166] In one embodiment of the invention, the pMHC multimer can be used to visualize specific T cell populations of MHC class I peptide complexes or HLA corresponding to MHC class I / peptide complexes as described above. For example, the pMHC multimer can be a multimer in which the heavy chain of MHC is biotinylated, allowing it to combine with streptavidin to form a tetramer. Such pMHC tetramers have increased affinity for suitable TCR-carrier T lymphocytes and can therefore be used to visualize reactive populations by immunofluorescence. In another embodiment of the invention, the pMHC multimer can be used for detection and / or isolation of specific T cell populations of pMHC complexes as described above by screening (in flow cytometry or by immunomagnetic screening).

[0167] Antigen peptide-specific cytotoxic T lymphocytes (CTLs)

[0168] In a further aspect, the present invention also provides antigen peptide-specific cytotoxic T lymphocytes (CTLs) according to the present invention, specifically antigen peptide-specific activated cytotoxic T lymphocytes (CTLs) according to the present invention.

[0169] The present invention further provides a method for generating antigen peptide-specific cytotoxic T lymphocytes (CTLs) according to the invention, specifically, antigen peptide-specific activated cytotoxic T lymphocytes (CTLs) according to the invention, the method comprising contacting the CTLs in vitro with an antigen-loaded human class I or II MHC molecule expressed on the surface of an antigen-presenting cell or an artificial construct mimicking an antigen-presenting cell, wherein the antigen is an antigen peptide according to the invention. Preferred antigen-presenting cells include dendritic cells. The artificial construct mimicking an antigen-presenting cell can be, for example, a peptide-MHC multimer according to the invention. The step of contacting the CTLs with the antigen-loaded human class I or II MHC molecule expressed on the surface of the antigen-presenting cell or the artificial construct mimicking an antigen-presenting cell can be performed for a time sufficient to activate the CTLs in an antigen-specific manner. Preferably, the antigen peptide is a preferred antigen peptide as described above, such as an antigen peptide comprising or consisting of the amino acid sequences shown in any one of SEQ ID NO 65, 110, 114, and 220.

[0170] Activated T cells targeting the antigenic peptides of the present invention can be used for treatment. Specifically, activated T cells generated by the above method selectively recognize cells that abnormally express polypeptides (i.e., tumor antigens) including the amino acid sequences of SEQ ID NO:258-280, such as polypeptides including the amino acid sequences shown in any one of SEQ ID NO 264, 270, 271 and 279.

[0171] Preferably, the antigen peptide-specific (activated) cytotoxic T lymphocytes (CTLs) according to the present invention can have (displayed / expressed) memory markers. Such memory markers are preferably memory markers of intestinal memory cells, such as CCR9, CXCR3, CD103, CX3CR1, and α4β7+.

[0172] Compared to vaccination with peptides not derived from microbial communities, such as human (reference) sequences and / or synthetic peptides (e.g., including, for example, artificially introduced mutations), antigen peptide-specific (activated) cytotoxic T lymphocytes (CTLs) according to the invention are preferably amplified more / stronger after vaccination with the antigen peptide (derived from human microbial communities). In other words, vaccination with the antigen peptide of the invention preferably increases the number of antigen peptide-specific (activated) cytotoxic T lymphocytes (CTLs) according to the invention more than vaccination with corresponding human peptides or synthetic peptides (not derived from microbial communities) associated with the same reference epitope.

[0173] The antigen peptide-specific (activated) cytotoxic T lymphocytes (CTLs) of the present invention preferably expand more, stronger, and / or faster in subjects who possess the peptide (expressed by the subject's microbiome) in the gut after vaccination, for example, compared to subjects who do not possess the peptide (not expressed by the subject's microbiome), such as subjects in which the peptide is not detected in fecal samples, the peptide can be found in the subject's fecal samples. Specifically, subjects who possess the peptide (expressed by the subject's microbiome) in the gut may respond more quickly (faster T cell expansion) and / or have T cells of the desired Tc1 type.

[0174] Cells loaded with antigenic peptides or immunogenic compounds

[0175] In a further aspect, the present invention also provides cells loaded with the antigenic peptides according to the present invention as described above, or immunogenic compounds comprising the antigenic peptides according to the present invention. Specifically, the preferred embodiments of the antigenic peptides described above are also applicable to such cells according to the present invention. For example, the antigenic peptides loaded onto the cells or the antigenic peptides included in the immunogenic compounds loaded onto the cells preferably comprise or consist of the following: MHC class I common sequences shown in any of SEQ ID Nos. 304-326, such as antigenic peptides comprising or consisting of the following: amino acid sequences shown in any of SEQ ID Nos. 1-247 and 476-500. For example, it is even more preferred to include the antigenic peptides according to the present invention comprising or consisting of the following: amino acid sequences shown in any of SEQ ID Nos. 10, 21, 33, 35, 39, 40, 61, 65, 68, 72, 86, 110, 114, and 220. For example, it is even more preferred to include the antigenic peptides according to the invention comprising or consisting of the following: the amino acid sequences shown in any one of SEQ ID NO 10, 21, 33, 35, 39, 40, 110, 114, and 220. For example, it is even more preferred to include the antigenic peptides according to the invention comprising or consisting of the following: the amino acid sequences shown in any one of SEQ ID NO 10, 110, 114, and 220. For example, it is even more preferred to include the antigenic peptides according to the invention comprising or consisting of the following: the amino acid sequences shown in any one of SEQ ID NO 65, 110, 114, and 220. Moreover, combinations thereof are preferred, i.e., cells loaded with different antigenic peptides (or corresponding immunogenic compounds (one or more)) according to the invention.

[0176] Preferred cells loaded with the antigenic peptide or immunogenic compound according to the invention are antigen-presenting cells (APCs), more preferably dendritic cells (DCs).

[0177] APCs are of particular interest because their primary function is to process antigens and present them on the cell surface of T cells in the immune system, thereby initiating and regulating T cell responses in vivo. In the context of this invention, it is preferred that the APCs are loaded with one or more antigenic peptides and / or one or more immunogenic compounds according to the invention. This can be accomplished by exposing the APCs in vitro to said antigenic peptides and / or one or more immunogenic compounds (e.g., Rizzo MM, Alaniz L, Mazzolini G. Ex vivo loading of autologous dendritic cells with tumor antigens. Methods Mol Biol. 2014; 1139:41-4; Rolinski J, Hus I. Breaking immunootolerance of tumors: a new perspective for dendritic cell therapy. J Immunotoxicol. 2014 Oct; 11(4):311-8).

[0178] The preferred APC according to the invention is dendritic cell (DC). Combining at least one antigenic peptide or immunogenic compound according to the invention with DC is indeed advantageous, as it is the most effective APC and is reported to be frequently functionally deficient in cancer patients. Those skilled in the art can readily obtain DC from healthy compatible donors (i.e., DCs that are HLA-associated) or from the patient themselves—provided they are functional (i.e., DCs that are autologous), for example, by direct isolation from peripheral blood or by derivatization from peripheral blood cells (such as CD14+ monocytes or CD34+ hematopoietic precursors) (Figdor CG, de Vries IJ, Lesterhuis WJ, Melif CJ. Dendritic cell immunotherapy: mapping the way. Nat Med. May 2004; 10(5):475-80). In fact, DCs can be distinguished from other cells in peripheral blood by their surface markers (such as S100, p55, CD83 and / or OX62), and therefore can be isolated and purified based on said markers using cell culture techniques known in the art.

[0179] The nucleic acid encoding the antigenic peptide and the host cell containing the nucleic acid.

[0180] In a further aspect, the present invention also provides nucleic acids encoding antigenic peptides according to the invention, polypeptides of formula (I) defined above, or immunogenic compounds according to the invention, wherein the immunogenic compounds are peptides or proteins. Specifically, preferred embodiments of the above-described antigenic peptides are also applicable to such nucleic acids according to the invention. For example, the antigenic peptide encoded by the nucleic acid preferably comprises or consists of the following: MHC class I common sequences shown in any of SEQ ID Nos. 304-326, such as antigenic peptides comprising or consisting of the following: amino acid sequences shown in any of SEQ ID Nos. 1-247 and 476-500. For example, it is even more preferred to include antigenic peptides according to the invention comprising or consisting of the following: amino acid sequences shown in any of SEQ ID Nos. 10, 21, 33, 35, 39, 40, 61, 65, 68, 72, 86, 110, 114, and 220. For example, it is even more preferred to include the antigenic peptides according to the invention comprising or consisting of the following: the amino acid sequences shown in any one of SEQ ID NO 10, 21, 33, 35, 39, 40, 110, 114, and 220. For example, it is even more preferred to include the antigenic peptides according to the invention comprising or consisting of the following: the amino acid sequences shown in any one of SEQ ID NO 10, 110, 114, and 220. For example, it is even more preferred to include the antigenic peptides according to the invention comprising or consisting of the following: the amino acid sequences shown in any one of SEQ ID NO 65, 110, 114, and 220. Moreover, combinations thereof are preferred, i.e., nucleic acids encoding different antigenic peptides according to the invention.

[0181] Nucleic acids preferably include single-stranded, double-stranded, or partially double-stranded nucleic acids, preferably selected from gDNA, cDNA, RNA, antisense DNA, antisense RNA, complementary RNA / DNA sequences with or without expression elements, minigenes, gene fragments, regulatory elements, promoters, and combinations thereof. Further preferred examples of nucleic acids (molecules) and / or polynucleotides include, for example, recombinant polynucleotides, vectors, oligonucleotides, RNA molecules such as rRNA, mRNA, or tRNA, or DNA molecules, as described above. Therefore, nucleic acids (molecules) are preferably DNA or RNA molecules; preferably selected from gDNA; cDNA; rRNA; mRNA; antisense DNA; antisense RNA; complementary RNA and / or DNA sequences; RNA and / or DNA sequences with or without expression elements, regulatory elements, and / or promoters; vectors; and combinations thereof.

[0182] In the fields of therapeutics, diagnostics, reagents, and bioassays, the ability to deliver nucleic acids, such as ribonucleic acid (RNA), intracellularly—whether in vitro, in vivo, in situ, or ex vivo—to induce intracellular translation of nucleic acids and the production of target-encoded peptides is of great interest. The delivery and function of non-integrating polynucleotides are of particular importance. Therefore, nucleic acids, such as mRNA, that do not integrate into the host chromosome are preferred. Generally, nucleic acids, such as mRNA, can be optimized to express the antigenic peptides of the present invention, for example, by methods known in the art, such as codon optimization. Additionally, nucleic acids can be modified, for example, to enhance their stability, prolong their lifespan, and / or increase the expression of the antigenic peptides of the present invention. Therefore, optimized or modified mRNAs (mmRNAs) encoding the antigenic peptides according to the present invention are preferred. mmRNAs are distinguished from wild-type mRNAs in terms of their functional and / or structural design features for optimal mRNA delivery and / or optimal expression of the antigenic peptides of the present invention (e.g., WO2013 / 151672A2, WO2013 / 101690A1, WO2013 / 052523A, which are incorporated herein by reference). In general, nucleic acids can be delivered "naked" or bound to a carrier (e.g., a cationic carrier). Cationic carriers (positively charged) generally bind readily to negatively charged nucleic acids. The carrier can be any kind of carrier, including, for example, polymers, proteins, lipids, and nanoparticles. Cationic lipids and nanoparticles (specifically, lipid nanoparticles, LNPs) are preferred for nucleic acid delivery. Therefore, the present invention also provides nucleic acids described herein bound to a carrier (e.g., lipids, specifically cationic lipids or LNPs).

[0183] In some embodiments, the nucleic acid molecule may be a vector. The term "vector," as used in the context of this invention, refers to a nucleic acid molecule, preferably an artificial nucleic acid molecule, i.e., a nucleic acid molecule that does not exist in nature. In the context of this invention, a vector is adapted to incorporate or contain a desired nucleic acid sequence. Such a vector may be a storage vector, an expression vector, a cloning vector, a transfer vector, etc. A storage vector is a vector that allows for convenient storage of nucleic acid molecules. Therefore, the vector may contain, for example, a sequence corresponding to a desired antigenic peptide according to the invention. An expression vector can be used to produce an expression product, such as RNA (e.g., mRNA), or a peptide, polypeptide, or protein. For example, an expression vector may contain a sequence required for transcription, such as a promoter sequence. A cloning vector is typically a vector containing a cloning site, which can be used to incorporate a nucleic acid sequence into the vector. A cloning vector may be, for example, a plasmid vector or a phage vector. A transfer vector may be a vector adapted to transfer nucleic acid molecules into cells or organisms, such as a viral vector. In the context of this invention, the vector may be, for example, an RNA vector or a DNA vector. Preferably, the vector is a DNA molecule. For example, a vector in the context of this application includes a cloning site, a selection marker such as an antibiotic resistance factor, and a sequence suitable for vector duplication such as an origin of replication. Preferably, the vector in the context of this application is a plasmid vector. Preferably, the vector in the context of this application is an expression vector. Preferred vectors are vectors for expression in bacterial cells. More preferably, the vector can be used for expression in so-called “live bacterial vaccine vectors”, wherein live bacterial cells (such as bacteria or bacterial spores, such as endospores, exospores, or microbial cysts) can act as a vaccine. A preferred example is described in da Silva et al., Live bacterial vaccine vectors: an overview; Braz J Microbiol. 2015 Mar 4; 45(4):1117-29.

[0184] The nucleic acid encoding the antigenic peptide according to the invention may be in the form of naked nucleic acid, or in the form of nucleic acid cloned into a plasmid or viral vector (Tregoning and Kinnear, Using Plasmids as DNA Vaccines for Infectious Diseases. Microbiol Spectr. Dec 2014; 2(6). doi:10.1128 / microbiolspec.PLAS-0028-2014), the latter being particularly preferred. Examples of suitable viral vectors according to the invention are not limited to retroviruses, adenoviruses, adeno-associated viruses (AAVs), herpesviruses, and poxvirus vectors. Cloning nucleic acids into plasmids or viral vectors using standard recombination techniques in the art is within the capabilities of those skilled in the art.

[0185] In a further aspect, the present invention also provides a host cell comprising nucleic acids according to the invention. Moreover, a combination thereof is preferred, i.e., a host cell comprising different nucleic acids according to the invention (e.g., encoding different antigenic peptides according to the invention).

[0186] Preferably, the nucleic acids included in the host cell are carriers. Preferably, the host cell is a bacterial cell. Such a host cell can preferably be used to produce the antigenic peptide according to the invention or the immunogenic compound according to the invention. Additionally, such a host cell can also be an active component in a vaccine.

[0187] Preferably, the host cell is a bacterial cell, more preferably an intestinal bacterial cell. The term "intestinal bacterial cell" refers to bacteria residing in the (human) intestine.

[0188] Such bacterial host cells can serve as “live bacterial vaccine vectors,” in which live bacterial cells (such as bacteria or bacterial spores, such as endospores, exospores, or microbial cysts) can act as vaccines. A preferred example is described in daSilva et al., Live bacterial vaccine vectors: an overview; Braz J Microbiol. 2015 Mar 4; 45(4):1117-29.

[0189] Bacterial cells (such as bacteria or bacterial spores, such as endospores, exospores, or microbial cysts), specifically (intact) gut bacterial species, can be advantageous because they have the potential to trigger a greater immune response than the (poly)peptides or nucleic acids they contain.

[0190] Optionally, the bacterial cells according to the invention, specifically intestinal bacteria, can be in the form of probiotics, i.e., live intestinal bacteria, which can therefore be used as a food additive due to the health benefits they can provide. They can be freeze-dried, for example, in granules, pills, or capsules, or consumed directly mixed with dairy products.

[0191] Nanoparticles including antigenic peptides or immunogenic compounds

[0192] In a further aspect, the present invention also provides nanoparticles comprising, specifically, nanoparticles loaded with the following:

[0193] - at least one antigenic peptide according to the invention, or

[0194] - At least one immunogenic compound according to the present invention;

[0195] Furthermore, the nanoparticles are optionally loaded with adjuvants.

[0196] Specifically, the preferred embodiments of the antigenic peptides described above also apply to nanoparticles according to the present invention. For example, the antigenic peptides loaded onto the nanoparticles or the antigenic peptides included in the immunogenic compounds preferably comprise or consist of the following: MHC class I common sequences shown in any of SEQ ID Nos. 304-326, such as antigenic peptides comprising or consisting of the following: amino acid sequences shown in any of SEQ ID Nos. 1-247 and 476-500. For example, it is even more preferred to include the following or consist of the antigenic peptides according to the present invention: amino acid sequences shown in any of SEQ ID Nos. 10, 21, 33, 35, 39, 40, 61, 65, 68, 72, 86, 110, 114, and 220. For example, it is even more preferred to include the following or consist of the antigenic peptides according to the present invention: amino acid sequences shown in any of SEQ ID Nos. 10, 21, 33, 35, 39, 40, 110, 114, and 220. For example, it is even more preferred to include the antigenic peptides according to the invention comprising or consisting of the following: the amino acid sequences shown in any one of SEQ ID NO 10, 110, 114 and 220. For example, it is even more preferred to include the antigenic peptides according to the invention comprising or consisting of the following: the amino acid sequences shown in any one of SEQ ID NO 65, 110, 114 and 220. Moreover, combinations thereof are preferred, i.e., nanoparticles loaded with different antigenic peptides (or corresponding immunogenic compounds (one or more)) according to the invention.

[0197] Nanoparticles, specifically used as vaccines, are known in the art and have been described, for example, in Shao et al., Nanoparticle-based immunotherapy for cancer, ACS Nano 2015, 9(1):16-30; Zhao et al., Nanoparticle vaccines, Vaccine 2014, 32(3):327-37; and Gregory et al., Vaccine delivery using nanoparticles, Front Cell Infect Microbiol. 2013, 3:13, doi:10.3389 / fcimb.2013.00013.eCollection 2013, Review. Specifically, nanoparticles are used to deliver antigenic peptides (or immunogenic compounds / peptides / proteins / nucleic acids containing antigenic peptides) and may optionally also act as adjuvants. Antigenic peptides (immunogenic compounds / peptides / proteins / nucleic acids containing antigenic peptides) are typically encapsulated within nanoparticles or attached to / bound to the surface of nanoparticles ("coating") ("coating"). Compared to conventional methods, nanoparticles can protect the payload (antigen / adjuvant) from the surrounding biological environment, increase half-life, minimize systemic toxicity, promote delivery to APCs, or even directly trigger the activation of TAA-specific T cells. Preferably, the size (diameter) of the nanoparticles is no greater than 300 nm, more preferably no greater than 200 nm, and most preferably no greater than 100 nm. Such nanoparticles are well evaded by phagocyte uptake, exhibit high structural integrity and long circulation time in circulation, can accumulate at tumor growth sites, and can penetrate deeply into tumor masses.

[0198] Examples of nanoparticles include polymer nanoparticles such as polyethylene glycol (PEG) and poly(D,L-lactic acid-co-glycolic acid) (PLGA); inorganic nanoparticles such as gold nanoparticles, iron oxide beads, iron oxide-zinc oxide nanoparticles, carbon nanotubes, and mesoporous silica nanoparticles; liposomes such as cationic liposomes; immunostimulatory complexes (ISCOM); virus-like particles (VLPs); and self-assembled proteins.

[0199] Polymer nanoparticles are nanoparticles based on / containing polymers such as poly(D,L-lactide-co-glycolic acid) (PLG), poly(D,L-lactic acid-co-glycolic acid) (PLGA), poly(γ-glutamic acid) (γ-PGA), poly(ethylene glycol) (PEG), and polystyrene. Polymer nanoparticles can encapsulate antigens (e.g., antigenic peptides or (poly)peptides containing them) or bind / conjugate to antigens (e.g., antigenic peptides or (poly)peptides containing them). Polymer nanoparticles can be used for delivery to, for example, certain cells, or to maintain antigen release through their slow biodegradation rate. For example, g-PGA nanoparticles can be used to encapsulate hydrophobic antigens. Polystyrene nanoparticles can be conjugated to a variety of antigens because they can be surface-modified with a variety of functional groups. Polymers such as poly(L-lactic acid) (PLA), PLGA, PEG, and natural polymers such as polysaccharides can also be used to synthesize hydrogel nanoparticles, which are a class of nanoscale hydrophilic three-dimensional polymer networks. Nanogels possess advantageous properties, including flexible pore size, large-area multivalent conjugation, high water content, and high antigen loading capacity. Therefore, preferred nanoparticles are nanogels, such as chitosan nanogels. Preferred polymer nanoparticles are those based on / containing PEG and PLGA.

[0200] Inorganic nanoparticles are nanoparticles based on / containing inorganic substances, and examples of such nanoparticles include gold nanoparticles, iron oxide beads, iron oxide and zinc oxide nanoparticles, carbon nanoparticles (e.g., carbon nanotubes), and mesoporous silica nanoparticles. Inorganic nanoparticles offer a rigid structure and controllable synthesis. For example, gold nanoparticles can be readily produced in various shapes, such as spheres, rods, and cubes. Inorganic nanoparticles can be surface-modified, for example, with carbohydrates. Carbon nanoparticles offer good biocompatibility and can be produced, for example, as nanotubes or (mesoporous) spheres. For example, multiple copies of the antigenic peptide (or (poly)peptide) according to the invention can be conjugated to carbon nanoparticles (e.g., carbon nanotubes). Mesoporous carbon nanoparticles are preferred for oral administration. Silica-based nanoparticles (SiNPs) are also preferred. SiNPs are biocompatible and exhibit excellent properties in selective tumor targeting and vaccine delivery. The abundant silanol groups on the surface of SiNPs can be used for further modification to introduce additional functions, such as cell recognition, absorption of specific biomolecules, improved cell interactions, and enhanced cellular uptake. Mesoporous silica nanoparticles are particularly preferred.

[0201] Liposomes are typically formed from phospholipids such as 1,2-dioleoyl-3-trimethylpropaneammonium (DOTAP). Generally, cationic liposomes are preferred. Liposomes self-assemble using a phospholipid bilayer shell and an aqueous core. Liposomes can be generated as monolayer vesicles (having one phospholipid bilayer) or multilayer vesicles (having several concentric phospholipid shells separated by an aqueous layer). Therefore, antigens can be encapsulated within the core or between different layers / shells. Preferred liposome systems are those approved for human use, such as... V and

[0202] Immunostimulatory complexes (ISCOMs) are cage-like particles approximately 40 nm in diameter that are micelles containing colloidal saponins, such as those composed of the saponin adjuvant Quil-A, cholesterol, phospholipids, and (poly)peptide antigens (such as antigenic peptides or polypeptides containing them). These spherical particles can capture antigens through nonpolar interactions. Two types of ISCOMs have been described, both consisting of cholesterol, phospholipids (typically phosphatidylethanolamine or phosphatidylcholine), and saponins (such as Quil-A).

[0203] Virus-like particles (VLPs) are self-assembled nanoparticles formed through the self-assembly of biocompatible capsid proteins. Due to their naturally optimized nanoparticle size and repeating structural sequence, VLPs can induce effective immune responses. VLPs can be derived from various viruses and range in size from 20 nm to 800 nm, typically in the 20–150 nm range. VLPs can be engineered to express other peptides or proteins—either by fusing these peptides / proteins into the particles or by expressing multiple antigens. Additionally, antigens can be chemically linked to the viral surface to create bioconjugated VLPs.

[0204] Examples of self-assembling proteins include ferritin and major cavern protein (MVP). Ferritin is a protein capable of self-assembling into a nearly spherical 10 nm structure. 96 MVP units can self-assemble into barrel-shaped cavern nanoparticles approximately 40 nm wide and 70 nm long. Antigens genetically fused with minimal interaction domains via self-assembly can be encapsulated into cavern nanoparticles when mixed with MVP. Therefore, antigens (such as antigenic peptides or polypeptides containing them according to the invention) can be fused with self-assembling proteins or fragments / domains thereof (such as the minimal interaction domains of MVP). Thus, the present invention also provides fusion proteins comprising self-assembling proteins (or fragments / domains thereof) and antigenic peptides according to the invention.

[0205] In general, preferred examples of nanoparticles (NPs) include iron oxide beads, polystyrene microspheres, poly(γ-glutamic acid)(γ-PGA)NPs, iron oxide-zinc oxide NPs, cationic gelatin NPs, Pluronic-stabilized poly(propylene sulfuride)(PPS)NPs, PLGA NPs, (cationic) liposomes, (pH-responsive) polymer micelles, PLGAs, PLGAs coated with cancer cell membranes, lipid-calcium phosphate (LCP) NPs, liposome-protamine-hyaluronic acid (LPH) NPs, polystyrene latex beads, magnetic beads, dextran iron particles, and quantum dot nanocrystals.

[0206] Preferably, the nanoparticles further comprise an adjuvant, such as a toll-like receptor (TLR) agonist. This allows the antigenic peptide (an immunogenic compound / peptide / protein / nucleic acid containing the antigenic peptide) to be delivered together with the adjuvant to, for example, antigen-presenting cells (APCs), such as dendritic cells (DCs). The adjuvant may be encapsulated by the nanoparticles or bound / conjugated to the surface of the nanoparticles, preferably similar to the antigenic peptide.

[0207] Particularly preferred adjuvants are polyinosinic acid:polycytidylic acid (also known as "polyI:C") and / or its derivative poly-ICLC. PolyI:C is a mismatched double-stranded RNA, where one strand is an inosinic acid polymer and the other is a cytidine polymer. PolyI:C is an immunostimulant known to interact with Toll-like receptor 3 (TLR3). PolyI:C is structurally similar to double-stranded RNA, which is a "natural" stimulant of TLR3. Therefore, polyI:C can be considered a synthetic double-stranded RNA analog. Poly-ICLC is a synthetic complex of carboxymethyl cellulose, polyinosinic acid-polycytidylic acid, and poly-L-lysine double-stranded RNA. Similar to polyI:C, poly-ICLC is also a ligand for TLR3. PolyI:C and poly-ICLC typically stimulate the release of cytotoxic cytokines. Preferred examples of poly-ICLC are...

[0208] Pharmaceutical Composition

[0209] In a further aspect, the present invention also provides a pharmaceutical composition comprising at least one of the following:

[0210] -The antigenic peptides according to the present invention described herein,

[0211] -The immunogenic compounds according to the present invention described herein,

[0212] -The nanoparticles according to the present invention described herein,

[0213] -The cells described herein are based on the present invention.

[0214] -The nucleic acid according to the present invention described herein,

[0215] -The host cell described herein according to the invention, and / or

[0216] -The cytotoxic T lymphocytes according to the present invention described herein,

[0217] And optionally one or more pharmaceutically acceptable excipients or carriers.

[0218] Therefore, the present invention provides pharmaceutical compositions comprising (at least) one antigenic peptide according to the invention as described herein. Furthermore, the present invention provides pharmaceutical compositions comprising (at least) one immunogenic compound according to the invention as described herein. Furthermore, the present invention provides pharmaceutical compositions comprising (at least) one nanoparticle according to the invention as described herein. Furthermore, the present invention provides pharmaceutical compositions comprising (at least) one cell according to the invention as described herein. Furthermore, the present invention provides pharmaceutical compositions comprising (at least) one nucleic acid according to the invention as described herein. Furthermore, the present invention provides pharmaceutical compositions comprising (at least) one host cell according to the invention as described herein. Furthermore, the present invention provides pharmaceutical compositions comprising (at least) one cytotoxic T lymphocyte according to the invention as described herein.

[0219] Specifically, the preferred embodiments of the antigenic peptides described above also apply to pharmaceutical compositions according to the present invention. For example, the antigenic peptides included in the pharmaceutical composition or in any of the immunogenic compounds, nanoparticles, cells, nucleic acids, or host cells included in the pharmaceutical composition preferably comprise or consist of the following amino acid sequences shown in any of SEQ ID NOs 1-247 and 476-500. For example, it is even more preferred to include the following or consist of the following antigenic peptides according to the present invention: the amino acid sequences shown in any of SEQ ID NOs 10, 21, 33, 35, 39, 40, 61, 65, 68, 72, 86, 110, 114, and 220. For example, it is even more preferred to include the following or consist of the following antigenic peptides according to the present invention: the amino acid sequences shown in any of SEQ ID NOs 10, 21, 33, 35, 39, 40, 110, 114, and 220. For example, it is even more preferred to include the antigenic peptide according to the invention comprising the amino acid sequences shown in any one of SEQ ID NO 10, 110, 114 and 220. For example, it is even more preferred to include the antigenic peptide according to the invention comprising the amino acid sequences shown in any one of SEQ ID NO 65, 110, 114 and 220.

[0220] Moreover, combinations thereof are preferred, i.e., pharmaceutical compositions comprising different antigenic peptides according to the invention. For example, the pharmaceutical composition may include

[0221] (i) at least two different antigenic peptides according to the invention;

[0222] (ii) at least two different immunogenic compounds according to the invention;

[0223] (iii) At least two different nanoparticles according to the invention;

[0224] (iv) at least two different nucleic acids according to the invention; and / or

[0225] (v) At least two different cytotoxic T lymphocytes according to the invention.

[0226] Therefore, a pharmaceutical composition may include at least "two different components" (according to the pharmaceutical composition of the invention). Preferably, three or four different components. Generally, as used herein, "different components" refers to...

[0227] (1) A first component, such as the antigenic peptide, immunogenic compound, nanoparticle, cell, nucleic acid, host cell, or cytotoxic T lymphocyte according to the invention as described herein; and

[0228] (2) At least one other component (which is different from the first component; and in the case of more than two different components, each component is different from each other component), such as the above-described anticancer therapeutic agent, different antigenic peptides according to the invention described herein, different immunogenic compounds according to the invention described herein, different nanoparticles according to the invention described herein, different cells according to the invention described herein, different nucleic acids according to the invention described herein, different host cells according to the invention described herein, different cytotoxic T lymphocytes according to the invention described herein, or any form (“naked”, as an immunogenic compound, as a nanoparticle, as a (host) cell, or as a nucleic acid) of one or more human tumor antigens (fragments).

[0229] Therefore, the “different components” are preferably active components (as described above) in the context of the disease to be prevented and / or treated (B-cell malignancy). In other words, each of the different components, if administered separately (rather than in combination as described herein), is also effective for the prevention and / or treatment of said cancer—although combination (i.e., combined administration) generally enhances its preventive and / or therapeutic effects (such as immune response), preferably in a synergistic manner.

[0230] Preferably, the “different components” are of the same type (e.g., different antigenic peptides, different immunogenic compounds, different nanoparticles, different cells, different nucleic acids, different host cells, or different cytotoxic T lymphocytes) and differ from each other only in that they involve different antigenic peptides of the present invention as described herein.

[0231] For example, at least three or four different active components are preferably of the same type, but differ (only) in that they each involve different antigenic peptides. More preferably...

[0232] - The first component involves (microbial cluster) sequence variants comprising or composed of the human tumor antigen CD22 fragment;

[0233] - The (different) second component involves (microbial cluster) sequence variants that include fragments of the human tumor antigen CD37 or antigenic peptides composed thereof;

[0234] - The (different) third component involves (microbial community) sequence variants comprising or composed of antigenic peptides containing fragments of the human tumor antigen TNFRSF13C; and

[0235] -Optionally, the (different) fourth component involves (microbial cluster) sequence variants comprising or composed of the human tumor antigen MS4A1 (CD20) fragment.

[0236] Even better

[0237] - The first component involves an antigenic peptide comprising a (microbial cluster) sequence variant of SEQ ID NO:270 or thereof;

[0238] - The (different) second component involves an antigenic peptide comprising a (microbial cluster) sequence variant of SEQ ID NO:271 or thereof;

[0239] - The (different) third component involves (microbial cluster) sequence variants including or composed of antigenic peptides of SEQ ID NO:279; and

[0240] -Optionally, the (different) fourth component involves an antigenic peptide comprising a (microbial cluster) sequence variant of SEQ ID NO:264 or composed thereof.

[0241] Even better

[0242] -The first component involves an antigenic peptide comprising the amino acid sequence shown in SEQ ID NO:110 or composed thereof;

[0243] - The (different) second component involves an antigenic peptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:114;

[0244] - (Different) third components involve antigenic peptides comprising or consisting of the amino acid sequence shown in SEQ ID NO:220; and

[0245] -Optionally, the (different) fourth component involves an antigenic peptide comprising the amino acid sequence shown in SEQ ID NO:65 or composed thereof.

[0246] Preferably, the pharmaceutical composition comprises at least two different antigenic peptides according to the invention.

[0247] Preferably, the pharmaceutical composition comprises a first antigenic peptide according to the invention, which comprises or consists of a (microbial community) sequence variant of a fragment of human tumor antigen CD22; and a second antigenic peptide according to the invention, which comprises or consists of a (microbial community) sequence variant of a fragment of human tumor antigen TNFRSF13C. Preferably, the first antigenic peptide comprises or consists of the following: a (microbial community) sequence variant of the CD22 fragment (human reference peptide) "WVFEHPETL" (SEQ ID NO: 270), such as an antigenic peptide comprising or consisting of the following: amino acid sequences shown in SEQ ID NO: 106-110, 316 and 387-390 and 485-488, for example, an antigenic peptide comprising or consisting of the following: amino acid sequences shown in any one of SEQ ID NO: 107, 108, 109 and 110, and the second antigenic peptide comprises or consists of the following: a (microbial community) sequence variant of the TNFRSF13C fragment (human reference peptide) "LLFGAPALL" (SEQ ID NO: 279), such as an antigenic peptide comprising or consisting of the following: amino acid sequences shown in SEQ ID NO: 210-225, 325 and 447-455 and 494-500, for example, an antigenic peptide comprising or consisting of the following: SEQ ID NO: 106-110, 316 and 387-390 and 485-48 ... As shown in any of SEQ ID NO: 212, 217, 220, and 224, specifically the amino acid sequences shown in SEQ ID NO: 220, 325, and 450. More preferably, the first antigenic peptide comprises or consists of the amino acid sequences shown in SEQ ID NO: 110, 387, and 390, and the second antigenic peptide comprises or consists of the amino acid sequences shown in SEQ ID NO: 220 and 450. Even more preferably, the pharmaceutical composition comprises an antigenic peptide comprising or consisting of SEQ ID NO: 110 and an antigenic peptide comprising or consisting of SEQ ID NO: 220.

[0248] More preferably, the pharmaceutical composition comprises at least three different antigenic peptides according to the invention.

[0249] Specifically, the pharmaceutical composition may include a first antigenic peptide according to the invention, which comprises or consists of a (microbial community) sequence variant of a fragment of human tumor antigen CD22; a second antigenic peptide according to the invention, which comprises or consists of a (microbial community) sequence variant of a fragment of human tumor antigen TNFRSF13C; and a third antigenic peptide according to the invention, which comprises or consists of a (microbial community) sequence variant of a fragment of human tumor antigen CD37. Preferably, the pharmaceutical composition comprises a first antigenic peptide comprising or consisting of the following: a (microbial community) sequence variant of the CD22 fragment (human reference peptide) "WVFEHPETL" (SEQ ID NO:270), such as an antigenic peptide comprising or consisting of the following: amino acid sequences shown in SEQ ID NO:106-110, 316 and 387-390 and 485-488, such as an antigenic peptide comprising or consisting of the following: amino acid sequences shown in any one of SEQ ID NO:107, 108, 109 and 110; and a second antigenic peptide comprising or consisting of the following: a (microbial community) sequence variant of the TNFRSF13C fragment (human reference peptide) "LLFGAPALL" (SEQ ID NO:279), such as an antigenic peptide comprising or consisting of the following: amino acid sequences shown in SEQ ID NO:210-225, 325 and 447-455 and 494-500, such as an antigenic peptide comprising or consisting of the following: SEQ ID NO: As shown in any of SEQ ID NO: 212, 217, 220, and 224, specifically the amino acid sequences shown in SEQ ID NO: 220, 325, and 450; and a third antigenic peptide comprising or consisting of the following: (microbial cluster) sequence variants of the CD37 fragment (human reference peptide) "GLAFVPLQI" (SEQ ID NO: 271), such as antigenic peptides comprising or consisting of the following: amino acid sequences shown in SEQ ID NO: 111-130, 317, and 391-402 and 489-493, for example, antigenic peptides comprising or consisting of the following: amino acid sequences shown in any of SEQ ID NO: 114, 117, 119, 120, 491, and 493, specifically the amino acid sequences shown in SEQ ID NO: 113-116, 324, and 392-393.

[0250] More preferably, the pharmaceutical composition comprises at least four different antigenic peptides according to the invention.

[0251] Specifically, the pharmaceutical composition may include a first antigenic peptide according to the invention, comprising or consisting of a (microbial community) sequence variant of a fragment of human tumor antigen CD22; a second antigenic peptide according to the invention, comprising or consisting of a (microbial community) sequence variant of a fragment of human tumor antigen TNFRSF13C; a third antigenic peptide according to the invention, comprising or consisting of a (microbial community) sequence variant of a fragment of human tumor antigen CD37; and a fourth antigenic peptide according to the invention, comprising or consisting of a (microbial community) sequence variant of a fragment of human tumor antigen CD19 or CD20.Preferably, the pharmaceutical composition comprises a first antigenic peptide comprising or consisting of the following: a (microbial community) sequence variant of the CD22 fragment (human reference peptide) "WVFEHPETL" (SEQ ID NO:270), such as an antigenic peptide comprising or consisting of the following: amino acid sequences shown in SEQ ID NO:106-110, 316 and 387-390 and 485-488, such as an antigenic peptide comprising or consisting of the following: amino acid sequences shown in any one of SEQ ID NO:107, 108, 109 and 110; and a second antigenic peptide comprising or consisting of the following: a (microbial community) sequence variant of the TNFRSF13C fragment (human reference peptide) "LLFGAPALL" (SEQ ID NO:279), such as an antigenic peptide comprising or consisting of the following: amino acid sequences shown in SEQ ID NO:210-225, 325 and 447-455 and 494-500, such as an antigenic peptide comprising or consisting of the following: SEQ ID NO: As shown in any of SEQ ID NO: 212, 217, 220, and 224, specifically the amino acid sequences shown in SEQ ID NO: 220, 325, and 450; a third antigenic peptide comprising or consisting of the following: a (microbial cluster) sequence variant of the CD37 fragment (human reference peptide) “GLAFVPLQI” (SEQ ID NO: 271), such as an antigenic peptide comprising or consisting of the following: amino acid sequences shown in SEQ ID NO: 111-130, 317, and 391-402 and 489-493, for example, an antigenic peptide comprising or consisting of the following: as shown in any of SEQ ID NO: 114, 117, 119, 120, 491, and 493, specifically the amino acid sequences shown in SEQ ID NO: 113-116, 324, and 392-393; and a fourth antigenic peptide comprising or consisting of the following: a CD20 (MS4A1) fragment (human reference peptide) “IMNSLSLFA” (SEQ ID NO: 220, 325, and 450); (NO:264) sequence variants of the (microbial community) such as those comprising or consisting of the following amino acid sequences shown in SEQ ID NO:65-70, 310 and 361-364 and 476-484, for example comprising or consisting of the following amino acid sequences shown in any of SEQ ID NO:65, 68, 70 and 477.

[0252] Preferably, the pharmaceutical composition comprises:

[0253] -The first antigenic peptide according to the invention comprises or is composed of a (microbial community) sequence variant of the human tumor antigen CD22 fragment.

[0254] -The second antigenic peptide according to the invention comprises or is composed of a (microbial cluster) sequence variant of the human tumor antigen TNFRSF13C fragment,

[0255] - The third antigenic peptide according to the invention comprises or is composed of a (microbial community) sequence variant of the human tumor antigen CD37 fragment, and

[0256] - Optionally, the fourth antigenic peptide according to the invention comprises or is composed of a (microbial cluster) sequence variant of the human tumor antigen CD20 fragment.

[0257] More preferably, the pharmaceutical composition comprises:

[0258] - A first antigenic peptide comprising or consisting of the following: a (microbial cluster) sequence variant of the CD22 fragment (human reference peptide) "WVFEHPETL" (SEQ ID NO: 270), such as an antigenic peptide comprising or consisting of the following: the amino acid sequences shown in SEQ ID NO: 106-110, 316 and 387-390.

[0259] - A second antigenic peptide comprising or consisting of the following: a (microbial cluster) sequence variant of the TNFRSF13C fragment (human reference peptide) "LLFGAPALL" (SEQ ID NO:279), such as an antigenic peptide comprising or consisting of the following: the amino acid sequences shown in SEQ ID NO:220, 325, and 450.

[0260] - A third antigenic peptide comprising or consisting of the following: a (microbial cluster) sequence variant of the CD37 fragment (human reference peptide) "GLAFVPLQI" (SEQ ID NO:271), such as an antigenic peptide comprising or consisting of the following: the amino acid sequences shown in SEQ ID NO:113-116, 324 and 392-393, and

[0261] - Optionally, a fourth antigenic peptide comprising or consisting of the following: a (microbial cluster) sequence variant of the CD20 fragment (human reference peptide) “IMNSLSLFA” (SEQ ID NO:264), such as an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO:65-70, 310 and 361-364.

[0262] More preferably, the pharmaceutical composition comprises an antigenic peptide according to the invention, said antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO 110, 114, 220 and optionally 65.

[0263] In some embodiments, the pharmaceutical composition does not further include antigenic peptides (other than the antigenic peptides of the present invention described above).

[0264] It should be understood that the pharmaceutical composition may also contain—instead of the preferred combination of the above-described antigenic peptides—a corresponding combination of the immunogenic compounds of the present invention, a corresponding combination of the nanoparticles of the present invention, or a corresponding combination of the nucleic acids of the present invention.

[0265] Preferably, the pharmaceutical composition further includes one or more pharmaceutically acceptable excipients or carriers.

[0266] The pharmaceutical compositions of the present invention may be in any form suitable for the purposes of the present invention. For example, the compositions may be in forms suitable for parenteral, enteral, or topical administration, such as liquid suspensions, solid dosage forms (granules, pills, capsules, or tablets), or pastes or gels. The selection of a suitable form of composition for the intended purpose is within the competence of those skilled in the art.

[0267] The compositions according to the invention may further include other active agents, such as those that enhance the effect of the antigenic peptide or immunogenic compound. Optionally, the compositions may not include any other active agents (i.e., in addition to the antigenic peptide according to the invention, the immunogenic compound according to the invention, the nanoparticles according to the invention, the cells according to the invention, the nucleic acids according to the invention, and / or the host cells according to the invention).

[0268] The pharmaceutical compositions defined herein are preferably immunogenic compositions, i.e., compositions capable of inducing, increasing, prolonging, or maintaining an immune response. This can be achieved by including an antigenic peptide according to the invention or an immunogenic compound according to the invention in the composition. Preferably, the pharmaceutical composition further includes one or more immune adjuvant substances. The pharmaceutical composition, specifically the immunogenic composition, may also be referred to herein as a "vaccine composition".

[0269] Preferably, the pharmaceutical composition further comprises at least one immunostimulant, specifically to increase, enhance, prolong, or maintain the antigen peptide-mediated immune response. Preferred immunostimulants according to the invention non-limitingly include immune adjuvants, antigen-presenting cells, and combinations thereof. Preferably, the immunostimulant is an immune adjuvant or an antigen-presenting cell (APC).

[0270] Preferably, the immunostimulant is an immune adjuvant. Some immune adjuvants can facilitate and prolong the duration of the interaction between the antigen and the immune system, while others can recruit and activate innate immune cells to induce an adaptive response. Adjuvants belonging to the former class include, without limitation, mineral compounds such as alum, aluminum hydroxide, aluminum phosphate, calcium phosphate hydroxide; and oil-based emulsions such as paraffin oil, starch oil, Freund's complete / incomplete adjuvant (FCA / FIA), and saponins (e.g., derived from plants such as Quillaja, soybean, and Polygala senega). Adjuvants belonging to the latter class include, without limitation, immunostimulatory complexes (ISCOMs), such as cytokines (e.g., GM-CSF; interleukins such as IL-1, IL-2, IL-6, IL-8, or IL-12; tumor necrosis factor (TNF), such as TNFα or TNFβ; interferon IFNS, such as IFNα, IFNβ, IFNγ, or IFNδ); ligands of Toll-like receptors (TLRs), such as imiquimod, resiquimod, or MPL; exogenous bodies, such as those derived from dendritic cells (DCs) or tumor cells; bacterial products, such as heat shock proteins (HSPs, such as gp96, hsp90, hsp70, calreticulin, hsp110, hsp170), pathogen-associated molecular patterns (PAMPs), trehalose dimethyl ester (TDM), muramyl dipeptide (MDP), and polysaccharides (PLS) (such as polysaccharide K).

[0271] More preferably, the immune adjuvant is a protein / peptide with immune adjuvant properties, such as providing stimulation to CD4+ Th1 cells, as described herein (“helper” peptides). Preferred examples are non-tumor antigens that restore immune memory or provide non-specific assistance, or can be tumor-specific helper peptides, such as tetanus helper peptides, pinworm hemocyanin peptides, or PADRE peptides, as described herein. Another preferred example is a tumor-specific helper peptide that can be presented by MHC II, specifically by HLA-DR, HLA-DP, or HLA-DQ, such as fragments of shared overexpressed tumor antigens, for example, HER2, NY-ESO-1, hTERT, or IL13RA2 as described above. Specifically, the immune adjuvant can be the HHD-DR3 peptide of the sequence MAKTIAYDEEARRGLERGLN (SEQ ID NO: 473). This peptide represents another example of a helper peptide (with immune adjuvant properties) that is preferred in the context of this invention. Another preferred example is h-pAg T13L (sequence: TPPAYRPPNAPIL; SEQ ID NO: 474; Bhasin M, Singh H, Raghava GP (2003) MHCBN: a comprehensive database of MHC binding and non-binding peptides. Bioinformatics 19:665–666). Further examples of preferred immune adjuvants (specifically, helper peptides) include UCP2 peptides (e.g., as described in WO 2013 / 135553A1 or Dosset et al., Clin Cancer Res. 2012 Nov 15; 18(22):6284-95) and BIRC5 peptides (e.g., as described in EP2119726 A1 or Wildenmeyer et al., Int J Cancer. 2012 July 1; 131(1):140-9). The preferred accessory peptide is the UCP2 peptide (amino acid sequence: KSVWSKLQSIGIRQH; SEQ ID NO: 475).

[0272] Preferably, the pharmaceutical composition comprises at least two different antigenic peptides and accessory peptides according to the invention, preferably UCP2 peptide (SEQ ID NO: 475).

[0273] Specifically, the pharmaceutical composition may include a first antigenic peptide according to the invention, which comprises or consists of a sequence variant of a fragment of human tumor antigen CD22; a second antigenic peptide according to the invention, which comprises or consists of a sequence variant of a fragment of human tumor antigen TNFRSF13C; and an accessory peptide, preferably a UCP2 peptide (SEQ ID NO:475). Preferably, the pharmaceutical composition comprises a first antigenic peptide comprising or consisting of the following: a sequence variant of the CD22 fragment (human reference peptide) “WVFEHPETL” (SEQ ID NO:270), such as an antigenic peptide comprising or consisting of the following amino acid sequences shown in SEQ ID NO:106-110, 316, and 387-390; a second antigenic peptide comprising or consisting of the following: a sequence variant of the TNFRSF13C fragment (human reference peptide) “LLFGAPALL” (SEQ ID NO:279), such as an antigenic peptide comprising or consisting of the following amino acid sequences shown in SEQ ID NO:220, 325, and 450; and an accessory peptide, preferably a UCP2 peptide (SEQ ID NO:475). More preferably, the first antigenic peptide comprises or consists of the following amino acid sequences shown in SEQ ID NO:110, 387, and 390, and the second antigenic peptide comprises or consists of the following amino acid sequences shown in SEQ ID NO:220 and 450. More preferably, the pharmaceutical composition comprises an antigenic peptide comprising or consisting of SEQ ID NO:110, an antigenic peptide comprising or consisting of SEQ ID NO:220, and a UCP2 helper peptide (SEQ ID NO:475).

[0274] More preferably, the pharmaceutical composition comprises at least three different antigenic peptides and accessory peptides according to the invention, preferably the UCP2 peptide (SEQ ID NO:475).

[0275] Specifically, the pharmaceutical composition may include a first antigenic peptide according to the invention, comprising or consisting of a (microbial community) sequence variant of a fragment of human tumor antigen CD22; a second antigenic peptide according to the invention, comprising or consisting of a (microbial community) sequence variant of a fragment of human tumor antigen TNFRSF13C; a third antigenic peptide according to the invention, comprising or consisting of a (microbial community) sequence variant of a fragment of human tumor antigen CD37; and an accessory peptide, preferably a UCP2 peptide (SEQ ID NO:475). Preferably, the pharmaceutical composition comprises a first antigenic peptide comprising or consisting of the following: a (microbial community) sequence variant of the CD22 fragment (human reference peptide) "WVFEHPETL" (SEQ ID NO:270), such as an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO:106-110, 316, and 387-390; a second antigenic peptide comprising or consisting of the following: a (microbial community) sequence variant of the TNFRSF13C fragment (human reference peptide) "LLFGAPALL" (SEQ ID NO:279), such as an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO:220, 325, and 450; and a third antigenic peptide comprising or consisting of the following: a (microbial community) sequence variant of the CD37 fragment (human reference peptide) "GLAFVPLQI" (SEQ ID NO:271), such as an antigenic peptide comprising or consisting of the following: SEQ ID NO: The amino acid sequences shown in NO:113-116, 324 and 392-393; and the accessory peptide, preferably the UCP2 peptide (SEQ ID NO:475).

[0276] Most preferably, the pharmaceutical composition comprises at least four different antigenic peptides and accessory peptides according to the invention, preferably the UCP2 peptide (SEQ ID NO:475).

[0277] Specifically, the pharmaceutical composition may include a first antigenic peptide according to the invention, comprising or consisting of a (microbial community) sequence variant of a fragment of human tumor antigen CD22; a second antigenic peptide according to the invention, comprising or consisting of a (microbial community) sequence variant of a fragment of human tumor antigen TNFRSF13C; a third antigenic peptide according to the invention, comprising or consisting of a (microbial community) sequence variant of a fragment of human tumor antigen CD37; a fourth antigenic peptide according to the invention, comprising or consisting of a (microbial community) sequence variant of a fragment of human tumor antigen CD19 or CD20; and an accessory peptide, preferably a UCP2 peptide (SEQ ID NO:475).

[0278] Preferably, the pharmaceutical composition comprises a first antigenic peptide according to the invention, comprising or consisting of a (microbial community) sequence variant of a fragment of human tumor antigen CD22; a second antigenic peptide according to the invention, comprising or consisting of a (microbial community) sequence variant of a fragment of human tumor antigen TNFRSF13C; a third antigenic peptide according to the invention, comprising or consisting of a (microbial community) sequence variant of a fragment of human tumor antigen CD37; (optionally) a fourth antigenic peptide according to the invention, comprising or consisting of a (microbial community) sequence variant of a fragment of human tumor antigen CD20; and an accessory peptide, preferably a UCP2 peptide (SEQ ID NO:475). More preferably, the pharmaceutical composition comprises a first antigenic peptide comprising or consisting of the following: a (microbial community) sequence variant of the CD22 fragment (human reference peptide) “WVFEHPETL” (SEQ ID NO:270), such as an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO:106-110, 316, and 387-390; a second antigenic peptide comprising or consisting of the following: a (microbial community) sequence variant of the TNFRSF13C fragment (human reference peptide) “LLFGAPALL” (SEQ ID NO:279), such as an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO:220, 325, and 450; and a third antigenic peptide comprising or consisting of the following: a (microbial community) sequence variant of the CD37 fragment (human reference peptide) “GLAFVPLQI” (SEQ ID NO:271), such as an antigenic peptide comprising or consisting of the following: SEQ ID NO: The amino acid sequences shown in NO:113-116, 324 and 392-393; (optionally) a fourth antigenic peptide comprising or consisting of the following: a (microbial cluster) sequence variant of the CD20 fragment (human reference peptide) “IMNSLSLFA” (SEQ ID NO:264), such as an antigenic peptide comprising or consisting of the following: the amino acid sequences shown in SEQ ID NO:65-70, 310 and 361-364; and an accessory peptide, preferably a UCP2 peptide (SEQ ID NO:475).

[0279] Particularly preferred immunoadjuvants are polyinosinic acid:polycytidylic acid (also known as "polyI:C") and / or its derivative poly-ICLC. PolyI:C is a mismatched double-stranded RNA, where one strand is a polymer of inosinic acid and the other is a polymer of cytidine. PolyI:C is an immunostimulant known to interact with Toll-like receptor 3 (TLR3). PolyI:C is structurally similar to double-stranded RNA, which is a "natural" stimulant of TLR3. Therefore, polyI:C can be considered a synthetic double-stranded RNA analog. Poly-ICLC is a synthetic complex of carboxymethyl cellulose, polyinosinic acid-polycytidylic acid, and poly-L-lysine double-stranded RNA. Similar to polyI:C, poly-ICLC is a ligand for TLR3. PolyI:C and poly-ICLC typically stimulate the release of cytotoxic cytokines. Preferred examples of poly-ICLC are...

[0280] Most preferably, the adjuvant is Montanide, such as Montanide ISA 51VG and / or Montanide ISA 720VG. These adjuvants provide a stable water-in-oil emulsion when mixed with an aqueous antigen medium. Montanide ISA51VG is based on a mixture of mannose monooleate surfactant and mineral oil, while Montanide ISA 720VG uses non-mineral oil (Aucouturier J, Dupuis L, Deville S, Ascarateil S, Ganne V. Montanide ISA720 and 51: a new generation of water in oil emulsions as adjuvants for human vaccines. Expert Rev Vaccines. June 2002; 1(1):111-8; Ascarateil S, Puget A, Koziol M-E. Safety data of Montanide ISA 51VG and Montanide ISA 720VG, two adjuvants dedicated to human therapeutic vaccines. Journal for Immunotherapy of Cancer. 2015; 3(Suppl 2):P428. doi:10.1186 / 2051-1426-3-S2-P428).

[0281] It is also preferred that the immunostimulant is an antigen-presenting cell (APC). APCs are of particular interest because their primary function is to process antigens and present them on the cell surface of T cells of the immune system, thereby initiating and regulating T cell responses in vivo. In the compositions of the present invention, it is preferred that the APCs are loaded with one or more antigenic peptides and / or one or more immunogenic compounds according to the present invention, which can be accomplished by exposing the APCs to said antigenic peptides and / or one or more immunogenic compounds in vitro (Rizzo et al., Methods Mol Biol. 2014; 1139:41-4; Rolinski and Hus, J Immunotoxicol. 2014 Oct; 11(4):311-8).

[0282] Preferably, the APC is dendritic cells (DC). DCs are the most potent APCs and are reported to be frequently functionally deficient in cancer patients. DCs can be readily obtained by those skilled in the art from healthy, compatible donors (i.e., the dendritic cells are HLA-associated) or from the patient themselves—provided they are functional (i.e., the DCs are autologous)—for example, by direct isolation from peripheral blood or by derivatization from peripheral blood cells such as CD14+ monocytes or CD34+ hematopoietic precursors (Emens et al., 2008). In fact, DCs can be distinguished from other cells in peripheral blood by their surface markers (such as S100, p55, CD83, and / or OX62) and can therefore be isolated and purified based on said markers using cell culture techniques known in the art.

[0283] According to a preferred embodiment, the pharmaceutical composition may further comprise at least one anticancer therapeutic agent. Therefore, the therapeutic agent is preferably capable of preventing and / or treating the same type of cancer as that for which the antigenic peptide according to the invention is used. Preferably, the anticancer therapeutic agent is selected from antibodies, CAR-T cells, tumor cell lysates, chemotherapy agents, radiotherapy agents, immune checkpoint modulators, and combinations thereof.

[0284] Antibodies are particularly advantageous in cancer treatment because they can bind to specific antigens on the surface of cancer cells, thereby directing treatment towards the tumor (i.e., these are called tumor-targeting antibodies), or block dysregulated immune checkpoints in cancer (i.e., these are referred to herein as immunomodulatory antibodies). The latter type of antibody aims to suppress cancer immune resistance, which can be significantly observed in tumor antigen-specific T cells. Indeed, as is well known in the art, under normal physiological conditions, immune checkpoints are essential for maintaining self-tolerance (i.e., preventing autoimmunity) and protecting tissues from damage when the immune system responds to pathogenic infections. However, in cancer, immune checkpoint expression can be dysregulated as an important mechanism of immune resistance. Regarding the PD-L1 checkpoint, the resistance has been observed in melanoma, ovarian cancer, lung cancer, glioblastoma, breast cancer, and pancreatic cancer (Konishi et al., B7-H1 expression on non-small cell lung cancer cells and its relationship with tumor-infiltrating lymphocytes and their PD-1 expression. Clin Cancer Res. 2004 Aug 1; 10(15):5094-100; Ghebeh et al., The B7-H1 (PD-L1) Tlymphocyte-inhibitory molecule is expressed in breast cancer patients with infiltrating ductal carcinoma: correlation with important high-risk prognostic factors. Neoplasia. 2006 Mar; 8(3):190-8; Hino et al., Tumor cell expression of programmed cell death-1 ligand 1 is a prognostic factor formalignant). melanoma. Cancer. 2010 Apr 1; ​​116(7):1757-66). Other examples of immune checkpoints include, without limitation, PD-L2, PD-1, CD80, CD86, CTLA-4, B7H3, B7H4, PVR, TIGIT, GAL9, LAG-3, GITR, CD137, TIM3, VISTA, VISTA-R (Pico de et al., Checkpoint blockade for cancer therapy: revitalizing a suppressed immune system. Trends Mol Med. 2015 Aug;21(8):482-91; Pardoll DM. The blockade of immune checkpoints in cancerimmunotherapy. Nat Rev Cancer. 2012 Mar 22;12(4):252-64).

[0285] Antibodies are typically used for the above purposes in the form of naked monoclonal antibodies (i.e., unconjugated) or conjugated with other molecules that are cytotoxic or radioactive.

[0286] Examples of known monoclonal tumor-targeting antibodies used in cancer immunotherapy, without limitation, include alematuzumab (chronic lymphocytic leukemia), bevacizumab (colorectal cancer, glioblastoma, cervical cancer, lung cancer, renal cancer), brentuximab / vedotin (lymphoma), blinatumumab (acute lymphoblastic leukemia), catumaxomab (EPCAM+ malignant ascites in cancer), cetuximab (head and neck cancer, colorectal cancer), denosumab (breast cancer, prostate cancer, and bone cancer), and gemtuzumab / ozogamicin. n) Acute myeloid leukemia, ibritumomab / tiuxetan (non-Hodgkin's lymphoma), panitumumab (colorectal cancer), pertuzumab (breast cancer), obinutuzumab (chronic lymphocytic leukemia), ofatumumab (chronic lymphocytic leukemia), opilimumab (melanoma), ramucirumab (gastric and gastroesophageal cancer), rituximab (chronic lymphocytic leukemia and non-Hodgkin's lymphoma), siltuximab (multicentric Carterman's disease). Catsleman's disease), tositumomab (non-Hodgkin's lymphoma), and trastuzumab (breast cancer, gastric cancer, and gastroesophageal cancer); while examples of immunomodulatory antibodies include, without limitation, ipilimumab (melanoma) – which blocks CTLA4-dependent immune checkpoints, nivolumab (melanoma, lung cancer) and prembrolizubmab (melanoma) – both of which block PDCD1-dependent immune checkpoints, and MPDL3280A, MEDI4736, MEDI0680, and MSB0010718C – all of which block PD-L1-dependent immune checkpoints (Sharma and Allison, The future of immune checkpoint therapy. Science. April 3, 2015; 348(6230):56-61).

[0287] Other antibodies used in cancer immunotherapy have been described in Buqué et al., Trial Watch: Immunomodulatory monoclonal antibodies for oncological indications. Oncoimmunology. 2015 Mar 2; 4(4):e1008814.eCollection 2015 Apr; Redman et al., Mechanisms of action of therapeutic antibodies for cancer. MolImmunol. 2015 Oct; 67(2Pt A):28-45; Simpson and Caballero, Monoclonal antibodies for the therapy of cancer MC Proc. 2014; 8(Suppl 4):O6 and the Antibody Society website (a list of therapeutic monoclonal antibodies approved or under review by the European Union or United States is available at the website link http: / / www.antibodysociety.org / news / approved_mabs.php).

[0288] Adoptive cellular immunotherapy using chimeric antigen receptor (CAR) T cells has transformed the treatment prospects for B-cell non-Hodgkin lymphoma (NHL), particularly aggressive B-cell lymphoma. For example, CD19-targeted CAR T cells represent a new standard of care for patients with DLBCL who are refractory to at least two prior lines of therapy. Two CAR T-cell products are axi-cel (KTE-019) and YESCARTA. TM ) and tisagenlecleucel(CTL019)(KYMRIAH TM This product has been approved by the U.S. Food and Drug Administration for the treatment of refractory DLBCL after two lines of therapy. A third product, lisocabtagene maraleucel (liso-cel) (JCAR017), is currently being evaluated in clinical trials. Other CAR T-cells include CD20-CAR-T cells.

[0289] Tumor cell lysates can also be combined with one or more antigenic peptides according to the invention. Tumor cells are indeed able to elicit an immune response by presenting an endogenous peptide-MHC complex, and via host dendritic cells (DCs) capable of processing and presenting the antigens delivered by the lysates. This increases the range of antigens that can induce an immune response. Tumor cell lysates can be readily obtained by treating tumor cells with heat shock and / or chemical treatment, and can be autologous (i.e., isolated from a patient) or allogeneic (i.e., isolated from another subject).

[0290] Standard chemotherapy drugs and radiation therapy agents need not be further described in this article, as they have been extensively described in the literature, especially by Baskar et al. (Baskar et al., Cancer and radiation therapy: current advances and future directions. Int J Med Sci. 2012; 9(3):193-9), Paci et al. (Paci et al., Review of therapeutic drug monitoring of anticancer drugs part 1--cytotoxics. Eur J Cancer. 2014 Aug; 50(12):2010-9), and Widmer et al. (Widmer et al., Review of therapeutic drug monitoring of anticancer drugs part two--targetedtherapies. Eur J Cancer. 2014 Aug; 50(12):2020-36). A list of these drugs and agents is also available on cancer.gov (http: / / www.cancer.gov / about-cancer / treatment / drugs).

[0291] Preferably, the immune checkpoint modulator used in combination with the antigenic peptide defined herein is an activator or inhibitor of one or more immune checkpoint molecules selected from the following: CD27, CD28, CD40, CD122, CD137, OX40, GITR, ICOS, A2AR, B7-H3, B7-H4, BTLA, CD40, CTLA-4, IDO, KIR, LAG3, PD-1, TIM-3, VISTA, CEACAM1, GARP, PS, CSF1R, CD94 / NKG2A, TDO, GITR, TNFR and / or FasR / DcR3; or an activator or inhibitor of one or more ligands thereof.

[0292] More preferably, the immune checkpoint modulator is an activator of (co)stimulatory checkpoint molecules or an inhibitor of inhibitory checkpoint molecules, or a combination thereof. Therefore, the immune checkpoint modulator is more preferably an activator of (i) CD27, CD28, CD40, CD122, CD137, OX40, GITR and / or ICOS or (ii) an inhibitor of A2AR, B7-H3, B7-H4, BTLA, CD40, CTLA-4, IDO, KIR, LAG3, PD-1, PDL-1, PD-L2, TIM-3, VISTA, CEACAM1, GARP, PS, CSF1R, CD94 / NKG2A, TDO, TNFR and / or FasR / DcR3.

[0293] More preferably, the immune checkpoint modulator is an inhibitor of inhibitory checkpoint molecules (but preferably not an inhibitor of stimulatory checkpoint molecules). Therefore, the immune checkpoint modulator is even more preferably an inhibitor of A2AR, B7-H3, B7-H4, BTLA, CTLA-4, IDO, KIR, LAG3, PD-1, PDL-1, PD-L2, TIM-3, VISTA, CEACAM1, GARP, PS, CSF1R, CD94 / NKG2A, TDO, TNFR, and / or DcR3 or their ligands.

[0294] It is also preferred that the immune checkpoint modulator is an activator of stimulatory or co-stimulatory checkpoint molecules (but preferably not an activator of inhibitory checkpoint molecules). Therefore, the immune checkpoint modulator is more preferably an activator of CD27, CD28, CD40, CD122, CD137, OX40, GITR and / or ICOS or their ligands.

[0295] More preferably, the immune checkpoint modulator is a modulator of the CD40 pathway, IDO pathway, LAG3 pathway, CTLA-4 pathway, and / or PD-1 pathway. Specifically, the immune checkpoint modulator is preferably a modulator of CD40, LAG3, CTLA-4, PD-L1, PD-L2, PD-1, and / or IDO; more preferably, the immune checkpoint modulator is an inhibitor of CTLA-4, PD-L1, PD-L2, PD-1, LAG3, and / or IDO, or an activator of CD40; even more preferably, the immune checkpoint modulator is an inhibitor of CTLA-4, PD-L1, PD-1, LAG3, and / or IDO; even more preferably, the immune checkpoint modulator is an inhibitor of LAG3, CTLA-4, and / or PD-1; and most preferably, the immune checkpoint modulator is an inhibitor of CTLA-4 and / or PD-1.

[0296] Therefore, the checkpoint modulator used in combination with the antigenic peptide can be selected from known modulators of the CTLA-4 or PD-1 pathways. Preferably, the checkpoint modulator used in combination with the antigenic peptide defined herein can be selected from known modulators of the CTLA-4 or PD-1 pathways. Particularly preferably, the immune checkpoint modulator is a PD-1 inhibitor. Preferred inhibitors of the CTLA-4 or PD-1 pathways include monoclonal antibodies. (Ipilimumab; Bristol Myers Squibb) and tremelimumab (Pfizer / MedImmune) and (Nivolumab; Bristol Myers Squibb) (Pembrolizumab, also known as Lambrolizumab or MK-3475; Merck) (Durvalumab, also known as MEDI4736; MedImmune / AstraZeneca) (Atezolizumab, also known as MPDL3280A; Roche / Genentech), Pidilizumab (CT-011; CureTech), MEDI0680 (AMP-514; AstraZeneca) (Avelumab; Merck KGaA / Pfizer, also known as MSB-0010718C), MIH1 (Affymetrix), LY3300054 (EliLilly), and Spartalizumab (also known as PDR001; Novartis). More preferred checkpoint inhibitors include CTLA-4 inhibitors. (Ipilimumab; Bristol Myers Squibb) and teximumab (Pfizer / MedImmune) and PD-1 inhibitors (Nivolumab; Bristol Myers Squibb) (Pembrolizumab; Merck), Pidilizumab (CT-011; CureTech), MEDI0680 (AMP-514; AstraZeneca), AMP-224 (PD-L2Fc fusion protein; MedImmune).

[0297] Preferably, the immune checkpoint modulator used in combination with the antigenic peptides defined herein is selected from pembrolizumab, ipilimumab, nivolumab, atezolizumab, durvalumab, tesimumab, avelumab, spartalizumab, LAG525 (anti-LAG-3 monoclonal antibody), epadostat (also known as INCB24360; IDO inhibitor), varlilumab (anti-CD27 monoclonal antibody), ureilumab (anti-CD137 monoclonal antibody), AMP-224, and CM-24 (anti-CEACAM1 monoclonal antibody).

[0298] The selection of an immunotherapeutic agent suitable for the purposes of this invention is within the capabilities of those skilled in the art. For example, if it is desired to prevent or treat melanoma, it is preferable to use melanoma cell lysate and / or the antibody ipilimumab together with a suitable antigenic peptide. A suitable antigenic peptide can be selected by: (i) selecting tumor antigens known in the art and / or those described in Table 1B herein suitable for a certain type of cancer, and (ii) selecting antigenic peptides according to the invention that are suitable for the selected tumor antigens (e.g., Table 1A).

[0299] The anticancer therapeutic agent can also be administered in combination with the composition of the present invention, simultaneously, separately, or sequentially. If the composition and the therapeutic agent are administered separately or sequentially, they can be administered in different pharmaceutical forms.

[0300] Therefore, in another aspect, the present invention relates to the compositions of the present invention and at least one anticancer therapeutic agent as described above, as a combination formulation for simultaneous, separate, or sequential administration. In other words, the present invention proposes the combined use of the compositions of the present invention and at least one anticancer therapeutic agent as described above for simultaneous, separate, or sequential administration.

[0301] Multipart kits

[0302] In a further aspect, the present invention also provides a multi-component kit (also referred to herein as a "kit") comprising at least one of the following:

[0303] -The antigenic peptides according to the present invention described herein,

[0304] -The immunogenic compounds according to the present invention described herein,

[0305] -The nanoparticles according to the present invention described herein,

[0306] -The cells described herein are based on the present invention.

[0307] -The nucleic acid according to the present invention described herein,

[0308] -The host cell described herein according to the present invention,

[0309] -The cytotoxic T lymphocytes and / or T lymphocytes according to the present invention described herein

[0310] - The pharmaceutical compositions according to the present invention described herein.

[0311] Specifically, the preferred embodiments of the antigenic peptides described above are also applicable to this kit according to the present invention. For example, the antigenic peptides included in the kit, or the antigenic peptides included in any of the immunogenic compounds, nanoparticles, cells, nucleic acids, host cells, or pharmaceutical compositions included in the kit, preferably comprise or consist of the following amino acid sequences shown in any of SEQ ID NOs 1-247 and 476-500. For example, it is even more preferred to include the following or consist of the following antigenic peptides according to the present invention: the amino acid sequences shown in any of SEQ ID NOs 10, 21, 33, 35, 39, 40, 61, 65, 68, 72, 86, 110, 114, and 220. For example, it is even more preferred to include the following or consist of the following antigenic peptides according to the present invention: the amino acid sequences shown in any of SEQ ID NOs 10, 21, 33, 35, 39, 40, 110, 114, and 220. For example, it is even more preferred to include the antigenic peptide according to the invention comprising the amino acid sequences shown in any one of SEQ ID NO 10, 110, 114 and 220. For example, it is even more preferred to include the antigenic peptide according to the invention comprising the amino acid sequences shown in any one of SEQ ID NO 65, 110, 114 and 220.

[0312] Moreover, a combination thereof is preferred, i.e., a kit comprising different antigenic peptides according to the present invention. Specifically, the multi-part kit of the present invention may comprise more than one of the above-mentioned components, for example, 2, 3, 4, 5, 6, 7, 8, 9 or 10 different components. For example, the multipart kit according to the present invention may include at least two (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10) different immunogenic compounds, at least two (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10) different antigenic peptides, at least two (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10) different nanoparticles, at least two (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10) different cells, at least two (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10) different nucleic acids, at least two (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10) different host cells, and / or at least two (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10) different pharmaceutical compositions. Preferably, the difference between the different components included in the multi-part kit lies in the antigenic peptide according to the invention; for example, one component relates to a first antigenic peptide, and another component relates to a second antigenic peptide (different from the first antigenic peptide). For example, the kit may include at least two different immunogenic compounds according to the invention. For example, the kit may include at least two different antigenic peptides according to the invention. For example, the kit may include at least two different nanoparticles according to the invention. For example, the kit may include at least two different nucleic acids according to the invention. For example, the kit may include at least two different cytotoxic T lymphocytes according to the invention.

[0313] The preferred combination of components according to the invention included in the kit, such as antigenic peptides, corresponds to the preferred combination of components according to the invention included in the above-described pharmaceutical composition.

[0314] Therefore, the present invention provides a kit comprising (at least one) antigenic peptide according to the invention described herein. Additionally, the present invention provides a kit comprising (at least one) immunogenic compound according to the invention described herein. Additionally, the present invention provides a kit comprising (at least one) nanoparticles according to the invention described herein. Additionally, the present invention provides a kit comprising (at least one) cells according to the invention described herein. Additionally, the present invention provides a kit comprising (at least one) nucleic acid according to the invention described herein. Additionally, the present invention provides a kit comprising (at least one) host cells according to the invention described herein.

[0315] The various components of the multipart kit may be packaged in one or more containers. These components may be provided in lyophilized or dried form or dissolved in a suitable buffer. The kit may also include additional reagents, including, for example, preservatives, growth media, and / or buffers, washing solutions, etc., for the storage and / or reconstitution of the aforementioned components.

[0316] Therefore, the present invention provides a kit comprising at least two, preferably three, different antigenic peptides (or the aforementioned immunogenic compounds, nanoparticles, nucleic acids, cells, etc., which differ in terms of antigenic peptides) described herein, and optionally an accessory peptide, such as UCP2 peptide, and / or an adjuvant, such as MONTANIDE ISA 51. The different antigenic peptides (or the aforementioned immunogenic compounds, nanoparticles, nucleic acids, cells, etc., which differ in terms of antigenic peptides) may be contained in the same or different containers. For example, the kit may comprise a (single) container containing a first antigenic peptide and a second antigenic peptide described herein. The (single) container may additionally include an accessory peptide, such as UCP2. Optionally, the first and second antigenic peptides (and optionally the accessory peptides) contained in the (single) container may be formulated together, for example, with water for injection and / or dimethyl sulfoxide (DMSO). Additionally, the kit may comprise a further container (different from the container containing the antigenic peptides) containing an adjuvant, such as MONTANIDE ISA 51.

[0317] Therefore, the preferred kit includes

[0318] (i) A first vial comprising one or more antigenic peptides of the present invention (e.g., at least 200 or 300 μg of each antigenic peptide), and optionally an accessory peptide, such as UCP2 (e.g., at least 200 or 300 μg of the accessory peptide), optionally formulated in water for injection and dimethyl sulfoxide (DMSO); and

[0319] (ii) A second vial containing MONTANIDE ISA 51 (e.g., at least 0.4 or 0.5 ml).

[0320] Additionally, the kit may include one or more (e.g., two or three) syringes, such as silicone-free and rubber-free syringes. The kit may also include connectors, such as Type I connectors.

[0321] A non-limiting example of such a connector is:

[0322] The Type I connector developed by Green Peptide (Japan)

[0323] Connector from Didanorm (France), part number DIDRACDLLFT.

[0324] Promepla (Monaco) Type I connector (part number: ODG0015ST), and

[0325] Smiths Medical (US) Type I connector (part number: MX494).

[0326] The syringe is preferably suitable for Montanaide, i.e., silicone-free and rubber-free (i.e., without any rubber tip on the plunger), and preferably also latex-free. Non-limiting examples of such a syringe are:

[0327] 2ml INKJET (product number: 4606701V, from B-Braun, Germany)

[0328] 5ml INKJET (product number: 4606710V, from B-Braun, Germany)

[0329] 2ml Norm-Ject (serial number: 4020.000V0, from Henke Sass Wolf GmbH, Germany), and

[0330] 5ml Norm-Ject (serial number: 4050.000V0, from Henke Sass Wolf GmbH, Germany).

[0331] For example, the kit may include (i) a first vial containing at least 300 μg of the antigenic peptide of the present invention (or two or three antigenic peptides, each at least 300 μg), and optionally at least 300 μg of UCP2, formulated in water for injection and dimethyl sulfoxide (DMSO); (ii) a second vial containing at least 0.5 ml of MONTANIDE ISA 51; (iii) two silicone-free and rubber-free syringes; and (iv) a Type I connector.

[0332] Optionally, the kit may also include vials of water for injection and / or vial adapters. Sterile needles may also be included, for example, for administering a vaccine to a patient after obtaining the emulsion. The syringe in the kit may be, for example, a 2 ml syringe.

[0333] Additionally, the multi-part kit according to the invention may optionally include instructions for use. Therefore, it is preferred that the kit includes a package insert or instruction manual with guidance on preventing or treating B-cell malignancies using the immunogenic compound according to the invention, the antigenic peptide according to the invention, the nanoparticles according to the invention, the cells according to the invention, the nucleic acid according to the invention, the host cell according to the invention, or the pharmaceutical composition according to the invention.

[0334] Preferably, in addition to any of the components described above, the kit also includes the anticancer therapeutic agent described herein.

[0335] In addition, the present invention provides a vaccine kit for treating, preventing and / or stabilizing B-cell malignancies, comprising the pharmaceutical composition or vaccine described herein and instructions for use of the pharmaceutical composition or vaccine in the prevention and / or treatment of B-cell malignancies.

[0336] Medical treatment and uses

[0337] As described above, the compositions of the present invention are particularly effective for therapeutic use (as medicines), especially for triggering specific immune responses against specific tumor antigens / proteins, such as for the prevention or treatment of B-cell malignancies (e.g., B-cell lymphoma) in patients who require this.

[0338] In view of this, the present invention provides

[0339] -The antigenic peptides according to the present invention described herein,

[0340] -The immunogenic compounds according to the present invention described herein,

[0341] -The nanoparticles according to the present invention described herein,

[0342] -The cytotoxic T lymphocytes (CTLs) according to the present invention described herein,

[0343] -The cells described herein are based on the present invention.

[0344] -The nucleic acid according to the present invention described herein,

[0345] -The host cell described herein according to the present invention,

[0346] -The pharmaceutical compositions according to the present invention described herein,

[0347] -The kit according to the present invention described herein, or

[0348] -The combination according to the present invention described herein,

[0349] For medical use, specifically for the prevention and / or treatment of B-cell malignancies.

[0350] Specifically, the preferred embodiments of the above-described antigenic peptides are also applicable to their use in the prevention and / or treatment of B-cell malignancies according to the present invention. For example, the antigenic peptide included in any of the following—antigenic compounds, nanoparticles, cells, nucleic acids, host cells, or pharmaceutical compositions for the prevention and / or treatment of cancer—preferably comprises or consists of the following amino acid sequences shown in any of SEQ ID NO 1-257 and 476-500. In some embodiments, the antigenic peptide according to the invention comprises or consists of the following amino acid sequences shown in any one of SEQ ID NO 1-12, 34-35, 36-39, 40, 41-64, 65-70, 476-484, 71-80, 81-87, 106-110, 485-488, 111-130, 489-493, 210-225, 494-500, and 226-257; preferably, the antigenic peptide comprises or consists of the following amino acid sequences shown in any one of SEQ ID NO 34-35, 65-70, 476-484, 106-110, 485-488, 111-130, 489-493, 210-225, 494-500, and 226-257; more preferably, the antigenic peptide comprises or consists of the following: SEQ ID NO The amino acid sequence represented by any one of SEQ ID NOs 65-70, 476-484, 106-110, 485-488, 111-130, 489-493, 210-225, and 494-500. More preferably, the antigenic peptide comprises or consists of the following: amino acid sequences represented by any one of SEQ ID NOs 10, 34, 35, 39, 40, 61, 68, 70, 72, 86, 107-110, 114, 117, 119, 120, 212, 217, 220, 224, 227, 231, 477, 491, and 493. More preferably, the antigenic peptide according to the invention comprises the following or a combination thereof: the amino acid sequence shown in any one of SEQ ID NO 10, 21, 33, 35, 39, 40, 61, 65, 68, 72, 86, 110, 114, and 220. For example, more preferably, the antigenic peptide according to the invention comprises the following or a combination thereof: the amino acid sequence shown in any one of SEQ ID NO 10, 21, 33, 35, 39, 40, 110, 114, and 220. For example, more preferably, the antigenic peptide according to the invention comprises the following or a combination thereof: the amino acid sequence shown in any one of SEQ ID NO 10, 110, 114, and 220. For example, more preferably, the antigenic peptide according to the invention comprises the following or a combination thereof: the amino acid sequence shown in any one of SEQ ID NO 65, 110, 114, and 220.

[0351] Moreover, combinations thereof are preferred, i.e., different antigenic peptides according to the invention, for the prevention and / or treatment of B-cell malignancies. Specifically, more than one of the above-mentioned components can be used for the prevention and / or treatment of B-cell malignancies. For example, at least two different antigenic peptides, at least two different immunogenic compounds, at least two different nanoparticles, at least two different cells, at least two different nucleic acids, at least two different host cells, and / or at least two different pharmaceutical compositions can be used for the prevention and / or treatment of B-cell malignancies. Preferably, the difference between these different components for the prevention and / or treatment of B-cell malignancies lies in the antigenic peptide according to the invention, for example, one component relates to a first antigenic peptide, and one component relates to a second antigenic peptide (different from the first antigenic peptide). For example, at least two different immunogenic compounds according to the invention can be used for the prevention and / or treatment of B-cell malignancies. For example, at least two different antigenic peptides according to the invention can be used for the prevention and / or treatment of B-cell malignancies. For example, at least two different nanoparticles according to the invention can be used for the prevention and / or treatment of B-cell malignancies. For example, at least two different nucleic acids according to the invention can be used for the prevention and / or treatment of B-cell malignancies.

[0352] Therefore, the present invention provides at least one antigenic peptide according to the invention described herein for the prevention and / or treatment of B-cell malignancies. Additionally, the present invention provides at least one immunogenic compound according to the invention described herein for the prevention and / or treatment of B-cell malignancies. Furthermore, the present invention provides at least one nanoparticle according to the invention described herein for the prevention and / or treatment of B-cell malignancies. Additionally, the present invention provides at least one cell according to the invention described herein for the prevention and / or treatment of B-cell malignancies. Furthermore, the present invention provides at least one nucleic acid according to the invention described herein for the prevention and / or treatment of B-cell malignancies. Furthermore, the present invention provides at least one host cell according to the invention described herein for the prevention and / or treatment of B-cell malignancies. Furthermore, the present invention provides at least one pharmaceutical composition according to the invention described herein for the prevention and / or treatment of B-cell malignancies. Additionally, the present invention provides a kit according to the invention described herein for the prevention and / or treatment of B-cell malignancies.

[0353] Therefore, the present invention also provides a method for preventing (reducing the occurrence) and / or treating B-cell malignancies or inducing, enhancing, or prolonging an antitumor response against B-cell malignancies in a subject in need, comprising administering to the subject an antigenic peptide according to the present invention.

[0354] -The immunogenic compound according to the present invention,

[0355] -The nanoparticles according to the present invention,

[0356] -The cell according to the invention,

[0357] -The cytotoxic T lymphocytes (CTLs) according to the present invention described herein,

[0358] -Acrylic acid according to the present invention,

[0359] -According to the host cell of the present invention,

[0360] -The pharmaceutical composition according to the present invention,

[0361] -The kit according to the present invention, or

[0362] - The combination according to the invention, as described herein.

[0363] Preferably, the B-cell malignancies to be treated include leukemia and lymphoma, such as acute myeloid leukemia (AML), chronic myeloid leukemia (CML), acute lymphoblastic leukemia (ALL), B-cell chronic lymphocytic leukemia (BCLL), chronic lymphocytic leukemia (CLL, Richter's), hairy cell leukemia (HCL), lymphoplasmacytic lymphoma (LPC) or Woldanström's macroglobulinemia, prolymphocytic leukemia (PLL), small lymphocytic lymphoma (SLL), mantle cell lymphoma (MCL), marginal zone lymphoma (MZL), Burkitt lymphoma (BL), Hodgkin's lymphoma (HL), non-Hodgkin's lymphoma (NHL), and anaplastic large cell lymphoma. Follicular lymphoma (ALCL), follicular lymphoma (FL), refractory follicular lymphoma, diffuse large B-cell lymphoma (DLBCL), and multiple myeloma (MM). In some embodiments, the B-cell malignancy is selected from acute lymphoblastic leukemia (ALL), adult ALL, chronic lymphoblastic leukemia (CLL), non-Hodgkin lymphoma (NHL), and diffuse large B-cell lymphoma (DLBCL). In some embodiments, the disease or condition is NHL, and the NHL is selected from: indolent (slow-growing) NHL, aggressive NHL, diffuse large B-cell lymphoma (DLBCL), NOS (derived from de novo synthesis and indolent conversion), primary mediastinal large B-cell lymphoma (PMBCL), T-cell / histocyte-rich large B-cell lymphoma (TCHRBCL), Burkitt lymphoma, mantle cell lymphoma (MCL), and / or follicular lymphoma (FL), optionally follicular lymphoma grade 3B (FL3B).

[0364] Additionally, this invention provides a method for raising or raising CD8-dependent objects. + A method for an immune response of cytotoxic T cells against one or more epitopes, wherein the method comprises administering to the subject any of the following:

[0365] -According to the antigenic peptide of the present invention,

[0366] -The immunogenic compound according to the present invention,

[0367] -The nanoparticles according to the present invention,

[0368] -Cytotoxic T lymphocytes (CTLs) according to the present invention,

[0369] -The cell according to the invention,

[0370] -Acrylic acid according to the present invention,

[0371] -According to the host cell of the present invention,

[0372] -The pharmaceutical composition according to the present invention,

[0373] -The kit according to the present invention, or

[0374] - The combination according to the invention, as described herein.

[0375] Dependent on CD8 + The immune response can be determined by evaluating inflammatory responses and pro-inflammatory cytokine responses, including an increase in the expression of one or more of IFN-γ, TNF-α, and IL-2 mRNA or protein relative to levels prior to administration of the compound of the present invention. It can also be measured by an increase in the frequency or absolute number of antigen-specific T cells following administration of the compound of the present invention, measured by HLA-peptide multimer staining, ELISPOT assay, and delayed-type hypersensitivity test. It can also be indirectly measured by an increase in antigen-specific serum antibodies dependent on antigen-specific T helper cells.

[0376] The present invention also provides a method for inducing or enhancing an immune response in a subject that is restricted by a variety of MHC class I molecules against one or more antigens or antigenic epitopes, wherein the method comprises administering to the subject any of the following:

[0377] -According to the antigenic peptide of the present invention,

[0378] -The immunogenic compound according to the present invention,

[0379] -The nanoparticles according to the present invention,

[0380] -Cytotoxic T lymphocytes (CTLs) according to the present invention,

[0381] -The cell according to the invention,

[0382] -Acrylic acid according to the present invention,

[0383] -According to the host cell of the present invention,

[0384] -The pharmaceutical composition according to the present invention,

[0385] -The kit according to the present invention, or

[0386] - The combination according to the invention, as described herein.

[0387] The methods described herein for inducing or enhancing immune responses against multiple epitopes restricted by various MHC class I molecules can be determined by evaluating cytokine responses, including an increase in the expression of one or more of IFN-γ, TNF-α, and IL-2 mRNA or protein relative to pre-administration levels after in vitro stimulation of T cells with individual peptides of discrete MHC class I molecules bound to antigen-presenting cells. Restriction of MHC class I molecules can also be verified using antigen-presenting cells expressing MHC class I molecules or by using MHC class I blocking antibodies. It can also be measured by an increase in the frequency or absolute number of antigen-specific T cells after administration of the compounds of the present invention, measured using HLA-peptide multimer staining with multimers assembled with MHC class I molecules.

[0388] Therefore, in another aspect, the present invention also provides

[0389] -According to the antigenic peptide of the present invention,

[0390] -The immunogenic compound according to the present invention,

[0391] -The nanoparticles according to the present invention,

[0392] -Cytotoxic T lymphocytes (CTLs) according to the present invention,

[0393] -The cell according to the invention,

[0394] -Acrylic acid according to the present invention,

[0395] -According to the host cell of the present invention,

[0396] -The pharmaceutical composition according to the present invention,

[0397] -The kit according to the present invention, or

[0398] - According to the combination of the invention, as described herein,

[0399] Used as a medicine.

[0400] More specifically, this invention relates to compositions as defined above, used as vaccines for immunotherapy. Additionally,

[0401] -According to the antigenic peptide of the present invention,

[0402] -The immunogenic compound according to the present invention,

[0403] -The nanoparticles according to the present invention,

[0404] -Cytotoxic T lymphocytes (CTLs) according to the present invention,

[0405] -The cell according to the invention,

[0406] -Acrylic acid according to the present invention,

[0407] -According to the host cell of the present invention,

[0408] -The pharmaceutical composition according to the present invention,

[0409] -The kit according to the present invention, or

[0410] - According to the combination of the invention, as described herein,

[0411] It can be used as a vaccine, specifically for (cancer) immunotherapy.

[0412] As used in the context of this invention, the term "vaccine" refers to a (biological) preparation that provides innate and / or adaptive immunity, generally targeting a specific disease, preferably B-cell malignancy. Thus, a vaccine specifically supports an innate and / or adaptive immune response in the immune system of a patient to be treated. For example, the antigenic peptide according to the invention generally induces or supports an adaptive immune response in patients to be treated.

[0413] In the context of this invention, the vaccine (composition) can induce a specific immune response against a tumor antigen and is therefore preferably used for the prevention or treatment of B-cell malignancies.

[0414] Therefore, in a preferred embodiment, the present invention relates to the composition as defined above for the prevention and / or treatment of cancer in a subject with this need. More preferably, the present invention relates to the use of the composition of the present invention in the preparation of a medicament for the prevention or treatment of cancer in a subject with this need. In other words, the present invention relates to a method for the prevention or treatment of cancer in a subject with this need, comprising administering an effective amount of the composition of the present invention to said subject.

[0415] Preferably, the method described herein is preferred.

[0416] -According to the antigenic peptide of the present invention,

[0417] -The immunogenic compound according to the present invention,

[0418] -The nanoparticles according to the present invention,

[0419] -Cytotoxic T lymphocytes (CTLs) according to the present invention,

[0420] -The cell according to the invention,

[0421] -Acrylic acid according to the present invention,

[0422] -According to the host cell of the present invention,

[0423] -The pharmaceutical composition according to the present invention,

[0424] -The kit according to the present invention, or

[0425] -The combination according to the invention

[0426] Cancer prevention and / or treatment involve (reference) tumor antigens of the antigenic peptides described herein. That is, a suitable antigenic peptide can be selected by: (i) selecting tumor antigens known in the art suitable for a certain type of cancer; and (ii) selecting antigenic peptides according to the invention that are suitable for the selected tumor antigens (e.g., Table 1A). Those skilled in the art will readily understand that the antigenic peptides of the present invention can be selected based on the properties of the B-cell malignancy to be prevented or treated and / or the human genes / human tumor antigens involved in said B-cell malignancy.

[0427] In general, the antigenic peptides of the present invention can be administered "naked" or in the form of immunogenic compounds according to the present invention, their loaded cells according to the present invention, nanoparticles according to the present invention, nucleic acids according to the present invention, host cells according to the present invention, and / or pharmaceutical compositions according to the present invention.

[0428] In a preferred embodiment, it can be given in the form of microorganisms such as gut bacteria species. Whole gut bacteria species may also be advantageous because they have the potential to trigger a greater immune response than the (poly)peptides or nucleic acids they contain. Optionally, the gut bacteria according to the invention can be in the form of probiotics, i.e., live gut bacteria, which can therefore be used as a food additive based on the health benefits they provide. They can be, for example, freeze-dried into granules, pills, or capsules, or mixed directly with dairy products for consumption.

[0429] Administration methods are well known to those skilled in the art. The compositions of the present invention can be administered directly to the subject, to a diseased organ (i.e., local administration), or systemically (i.e., enteric or parenteral administration), or even ex vivo to cells derived from the subject or a human cell line, which are subsequently administered to the subject, or even used in vitro to select a subset of immune cells derived from the subject, which is then re-administered to the subject. Enteric administration includes oral and rectal administration, as well as administration via a gastric feeding tube, duodenal feeding tube, or gastrostomy, while parenteral administration includes subcutaneous, intravenous, intramuscular, intra-arterial, intradermal, intraosseous, intracerebral, and intrathecal injections. The administration method will generally depend on the antigenic peptides (one or more) and / or immunogenic compounds (one or more) present in the composition, the type of cancer to be treated, and other active agents that may be included in the composition. For example, if the immunogenic compound is a nucleic acid as defined above, administration is preferably intramuscular or intradermal injection; if the nucleic acid is cloned into a viral vector, oral / nasal administration is particularly preferred. Optionally, if the antigenic peptide and / or immunogenic compound is a (poly)peptide as defined above or is loaded in / on nanoparticles as described herein, administration is preferably intramuscular, intradermal, or oral. However, alternatively, if the antigenic peptide and / or immunogenic compound is delivered in the form of intestinal bacteria as defined above, especially if the intestinal bacteria are in the form of probiotics, administration is preferably oral.

[0430] The antigenic peptides, immunogenic compounds, and nucleic acids according to the present invention can be further encapsulated, thereby facilitating their delivery to subjects in need. For example, these can be encapsulated into peptide nanocarriers (preferably if the immunogenic compound is a nucleic acid or (poly)peptide), virions (preferably if the immunogenic compound is a nucleic acid or (poly)peptide), or lipid-based carrier systems such as liposome-polycation-DNA complexes (preferably if the immunogen is a nucleic acid or (poly)peptide) (Trovato M, De Berardinis P. Novel antigen delivery systems. World J Virol. 2015 Aug 12; 4(3):156-68; Saade F, Petrovsky N. Technologies for enhanced efficacy of DNA vaccines. Expert Rev Vaccines. 2012 Feb; 11(2):189-209; Li et al., PeptideVaccine: Progress and Challenges. Vaccines (Basel). 2014 Jul 2; 2(3):515-36).

[0431] The composition may also be administered more than once to achieve the desired effect. In a preferred embodiment, the composition is administered repeatedly, at least twice, and preferably more than twice. This can be done over a long period of time, such as weekly, every other week, monthly, annually, or for several years after the first administration, to ensure that the subject is properly immunized.

[0432] Combination therapy

[0433] The administration of the antigenic peptides, immunogenic compounds, nanoparticles, cells, nucleic acids, host cells, and pharmaceutical compositions according to the invention, specifically in the methods and uses according to the invention, can be carried out alone or in combination with adjuvants such as anticancer therapeutic agents that can be used to treat and / or prevent cancer.

[0434] The therapeutic agent is therefore preferably capable of preventing and / or treating the same type of cancer as that used with the antigenic peptide according to the invention. Particularly preferred anticancer therapeutic agents according to the invention include, without limitation, antibodies, CAR-T cells, tumor cell lysates, chemotherapy agents, radiotherapy agents, immune checkpoint modulators, and combinations thereof.

[0435] Antibodies are particularly advantageous in cancer treatment because they can bind to specific antigens on the surface of cancer cells, thereby directing treatment towards the tumor (i.e., these are called tumor-targeting antibodies), or block dysregulated immune checkpoints in cancer (i.e., these are referred to herein as immunomodulatory antibodies). The latter type of antibody aims to suppress cancer immune resistance, which has been observed particularly against tumor antigen-specific T cells. Indeed, as is well known in the art, under normal physiological conditions, immune checkpoints are essential for maintaining self-tolerance (i.e., preventing autoimmunity) and protecting tissues from damage when the immune system responds to pathogen infection. However, in cancer, immune checkpoint expression can be dysregulated as an important mechanism of immune resistance. Regarding the PD-L1 checkpoint, the resistance has been observed particularly in melanoma, ovarian cancer, lung cancer, glioblastoma, breast cancer, and pancreatic cancer (Konishi et al., B7-H1 expression on non-small cell lung cancer cells and its relationship with tumor-infiltrating lymphocytes and their PD-1 expression. Clin Cancer Res. 2004 Aug 1; 10(15):5094-100; Ghebeh et al., The B7-H1 (PD-L1) Tlymphocyte-inhibitory molecule is expressed in breast cancer patients with infiltrating ductal carcinoma: correlation with important high-risk prognostic factors. Neoplasia. 2006 Mar; 8(3):190-8; Hino et al., Tumor cell expression of programmed cell death-1 ligand 1 is a prognostic factor formalignant). melanoma. Cancer. 2010 Apr 1; ​​116(7):1757-66). Other examples of immune checkpoints include, without limitation, PD-L2, PD-1, CD80, CD86, CTLA4, B7H3, B7H4, PVR, TIGIT, GAL9, LAG-3, GITR, CD137, TIM3, VISTA, VISTA-R (Pico de et al., Checkpoint blockade for cancer therapy: revitalizing a suppressed immune system. Trends Mol Med. 2015 Aug;21(8):482-91; Pardoll DM1. The blockade of immune checkpoints in cancerimmunotherapy. Nat Rev Cancer. 2012 Mar 22;12(4):252-64).

[0436] Antibodies are typically used for the above purposes in the form of naked monoclonal antibodies (i.e., unconjugated) or in the form of conjugated with other molecules that are cytotoxic or radioactive.

[0437] Examples of well-known monoclonal tumor-targeting antibodies used in cancer immunotherapy include, without limitation, aleizumab (chronic lymphocytic leukemia), bevacizumab (colorectal cancer, glioblastoma, cervical cancer, lung cancer, kidney cancer), vedotin / bentoximab (lymphoma), blinatumumab (acute lymphoblastic leukemia), caputuzumab (malignant ascites in EPCAM+ cancer), cetuximab (head and neck cancer, colorectal cancer), denosumab (breast cancer, prostate cancer, and bone cancer), gemtuzumab / oxozamicin (acute myeloid leukemia), tiuxetan / tiimumab (non-Hodgkin lymphoma), panitumumab (colorectal cancer), pertuzumab (breast cancer), obbituzumab (chronic lymphocytic leukemia), and olfam. Monoclonal antibodies (chronic lymphocytic leukemia), oxipimab (melanoma), lamoximab (gastric and gastroesophageal cancer), rituximab (chronic lymphocytic leukemia and non-Hodgkin's lymphoma), cetuximab (multicentric Carterman's disease), tocilizumab (non-Hodgkin's lymphoma), and trastuzumab (breast cancer, gastric cancer, and gastroesophageal cancer); while examples of immunomodulatory antibodies include, without limitation, ipilimumab (melanoma) – which blocks CTLA4-dependent immune checkpoints, nivolumab (melanoma, lung cancer) and pembrolizumab (melanoma) – both of which block PDCD1-dependent immune checkpoints, and MPDL3280A, MEDI4736, MEDI0680, and MSB0010718C – all of which block PD-L1-dependent immune checkpoints (Sharma). and Allison, The future of immune checkpoint therapy. Science. 2015 April 3;348(6230):56-61).

[0438] Other antibodies used in cancer immunotherapy have been described in Buqué et al., Trial Watch: Immunomodulatory monoclonal antibodies for oncological indications. Oncoimmunology. 2015 Mar 2; 4(4):e1008814.eCollection 2015 Apr; Redman et al., Mechanisms of action of therapeutic antibodies for cancer. MolImmunol. 2015 Oct; 67(2Pt A):28-45; and Simpson and Caballero, Monoclonal antibodies for the therapy of cancer MC Proc. 2014; 8(Suppl 4):O6 and the Antibody Society website (a list of therapeutic monoclonal antibodies approved or under review by the European Union or United States is available at the website link http: / / www.antibodysociety.org / news / approved_mabs.php).

[0439] Adoptive cellular immunotherapy using chimeric antigen receptor (CAR) T cells has transformed the treatment prospects for B-cell non-Hodgkin lymphoma (NHL), particularly aggressive B-cell lymphoma. For example, CD19-targeted CAR T cells represent a new standard of care for patients with DLBCL who are refractory to at least two prior lines of therapy. Two CAR T-cell products are axi-cel (KTE-019) and YESCARTA. TM ) and tisagenlecleucel(CTL019)(KYMRIAH TM This product has been approved by the U.S. Food and Drug Administration for the treatment of refractory DLBCL after two lines of therapy. A third product, lisocabtagene maraleucel (liso-cel) (JCAR017), is currently being evaluated in clinical trials. Other CAR T-cells include CD20-CAR-T cells.

[0440] Tumor cell lysates can also be combined with one or more antigenic peptides according to the invention. Tumor cells are indeed capable of inducing an immune response—by presenting endogenous peptide-MHC complexes, and via host dendritic cells (DCs) capable of processing and presenting antigens delivered by the lysates. The range of antigens that can induce an immune response is thus increased. Tumor cell lysates can be readily obtained by treating tumor cells with heat shock and / or chemical treatment, and can be autologous (i.e., isolated from a patient) or allogeneic (i.e., isolated from another subject).

[0441] Standard chemotherapy drugs and radiation therapy agents need not be further described in this article, as they have been extensively described in the literature, especially by Baskar et al. (Baskar et al., Cancer and radiation therapy: current advances and future directions. Int J Med Sci. 2012; 9(3):193-9), Paci et al. (Paci et al., Review of therapeutic drug monitoring of anticancer drugs part 1--cytotoxics. Eur J Cancer. 2014 Aug; 50(12):2010-9), and Widmer et al. (Widmer et al., Review of therapeutic drug monitoring of anticancer drugs part two--targetedtherapies. Eur J Cancer. 2014 Aug; 50(12):2020-36). A list of these drugs and agents is also available on cancer.gov (http: / / www.cancer.gov / about-cancer / treatment / drugs).

[0442] Preferably, the immune checkpoint modulator used in combination with the antigenic peptide defined herein is an activator or inhibitor of one or more immune checkpoint molecules selected from CD27, CD28, CD40, CD122, CD137, OX40, GITR, ICOS, A2AR, B7-H3, B7-H4, BTLA, CD40, CTLA-4, IDO, KIR, LAG3, PD-1, TIM-3, VISTA, CEACAM1, GARP, PS, CSF1R, CD94 / NKG2A, TDO, GITR, TNFR and / or FasR / DcR3; or an activator or inhibitor of one or more ligands thereof.

[0443] More preferably, the immune checkpoint modulator is an activator of (co)stimulatory checkpoint molecules or an inhibitor of inhibitory checkpoint molecules, or a combination thereof. Therefore, the immune checkpoint modulator is more preferably an activator of (i) CD27, CD28, CD40, CD122, CD137, OX40, GITR and / or ICOS or (ii) an inhibitor of A2AR, B7-H3, B7-H4, BTLA, CD40, CTLA-4, IDO, KIR, LAG3, PD-1, PDL-1, PD-L2, TIM-3, VISTA, CEACAM1, GARP, PS, CSF1R, CD94 / NKG2A, TDO, TNFR and / or FasR / DcR3.

[0444] More preferably, the immune checkpoint modulator is an inhibitor of inhibitory checkpoint molecules (but preferably not an inhibitor of stimulatory checkpoint molecules). Therefore, the immune checkpoint modulator is even more preferably an inhibitor of A2AR, B7-H3, B7-H4, BTLA, CTLA-4, IDO, KIR, LAG3, PD-1, PDL-1, PD-L2, TIM-3, VISTA, CEACAM1, GARP, PS, CSF1R, CD94 / NKG2A, TDO, TNFR, and / or DcR3 or their ligands.

[0445] It is also preferred that the immune checkpoint modulator is an activator of stimulatory or co-stimulatory checkpoint molecules (but preferably not an activator of inhibitory checkpoint molecules). Therefore, the immune checkpoint modulator is more preferably an activator of CD27, CD28, CD40, CD122, CD137, OX40, GITR and / or ICOS or their ligands.

[0446] More preferably, the immune checkpoint modulator is a modulator of the CD40 pathway, IDO pathway, LAG3 pathway, CTLA-4 pathway, and / or PD-1 pathway. Specifically, the immune checkpoint modulator is preferably a modulator of CD40, LAG3, CTLA-4, PD-L1, PD-L2, PD-1, and / or IDO; more preferably, the immune checkpoint modulator is an inhibitor of CTLA-4, PD-L1, PD-L2, PD-1, LAG3, and / or IDO, or an activator of CD40; even more preferably, the immune checkpoint modulator is an inhibitor of CTLA-4, PD-L1, PD-1, LAG3, and / or IDO; even more preferably, the immune checkpoint modulator is an inhibitor of LAG3, CTLA-4, and / or PD-1; and most preferably, the immune checkpoint modulator is an inhibitor of CTLA-4 and / or PD-1.

[0447] Therefore, the checkpoint modulator used in combination with the antigenic peptide can be selected from known modulators of the CTLA-4 or PD-1 pathways. Preferably, the checkpoint modulator used in combination with the antigenic peptide defined herein can be selected from known modulators of the CTLA-4 or PD-1 pathways. Particularly preferably, the immune checkpoint modulator is a PD-1 inhibitor. Preferred inhibitors of the CTLA-4 and PD-1 pathways include monoclonal antibodies. (Ipilimumab; Bristol Myers Squibb) and tesimumab (Pfizer / Medlmmune) and (Nivolumab; Bristol Myers Squibb) (Pembrolizumab; also known as pembrolizumab or MK-3475; Merck) (Dvalumab, also known as MEDI4736; Medlmmune / AstraZeneca) (Atezolizumab, also known as MPDL3280A; Roche / Genentech), Pidilizumab (CT-011; CureTech), MEDI0680 (AMP-514; AstraZeneca) (Aviruzumab; Merck KGaA / Pfizer, also known as MSB-0010718C), MIH1 (Affymetrix), LY3300054 (Eli Lilly), and Spartalizumab (also known as PDR001; Novartis). More preferred checkpoint inhibitors include CTLA-4 inhibitors. (Ipilimumab; Bristol Myers Squibb) and teximumab (Pfizer / Medlmmune) and PD-1 inhibitors (Nivolumab; Bristol Myers Squibb) (Pembrolizumab; Merck), Pidilizumab (CT-011; CureTech), MEDI0680 (AMP-514; AstraZeneca), AMP-224 (PD-L2Fc fusion protein; Medlmmune).

[0448] Preferably, the immune checkpoint modulator used in combination with the antigenic peptides defined herein is selected from pembrolizumab, ipilimumab, nivolumab, atezolizumab, MEDI4736, tesimumab, avelumab, spartalizumab, LAG525 (anti-LAG3 monoclonal antibody), Epacadostat (formerly INCB24360; IDO inhibitor), Varlilumab (anti-CD27 monoclonal antibody), Urelumab (anti-CD137 monoclonal antibody), AMP-224, and CM-24 (anti-CEACAM1 monoclonal antibody).

[0449] The selection of an immunotherapeutic agent suitable for the purposes of this invention is within the capabilities of a person skilled in the art.

[0450] The anticancer therapeutic agent can also be administered in combination with the antigenic peptide according to the invention, the immunogenic compound according to the invention, the nanoparticle according to the invention, the cell according to the invention, the nucleic acid according to the invention, the host cell according to the invention, or the pharmaceutical composition according to the invention, as described herein, approximately simultaneously or sequentially, and in the same or different pharmaceutical forms. Therefore, the present invention proposes the combined use of the compositions of the invention and at least one of the aforementioned anticancer therapeutic agents for simultaneous, separate, or sequential administration as described herein.

[0451] Furthermore, the present invention relates to combinations of at least two different antigenic peptides according to the invention, for example, for the prevention and / or treatment of B-cell malignancies. Furthermore, the present invention relates to combinations of at least two different immunogenic compounds according to the invention, for example, for the prevention and / or treatment of B-cell malignancies. Furthermore, the present invention relates to combinations of at least two different nanoparticles according to the invention, for example, for the prevention and / or treatment of B-cell malignancies. Furthermore, the present invention relates to combinations of at least two different nucleic acids according to the invention, for example, for the prevention and / or treatment of B-cell malignancies.

[0452] Therefore, according to a preferred embodiment, at least two antigenic peptides according to the invention may be administered in combination, for example, in the same pharmaceutical composition. For example, at least 3 antigenic peptides, at least 4 antigenic peptides, at least 5 antigenic peptides, at least 6 antigenic peptides, at least 7 antigenic peptides, at least 8 antigenic peptides, at least 9 antigenic peptides, at least 10 antigenic peptides, at least 11 antigenic peptides, at least 12 antigenic peptides, at least 13 antigenic peptides, at least 14 antigenic peptides, at least 15 antigenic peptides, at least 20 antigenic peptides, at least 25 antigenic peptides, at least 50 antigenic peptides, and at least 100 antigenic peptides may be administered in combination, for example, in the same pharmaceutical composition. Selecting a combination of antigenic peptides suitable for the intended purpose is within the capabilities of those skilled in the art.

[0453] In a particularly preferred embodiment, two different antigenic peptides according to the invention (e.g., relating to the same B-cell malignancy type and / or the same reference antigen) are combined. For example,

[0454] (i) at least two different immunogenic compounds according to the invention;

[0455] (ii) at least two different antigenic peptides according to the invention;

[0456] (iii) at least two different nanoparticles according to the invention; or

[0457] (iv) at least two different nucleic acids according to the invention

[0458] It can be combined.

[0459] For example, the present invention provides a combination of at least two different antigenic peptides according to the present invention, specifically

[0460] (i) the first antigenic peptide according to the invention, and

[0461] (ii) The second antigenic peptide according to the present invention (different from the first (antigenic peptide)),

[0462] Preferably used for the prevention and / or treatment of B-cell malignancies.

[0463] For example, the present invention provides a combination of at least two different immunogenic compounds according to the present invention, specifically

[0464] (i) An immunogenic compound according to the invention, comprising a first antigenic peptide according to the invention, and

[0465] (ii) An immunogenic compound according to the invention, comprising a second antigenic peptide according to the invention (different from the first (antigenic peptide)),

[0466] Preferably used for the prevention and / or treatment of B-cell malignancies.

[0467] For example, the present invention provides a combination of at least two different nanoparticles according to the present invention, specifically

[0468] (i) Nanoparticles according to the invention, comprising the first antigenic peptide according to the invention, and

[0469] (ii) Nanoparticles according to the invention, comprising a second antigenic peptide according to the invention (different from the first antigenic peptide)

[0470] Preferably used for the prevention and / or treatment of B-cell malignancies.

[0471] For example, the present invention provides combinations of at least two different nucleic acids according to the present invention, specifically

[0472] (i) A nucleic acid according to the invention, comprising a polynucleotide encoding a first antigenic peptide according to the invention, and

[0473] (ii) The nucleic acid according to the invention comprises a polynucleotide encoding a second antigenic peptide (different from the first (antigenic peptide)) according to the invention.

[0474] Preferably used for the prevention and / or treatment of B-cell malignancies.

[0475] For example, the present invention provides a combination of at least two different cytotoxic T lymphocytes according to the invention, specifically (i) cytotoxic T lymphocytes according to the invention that are specific to the first antigenic peptide of the invention, and

[0476] (ii) Cytotoxic T lymphocytes according to the invention specifically derived from the second antigenic peptide (different from the first antigenic peptide) according to the invention.

[0477] It is preferably used for the prevention and / or treatment of B-cell malignancies.

[0478] In the combination according to the invention, such a combination of components according to the invention, such as antigenic peptides, is preferred, which corresponds to the preferred combination of components according to the invention, such as antigenic peptides, included in the pharmaceutical composition as described above.

[0479] Furthermore, the antigenic peptide according to the invention can also be combined with a corresponding (human) tumor antigenic epitope (as described above with respect to the peptide "family"). Thus, the selection of highly effective T-cell clones against tumors is obtained / supported. Specifically, the antigenic peptide according to the invention and the corresponding (human) tumor antigenic epitope can be co-administered. This co-administration can be approximately simultaneous or sequential, wherein, in the case of sequential administration, it is preferred that the antigenic peptide according to the invention is administered first and the corresponding (human) tumor antigenic epitope is then administered. Specifically, the antigenic peptide according to the invention can be administered first, and the corresponding (human) tumor antigenic epitope can be used as a (re)boost. For example, the antigenic peptide according to SEQ ID NO:10 can be combined with the reference peptide according to SEQ ID NO:258. In another example, the antigenic peptide according to SEQ ID NO:110 can be combined with the reference peptide according to SEQ ID NO:270. In another example, the antigenic peptide according to SEQ ID NO:114 can be combined with the reference peptide according to SEQ ID NO:271. In another example, the antigenic peptide according to SEQ ID NO:220 may be combined with the reference peptide according to SEQ ID NO:279.

[0480] The peptides to be combined, such as (a) the antigenic peptide according to the invention and the corresponding (human) tumor antigen epitope or (b) two different antigenic peptides according to the invention, can be given as follows:

[0481] - In the same immunogenic compound according to the invention or in different immunogenic compounds according to the invention,

[0482] -(loaded) in the same nanoparticles according to the invention or in different nanoparticles according to the invention

[0483] -(loaded) in the same cell according to the invention or in different cells according to the invention

[0484] - Encoded by the same nucleic acid according to the invention or by different nucleic acids according to the invention.

[0485] -Expressed by the same host cell according to the invention or by a different host cell according to the invention, or

[0486] - (included) in the same pharmaceutical composition according to the invention or in different pharmaceutical compositions according to the invention.

[0487] In general, the expression "two different components" in the context of combination—for example, for use according to the invention (combination therapy)—is as defined above (in the context of pharmaceutical compositions). Specifically, it refers to...

[0488] (1) A first component, such as the antigenic peptide, immunogenic compound, nanoparticle, cell, nucleic acid, host cell, or pharmaceutical composition according to the invention as described herein; and

[0489] (2) A second component (which is different from the first component), such as the above-described anticancer therapeutic agent, different antigenic peptides according to the invention described herein, different immunogenic compounds according to the invention described herein, different nanoparticles according to the invention described herein, different cells according to the invention described herein, different nucleic acids according to the invention described herein, different host cells according to the invention described herein, different pharmaceutical compositions according to the invention described herein, or any form (“naked”, as an immunogenic compound, as a nanoparticle, as a (host) cell, as a nucleic acid, or as a pharmaceutical composition described herein) of one or more human tumor antigens (fragments).

[0490] Therefore, when referred to herein as “two different components” in the context of a combination (combination therapy) for use according to the invention, it is preferably the active component in the context of the disease to be prevented and / or treated (B-cell malignancy). In other words, each of the at least two different components, if administered separately (rather than in the combination described herein), may also be effective for the prevention and / or treatment of said cancer—although the combination (i.e., combined administration) generally enhances its preventive and / or therapeutic effects (such as immune response), specifically in a synergistic manner.

[0491] Therefore, the present invention also provides a combination of (at least) two different antigenic peptides according to the invention described herein. In this context, the (at least) two different antigenic peptides can be in any form, for example, "naked," included in an immunogenic compound, nanoparticles, (pharmaceutical) composition or its loaded cells, or encoded by a nucleic acid (e.g., a vector). Thus, the (at least) two different antigenic peptides can be included in (at least) two different components (to be combined). In a preferred embodiment, the at least two different components of the combination according to the invention are at least different antigenic peptides according to the invention (in any form, for example, included in an immunogenic compound, nanoparticles, cells, pharmaceutical composition, encoded by a nucleic acid, etc.).

[0492] Preferably, the at least two different components used in the combination according to the invention relate to the same type of cancer, such as the same or different antigens associated with this cancer and / or the same or different (reference) epitopes within this cancer-associated antigen. More preferably, the at least two different components relate to the same tumor antigen.

[0493] In some embodiments, the at least two different components for use according to the invention are included in the same or different compositions. In some embodiments, the at least two different components for use according to the invention are administered via the same or different administration routes. In some embodiments, the at least two different components for use according to the invention are administered approximately simultaneously or sequentially.

[0494] Preferably, the at least two different components for use according to the invention are administered approximately simultaneously. More generally, it is preferred that the first component and the second component are administered approximately simultaneously, wherein the at least two different components for use according to the invention are preferably administered in the same form (i.e., in the same type of formulation, for example, as nanoparticles, as a pharmaceutical composition, etc.).

[0495] "Approximately simultaneously," as used herein, specifically means that the second component is administered either simultaneously or immediately following the administration of the first component, or immediately following the administration of the second component. Those skilled in the art will understand that "immediately following" includes the time required to prepare the second administration—specifically, the time required to expose and disinfect the second administration site and to properly prepare the "administration device" (e.g., syringe, pump, etc.). Simultaneous administration also includes situations where the administration times of the first and second components overlap, or, for example, where one component is administered over a longer period such as 30 minutes, 1 hour, 2 hours, or longer (e.g., intravenous infusion), while the other component is administered at a certain point within this longer period. Approximately simultaneous administration of the first and second components is particularly preferred if different routes of administration and / or different administration sites are used.

[0496] It is also preferred that the at least two different components for use according to the invention are administered sequentially. More generally, it is preferred that the first component and the second component are administered sequentially, wherein the at least two different components for use according to the invention are preferably administered in the same form (i.e., in the same type of formulation, for example, as nanoparticles, as a pharmaceutical composition, etc.).

[0497] This means that the first component is given before or after the second component. When given sequentially, the time between the giving of the first component and the giving of the second component preferably does not exceed one week, more preferably not more than three days, even more preferably not more than two days, and most preferably not more than 24 hours. Particularly preferred is that the first component and the second component are given on the same day, wherein the time between the giving of the first component and the giving of the second component preferably does not exceed six hours, more preferably not more than three hours, even more preferably not more than two hours, and most preferably not more than one hour.

[0498] Preferably, the first and second components are administered via the same route of administration. More generally, it is preferred that the first and second components are administered via the same route of administration, wherein the at least two different components used in the combination according to the invention are preferably administered in the same form (i.e., in the same type of formulation, for example, as nanoparticles, as a pharmaceutical composition, etc.).

[0499] It is also preferred that the at least two different components of the combination for use according to the invention are administered via different routes of administration. More generally, it is preferred that the first component and the second component are administered via different routes of administration, wherein the at least two different components of the combination for use according to the invention are preferably administered in the same form (i.e., in the same type of formulation, for example, as nanoparticles, as a pharmaceutical composition, etc.).

[0500] Preferably, the at least two different components for use according to the invention are included in the same composition. More generally, it is preferred that the first component and the second component are included in the same composition, wherein the at least two different components for use according to the invention are preferably given in the same form (i.e., in the same type of formulation, for example, as nanoparticles, etc.).

[0501] It is also preferred that the at least two different components for use according to the invention are included in different compositions. More generally, it is preferred that the first component and the second component are included in different compositions, wherein the at least two different components for use according to the invention are preferably given in the same form (i.e., in the same type of formulation, for example, as nanoparticles, etc.).

[0502] Specifically, the present invention provides a pharmaceutical composition comprising a first antigenic peptide according to the present invention, wherein the first antigenic peptide comprises or is composed of a sequence variant of a fragment of a human tumor antigen.

[0503] Specifically, the present invention provides a combination, for example for the prevention and / or treatment of B-cell malignancies, comprising a first antigenic peptide according to the invention, comprising or consisting of a microbial cluster sequence variant of a fragment of the human tumor antigen CD22; and a second antigenic peptide according to the invention, comprising or consisting of a sequence variant of a fragment of the human tumor antigen TNFRSF13C. Preferably, the first antigenic peptide comprises or consists of a sequence variant of the CD22 fragment (human reference peptide) “WVFEHPETL” (SEQ ID NO:270), such as an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO:106-110, 316, and 387-390, and the second antigenic peptide comprises or consists of a sequence variant of the TNFRSF13C fragment (human reference peptide) “LLFGAPALL” (SEQ ID NO:279), such as an antigenic peptide comprising or consisting of the amino acid sequences shown in SEQ ID NO:220, 325, and 450. More preferably, the first antigenic peptide comprises or is composed of the amino acid sequences shown in SEQ ID NO:110, 387, and 390, and the second antigenic peptide comprises or is composed of the amino acid sequences shown in SEQ ID NO:220 and 450. Even more preferably, the pharmaceutical composition comprises an antigenic peptide comprising or composed of SEQ ID NO:110 and an antigenic peptide comprising or composed of SEQ ID NO:220.

[0504] More preferably, the combination according to the invention (e.g., for the prevention and / or treatment of B-cell malignancies) comprises at least three different components as described above, specifically at least three different antigenic peptides according to the invention. The above description of combinations of two different components applies accordingly to combinations of three different components.

[0505] Most preferably, the combination according to the invention (e.g., for the prevention and / or treatment of B-cell malignancies) comprises at least four different components as described above, specifically at least four different antigenic peptides according to the invention. The above description regarding combinations of two different components applies accordingly to combinations of four different components.

[0506] It should be understood that the combination (e.g. for the prevention and / or treatment of B-cell malignancies) may also contain—instead of the preferred combination of the above-described antigenic peptides—a corresponding combination of the immunogenic compounds of the present invention, a corresponding combination of the nanoparticles of the present invention, or a corresponding combination of the nucleic acids of the present invention. Attached Figure Description

[0507] A brief description of the accompanying drawings is given below. The drawings are intended to illustrate the invention in more detail. However, they are not intended to limit the subject matter of the invention in any way.

[0508] Figure 1 : Showing the in vitro affinity of the antigenic peptide CD22-B1 of Example 1 compared to the corresponding human CD22 epitope CD22-H1.

[0509] Figure 2 : Showing the in vitro affinity of the antigenic peptide CD37-B1 of Example 1 compared to the corresponding human CD37 epitope CD37-H1.

[0510] Figure 3 : Showing the in vitro affinity of the antigenic peptide CD19-B1 of Example 1 compared to the corresponding human CD19 epitope CD19-H1.

[0511] Figure 4 : Showing the in vitro affinity of the antigenic peptide CD19-B2 of Example 1 compared to the corresponding human CD19 epitope CD19-H2.

[0512] Figure 5 : Showing the in vitro affinity of the antigenic peptide TNFRSF13C-B1 of Example 1 compared to the corresponding human TNFRSF13C epitope TNFRSF13C-H1.

[0513] Figure 6The figure shows the ELISPOT results and cross-reactivity with the human corresponding peptide CD22-H1 in HHD DR1 HLA-A2 transgenic mice vaccinated with the antigenic peptide CD22-B1 in Example 2. Data are expressed in terms of 1.10... 6 The total number of spots for T cells is provided.

[0514] Figure 7 The figure shows the ELISPOT results and cross-reactivity with the human corresponding peptide TNFRSF13C-H1 in HHD DR1HLA-A2 transgenic mice vaccinated with the antigenic peptide TNFRSF13C-B1 in Example 2. Data are expressed in terms of 1.10... 6 The total number of spots for T cells is provided.

[0515] Figure 8 The figure shows the ELISPOT results and cross-reactivity with the human corresponding peptide CD37-H1 in HHD DR1 HLA-A2 transgenic mice vaccinated with the antigenic peptide CD37-B1 in Example 2. Data are expressed in terms of 1.10... 6 The total number of spots for T cells is provided.

[0516] Figure 9 The figure shows the ELISPOT results and cross-reactivity with the human corresponding peptide CD19-H2 in HHD DR1 HLA-A2 transgenic mice vaccinated with the antigenic peptide CD19-B2 in Example 2. Data are expressed in 1.10... 6 The total number of spots for T cells is provided.

[0517] Figure 10 The figure shows the ELISPOT results and cross-reactivity with the human corresponding peptide CD19-H1 in HHD DR1 HLA-A2 transgenic mice vaccinated with the antigenic peptide CD19-B1 in Example 2. Data are expressed in terms of 1.10... 6 The total number of spots for T cells is provided.

[0518] Figure 11 The figure shows the ELISPOT results and cross-reactivity with the human corresponding peptide CD22-H1 in HHD DR3 HLA-A2 transgenic mice vaccinated with the antigenic peptide CD22-B1 in Example 2. Data are expressed in terms of 1.10... 6 The total number of spots for T cells is provided.

[0519] Figure 12The figure shows the ELISPOT results and cross-reactivity with the human corresponding peptide TNFRSF13C-H1 in HHD DR3HLA-A2 transgenic mice vaccinated with the antigenic peptide TNFRSF13C-B1 in Example 2. Data are expressed in terms of 1.10... 6 The total number of spots for T cells is provided.

[0520] Figure 13 The figure shows the ELISPOT results and cross-reactivity with the human corresponding peptide CD37-H1 in HHD DR3 HLA-A2 transgenic mice vaccinated with the antigenic peptide CD37-B1 in Example 2. Data are expressed in terms of 1.10... 6 The total number of spots for T cells is provided.

[0521] Figure 14 : Showing the in vitro affinity of the antigenic peptide MS4A1-B4 of Example 1 compared to the corresponding human MS4A1 epitope MS4A1-H4.

[0522] Figure 15 The figure shows the ELISPOT results and cross-reactivity with the human corresponding peptide MS4A1-H4 in HHD DR1HLA-A2 transgenic mice vaccinated with the antigenic peptide MS4A1-B4 in Example 2. Data are expressed in terms of 1.10... 6 The total number of spots for T cells is provided.

[0523] Figure 16 The figure shows the ELISPOT results and cross-reactivity with the human corresponding peptide MS4A1-H4 in HHD DR3HLA-A2 transgenic mice vaccinated with the antigenic peptide MS4A1-B4 in Example 2. Data are expressed in terms of 1.10... 6 The total number of spots for T cells is provided.

[0524] Figure 17 Example 3 shows the detection of CD22-B1, CD37B1, TNFRSF13C-B1, and MS4A1-B4 peptide-specific CD8+ T cells in peripheral blood from healthy donors (HLA-A2 positive).

[0525] Figure 18 This demonstrates the cytotoxicity of human T cell clones specifically targeting the peptides CD22-B1, CD37B1, TNFRSF13C-B1, and MS4A1-B4, expanded in vitro via stimulation with microbial cluster-derived peptides, as described in Example 3. These CD22-B1, CD37B1, TNFRSF13C-B1, and MS4A1-B4 peptide-specific T cells exhibited the ability to kill T2 cells loaded with bacteria or human peptides.

[0526] Figure 19 The in vitro affinity of the antigenic peptides CD22-B1, CD22-B12, and CD22-B13 of Example 4 is shown compared to the corresponding human CD22 epitope CD22-H1.

[0527] Figure 20 The in vitro affinity of the antigenic peptides CD37-B1, CD37-B12, CD37-B13, CD37-B14, and CD37-B15 of Example 4 is shown compared to the corresponding human CD37 epitope CD37-H1.

[0528] Figure 21 The in vitro affinity of the antigenic peptides TNFRSF13C-B1, TNFRSF13C-B11, TNFRSF13C-B12 and TNFRSF13C-B13 of Example 4 is shown compared to the corresponding human TNFRSF13C epitope TNFRSF13C-H1.

[0529] Figure 22 The in vitro affinity of the antigenic peptides MS4A1-B4, MS4A1-B42, and MS4A1-B43 of Example 4 is shown compared to the corresponding human MS4A1 epitope MS4A1-H4. Example

[0530] The following describes specific embodiments of various methods and aspects of the present invention. However, the scope of the invention is not limited to the specific embodiments described herein. The following formulations and examples are given to enable those skilled in the art to more clearly understand and practice the invention. However, the scope of the invention is not limited to the exemplary embodiments, which are intended only as examples of a single aspect of the invention, and functionally equivalent methods are within the scope of the invention. In fact, in addition to those described herein, various modifications of the invention will also be apparent to those skilled in the art based on the foregoing description, the accompanying drawings, and the following embodiments. All such modifications fall within the scope of the appended claims.

[0531] Example 1: The antigenic peptide has a superior affinity for the HLA-A*0201 allele.

[0532] Then, the binding affinity of various selected antigenic peptides and corresponding human tumor antigen (human reference peptide) fragments to the HLA-A*0201 allele was confirmed in vitro. Specifically, the antigenic peptide of sequence SEQ ID NO:110 (《YIFEHPELL》, also referred to herein as CD22-B1) was compared with the corresponding human reference peptide derived from CD22 (《WVFEHPETL》, SEQ ID NO:270, also referred to herein as CD22-H1). Furthermore, the antigenic peptide of sequence SEQ ID NO:109 (《YVFEHPELL》, also referred to herein as CD22-B11) was compared with the corresponding human reference peptide derived from CD22 (《WVFEHPETL》, SEQ ID NO:270, also referred to herein as CD22-H1). Furthermore, the antigenic peptide of sequence SEQ ID NO:114 (FLAFVPLQL, also referred to herein as CD37-B1) was compared with the corresponding reference human peptide derived from CD37 (GLAFVPLQI, SEQ ID NO:271, also referred to herein as CD37-H1). Additionally, the antigenic peptide of sequence SEQ ID NO:117 (GMAFVPLLL, also referred to herein as CD37-B11) was compared with the corresponding reference human peptide derived from CD37 (GLAFVPLQI, SEQ ID NO:271, also referred to herein as CD37-H1). Furthermore, the antigenic peptide of sequence SEQ ID NO:34 (LLVGILHLV, also referred to herein as CD19-B1) was compared with the corresponding reference human peptide derived from CD19 (SLVGILHLQ, SEQ ID NO:260, also referred to herein as CD19-H1). Furthermore, the antigenic peptide of sequence SEQ ID NO:10 (《TLLFLTPML》, also referred to herein as CD19-B2) was compared with the corresponding reference human peptide derived from CD19 (《FLLFLTPME》, SEQ ID NO:258, also referred to herein as CD19-H2). Additionally, the antigenic peptide of sequence SEQ ID NO:39 (《YLAYLIFEL》, also referred to herein as CD19-B6) was compared with the corresponding reference human peptide derived from CD19 (《TLAYLIFCL》, SEQ ID NO:261, also referred to herein as CD19-H6). Furthermore, the antigenic peptide of sequence SEQ ID NO:40 (《LQMGGFYLL》, also referred to herein as CD19-B7) was compared with the corresponding reference human peptide derived from CD19 (《QQMGGFYLC》, SEQ ID NO:262, also referred to herein as CD19-H7).Furthermore, the antigenic peptide of sequence SEQ ID NO:220 (《LMFGAPALV》, also referred to herein as TNFRSF13C-B1) was compared with the corresponding reference human peptide derived from TNFRSF13C (《LLFGAPALL》, SEQ ID NO:279, also referred to herein as TNFRSF13C-H1). Additionally, the antigenic peptide of sequence SEQ ID NO:231 (《ILPGLLFGL》, also referred to herein as TNFRSF13C-B31) was compared with the corresponding reference human peptide derived from TNFRSF13C (《PLPGLLFGA》, SEQ ID NO:280, also referred to herein as TNFRSF13C-H3). Furthermore, the antigenic peptide of sequence SEQ ID NO:227 (《FMPGLLFGA》, also referred to herein as TNFRSF13C-B33) was compared with the corresponding reference human peptide derived from TNFRSF13C (《PLPGLLFGA》, SEQ ID NO:280, also referred to herein as TNFRSF13C-H3). Furthermore, the antigenic peptide of sequence SEQ ID NO:61 (《YILGGLLMV》, also referred to herein as MS4A1-B12) was compared with the corresponding reference human peptide derived from MS4A1 (also referred to as CD20) (《IALGGLLMI》, SEQ ID NO:263, also referred to herein as MS4A1-H1). Additionally, the antigenic peptide of sequence SEQ ID NO:72 (《ILIPAGIYL》, also referred to herein as MS4A1-B3) was compared with the corresponding reference human peptide derived from MS4A1 (also referred to as CD20) (《LMIPAGIYA》, SEQ ID NO:265, also referred to herein as MS4A1-H3). Furthermore, the antigenic peptide of sequence SEQ ID NO:65 (《AMNSLSLYI》, also referred to herein as MS4A1-B4) was compared with the corresponding reference human peptide derived from MS4A1 (also referred to as CD20) (《IMNSLSLFA》, SEQ ID NO:264, also referred to herein as MS4A1-H4). In addition, the antigenic peptide of sequence SEQ ID NO:86 (《YLFLGILSL》, also referred to herein as MS4A1-B5) was compared with the corresponding reference human peptide derived from MS4A1 (also referred to as CD20) (《SLFLGILSV》, SEQ ID NO:266, also referred to herein as MS4A1-H5).

[0533] A. Materials and Methods

[0534] A1. Measure the affinity of the peptide for the T2 cell line.

[0535] The experimental protocol was similar to that used for validating HLA-A*0201-presented peptides (Tourdot et al., A general strategy to enhance immunogenicity of low-affinity HLA-A2.1-associated peptides: implication in the identification of cryptic tumor epitopes. Eur J Immunol. Dec. 2000; 30(12):3411-21). Peptide affinity measurements were performed using T2 human tumor cells that expressed HLA-A*0201 molecules but were TAP1 / 2 negative and unable to present endogenous peptides.

[0536] T2 cells (5.10) 4 Cells / well were incubated with peptides at decreasing concentrations from 100 μM to 0.1 μM (4 spots: 100 μM, 10 μM, 1 μM, 0.1 μM) in serum-free medium (TexMacs) supplemented with 100 ng / μl β2 microglobulin at 37°C for 16 h. Cells were then washed twice and labeled with an anti-HLA-A2 antibody (clone BB7.2, BD Pharmagen) conjugated to PE.

[0537] The analysis was performed using FACS (Macsquant Analyzer 10-Miltenyi).

[0538] For each peptide concentration, the geometric mean of the target peptide-related markers was subtracted from the background noise and reported as a percentage of the geometric mean of the HLA-A*0202 markers obtained for the reference peptide HIV pol 589-597 at a concentration of 100 μM. Relative affinity was then determined as follows:

[0539] Relative affinity = concentration of each peptide that induces 20% HLA-A*0201 expression / concentration of the reference peptide that induces 20% HLA-A*0201 expression.

[0540] A2. Peptide solubilization

[0541] Solubilize each peptide by taking into account its amino acid composition. For peptides that do not contain any cysteine, methionine, or tryptophan, DMSO can be added up to 10% of the total volume. Other peptides are resuspended in water or PBS at pH 7.4.

[0542] B. Result

[0543] Table 2 below shows the mean relative fluorescence intensity values ​​of T2 cells obtained with different concentrations of each peptide (the data are normalized relative to the mean fluorescence value of the HIV peptide, i.e., a value of 100 is equivalent to the best observed binding of the HIV peptide):

[0544]

[0545]

[0546] Table 2.

[0547] Table 3 below summarizes the concentrations and in vitro binding affinity of each test peptide required to induce 20% HLA-A2 expression (*normalized for HIV-pol concentrations of peptides used to induce 20% HLA-A2 expression in the same experiment).

[0548]

[0549]

[0550] Table 3. ND - Undetermined

[0551] in addition, Figure 1-5 Examples 1 and 14 illustrate the results of the selected instance, namely the antigenic peptide CD22-B1—compared to the corresponding human CD22 fragment CD22-H1 ( Figure 1 ); Antigen peptide CD37-B1—compared to the corresponding human CD37 fragment CD37-H1 ( Figure 2 ); Antigen peptide CD19-B1 — compared with the corresponding human CD19 fragment CD19-H1 ( Figure 3 ); Antigen peptide CD19-B2 — compared with the corresponding human CD19 fragment CD19-H2 ( Figure 4 ); antigenic peptide TNFRSF13C-B1 — compared with the corresponding human TNFRSF13C fragment TNFRSF13C-H1 ( Figure 5 ); and antigenic peptide MS4A1-B4—compared to the corresponding human MS4A1 fragment MS4A1-B4 ( Figure 14 ).

[0552] In summary, the results show that the antigenic peptides according to the invention exhibit binding affinity for HLA-A*0201 that is at least similar to that of the corresponding human tumor antigen fragment. In most cases, the binding affinity observed for the antigenic peptides according to the invention is stronger than that for the corresponding human epitopes. Without being bound by any theory, it is assumed that this strong binding affinity of the antigenic peptides according to the invention reflects their ability to elicit an immune response (i.e., their immunogenicity).

[0553] Example 2: CD22-B1, CD19B1, CD19-B2, CD37B1, and TNFRSF13C-B1 in HLA-A2 transgenic mice Immunogenicity of MS4A1-B4 and cross-reactivity with corresponding human peptides.

[0554] A. Materials and Methods

[0555] A.1 Mouse Model

[0556] In short, HLA-A2 HHD-DR1 humanized mice (C57BL / 6JB2mtm1UncIAb- / -Tg(HLA-DRA,HLA-DRB1*0101)#GjhTg(HLA-A / H2-D / B2M)1Bpe) or HHD-DR3 humanized mice (C57BL / 6JB2mtm1UncIAb- / -Tg(HLA-DRA,HLA-DRB1*0301)#GjhTg(HLA-A / H2-D / B2M)1Bpe) were randomly assigned (based on mouse sex and age) to experimental groups. Each group received the same accessory peptide (h-pAg UCP2; sequence: KSVWSKLQSIGIRQH; SEQ ID NO:475; for HHD DR1 mice or h-pAg) DR3; Sequence MAKTIAYDEEARRGLERGLN; SEQ ID NO473; Immunization of a specific vaccine peptide (vacc-pAg) combination for HHD DR3 mice (summarized in Table 4 below).

[0557] Table 4. Experimental group compositions. h-pAg: 'helper' peptide; vacc-pAg: vaccine peptide. Boost injection numbers are shown in parentheses.

[0558]

[0559]

[0560] The peptides are provided as follows:

[0561] • vacc-pAg: CD22-B1, CD19-B1, CD19-B2, CD37-B1, TNFRSF13C-B1, and MS4A1-B4 are all generated and provided at a concentration of 4 mg / ml (4 mM);

[0562] h-pAg: DR3 or UCP2 was resuspended in pure distilled water at a concentration of 10 mg / mL.

[0563] Fresh peptide formulations (emulsions) were prepared for injection on each day and for each group. Mixtures for 10 animals were prepared using 2 mL Luer lock syringes (4606701V, B BRAUN) and Luer connectors (Cole-Parmer, 45502-22): 500 μL of the peptide mixture in syringe 1 was emulsified as quickly as possible with 500 μL of IFA in syringe 2 until a thick (white foam) emulsion was formed. Each emulsion was prepared in excess to compensate for the dead volume at the time of injection.

[0564] Animals were immunized with a primary immunization on day 0 (d0) and a booster immunization on day 14. Each mouse was injected subcutaneously at the tail base with 100 μL of an oil-based emulsion containing:

[0565] • 60 nM vacc-pAg; 105 nM UCP2 helper peptide (for HHD-DR1 mice) or 65 nM DR3 helper peptide (for HHD-DR3 mice)

[0566] • Add 10 μL of PBS to achieve a total volume of 50 μL (per mouse);

[0567] • Incomplete Freund's adjuvant (IFA), added at a 1:1 (v:v) ratio (50 μL / mouse).

[0568] A.2 Analysis

[0569] Seven days after the booster injection (i.e., on day 21), the animals were euthanized and the spleens were obtained. Spleen cells were prepared by mechanically destroying the organ and then purifying it using 70 μm filtration and Ficoll density gradient.

[0570] Cell suspensions were further used for the ELISPOT-IFNγ assay (Table 5). Cells were cultured in 200 μL of complete T cell culture medium. Experimental conditions (reproduced) were as follows: 2 x 10⁵ total cells per well when cultured with various pAg (10 μM) or in the presence of culture medium alone; 2 x 10⁴ total cells when cultured with beads loaded with CD3 / CD28 (T Cell Activation / Expansion Kit, 130-093-627, Miltenyi) (bead to cell ratio 1:1). The ability of the culture to secrete IFNγ was evaluated according to the manufacturer's instructions (incubated for 16-18 hours prior to assay) (Diaclone Kit Murine IFNγELISpot, 862.031-005PC). The peptides used for restimulation are described in Table 5.

[0571] Table 5. Setup for ELISPOT-IFNγ assay.

[0572]

[0573] The spots were counted on a CTL ELISpot reader. Data plotting and statistical analysis were performed using Prism-5 software (GraphPad Software Inc.).

[0574] B. Result

[0575] At the start of the experiment, all mice were 8 to 13 weeks old. Both males and females were used in the study. A maximum of 6 animals were housed per cage. At sacrifice, flow cytometry analysis of the spleen T cell population showed that the majority belonged to the CD4+ T cell subset.

[0576] After plating and incubation with appropriate stimulation, cells producing IFNγ were displayed and counted. Data are presented in increments of 1.10. 6 The number of spots for total T cells was provided. The group mean was then plotted using the single mean (obtained from triplicate). Statistical analysis was performed using unpaired nonparametric tests (Mann-Whitney) (**: p < 0.01; *: p < 0.05) for comparisons (with culture conditions).

[0577] In summary, vaccination with the antigenic peptides (CD19-B1, CD19-B2, CD22-B1, CD37-B1, TNFRSF13C-B1, and MS4A1-B4) according to the present invention induced a significant T cell response in HHD DR1 mice in an ELISPOT-IFNγ assay. Figure 6-10 The immunogenicity of CD22-B1, CD37-B1, TNFRSF13C-B1, and MS4A1-B4 was confirmed in HHD DR1 mice. Figure 11-13 ).

[0578] result( Figure 6 The results showed that immunization of HHD-DR1 mice with CD22-B1 allowed the induction of T cells, which were able to respond strongly upon challenge with either CD22-B1 or its human counterpart, CD22-H1. Therefore, CD22-B2 possesses strong immunogenicity and can drive an effective immune response against its corresponding human peptide.

[0579] These results were confirmed in HHD DR3 mice that expressed human HLA-A2 and HLA-DR3 MHC and lacked mouse H-2 class I and II MHC. Figure 11 ).

[0580] result( Figure 7The results showed that immunization of HHD-DR1 mice with TNFRSF13C-B1 allowed the induction of T cells, which were able to respond strongly upon challenge with TNFRSF13C-B1 or its human counterpart, TNFRSF13-H1. Therefore, TNFRSF13C-B1 is highly immunogenic and can drive an effective immune response against its corresponding human peptide.

[0581] These results were confirmed in HHD DR3 mice that expressed human HLA-A2 and HLA-DR3 MHC and lacked mouse H-2 class I and II MHC. Figure 12 ).

[0582] result( Figure 8 The results showed that immunization of HHD-DR1 mice with CD37-B1 allowed the induction of T cells, which were able to respond strongly upon challenge with either CD37-B1 or the human counterpart, CD37-H1. Therefore, CD37-B2 is highly immunogenic and can drive an effective immune response against its corresponding human peptide.

[0583] These results were confirmed in HHD DR3 mice that expressed human HLA-A2 and HLA-DR3 MHC and lacked mouse H-2 class I and II MHC. Figure 13 ).

[0584] result( Figure 9 The results showed that immunization of HHD-DR1 mice with CD19-B2 allowed the induction of T cells, which were able to respond strongly upon challenge with either CD19-B2 or its human counterpart, CD19-H2. Therefore, CD19-B2 is highly immunogenic and can drive an effective immune response against its corresponding human peptide.

[0585] result( Figure 10 The results showed that immunization of HHD-DR1 mice with CD19-B1 allowed the induction of T cells, which were able to respond strongly upon challenge with either CD19-B1 or the human counterpart, CD19-H1. Therefore, CD19-B1 is highly immunogenic and can drive an effective immune response against the corresponding human peptide.

[0586] result( Figure 15 The results showed that immunization of HHD-DR1 mice with MS4A1-B4 allowed the induction of T cells, which were able to respond strongly upon challenge with MS4A1-B4 or its human counterpart, MS4A1-H4. Therefore, MS4A1-B4 is highly immunogenic and can drive an effective immune response against its corresponding human peptide.

[0587] These results were confirmed in HHD DR3 mice that expressed human HLA-A2 and HLA-DR3 MHC and lacked mouse H-2 class I and II MHC. Figure 16 ).

[0588] In summary, the immunogenicity studies described in Example 2, conducted in HHD DR3 and HHD DR1 mice, demonstrated that the six antigenic peptides of the present invention—CD19-B1, CD19-B2, CD22-B1, CD37-B1, TNFRSF13C-B1, and MS4A1-B4—induce a strong immune response. Cross-reactivity with T cells generated by the corresponding human peptides against CD19-B1, CD19-B2, CD22-B1, CD37-B1, TNFRSF13C-B1, and MS4A1-B4 was also demonstrated in HHD DR3 and HHD DR1 mice.

[0589] Therefore, these results provide experimental evidence that antigen-based immunotherapy can improve T-cell responses in vivo and that the antigenic peptides according to the invention are particularly effective for this purpose.

[0590] Example 3: In vitro cell culture of CD22-B1, CD37B1, TNFRSF13C-B1 and MS4A1-B4 specific CD8 human T cells Cytotoxic effects.

[0591] Multiple studies support the concept of a specific T-cell pool targeting microbial peptides. The number of peptide-specific microbe-specific T cells is expected to be low, but sufficient to be reactivated through vaccine challenges.

[0592] To identify and functionally characterize circulating CD22-B1, CD37B1, TNFRSF13C-B1, and MS4A1-B4-specific T cells in humans, in vitro expansion protocols have been developed to detect T cells specific to each antigenic peptide and investigate their cytotoxicity.

[0593] 3.1 Identifying antigenic peptide-specific CD8 T cells in the human body

[0594] In vitro expansion methods and specific pMHC multimers have been used to identify CD22-B1, CD37-B1, TNFRSF13C-B1, and MS4A1-B4 specific T cells. pMHC multimers were generated for all bacterial peptides and their corresponding human counterparts. PBMCs from several HLA-A*02 healthy donors (up to 19 donors) were collected, enriched after CD137 and CD8 selection, and subjected to multiple rounds of in vitro expansion using T2 cells loaded with the EO2463 peptide to increase the number of specific T cell clones. OMP peptide-specific CD8 T cells were detected using cell counting analysis with fluorescent multimers on the enriched CD8 T cell population.

[0595] Figure 17The results obtained using a healthy HLA-A2 donor are illustrated. For this donor, cell expansion allowed for the detection of MS4A1-B4 specific cells (19.7%), TNFRSF13C-B1 specific cells (13%), CD22-B1 specific cells (4.6%), and CD37-B1 specific cells (2.5%).

[0596] In summary, these results demonstrate the presence of CD8 T cells in the blood of healthy HLA-A2 donors, which can recognize peptides derived from the microbiome and, importantly, human counterparts.

[0597] 3.2 Antigen peptide-specific CD8 T cell cytotoxicity function

[0598] The amplified CD8+ T cells described above were used for cytotoxicity assays in the presence of different proportions of target and effector cells, with flow cytometry readings used to evaluate their cytotoxicity. Target cells were T2 cell lines loaded with bacterial peptides or their human counterparts. Negative controls included unloaded T2 cells and T2 cells loaded with irrelevant peptides. Figure 18 As shown, the in vitro expanded antigen peptide-specific human T cell clones possessed the ability to kill T2 cells loaded with all bacterial peptides CD22-B1, CD37B1, TNFRSF13C-B1, and MS4A1-B4. More importantly, when cross-reactivity was observed by staining, the in vitro expanded CD22-B1, CD37B1, TNFRSF13C-B1, and MS4A1-B4-specific human T cell clones were able to kill T2 cells loaded with human TNFR13C (BAFF-R) or MS4A1 (CD20) peptides. Figure 17 ).

[0599] Overall, these results demonstrate that T-cell clones exist in healthy volunteers capable of recognizing microbial peptides and killing targets using both the microbial peptides and their human counterparts. These data are particularly encouraging because T-cell clones have been obtained from healthy donors, thus we can expect that specific T-cell clones can be effectively expanded in patients exposed to the antigenic peptides of this invention.

[0600] Example 4: The further antigenic peptide has excellent affinity for the HLA-A*0201 allele.

[0601] Next, the binding affinity of the selected antigenic peptide and the corresponding fragment of the human tumor antigen (human reference peptide) to the HLA-A*0201 allele was confirmed in vitro.

[0602] That is, the antigenic peptides of sequences SEQ ID NO:110 (《YIFEHPELL》, also referred to herein as CD22-B1), SEQ ID NO:107 (《LIFEHPERV》, also referred to herein as CD22-B12), and SEQ ID NO:108 (《RVFEHPELV》, also referred to herein as CD22-B13) are compared with the corresponding reference human peptides derived from CD22 (《WVFEHPETL》, SEQ ID NO:270, also referred to herein as CD22-H1).

[0603] In addition, the antigenic peptides of sequences SEQ ID NO:114 (FLAFVPLQL, also referred to herein as CD37-B1), SEQ ID NO:119 (ILAFVPLYL, also referred to herein as CD37-B12), SEQ ID NO:120 (IMAFVPLAV, also referred to herein as CD37-B13), SEQ ID NO:491 (FLAFVPLDV, also referred to herein as CD37-B14), and SEQ ID NO:493 (VLAFVPLGV, also referred to herein as CD37-B15) were compared with their corresponding reference human peptides derived from CD37 (GLAFVPLQI, SEQ ID NO:271, also referred to herein as CD37-H1).

[0604] In addition, the antigenic peptides of sequences SEQ ID NO:220 (《LMFGAPALV》, also referred to herein as TNFRSF13C-B1), SEQ ID NO:212 (《FLFGAPASA》, also referred to herein as TNFRSF13C-B11), SEQ ID NO:217 (《LLFGAPAGV》, also referred to herein as TNFRSF13C-B12), and SEQ ID NO:224 (《VLFGAPAYL》, also referred to herein as TNFRSF13C-B13) were compared with their corresponding reference human peptides derived from TNFRSF13C (《LLFGAPALL》, SEQ ID NO:279, also referred to herein as TNFRSF13C-H1).

[0605] In addition, the antigenic peptides of sequences SEQ ID NO:65 (《AMNSLSLYI》, also referred to herein as MS4A1-B4), SEQ ID NO:70 (《YMNSLSLAL》, also referred to herein as MS4A1-B42), and SEQ ID NO:477 (《AMNSLSLTV》, also referred to herein as MS4A1-B43) were compared with their corresponding reference human peptides derived from MS4A1 (also referred to herein as CD20) (《IMNSLSLFA》, SEQ ID NO:264, also referred to herein as MS4A1-H4).

[0606] A. Materials and Methods

[0607] A1. Measure the affinity of the peptide for the T2 cell line.

[0608] The experimental protocol was similar to that used for validating HLA-A*0201-presented peptides (Tourdot et al., A general strategy to enhance immunogenicity of low-affinity HLA-A2.1-associated peptides: implication in the identification of cryptic tumor epitopes. Eur J Immunol. Dec. 2000; 30(12):3411-21). Peptide affinity measurements were performed using T2 human tumor cells that expressed HLA-A*0201 molecules but were TAP1 / 2 negative and unable to present endogenous peptides.

[0609] T2 cells (5.10) 4 Cells / well were incubated with peptides at decreasing concentrations from 100 μM to 0.1 μM (4 spots: 100 μM, 10 μM, 1 μM, 0.1 μM) in serum-free medium (TexMacs) supplemented with 100 ng / μl β2 microglobulin at 37°C for 16 h. Cells were then washed twice and labeled with an anti-HLA-A2 antibody (clone BB7.2, BD Pharmagen) conjugated to PE.

[0610] The analysis was performed using FACS (Macsquant Analyzer 10-Miltenyi).

[0611] For each peptide concentration, the geometric mean of the target peptide-related label was subtracted from the background noise and reported as a percentage of the geometric mean of the HLA-A*0202 label obtained for the reference peptide HIV pol 589-597 at a concentration of 100 μM.

[0612] A2. Peptide solubilization

[0613] Solubilize each peptide by taking into account its amino acid composition. For peptides that do not contain any cysteine, methionine, or tryptophan, DMSO can be added up to 10% of the total volume. Other peptides are resuspended in water or PBS at pH 7.4.

[0614] B. Result

[0615] The results show Figure 19-22 In the middle. For each test, the human reference epitope CD22-H1 ( Figure 19 ), CD37-H1( Figure 20 ), TNFRSF13C-H1 ( Figure 21 ) and MS4A1-H4 ( Figure 22 According to the present invention, the corresponding antigen peptide shows a strong binding affinity for HLA-A*0201.

[0616] In summary, the results show that the antigenic peptide according to the invention exhibits a stronger binding affinity for HLA-A*0201 than the corresponding human tumor antigen fragment. As outlined above, without being bound by any theory, it is assumed that this strong binding affinity of the antigenic peptide according to the invention reflects its ability to elicit an immune response (i.e., its immunogenicity). sequence list <110> Entron Company <120> Antigen peptides used for the prevention and treatment of B-cell malignancies <130> EB01P016WO1 <150> EP19306475.5 <151> 2019-11-15 <150> PCT / EP2020 / 079257 <151> 2020-10-16 <160> 509 <170> PatentIn version 3.5 <210> 1 <211> 9 <212> PRT <213> Artificial sequence <220> <223> peptides <400> 1 Phe Leu Leu Phe Leu Thr Pro Ile Leu 1 5 <210> 2 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 2 Phe Leu Leu Phe Leu Thr Pro Leu Leu 1 5 <210> 3 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 3 Phe Met Leu Phe Leu Thr Pro Arg Ile 1 5 <210> 4 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 4 Gly Leu Leu Phe Leu Thr Pro Leu Ala 1 5 <210> 5 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 5 Gly Leu Leu Phe Leu Thr Pro Leu Leu 1 5 <210> 6 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 6 Gly Leu Leu Phe Leu Thr Pro Leu Met 1 5 <210> 7 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 7 Ile Leu Leu Phe Leu Thr Pro Leu Leu 1 5 <210> 8 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 8 Ser Leu Leu Phe Leu Thr Pro Leu Leu 1 5 <210> 9 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 9 Thr Leu Leu Phe Leu Thr Pro Leu Ile 1 5 <210> 10 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 10 Thr Leu Leu Phe Leu Thr Pro Met Leu 1 5 <210> 11 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 11 Val Leu Leu Phe Leu Thr Pro Met Leu 1 5 <210> 12 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 12 Tyr Leu Leu Phe Leu Thr Pro Val Leu 1 5 <210> 13 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 13 Ala Leu Ser Leu Gly Leu Pro Gly Leu 1 5 <210> 14 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 14 Ala Leu Ser Leu Gly Leu Pro Leu Leu 1 5 <210> 15 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 15 Ala Leu Ser Leu Gly Leu Pro Met Leu 1 5 <210> 16 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 16 Ala Leu Ser Leu Gly Leu Pro Gln Leu 1 5 <210> 17 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 17 Ala Leu Ser Leu Gly Leu Pro Arg Leu 1 5 <210> 18 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 18 Ala Met Ser Leu Gly Leu Pro Cys Leu 1 5 <210> 19 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 19 Ala Met Ser Leu Gly Leu Pro Met Leu 1 5 <210> 20 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 20 Phe Leu Ser Leu Gly Leu Pro Ile Leu 1 5 <210> twenty one <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> twenty one Phe Leu Ser Leu Gly Leu Pro Lys Leu 1 5 <210> twenty two <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> twenty two Ile Leu Ser Leu Gly Leu Pro Ile Leu 1 5 <210> twenty three <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> twenty three Lys Leu Ser Leu Gly Leu Pro Val Leu 1 5 <210> twenty four <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> twenty four Leu Leu Ser Leu Gly Leu Pro Phe Leu 1 5 <210> 25 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 25 Leu Leu Ser Leu Gly Leu Pro Gly Leu 1 5 <210> 26 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 26 Met Leu Ser Leu Gly Leu Pro Ile Leu 1 5 <210> 27 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 27 Arg Leu Ser Leu Gly Leu Pro Gly Leu 1 5 <210> 28 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 28 Ser Leu Ser Leu Gly Leu Pro Ile Leu 1 5 <210> 29 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 29 Ser Leu Ser Leu Gly Leu Pro Lys Leu 1 5 <210> 30 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 30 Val Leu Ser Leu Gly Leu Pro Leu Leu 1 5 <210> 31 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 31 Val Leu Ser Leu Gly Leu Pro Thr Ala 1 5 <210> 32 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 32 Val Leu Ser Leu Gly Leu Pro Thr Val 1 5 <210> 33 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 33 Tyr Leu Ser Leu Gly Leu Pro Ile Leu 1 5 <210> 34 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 34 Leu Leu Val Gly Ile Leu His Leu Val 1 5 <210> 35 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 35 Ser Leu Val Gly Ile Leu His Ile Ile 1 5 <210> 36 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 36 Phe Leu Ala Tyr Leu Ile Phe Gly Leu 1 5 <210> 37 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 37 Phe Leu Ala Tyr Leu Ile Phe Thr Leu 1 5 <210> 38 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 38 Arg Leu Ala Tyr Leu Ile Phe Leu Leu 1 5 <210> 39 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 39 Tyr Leu Ala Tyr Leu Ile Phe Glu Leu 1 5 <210> 40 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 40 Leu Gln Met Gly Gly Phe Tyr Leu Leu 1 5 <210> 41 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 41 Ala Ile Leu Gly Gly Leu Leu Leu Ile 1 5 <210> 42 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 42 Ala Leu Leu Gly Gly Leu Leu Leu Ile 1 5 <210> 43 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 43 Ala Leu Leu Gly Gly Leu Leu Met Leu 1 5 <210> 44 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 44 Phe Ala Leu Gly Gly Leu Leu Thr Val 1 5 <210> 45 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 45 Phe Leu Leu Gly Gly Leu Leu Leu Ile 1 5 <210> 46 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 46 Phe Leu Leu Gly Gly Leu Leu Met Val 1 5 <210> 47 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 47 Gly Met Leu Gly Gly Leu Leu Leu Ile 1 5 <210> 48 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 48 Gly Met Leu Gly Gly Leu Leu Met Leu 1 5 <210> 49 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 49 His Ile Leu Gly Gly Leu Leu Met Val 1 5 <210> 50 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 50 Ile Ile Leu Gly Gly Leu Leu Val Val 1 5 <210> 51 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 51 Ile Leu Leu Gly Gly Leu Leu Leu Ile 1 5 <210> 52 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 52 Leu Leu Leu Gly Gly Leu Leu Leu Ile 1 5 <210> 53 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 53 Leu Leu Leu Gly Gly Leu Leu Met Ile 1 5 <210> 54 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 54 Asn Leu Leu Gly Gly Leu Leu Leu Ile 1 5 <210> 55 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 55 Ser Ile Leu Gly Gly Leu Leu Leu Ile 1 5 <210> 56 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 56 Ser Leu Leu Gly Gly Leu Leu Leu Ile 1 5 <210> 57 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 57 Ser Leu Leu Gly Gly Leu Leu Met Leu 1 5 <210> 58 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 58 Ser Met Leu Gly Gly Leu Leu Leu Ile 1 5 <210> 59 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 59 Thr Leu Leu Gly Gly Leu Leu Met Ile 1 5 <210> 60 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 60 Tyr Ala Leu Gly Gly Leu Leu Glu Val 1 5 <210> 61 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 61 Tyr Ile Leu Gly Gly Leu Leu Met Val 1 5 <210> 62 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 62 Tyr Met Leu Gly Gly Leu Leu Leu Ile 1 5 <210> 63 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 63 Tyr Val Leu Gly Gly Leu Leu Met Ile 1 5 <210> 64 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 64 Tyr Val Leu Gly Gly Leu Leu Met Val 1 5 <210> 65 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 65 Ala Met Asn Ser Leu Ser Leu Tyr Ile 1 5 <210> 66 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 66 Ile Leu Asn Ser Leu Ser Leu Lys Ile 1 5 <210> 67 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 67 Ile Leu Asn Ser Leu Ser Leu Lys Leu 1 5 <210> 68 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 68 Ile Leu Asn Ser Leu Ser Leu Leu Leu 1 5 <210> 69 <211> 9 <212> PRT <213> artificial <220> <223> artificial <400> 69 Leu Leu Asn Ser Leu Ser Leu Phe Leu 1 5 <210> 70 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 70 Tyr Met Asn Ser Leu Ser Leu Ala Leu 1 5 <210> 71 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 71 Phe Leu Ile Pro Ala Gly Ile Phe Leu 1 5 <210> 72 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 72 Ile Leu Ile Pro Ala Gly Ile Tyr Leu 1 5 <210> 73 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 73 Leu Leu Ile Pro Ala Gly Ile Ala Val 1 5 <210> 74 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 74 Leu Leu Ile Pro Ala Gly Ile Glu Leu 1 5 <210> 75 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 75 Leu Leu Ile Pro Ala Gly Ile Gly Leu 1 5 <210> 76 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 76 Leu Leu Ile Pro Ala Gly Ile Leu Ile 1 5 <210> 77 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 77 Leu Leu Ile Pro Ala Gly Ile Leu Leu 1 5 <210> 78 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 78 Met Leu Ile Pro Ala Gly Ile Pro Ala 1 5 <210> 79 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 79 Met Met Ile Pro Ala Gly Ile Ala Val 1 5 <210> 80 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 80 Val Met Ile Pro Ala Gly Ile Phe Leu 1 5 <210> 81 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 81 Phe Leu Phe Leu Gly Ile Leu Gly Leu 1 5 <210> 82 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 82 Phe Leu Phe Leu Gly Ile Leu Pro Leu 1 5 <210> 83 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 83 Met Leu Phe Leu Gly Ile Leu Ser Val 1 5 <210> 84 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 84 Tyr Leu Phe Leu Gly Ile Leu Gly Ile 1 5 <210> 85 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 85 Tyr Leu Phe Leu Gly Ile Leu Gly Leu 1 5 <210> 86 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 86 Tyr Leu Phe Leu Gly Ile Leu Ser Leu 1 5 <210> 87 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 87 Tyr Leu Phe Leu Gly Ile Leu Tyr Leu 1 5 <210> 88 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 88 Phe Leu Ser Asn Asp Thr Val Leu Leu 1 5 <210> 89 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 89 Phe Leu Ser Asn Asp Thr Val Pro Leu 1 5 <210> 90 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 90 Phe Leu Ser Asn Asp Thr Val Ser Ala 1 5 <210> 91 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 91 Phe Met Ser Asn Asp Thr Val Lys Val 1 5 <210> 92 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 92 Ile Leu Ser Asn Asp Thr Val Trp Leu 1 5 <210> 93 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 93 Lys Met Ser Asn Asp Thr Val Val Leu 1 5 <210> 94 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 94 Arg Leu Ser Asn Asp Thr Val Gly Leu 1 5 <210> 95 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 95 Arg Met Ser Asn Asp Thr Val Glu Ile 1 5 <210> 96 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 96 Thr Leu Ser Asn Asp Thr Val Trp Leu 1 5 <210> 97 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 97 Ala Leu Leu Gly Pro Trp Leu Ile Val 1 5 <210> 98 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 98 Phe Leu Leu Gly Pro Trp Leu Cys Leu 1 5 <210> 99 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 99 Lys Leu Leu Gly Pro Trp Leu Ser Val 1 5 <210> 100 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 100 Leu Leu Leu Gly Pro Trp Leu Leu Leu 1 5 <210> 101 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 101 Tyr Leu Leu Gly Pro Trp Leu Leu Val 1 5 <210> 102 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 102 Ile Leu Ile Leu Ala Ile Cys Gly Val 1 5 <210> 103 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 103 Ile Met Ile Leu Ala Ile Cys Leu Val 1 5 <210> 104 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 104 Arg Leu Ile Leu Ala Ile Cys Gly Leu 1 5 <210> 105 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 105 Tyr Leu Ile Leu Ala Ile Cys Gly Val 1 5 <210> 106 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 106 Lys Ile Phe Glu His Pro Glu Leu Leu 1 5 <210> 107 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 107 Leu Ile Phe Glu His Pro Glu Arg Val 1 5 <210> 108 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 108 Arg Val Phe Glu His Pro Glu Leu Val 1 5 <210> 109 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 109 Tyr Val Phe Glu His Pro Glu Leu Leu 1 5 <210> 110 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 110 Tyr Ile Phe Glu His Pro Glu Thr Ala 1 5 <210> 111 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 111 Ala Leu Ala Phe Val Pro Leu Ala Val 1 5 <210> 112 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 112 Ala Leu Ala Phe Val Pro Leu Ser Val 1 5 <210> 113 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 113 Phe Leu Ala Phe Val Pro Leu Ile Leu 1 5 <210> 114 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 114 Phe Leu Ala Phe Val Pro Leu Gln Leu 1 5 <210> 115 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 115 Phe Leu Ala Phe Val Pro Leu Val Leu 1 5 <210> 116 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 116 Phe Met Ala Phe Val Pro Leu Gln Leu 1 5 <210> 117 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 117 Gly Met Ala Phe Val Pro Leu Leu Leu 1 5 <210> 118 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 118 His Leu Ala Phe Val Pro Leu Leu Val 1 5 <210> 119 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 119 Ile Leu Ala Phe Val Pro Leu Tyr Leu 1 5 <210> 120 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 120 Ile Met Ala Phe Val Pro Leu Ala Val 1 5 <210> 121 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 121 Ile Met Ala Phe Val Pro Leu Ile Val 1 5 <210> 122 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 122 Ile Met Ala Phe Val Pro Leu Val Val 1 5 <210> 123 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 123 Leu Leu Ala Phe Val Pro Leu Ala Leu 1 5 <210> 124 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 124 Leu Leu Ala Phe Val Pro Leu Asp Val 1 5 <210> 125 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 125 Leu Leu Ala Phe Val Pro Leu Met Leu 1 5 <210> 126 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 126 Leu Leu Ala Phe Val Pro Leu Ser Leu 1 5 <210> 127 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 127 Leu Met Ala Phe Val Pro Leu Thr Leu 1 5 <210> 128 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 128 Thr Leu Ala Phe Val Pro Leu Ala Val 1 5 <210> 129 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 129 Val Leu Ala Phe Val Pro Leu Leu Val 1 5 <210> 130 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 130 Val Met Ala Phe Val Pro Leu Val Val 1 5 <210> 131 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 131 Phe Leu Tyr Phe Gly Met Leu Leu Leu 1 5 <210> 132 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 132 Gly Leu Tyr Phe Gly Met Leu His Met 1 5 <210> 133 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 133 Leu Leu Tyr Phe Gly Met Leu Gly Leu 1 5 <210> 134 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 134 Leu Leu Tyr Phe Gly Met Leu Leu Leu 1 5 <210> 135 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 135 Thr Met Tyr Phe Gly Met Leu Tyr Leu 1 5 <210> 136 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 136 Val Leu Tyr Phe Gly Met Leu Leu Ile 1 5 <210> 137 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 137 Cys Leu Ile Asp Lys Thr Ser Val Val 1 5 <210> 138 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 138 Phe Leu Ile Asp Lys Thr Ser Ala Ala 1 5 <210> 139 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 139 Ile Leu Ile Asp Lys Thr Ser Gly Ala 1 5 <210> 140 <211> 9 <212> PRT <213> artificial <220> <223> peptides <400> 140 Ile Leu Ile Asp Lys Thr Ser Gly Val 1 5 <210> 141 <211> 9 <212> PRT <213> artificial &l...

Claims

1. An antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO:

65.

2. An immunogenic compound comprising the antigenic peptide according to claim 1.

3. The immunogenic compound according to claim 2, wherein the antigenic peptide is linked to a carrier molecule.

4. The immunogenic compound according to claim 3, wherein the carrier molecule is a carrier protein or a carrier peptide.

5. Nanoparticles, which are loaded with - The antigenic peptide according to claim 1, or - An immunogenic compound according to any one of claims 2-4.

6. The nanoparticles according to claim 5 are further loaded with an adjuvant.

7. Cells loaded with the antigenic peptide according to claim 1 or the immunogenic compound according to any one of claims 2-4.

8. The cell according to claim 7, wherein the cell is an antigen-presenting cell.

9. The cell according to claim 8, wherein the cell is a dendritic cell.

10. A nucleic acid encoding an antigenic peptide according to claim 1, or an immunogenic compound according to any one of claims 2-4, wherein the immunogenic compound is a peptide or a protein.

11. The nucleic acid according to claim 10, wherein the nucleic acid is a DNA molecule or an RNA molecule.

12. The nucleic acid according to claim 10 or 11, wherein the nucleic acid is selected from genomic DNA; cDNA; mRNA; vector; and combinations thereof.

13. A host cell comprising the nucleic acid according to any one of claims 10 to 12.

14. The host cell according to claim 13, wherein the nucleic acid is a carrier.

15. The host cell according to claim 13 or 14, wherein the host cell is a bacterial cell.

16. The host cell according to claim 15, wherein the host cell is an intestinal bacterial cell.

17. Cytotoxic T lymphocytes (CTLs) that are specific to the antigenic peptide according to claim 1.

18. A pharmaceutical composition, comprising - The antigenic peptide according to claim 1, - The immunogenic compound according to any one of claims 2-4, - The nanoparticles according to claim 5 or 6 -The cell according to any one of claims 7 to 9, - Nucleic acid according to any one of claims 10 to 12, - The host cell according to any one of claims 13 to 16, or - The cytotoxic T lymphocytes according to claim 17 And one or more pharmaceutically acceptable excipients or carriers.

19. The pharmaceutical composition of claim 18, wherein the composition comprises (i) The antigenic peptide according to claim 1, and the composition further comprising at least one antigenic peptide selected from the following: an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 110, an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 114, and an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 220; (ii) The immunogenic compound according to any one of claims 2-4, and the composition further comprising at least one immunogenic compound comprising an antigenic peptide selected from the following: an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 110, an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 114, and an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 220; (iii) The nanoparticles according to claim 5, and the composition further comprising at least one nanoparticle comprising an antigenic peptide selected from the following: an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 110, an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 114, and an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 220; (iv) The nucleic acid according to claim 10 or 11, wherein the composition further comprises at least one nucleic acid encoding an antigenic peptide selected from the following: an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 110, an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 114, and an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 220; or (v) The cytotoxic T lymphocytes according to claim 17, wherein the composition further comprises at least one cytotoxic T lymphocyte specific to an antigenic peptide selected from the following: an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 110, an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 114, and an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO:

220.

20. The pharmaceutical composition according to claim 18, comprising: (i) The antigenic peptide according to claim 1, and the composition further comprising at least two antigenic peptides selected from the following: an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 110, an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 114, and an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 220; (ii) The immunogenic compound according to any one of claims 2-4, and the composition further comprising at least two immunogenic compounds comprising an antigenic peptide selected from the following: an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 110, an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 114, and an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 220; (iii) The nanoparticles according to claim 5, wherein the composition further comprises at least two nanoparticles comprising an antigenic peptide selected from the following: an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 110, an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 114, and an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO:

220. (iv) The nucleic acid according to claim 10 or 11, wherein the composition further comprises at least two nucleic acids encoding an antigenic peptide selected from the following: an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 110, an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 114, and an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 220; or (v) The cytotoxic T lymphocytes according to claim 17, wherein the composition further comprises at least two cytotoxic T lymphocytes specific to an antigenic peptide selected from the following: an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 110, an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 114, and an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO:

220.

21. The pharmaceutical composition of claim 18, comprising the antigenic peptide of claim 1, and further comprising at least one antigenic peptide selected from the following: an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 110, an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 114, and an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO:

220.

22. The pharmaceutical composition of claim 18, comprising the antigenic peptide of claim 1, and further comprising... An antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO:

110. An antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 114, and An antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO:

220.

23. The pharmaceutical composition according to claim 18, further comprising an accessory peptide.

24. The pharmaceutical composition of claim 23, wherein the accessory peptide comprises the amino acid sequence according to SEQ ID NO:

475.

25. Reagent kit, including - The antigenic peptide according to claim 1, - The immunogenic compound according to any one of claims 2 to 4, - The nanoparticles according to claim 5 or 6 -The cell according to any one of claims 7 to 9, - Nucleic acid according to any one of claims 10 to 12, -The host cell according to any one of claims 13 to 16, - The cytotoxic T lymphocytes according to claim 17, or - The pharmaceutical composition according to any one of claims 18 to 24.

26. The kit of claim 25, further comprising a packaging insert or instruction manual with guidance on the prevention or treatment of B-cell malignancies using the antigenic peptide, the immunogenic compound, the nanoparticles, the cells, the nucleic acid, the host cells, the cytotoxic T lymphocytes and / or the pharmaceutical composition.

27. The kit according to claim 25 or 26, wherein the kit comprises the antigenic peptide according to claim 1, and further comprises at least one antigenic peptide selected from the following: an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 110, an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 114, and an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO:

220.

28. The kit according to claim 25 or 26, wherein the kit comprises the immunogenic compound according to any one of claims 2-4, and further comprises at least one immunogenic compound comprising an antigenic peptide selected from the following: an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 110, an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 114, and an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO:

220.

29. The kit according to claim 25 or 26, wherein the kit comprises the nanoparticles according to claim 5, and further comprises at least one nanoparticle comprising an antigenic peptide selected from the following: an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 110, an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 114, and an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO:

220.

30. The kit according to claim 25 or 26, wherein the kit comprises the nucleic acid according to claim 10 or 11, and further comprises at least two nucleic acids encoding an antigenic peptide selected from the following: an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 110, an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 114, and an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO:

220.

31. The kit according to claim 25 or 26, wherein the kit comprises the cytotoxic T lymphocytes according to claim 17, and further comprises at least two types of cytotoxic T lymphocytes specific to an antigenic peptide selected from the following: an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 110, an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 114, and an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO:

220.

32. A combination of at least two different components, said at least two different components being the antigenic peptide according to claim 1 and at least one antigenic peptide selected from the following: an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 110, an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 114, and an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO:

220.

33. A combination of at least two different components, said at least two different components being an immunogenic compound according to any one of claims 2-4 and at least one immunogenic compound comprising an antigenic peptide selected from the following: an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 110, an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 114, and an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO:

220.

34. A combination of at least two different components, said at least two different components being the nanoparticles according to claim 5 and at least one nanoparticle comprising an antigenic peptide selected from the following: an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 110, an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 114, and an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO:

220.

35. A combination of at least two different components, said at least two different components being a nucleic acid according to any one of claims 10 to 12 and at least two nucleic acids encoding an antigenic peptide selected from the following: an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 110, an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 114, and an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO:

220.

36. A combination of at least two different components, said at least two different components being the cytotoxic T lymphocytes according to claim 17 and at least two cytotoxic T lymphocytes specific to an antigenic peptide selected from the following: an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 110, an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO: 114, and an antigenic peptide consisting of the amino acid sequence shown in SEQ ID NO:

220.

37. The combination according to any one of claims 32-36, wherein the at least two different components are included in different compositions.

38. The combination according to any one of claims 32-36, wherein the at least two different components are included in the same composition.

39. The combination according to any one of claims 32-36, wherein the at least two different components are administered via different routes.

40. The combination according to any one of claims 32-36, wherein the at least two different components are administered via the same route.

41. The combination according to any one of claims 32-36, wherein the at least two different components are given sequentially.

42. The combination according to any one of claims 32-36, wherein the at least two different components are given approximately simultaneously.

43. The antigenic peptide according to claim 1, The immunogenic compound according to any one of claims 2 to 4, The nanoparticles according to claim 5 or 6, The cell according to any one of claims 7 to 9, Nucleic acid according to any one of claims 10 to 12, The host cell according to any one of claims 13 to 16, The cytotoxic T lymphocytes according to claim 17, The pharmaceutical composition according to any one of claims 18 to 24, or The kit according to any one of claims 25 to 31 Use in the preparation of drugs for the treatment of B-cell lymphoma.

44. The use according to claim 43, wherein the B-cell lymphoma is selected from: non-Hodgkin lymphoma (NHL), diffuse large B-cell lymphoma (DLBCL), de novo NOS, indolent NOS, primary mediastinal large B-cell lymphoma (PMBCL), T-cell / histocyte-rich large B-cell lymphoma (TCHRBCL), Burkitt lymphoma, mantle cell lymphoma (MCL), and follicular lymphoma (FL).

45. Peptide-MHC (pMHC) multimers, including the antigenic peptide according to claim 1.

Images