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

Abstract

We provide antigen-based immunotherapy, particularly cancer immunotherapy. [Solution] The present invention provides antigenic peptides that are different from human tumor antigen epitopes but have amino acid similarities thereto, and in particular share the same core sequence. The present invention further provides immunogenic compounds, nanoparticles, cells, and pharmaceutical compositions comprising such antigenic peptides and nucleic acids encoding such antigenic peptides.

Description

[Technical Field]

[0001] This invention relates to the field of cancer treatment, and more particularly to the field of cancer treatment by immunotherapy. In particular, this invention provides various peptides useful for cancer immunotherapy, especially for the prevention and treatment of B-cell malignancies. [Background technology]

[0002] Of all B-cell malignancies, including B-cell lymphoma, non-Hodgkin lymphoma (NHL) is the seventh leading cause of new cancer cases and accounts for approximately 3% of cancer-related deaths in the United States. Of all NHLs, diffuse large B-cell lymphoma (DLBCL) is the most common lymphoma subtype, accounting for 32.5% of all newly diagnosed cases, 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 of 6.9 per 100,000 people. Adding an anti-CD20 monoclonal antibody, rituximab, to the standard chemotherapy regimen of R-CHOP significantly improved the complete response (CR), event-free (EFS), and overall (OS) survival rates of DLBCL. Unfortunately, approximately 30–40% of cases relapse or progress after R-CHOP. There are certain subgroups of patients who do not respond well to and have poor outcomes with standard R-CHOP, such as MYC rearranged DLBCL, MYC, BCL2, or high-grade B-cell lymphoma with BCL rearrangement, and activated B-cell (ABC) DLBCL, which may benefit from a new approach (Non-Patent Literature 1).

[0003] Among the new approaches, CAR T cells, such as CD19-targeted chimeric antigen receptor (CAR) T cells, represent a new standard of treatment for DLBCL patients resistant to at least two existing lines of therapy. Two CAR T cell products, axicabtagene ciloleucel (axi-cel) (KTE-019) and tisagenlecleucel (CTL019), have received approval from the U.S. Food and Drug Administration for the treatment of DLBCL resistant to two lines of therapy. While this represents a significant add-on to DLBCL treatment options, approximately 50% of cases still do not recover from the disease. As a result, future research needs to focus on identifying disease, treatment, or patient-related factors that may help predict successful treatment outcomes.

[0004] Therefore, tumor antigen-based vaccination is a unique approach to cancer treatment that has gathered significant interest because it can mobilize a patient's own immune system to recognize, attack, and destroy tumors in a specific and sustainable manner. In fact, tumor cells are known to express numerous peptide antigens that are easily recognized by the immune system. Therefore, vaccines based on such antigens not only improve the overall survival rate of patients but also provide significant opportunities for monitoring immune responses and preparing GMP-grade products due to the low toxicity and low molecular weight of tumor antigens. Examples of tumor antigens include, among others, by-products of proteins transcribed from normally silent genes or overexpressed genes, and by-products of proteins transcribed from proteins expressed by oncoviruses (Non-Patent Document 2), as well as 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 self-antigens. During the thymic selection process, T cells that recognize peptide / self-MHC complexes with sufficient affinity are clonally deleted. By providing protection against autoimmune diseases, this mechanism of T cell repertoire selection also reduces the likelihood of developing immunity against TAAs and TSAs. This is demonstrated by the fact that cancer-reactive TCRs generally have low affinity. Furthermore, most vaccine trials conducted to date using selected TAAs and TSAs with high binding affinity to MHC have not been shown to induce strong immunity and are thought to reflect the results of thymic selection. Therefore, a strong anti-tumor response depends on the presentation of immunoreactive peptides and the presence of a sufficient number of reactive cells "trained" to recognize these antigens. Therefore, there is a need in the art to identify alternative antigenic peptides that can overcome the limitations encountered in the field.

Prior Art Documents

Non-Patent Documents

[0005]

Non-Patent Document 1

[0006] The present invention aims to meet the above-mentioned needs. This object is achieved by the subject matter described below, particularly by the items provided by the present invention and the appended claims.

[0007] Hereinafter, the present invention will be described in more detail.

[0008] Definitions Unless otherwise specified herein, scientific and technical terms used in the present application shall have the meanings commonly understood by those skilled in the art. Further, unless a different method is required from the context, the nomenclature used in this specification, as well as the techniques of cell and tissue culture, are those commonly used and well-known in the art.

[0009] Such techniques are fully described in the literature, for example, in Owen et al. (Kuby Immunology, 7 th , edition, 2013 - W. H. Freeman) and Sambrook et al. (Molecular cloning: A laboratory manual 4th edition, Cold Spring Harbor Laboratory Press - Cold Spring Harbor, NY, USA, 2012).

[0010] Nevertheless, the following definitions apply more specifically to the use of various terms in this specification.

[0011] The terms “peptide,” “polypeptide,” and “protein,” and variations thereof, mean peptides, oligopeptides, polypeptides, or proteins comprising at least two amino acids linked to one another, preferably by ordinary peptide bonds or by modified peptide bonds, such as in the case of isosteric peptides. The term “(poly)peptide” means peptides and / or polypeptides. In particular, the terms “peptide,” “polypeptide,” and “protein” mean a continuous chain of amino acids of any length linked to one another via peptide bonds (-NHCO-). Peptides, polypeptides, and proteins can play structural and / or functional roles in cells in vitro and / or in vivo. The terms “peptide,” “polypeptide,” and “protein” preferably encompass amino acid chains of size from 2 to at least about 1000 amino acid residues. The term “peptide” preferably, as used herein, encompasses amino acid chains of size less than about 30 amino acids, while the terms “polypeptide” and “protein” preferably encompass amino acid chains of size at least 30 amino acids. The terms “polypeptide” and “protein” are used herein in an interconvertible manner. In preferred embodiments, the terms “peptide,” “polypeptide,” and “protein” also include “peptide mimes,” defined as peptide analogs containing non-peptide structural elements, which can mimic or antagonize the biological effects of native parent peptides. Peptide mimes lack classical peptide properties, such as enzymatically cleavable peptide bonds. In particular, peptides, polypeptides, or proteins may contain, or be composed of, amino acids other than the 20 amino acids defined by the genetic code, in addition to these amino acids. In particular, peptides, polypeptides, or proteins in the context of the present invention may equally consist of amino acids modified by natural processes such as post-translational maturation processes or chemical processes, which are well known to those skilled in the art. Such modifications are well described in the literature.These modifications may appear at any location in the polypeptide, i.e., in the peptide backbone, in the amino acid chain, or even at the carboxy or amino terminus. In particular, peptides or polypeptides may be branched following ubiquitination, or cyclic with or without branching. This type of modification may be the result of natural or synthetic post-translational processes well known to those skilled in the art. In the context of this invention, the terms “peptide,” “polypeptide,” and “protein” also include, in particular, modified peptides, polypeptides, and proteins. For example, modifications of peptides, polypeptides, or proteins may include amino acid addition such as 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, iodization, methylation, myristoylation, oxidation, proteolytic processes, phosphorylation, prenylation, racemization, seneloylation, sulfated, arginylation, 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, glycoproteins, and so on.

[0012] In a preferred embodiment, the (poly)peptide or protein is a “classical” (poly)peptide or protein, thereby being composed of amino acids, typically selected from 20 amino acids defined by the genetic code, linked together by conventional peptide bonds.

[0013] As is well 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 herein in an interconvertible manner and mean an exact continuum of natural nucleotides (e.g., A, T, G, C, and U) or synthetic nucleotides, i.e., a chain of at least two nucleotides. In particular, the terms “nucleic acid,” “nucleic acid molecule,” “nucleic acid sequence,” “polynucleotide,” and “nucleotide sequence” mean DNA or RNA. Nucleic acids preferably include single-stranded, double-stranded, or partially double-stranded DNA or RNA, and are preferably selected from transcripts of such DNA, such as genomic DNA (gDNA), complementary DNA (cDNA), ribosomal DNA (rDNA), and 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 DNA molecules described above. Preferably, the nucleic acid (molecule) is selected from DNA molecules or RNA molecules; preferably 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 capable of encoding a specific amino acid sequence is within the skill of those skilled in the art.

[0014] The (poly)peptides and / or nucleic acids according to the present invention can be prepared by any known method in the art, including, but not limited to, any synthetic method, any recombinant method, any ex vivo production method, and any combination thereof. Such techniques are well described in the above-mentioned literature.

[0015] As used herein, the term “antigenic peptide” means a peptide that tends to induce, increase, prolong, or maintain an immune response in the target to which it is administered. In particular, the antigenic peptide is a sequence variant of a (human) tumor antigen (fragment / epitope). In other words, the antigenic peptide is preferably different from, but preferably amino acid similar to, the (human) tumor antigen (fragment / epitope). Importantly, the antigenic peptide shares the same core sequence as each (fragment / epitope) of the (human) tumor antigen. Preferably, the immune response induced, increased, prolonged, or maintained by the antigenic peptide also targets each (fragment / epitope) of the (human) tumor antigen.

[0016] As used herein, the term “tumor antigen” includes tumor-specific antigens (TSAs) and tumor-associated antigens (TAAs). Generally, the terms “tumor antigen” or “tumor protein” as used herein mean antigenic substances produced in tumor cells, and sometimes in normal cells, that can induce an immune response upon administration to a subject. In humans, these are classified according to their expression pattern, function, or genetic origin, and are not limited to, but include: overexpressed autoantigens (such as BIRC5); carcinomatous (CT) antigens (such as MAGE-1); mutant antigens also known as neoantigens (such as variants derived from p53); tissue-specific differentiation antigens (such as melanoma antigen Melan A / MART-1); viral antigens expressed by oncoviruses (such as HPV and EBV); carcinoembryonic antigens (such as alpha-fetoprotein AFP and carcinoembryonic antigen CEA); and ubiquitous antigens (such as telomerase).

[0017] As used herein, the term “B-cell tumor antigen” means an antigen that is related to and / or involved in the pathogenesis of the B-cell malignancy, for example, an antigen that is involved in or expressed in the B-cell malignancy. In other words, the antigen is one of several known B-cell markers that is expressed by or on B cells, including human B cells. Preferably, the B-cell tumor antigen is CD19, CD20, CD22, CD37, or TNFRSF13C, which are highly expressed (overexpressed) in B-cell lymphoma. Antigenic peptides “derived” from the B-cell tumor antigen typically share the same core sequence as the epitope of the B-cell tumor antigen ("reference epitope").

[0018] As used herein, the term “core sequence” means the amino acids located in the middle of a sequence, for example, in the middle of an antigenic peptide and / or (reference) epitope (also called the “central amino acids” of the sequence). Thus, the core sequence consists of all amino acids except the two most N-terminal and two most C-terminal amino acids. For example, in a 9-amino acid peptide (e.g., each (fragment / epitope) of the antigenic peptide or (human) tumor antigen according to the present invention), the five middle amino acids constitute the core sequence, and changes may occur at any of the two N-terminal and two C-terminal amino acid positions. Thus, a “shared core sequence” (or “maintained” core sequence) typically means that mutations / differences are permitted only 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” means the cumulative frequency of each protein in the human microbiota in which the core sequence shared with each (fragment / epitope) of the (human) tumor antigen is found and present in the antigenic peptide. In fact, the core sequence of interest may be present in one or more different antigenic peptides, and each of such antigenic peptides may be present in one or more different proteins expressed in the human microbiota. Thus, the overall prevalence of the core sequence is derived from the frequency of each protein in the human microbiota in which the core sequence shared with each (fragment / epitope) of the (human) tumor antigen is found, by considering the frequency of each protein in the human microbiota in which similar peptides (i.e., different antigenic peptides) sharing the same core sequence are found.

[0020] As used herein, the term “microbiota” refers to symbiotic microorganisms present in or on any multicellular organism studied to date, from plants to animals. In particular, the microbiota has been found to be important for the immunological, hormonal, and metabolic homeostasis of the host. The microbiota includes bacteria, archaea, protists, fungi, and viruses. Thus, a “microbiota sequence variant” (or “microbiota variant”) is a sequence variant of a (human) reference sequence (in particular, an epitope / fragment of a human tumor antigen) that arises in the microbiota (e.g., bacteria (which may be contained in microbiota proteins such as bacterial proteins)). Preferably, the antigenic peptide of the present invention is a microbiota sequence variant (of a reference epitope / fragment of a human B-cell tumor antigen). Therefore, preferably, the antigenic peptide is present in (e.g., included in) at least one protein expressed by the human microbiota.

[0021] A “sequence manifold” typically shares at least 50% sequence identity with the (reference) tumor antigen fragment / epitope, particularly over the entire length of the sequence. Preferably, the sequence manifold shares at least 60%, preferably at least 70%, 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., the (reference) tumor antigen fragment / epitope. Sequence identity can be calculated as is known in the art, particularly as described below. Preferably, the sequence manifold retains a specific function of the reference sequence, e.g., its function as a tumor epitope and / or its ability to induce or maintain an immune response. The microbiota sequence manifold is preferably selected from the group consisting of bacterial sequence manifolds, archaeal sequence manifolds, protist sequence manifolds, fungal sequence manifolds, and viral sequence manifolds. More preferably, the microbiota sequence manifold is a bacterial sequence manifold.

[0022] Anatomically, the microbiota is present on or within any of the many tissues and fluids, including the skin, conjunctiva, mammary glands, vagina, placenta, semen, uterus, follicles, lungs, saliva, oral cavity (particularly the oral mucosa), and the gastrointestinal tract, particularly the intestines. In the context of the present invention, the microbiota sequence variety is preferably a sequence variety of the gastrointestinal microbiota (microorganisms present in the gastrointestinal tract), and more preferably a sequence variety of the intestinal microbiota (microorganisms present in the intestines). Therefore, it is most preferable that the microbiota sequence variety is a (human) intestinal bacterial sequence variety (i.e., a sequence variety of bacteria present in the (human) intestines).

[0023] Microbiota are found inside and on many multicellular organisms (from plants to animals, all multicellular organisms studied to date), but microbiota found inside and on humans are preferred. Such microbiota are referred to herein as “human microbiota” (where the term “human” specifically means the localization / resident of the microbiota). In the context of the present invention, the microbiota sequence variety is a human microbiota sequence variety.

[0024] The term “immunogenic compound” means a compound comprising an antigenic peptide according to the present invention. An “immunogenic compound” can induce, increase, prolong, or maintain an immune response to the antigenic peptide in a subject to which it is administered. In some embodiments, the immunogenic compound comprises at least one antigenic peptide bound to a protein such as a carrier protein, or at least one compound comprising such antigenic peptide.

[0025] A "carrier protein" is typically a protein capable of transporting cargo, such as an antigenic peptide, according to the present invention. For example, the carrier protein can transport the cargo across a membrane. In the context of the present invention, the carrier protein particularly includes peptides or polypeptides capable of inducing an immune response to the antigenic peptide bound thereto. Carrier proteins are known in the art.

[0026] Alternatively, such carrier peptides or polypeptides may be co-administered in the form of immunoadjuvants.

[0027] Preferably, the antigenic peptides described herein may be co-administered or conjugated (e.g., by covalent or non-covalent bonding) with proteins / peptides having immunoadjuvant properties, such as providing stimulation of CD4+ Th1 cells. The antigenic peptides described herein preferably bind to MHC class I, but CD4+ helper epitopes can be further used to provide an efficient immune response. Th1 helper cells secrete interferon-gamma (IFN-γ), tumor necrosis factor alpha (TNF-α), and interleukin-2 (IL-2), thereby enhancing the expression of co-stimulatory signals in dendritic cells (DCs) and T cells, and maintaining efficient DC activation and specific CTL activation (Galaine et al., Interest of Tumor-Specific CD4 T Helper 1 Cells for Therapeutic Anticancer Vaccine. Vaccines (Basel). 2015 Jun 30;3(3):490-502).

[0028] For example, the adjuvant peptide / protein may preferably be different from the antigenic peptide according to the present invention. Preferably, the adjuvant peptide / protein may be a helper peptide that can evoke immunological memory, or that provides nonspecific help, or a specific helper peptide. Several helper peptides for providing nonspecific T cell help, such as tetanus helper peptide, keyhole limpet hemocyanin peptide, or PADRE peptide, have been described in the literature (Adotevi et al, Targeting antitumor CD4 helper T cells with universal tumor-reactive helper peptides derived from telomerase for cancer vaccine. Hum Vaccin Immunother. 2013 May;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. 2011 Sep-Oct;17(5):343-50). Therefore, tetanus helper peptide, keyhole limpet hemocyanin peptide, and PADRE peptide are preferred examples of such adjuvant peptides / proteins. HHD-DR3 peptide of sequence MAKTIAYDEEARRGLERGLN (SEQ ID NO: 473). This peptide is another preferred example of a helper peptide (having immunoadjuvant properties) in the context of the present 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 helper peptides include the UCP2 peptide (e.g., International Publication No. 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, Tartour E, Langlade-Demoyen P, Borg C, Adotevi O: Universal cancer peptide-based therapeutic vaccine breaks tolerance 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 2012 Oct 2) and BIRC5 peptide (for example, European Patent Publication No. 2119726A1 or Widenmeyer M, Griesemann H, Stevanovic 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 2011 Sep 14).The most preferred helper peptide is the UCP2 peptide (amino acid sequence: KSVWSKLQSIGIRQH; SEQ ID NO: 475, e.g., International Publication No. WO2013 / 135553A1 or Dosset et al., Clin Cancer Res. 2012 Nov 15;18(22):6284-95). In particular, the antigenic peptides described herein, or polypeptides containing said antigenic peptides, can be bound to the helper peptide, for example, by covalent or non-covalent bonds.

[0029] As used herein, the term “immunogenic composition” means a composition that can induce, trigger, increase, prolong, or maintain an immune response, in particular a composition that can induce, trigger, increase, prolong, or maintain an immune response when administered to a mammal, and especially when administered to a human organism. Preferably, the immunogenic composition further comprises one or more immunoadjuvant substances.

[0030] "Pharmacologically acceptable excipient or carrier" as used herein means a pharmaceutical-grade compound that improves the delivery, stability, or bioavailability of an active agent, is metabolizable by the target to which it is administered, and is non-toxic. Preferred excipients and carriers according to the present invention include any excipient or carrier commonly used in pharmaceuticals, such as water, saline, phosphate-buffered saline, dextrose, glycerol, ethanol, and combinations thereof. Often, it is preferable to include isotonic agents, such as sugars, polyhydric alcohols such as mannitol and sorbitol, or sodium chloride in the composition. Pharmaceutically acceptable excipients or carriers may further contain small amounts of auxiliary substances, such as wetting agents or emulsifiers, or preservatives.

[0031] "Vaccine" as used herein means a composition that can stimulate the immune system of an organism so as to provide protection against a harmful antigen, either by prevention or treatment. Prophylactic vaccines are preferred. Preferably, the vaccine or vaccine composition further comprises one or more immunoadjuvant substances.

[0032] According to the different aspects and embodiments of the present invention described herein, “subject” or “host” preferably means a mammal, most preferably a human. The subject may have a B-cell malignancy, be suspected of having a B-cell malignancy, or be at risk of developing a B-cell malignancy.

[0033] The term "B-cell malignancy" refers to diseases associated with the transformation of B cells. This includes, among others, B-cell lymphoma, acute lymphoblastic (or lymphoblastic) leukemia (ALL), and chronic lymphocytic leukemia (CLL, Richter). In the context of this invention, B-cell lymphomas such as non-Hodgkin lymphoma (NHL) are preferred. For example, NHL is selected from the group consisting of low-grade (slowly progressive) NHL, high-grade NHL, diffuse large B-cell lymphoma (DLBCL), NOS (de novo and transformed from indolent low-grade lymphoma), primary mediastinal large B-cell lymphoma (PMBCL), T-cell / histiocyte-rich large B-cell lymphoma (TCHRBCL), Burkitt lymphoma, mantle cell lymphoma (MCL), and / or follicular lymphoma (FL), and optionally, follicular lymphoma grade 3B (FL3B).

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

[0035] As used herein, “effective amount” or “effective dose” is the amount that provides the desired effect. For therapeutic purposes, the effective amount is the amount sufficient to provide a beneficial or desired clinical outcome. A preferred effective amount for a given use can be readily determined by those skilled in the art, for example, by considering the size, age, weight of the subject, the type of disease / disorder being prevented or treated, and the time elapsed since the onset of the disease / disorder. In the context of the present invention, from the viewpoint of prevention or treatment, the effective amount of the composition is the amount sufficient to induce a humoral and / or cellular immune response to the disease / disorder.

[0036] Throughout this specification and the subsequent claims, unless the context requires a different interpretation, the terms “comprise” and variations such as “comprises” and “comprising” are understood to mean that they include the described components, integers, or processes, but do not exclude any other undescribed components, integers, or processes. The term “consist of” is a specific embodiment of the term “comprise,” excluding any other undescribed components, integers, or processes. In the context of the present invention, the term “comprise” encompasses the term “consist of.” Therefore, the term “comprising” encompasses “including” and “consisting,” for example, a composition “comprising” X may consist of X alone or may include something additional (e.g., X + Y).

[0037] The terms “a,” “an,” and “the,” as well as similar terms used in the description of the invention (particularly in the context of the claims), are to be interpreted as encompassing both singular and plural unless otherwise specified herein or unless the context clearly contradicts this interpretation. The descriptions of value ranges herein are intended merely as a convenient way to individually refer to each distinct value within that range. Unless otherwise specified herein, each distinct value is incorporated into the specification as described individually herein. No term herein should be interpreted as indicating an unclaimed element essential to the implementation of the invention.

[0038] The term "substantially" does not exclude "completely"; for example, a composition that "substantially does not contain" Y may not contain Y completely. Where necessary, the term "substantially" may be omitted from the definitions of this invention.

[0039] The term "approximately" in relation to the number x means x ± 10%.

[0040] Further definitions are provided throughout the specification.

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

[0042] Detailed explanation The present invention will now be described in detail, but it should be understood that the present invention is not limited to the specific methods, protocols, and reagents described herein, for they may differ from those described. Furthermore, it should be understood that the terms used herein do not limit the scope of the present invention, which is limited only by the appended claims. Unless otherwise specified, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art.

[0043] The elements of the present invention are described below. These elements are listed together with specific embodiments, but it is understood that they can be combined in any way and in any number to create further embodiments. The various examples and preferred embodiments described are not to be construed as limiting the invention to the explicitly described embodiments. This description should be understood as supporting and encompassing embodiments that combine the explicitly described embodiments with any number of disclosed and / or preferred elements. Furthermore, unless otherwise stated in different context, any order and combination of all elements described herein should be considered disclosed by this description.

[0044] Antigenic peptide according to the present invention In a first embodiment, the present invention provides an antigenic peptide derived from a tumor antigen, in particular a B-cell tumor antigen, wherein the antigenic peptide shares the same core sequence as a reference epitope of the tumor antigen, and the shared core sequence is highly abundant in the human microbiota.

[0045] The present invention also provides an antigenic peptide comprising or comprising an amino acid sequence represented by any of SEQ ID NOs: 316, 304-315, 317-472, and 501-509. Preferably, the antigenic peptide may comprise or comprise an amino acid sequence represented by any of SEQ ID NOs: 316, 304-315, and 317-326.

[0046] Furthermore, the present invention provides an antigenic peptide comprising or consisting of an amino acid sequence represented by any of SEQ ID NOs: 1-257 and 476-500, wherein one or two amino acid residues may optionally be substituted, deleted, or added. When one or two amino acid residues are substituted, deleted, or added, preferably the core sequence (of SEQ ID NOs: 1-257 and 476-500) is maintained. More preferably, the antigenic peptide comprises or consists of an amino acid sequence represented by any of SEQ ID NOs: 1-257 and 476-500 (without mutations). The amino acid sequences relating to SEQ ID NOs: 1-257 and 476-500 are microbiota sequence variants (microbiota variants, particularly bacterial sequence variants) of human tumor epitopes, particularly reference epitopes / fragments of human B-cell tumor antigens. In other words, the amino acid sequences corresponding to SEQ ID NOs: 1-257 and 476-500 are present in bacterial proteins and show sequence similarity to the reference epitopes / fragments of human B-cell tumor antigens, as detailed in Table 1A.

[0047] The inventors have identified a series of antigenic peptides that can be used to induce a specific immune response against tumor cells. These antigenic peptides are different from, but share amino acid similarities with, human tumor antigens (or fragments thereof) highly expressed on B-cell lymphocytes, such as CD19, CD20, CD22, CD37, or TNFRSF13C, as shown in Tables 1A, 1B, and 1C. Importantly, the antigenic peptides according to the present invention have the same core sequence as the core sequence of the epitope (fragment) of the reference tumor antigen. Furthermore, the core sequence exhibits a high abundance based on the frequency of proteins present in the human microbiota in which the core sequence is found.

[0048] In particular, the antigenic peptides according to the present invention are contained in polypeptides and proteins produced by commensal bacteria from the human gut. Therefore, the antigenic peptides according to the present invention are bacterial sequences, not human sequences. While we do not wish to be bound by any particular theory, we believe that the human immune repertoire includes T cell clones that are reactive to bacterial peptides (contained in proteins produced by commensal bacteria from the gut) that have amino acid similarity to fragments of human tumor antigens. In particular, the antigenic peptides according to the present invention can induce a stronger immune response than the corresponding human peptides. This is because T cells that can strictly recognize human peptides are eliminated during maturation as they recognize self-antigens (which is not the case with the antigenic peptides according to the present invention). This explains why, when these peptides are administered to a (human) individual, the antigenic peptides described herein can induce an immune response, in particular a T cell response.

[0049] Therefore, although not bound by theory, the inventors believe that proteins produced by intestinal symbiotic bacteria can "mimic" tumor antigens and can be used to induce a specific immune response against tumor cells. These findings provide further evidence that symbiotic bacteria may contribute to the eradication of tumor cells.

[0050] The antigenic peptides disclosed herein can be prepared using well-known techniques. For example, the peptides can be prepared synthetically by recombinant DNA techniques 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 a peptide and a non-peptide). The antigenic peptides can be isolated, i.e., purified to substantially free of other naturally occurring host cell proteins and their fragments, for example, by at least about 70%, 80%, or 90%. Preferably, the antigenic peptides according to the present invention are isolated antigenic peptides.

[0051] The aforementioned core sequences represent key features of the antigenic peptide according to the present invention. Therefore, the inventors identified core sequences of great interest with high abundance. This is because these core sequences are present in several sequence variants of (reference) tumor antigen fragments and / or in several human microbiota proteins with high frequency in many parts of the general human population. The inventors selected the antigenic peptide of the present invention that readily induces the best cross-reactive tumor-specific cytotoxic T-cell immune response for the prevention and treatment of B-cell malignancies.

[0052] Generally, the shared core sequence has a high abundance in the human microbiota when its abundance is higher than 30%, preferably higher than 40%, preferably higher than 50%, more preferably higher than 60%, even more preferably higher than 70%, even more preferably higher than 80%, particularly preferably higher than 90%, and most preferably higher than 95%.

[0053] The abundance of the core sequence is derived from the frequency of each protein in the human microbiota in which a core sequence shared with each (fragment / epitope) of the (human) tumor antigen is found, by considering the frequency of each protein in the human microbiota in which a similar peptide (i.e., a different antigenic peptide) sharing the same core sequence is found. For example, to assess whether the core sequence has a high abundance, the frequency of each protein present in the human microbiota in which the core sequence is found is calculated from a microbiota sequence database. The abundance is then derived from the cumulative frequency calculated for each protein in the human microbiota in which the core sequence of interest is found and present in the antigenic peptide. Such a database may preferably include microbiota (sequence) data from multiple individuals (subjects). 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, Integrated reference catalog of human gut microbiome reference genes, Nat Biotechnol. 2014 Aug;32(8):834-41 URL: http: / / meta.genomics.cn / meta / home), which includes data from major human microbiome profiling efforts, the National Institutes of Health's Human Microbiome Project (NIH-HMP), and the Human Gut Initiative's European Metagenomics (MetaHIT).

[0054] Accordingly, the present invention relates to antigenic peptides having amino acid similarity to tumor antigens. This is because these antigenic peptides are derived from this tumor antigen (or tumor epitope). As used herein, the expression “having amino acid similarity to tumor antigens” means, in particular, sequence varieties of fragments of (reference) human tumor antigens, such as CD22 or other exemplified human tumor antigens described below in Tables 1A, 1B, and 1C. A “sequence variety” typically shares at least 50% sequence identity with a reference sequence, i.e., a fragment of the (reference) tumor antigen, particularly over the entire length of the sequence. Preferably, the sequence variety 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 respect to the reference sequence, i.e., a fragment of the (reference) tumor antigen. Sequence identity can be calculated as is known in the art, in particular as described below. Preferably, the sequence variant retains a specific function of the reference sequence, such as function as a tumor epitope and / or the ability to induce or maintain an immune response. In particular, the amino acid sequence variant has a modified sequence in which one or more amino acids in the reference sequence are mutated (e.g., deleted or substituted) or one or more amino acids are inserted into the sequence of the reference amino acid sequence. For example, a variant sequence that is at least 90% identical has 10 or fewer changes per 100 amino acids of the reference sequence, i.e., any combination of deletions, insertions, or substitutions.

[0055] Methods for comparing the identity (similarity) of two or more sequences are well known in the art. The percentage of identical sequences can be determined, for example, using mathematical algorithms. A preferred but not limited example of a mathematical algorithm that can be used is the algorithm by Karlin et al. (1993), PNAS USA, 90:5873-5877. Such algorithms are integrated into BLAST or NBLAST programs, which are part of the BLAST family of programs (see also Altschul et al., 1990, J.Mol.Biol.215, 403-410 or Altschul et al. (1997), Nucleic Acids Res, 25:3389-3402 (these are accessible from the NCBI homepage on the World Wide Web, 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 are, to some extent, identical to other sequences. Furthermore, programs available in the Wisconsin Sequence Analysis 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 identity percentage between two polynucleotides and between two (poly)peptide sequences. BESTFIT uses the "local homology" algorithm from Smith and Waterman (1981), J.Mol.Biol.147, 195-197 to find the best single region of similarity between two sequences.

[0056] Generally, the antigenic peptide according to the present invention binds to MHC class I (major histocompatibility complex class I, MHC I) molecules.

[0057] MHC class I molecules present epitopes to killer T cells, also known as cytotoxic T lymphocytes (CTLs). CTLs express a CD8 receptor in addition to a TCR (T cell receptor). When the CD8 receptor of a CTL docks with an MHC class I molecule, and the TCR of the CTL fits into the epitope within the MHC class I molecule, the CTL induces programmed cell death (apoptosis) in the cell. This pathway is particularly useful for the prevention and / or treatment of cancer because cancer cells are directly attacked. In humans, MHC class I includes HLA-A, HLA-B, and HLA-C molecules. Typically, peptides (epitopes) 8-10 amino acids in length are presented by MHC class I molecules.

[0058] In general, the antigenic peptide according to the present invention can be of any length. Preferably, the length of the antigenic peptide according to the present invention does not exceed 350 amino acids. For example, the maximum length of the antigenic peptide according to the present invention may be 300 or 250 amino acids. More preferably, the maximum length of the antigenic peptide according to the present invention does not exceed 200 amino acids, for example, 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 or less. In particular, the length of the antigenic peptide according to the present invention is preferably up to 30 or 25 amino acids, more preferably up to 20 or 15 amino acids, and even more preferably up to 10 amino acids. Particularly preferably, the antigenic peptide according to the present invention contains at least 8 or 9 amino acids, for example, 10 amino acids. Even more preferably, the antigenic peptide has a length of 9 or 10 amino acids. In particular, the antigenic peptide is not a full-length protein produced by the human microbiota (from which the antigenic peptide may originate). In other words, the antigenic peptide of the present invention is preferably a fragment of a full-length protein (produced by the human microbiota).

[0059] Similarly, the “fragment / epitope” of the (reference) tumor antigen, which normally functions as a reference sequence, preferably contains nine, and even ten, consecutive amino acids of the tumor antigen. It is understood that the “fragment / epitope” of the (reference) tumor antigen is not the full-length tumor antigen (protein).

[0060] The “fragments” (of proteins or nucleic acids (sequences)) used herein preferably have a maximum length of 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.

[0061] More generally, the present invention provides antigenic peptides comprising or comprising sequence variants of human (reference) tumor antigen fragments / epitopes, particularly microbiota sequence variants of human tumor antigen fragments / epitopes. The human tumor antigen can be selected from the group consisting of CD19, CD20, CD22, CD37, and TNFRSF13C. The fragment / epitope of the human (reference) tumor antigen can be selected from the group consisting of any of SEQ ID NOs. 258 to 280.

[0062] Preferably, the antigenic peptide includes or comprises a microbiota variant of a human reference peptide corresponding to any of SEQ ID NOs. 258-280. In particular, SEQ ID NOs. 258-280 are reference epitopes / fragments of human tumor epitopes, especially human B-cell tumor antigens. Examples of microbiota variants of human reference peptides represented by any of SEQ ID NOs. 258-280 are the peptides represented by SEQ ID NOs. 1-257 and 476-500 (shown in Table 1A below). Preferred microbiota sequence variants (microbiota variants) are bacterial sequence variants. In other words, the amino acid sequences corresponding to SEQ ID NOs. 1-257 and 476-500 are found in bacterial proteins and exhibit sequence similarity to reference epitopes / fragments of human B-cell tumor antigens, as detailed in Table 1A. More preferably, the antigenic peptide comprises or consists of a microbiota variant of a human reference peptide relating to any of SEQ ID NOs. 258, 260-266, 270, 271, 279, and 280. Even more preferably, the antigenic peptide comprises or consists of a microbiota variant of a human reference peptide relating to any of SEQ ID NOs. 260, 264, 270, 271, 279, and 280. Even more preferably, the antigenic peptide comprises or consists of a microbiota variant of a human reference peptide relating to any of SEQ ID NOs. 264, 270, 271, and 279.

[0063] Accordingly, the present invention also provides antigenic peptides comprising or comprising a microbiota variant of a human reference peptide relating to any of SEQ ID NOs. 258 to 280; preferably any of SEQ ID NOs. 258, 260 to 266, 270, 271, 279, and 280; more preferably any of SEQ ID NOs. 260, 264, 270, 271, 279, and 280; and even more preferably any of SEQ ID NOs. 264, 270, 271, and 279.

[0064] In certain embodiments, the present invention provides antigenic peptides derived from CD19, CD20, CD22, CD37, or TNFRSF13C, wherein the antigenic peptides share the same core sequence as the (reference) tumor antigen fragment / epitope, and the shared core sequence has a high abundance in the human microbiota. Such abundance is derived from the frequency of at least one protein present in the human microbiota in which the core sequence is found. Preferably, the antigenic peptides according to the present invention include or consist of any of sequence numbers 304-326 (MHC I consensus sequences).

[0065] In one embodiment, the antigenic peptide binds moderately, strongly, or very strongly to MHC class I (major histocompatibility complex class I, MHC I) molecules.

[0066] The binding of the at least one antigenic peptide to an MHC class I (major histocompatibility complex class I) molecule can be tested by MHC I in silico or in vitro binding tests as described herein. Thus, strong and very strong conjugates can be selected. Preferably, binding to the MHC I molecule is tested (in silico and / or in vitro as described herein) for the at least one antigenic peptide to the MHC I molecule, and further for the (respective) tumor antigen (the fragment / epitope of the (reference) tumor antigen) to the MHC I molecule, and it is preferable that binding affinity is obtained for both (the fragment / epitope of the tumor antigen and the antigenic peptide).

[0067] After the binding test, preferably, only antigenic peptides that bind moderately, strongly, or very strongly to MHC I are selected. More preferably, only strong and very strong conjugates 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, and the epitope / fragment of the (reference) tumor antigen (the "corresponding" tumor antigen epitope sequence) binds weaker (e.g., weakly or moderately) to the MHC I molecule. Even more preferably, only microbiota sequence variants that bind very strongly to the MHC I molecule are selected, and the epitope / fragment of the (reference) tumor antigen binds weakly to the MHC I molecule.

[0068] Therefore, the antigenic peptide of the present invention preferably binds more strongly to MHC I molecules than each of the human reference epitopes of the B cell tumor antigens (which share the same core sequence) (i.e., has a higher binding affinity). In other words, the reference epitopes of the human B cell tumor antigens that share the same core sequence preferably bind less strongly to MHC I molecules than each of the antigenic peptides (i.e., have a lower binding affinity).

[0069] Prediction of MHC class I binding (MHC in silico binding assay) can be performed using publicly available tools such as "NetMHCpan," for example, "NetMHCpan 3.0 Server" or "NetMHCpan 4.0 Server" (Biological Sequence Analysis Center, Technical University of Denmark (DTU); URL: http: / / www.cbs.dtu.dk / services / NetMHCpan / ). The NetMHCpan method, particularly versions 3.0 and later, is trained on over 180,000 quantitative binding data covering 172 MHC molecules derived from humans (HLA-A, B, C, E) and other species. Generally, affinity can be predicted by leaving default thresholds for strong and weak binding. For example, HLA-A *In the case of 0201, a calculated affinity of less than 50 nM can indicate a "strong bind," while an affinity of 50-300 nM can indicate a "moderate bind." In NetMHCpan, for example, 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 % rank binding affinity. This value is not affected by the inherent bias of a particular molecule to the average predicted affinity level. For example (HLA-A * In the case of 0201, for example, very strong conjugates can be defined as having a % rank < 0.5, strong conjugates as having a % rank < 1.0, moderate conjugates as having a % rank of 1.0 to 2.0, and weak conjugates as having a % rank > 2.0. The methods for in vitro testing are well known to those skilled in the art. For example, those skilled in the art can refer to 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(1 2):341 1-21 HLA-A * The experimental protocol validated for the peptide presented by 0201 can be used. In this context, a reference peptide such as HIV pol 589-597 can be additionally used in the aforementioned test. This allows for the calculation of the in vitro affinity for the binding observed with the reference peptide, for example, by the following formula: relative affinity = HLA-A * Concentrations of each peptide that induce 20% of O201 expression / HLA-A * The concentration of the reference peptide that induces 20% of O201 expression (wherein 100% is the concentration of the reference peptide, e.g., HIV pol 589-597, detected when used at a concentration of, for example, 100 μM) *0201 (which is the level of expression). For example, a peptide showing a relative affinity of less than 1 can be regarded as a "strong binder", a peptide showing a relative affinity of 1 to 2 can be regarded as a "moderate binder", and a peptide showing a relative affinity exceeding 3 can be regarded as a "weak binder".

[0070] In humans, there are three different loci encoding MHC class I molecules (the human MHC molecules are also referred to as human leukocyte antigens (HLA)), namely, HLA-A, HLA-B, and HLA-C exist. HLA-A * 01, HLA-A * 02, HLA-A * 24, and HLA-B * 07 are examples of various MHC class I alleles that can be expressed from these loci. For example, the antigenic peptide according to the present invention can bind to HLA-A * 01 molecule, HLA-A * 02 molecule, HLA-A * 24 molecule, and HLA-B * 07 molecule. In particular, the antigenic peptide according to the present invention binds to HLA-A * 02.

[0071] In one embodiment, the antigenic peptide according to the present invention is a microbiota sequence variant of an epitope of a human tumor antigen.

[0072] In particular, the microbiota sequence variant of the epitope of the human tumor antigen is identified in at least one protein expressed in the human microbiota. Preferably, at least one protein present in the human microbiota is secreted or contains a transmembrane domain.

[0073] Cellular localization, particularly whether a protein is secreted or contains a transmembrane domain, can be tested in silico or in vitro by methods well known to those skilled in the art. For example, "SignalP 4.1 Server" (Biological Sequence Analysis Center, Technical University of Denmark (DTU); URL: www.cbs.dtu.dk / services / SignalP) and / or "Phobius" (Combination of transmembrane topology and signal peptide predictors, Stockholm Bioinformatics Centre; URL: phobius.sbc.su.se) can be used. Preferably, two predictive tools (e.g., SignalP 4.1 Server and Phobius) can be combined.

[0074] For example, to test whether a protein is secreted, the presence of a signal peptide may be evaluated. A signal peptide is a ubiquitous protein sorting signal that targets passenger (cargo) proteins for transmembrane cell membrane translocation in prokaryotes. To test for the presence of a signal peptide, for example, "SignalP 4.1 Server" (Biological Sequence Analysis Center, Technical University of Denmark (DTU); URL: www.cbs.dtu.dk / services / SignalP) and / or "Phobius" (Combination of transmembrane topology and signal peptide predictors, Stockholm Bioinformatics Centre; URL: phobius.sbc.su.se) can be used. Preferably, two predictive tools (e.g., SignalP 4.1 Server and Phobius) can be combined.

[0075] Furthermore, it may 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 4.1 Server and Phobius can distinguish signal peptides from transmembrane domains. Preferably, a minimum number of two predicted transmembrane helices is set to distinguish between membrane proteins and cytoplasmic proteins, and a final consensus list is obtained.

[0076] In one embodiment, the microbiota sequence variant of a human tumor antigen fragment / epitope is presented in its entirety after cleavage of a bacterial protein expressed in the human microbiota. In this context, the term “cleavage” means peptide processing for MHC presentation, particularly MHC-I processing. For example, cleavage prediction scores and / or affinity can be used to predict appropriate MHC binding antigen cleavage (for presentation to CD8 T cells). Such “cleavage probability scores” can be calculated using software (preferably higher than 70%, preferably higher than 80%, and more preferably higher than 90%). For example, the “IEDB” (Immune Epitope Database and Analysis Resource, IEDB Analysis Resource, supported by contract from the National Institute of Allergy and Infectious Diseases, part of the Department of Health and Human Services, U.S. National Institutes of Health) can be used, which provides MHC-I processing predictions (URL: http: / / tools.iedb.org / mhcnp / ). In another example, NetChop3 (The role of the proteasome in generating 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 with experimental data on in vitro proteasome cleavage and known HLA ligands. The output of the network was a score between 0.0 and 1.0. A score of 0.5 was given for random predictors, and a score of 1 was given for perfect predictors. Therefore, high scores for peptides (e.g., above 0.7, preferably above 0.8, more preferably above 0.9, as described above) suggest effective cleavage at their N-terminus and / or C-terminus (and not their center), while low scores are associated with peptides cleaved at their center. This allows for the prediction of peptide presentation by combining information on proteasome cleavage, TAP transport, and MHC class 1 analysis tools.

[0077] In one embodiment, the antigenic peptide induces T cell cross-reactivity to the human epitope of the (reference) tumor antigen. T cell cross-reactivity is an immune system phenomenon defined as the recognition of two or more peptide-MHC complexes (pMHCs) by the T cell receptor (TCR).

[0078] Epitope mimicry relates to the concept of sequence and structural similarity between foreign and self-antigens as an induction mechanism for triggering cross-reactive immune responses against self-antigens. Interestingly, such epitope mimicry offers a possible way to avoid the limitation of the human T cell repertoire caused by clonal removal of T cells that recognize self-antigens.

[0079] In particular, antigens that are different from autoantigens (e.g., human epitopes of tumor antigens) but share sequence similarity with said autoantigens (i.e., antigenic peptides according to the present invention) can (i) be recognized by cross-reactivity of T cell receptors, and (ii) such antigens are expected to be recognized by T cells / TCRs that are not eliminated during the T cell education process. Therefore, such antigens can induce a potent immune response that results in the clonal proliferation of T cells with potential cross-reactivity with autoantigens.

[0080] The T-cell receptor cross-reactivity of the human (reference) tumor antigen with the epitope can be measured by the ELISPOT-IFNγ assay, as shown in the Examples section. Briefly, 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 class II MHC, and / or HHD DR3 mice expressing human HLA-A2 and HLA-DR3 MHC) are immunized with a prime injection on day 0 (d0) and immunized with a boost injection of the antigenic peptide of the present invention and the epitope of the human (reference) tumor antigen on d14. Seven days after the boost injection (i.e., d21), the mice are euthanized, and splenocytes are stimulated in vitro with the antigenic peptide of the present invention to evaluate IFN-gamma secretion capacity by ELISPOT.

[0081] In particular, the present invention provides antigenic peptides which are microbiota sequence variants of human tumor antigen fragments / epitopes, the epitopes of human tumor antigens which may include or consist of any of SEQ ID NOs: 1-257 and 476-500.

[0082] Table 1A below outlines the antigenic peptides according to the present invention, along with their amino acid sequences, sequence numbers, and corresponding fragments / epitopes of human tumor antigens (also referred to herein as "human reference peptides"). Table 1A also provides information on which tumor antigen each antigenic peptide according to the present invention relates to. Sequence numbers 1-257 and 476-500 relate to HLA-A according to the present invention. * 02 shows antigenic peptide.

[0083] Table 1A. HLA-A according to the present invention * 02 Antigenic Peptide [Table 1A] TIFF2026136207000002.tif219170 TIFF2026136207000003.tif219170 TIFF2026136207000004.tif219170 TIFF2026136207000005.tif219170 TIFF2026136207000006.tif220170 TIFF2026136207000007.tif220170 TIFF2026136207000008.tif189170

[0084] Table 1B below outlines the consensus sequences (MHC class I consensus sequences) of the antigenic peptides according to the present invention, along with their amino acid sequences, sequence numbers, and corresponding fragments / epitopes of human tumor antigens (also referred to herein as "human reference peptides"), as well as their core sequences.

[0085] Table 1B. MHC class I consensus sequence of antigenic peptide according to the present invention. [Table 1B]

[0086] Table 1C below shows the HLA-A of the antigenic peptide according to the present invention. * Table 1C also shows an overview of the consensus sequences, along with their amino acid sequences, sequence numbers, and the corresponding core sequences of human tumor antigens. * This provides information regarding the 02 consensus sequence.

[0087] Therefore, the antigenic peptide may contain or consist of an amino acid sequence represented by any of SEQ ID NOs: 309 to 326. In some embodiments, the antigenic peptide may contain or consist of an amino acid sequence represented by SEQ ID NOs: 316, 304 to 315, 317 to 472, and 501 to 509, preferably SEQ ID NOs: 316, 304 to 315, and 317 to 326.

[0088] Table 1C. HLA-A of the antigenic peptide according to the present invention * 02 Consensus Array [Table 1C] TIFF2026136207000011.tif221170 TIFF2026136207000012.tif216170 TIFF2026136207000013.tif221170 TIFF2026136207000014.tif154170

[0089] As can be seen from Tables 1A to 1C, the antigenic peptides according to the present invention are each human tumor antigen, each core sequence, and each MHC class I and / or HLA-A * It can be classified according to the 02 consensus sequence.

[0090] Therefore, the antigenic peptide may contain a (core) sequence corresponding to any of SEQ ID NOs: 281 to 303. Preferably, the antigenic peptide contains a (core) sequence corresponding to any of SEQ ID NOs: 281, 283 to 289, 293, 294, 302, and 303. More preferably, the antigenic peptide contains a (core) sequence corresponding to any of SEQ ID NOs: 283, 287, 293, 294, 302, and 303. Even more preferably, the antigenic peptide contains a (core) sequence corresponding to any of SEQ ID NOs: 287, 293, 294, and 302.

[0091] In one embodiment, the antigenic peptide according to the present invention is a sequence variety of the tumor antigen CD19 (human reference peptide) fragment, 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 present invention is a sequence variety of the tumor antigen CD19 fragment, such as an antigenic peptide having a core sequence consisting of an amino acid sequence represented by any of SEQ ID NOs: 281 to 285. Therefore, the antigenic peptide according to the present invention is a sequence variety of the tumor antigen CD19 fragment containing or consisting of an MHC class I consensus sequence represented by any of SEQ ID NOs: 304 to 308. In a particular embodiment, the antigenic peptide according to the present invention is HLA-A2 represented by any of SEQ ID NOs: 327 to 346. * A specific HLA molecule (HLA-A2) such as an antigenic peptide containing or consisting of the 02 consensus sequence. *02) is a sequence variety of the tumor antigen CD19 fragment that binds to 02). More preferably, the antigenic peptide according to the present invention is a microbiota sequence variety of the tumor antigen CD19 fragment, such as an antigenic peptide containing or consisting of an amino acid sequence represented by any of SEQ ID NOs: 1 to 40. More preferably, the antigenic peptide according to the present invention is a sequence variety of the CD19 fragment (human reference peptide) "FLLFLTPME" (SEQ ID NO: 258), such as an antigenic peptide containing or consisting of an amino acid sequence represented by SEQ ID NOs: 1 to 12, 304, and 327 to 333, for example, an antigenic peptide containing or consisting of an amino acid sequence represented by SEQ ID NO: 10. The antigenic peptide according to the present invention is also more preferably a sequence variety of the CD19 fragment (human reference peptide) "KLSLGLPGL" (SEQ ID NO: 259), such as an antigenic peptide containing or consisting of an amino acid sequence represented by SEQ ID NOs: 13 to 33, 305, and 334 to 343. The antigenic peptide according to the present invention is more preferably a sequence variant of the CD19 fragment (human reference peptide) "SLVGILHLQ" (SEQ ID NO: 260), such as an antigenic peptide containing or consisting of the amino acid sequences represented by SEQ ID NOs: 34-35, 306, and 344, for example, an antigenic peptide containing or consisting of the amino acid sequence represented by SEQ ID NO: 34 or 35. The antigenic peptide according to the present invention is more preferably a sequence variant of the CD19 fragment (human reference peptide) "TLAYLIFCL" (SEQ ID NO: 261), such as an antigenic peptide containing or consisting of the amino acid sequences represented by SEQ ID NOs: 36-39, 307, and 345-346, for example, an antigenic peptide containing or consisting of the amino acid sequence represented by SEQ ID NO: 39. The antigenic peptide according to the present invention is more preferably a sequence variant of the CD19 fragment (human reference peptide) "QQMGGFYLC" (SEQ ID NO: 262), such as an antigenic peptide containing or consisting of the amino acid sequences represented by SEQ ID NOs: 40 and 308, for example, an antigenic peptide containing or consisting of the amino acid sequence represented by SEQ ID NO: 40.

[0092] In one embodiment, the antigenic peptide according to the present invention is a sequence variety 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 present invention is a sequence variety of the tumor antigen CD20, such as an antigenic peptide having a core sequence consisting of an amino acid sequence represented by any of SEQ ID NOs. 286 to 289. Therefore, the antigenic peptide according to the present invention is a sequence variety of the tumor antigen CD20, containing or consisting of an MHC class I consensus sequence represented by any of SEQ ID NOs. 309 to 312. In a particular embodiment, the antigenic peptide according to the present invention is an HLA-A2 represented by any of SEQ ID NOs. 347 to 374. * A specific HLA molecule (HLA-A2) such as an antigenic peptide containing or consisting of the 02 consensus sequence. *02) is a sequence variant of the tumor antigen CD20 fragment that binds to 02). More preferably, the antigenic peptide according to the present invention is a microbiota sequence variant of the tumor antigen CD20 fragment, such as an antigenic peptide containing or consisting of an amino acid sequence represented by any of SEQ ID NOs: 41 to 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 containing or consisting of an amino acid sequence represented by SEQ ID NOs: 41 to 64, 309, and 347 to 360, for example, an antigenic peptide containing or consisting of an amino acid sequence represented by SEQ ID NO: 61. The antigenic peptide according to the present invention is more preferably a sequence variant of the CD20 fragment (human reference peptide) "IMNSLSLFA" (SEQ ID NO: 264), such as an antigenic peptide containing or comprising the amino acid sequences represented by SEQ ID NOs. 65-70, 310, 361-364, and 476-484, for example, an antigenic peptide containing or comprising the amino acid sequence represented by any of SEQ ID NOs. 65, 68, 70, and 477. The antigenic peptide according to the present invention is also more preferably a sequence variant of the CD20 fragment (human reference peptide) "LMIPAGIYA" (SEQ ID NO: 265), such as an antigenic peptide containing or comprising the amino acid sequences represented by SEQ ID NOs. 71-80, 311, and 365-369, for example, an antigenic peptide containing or comprising the amino acid sequence represented by SEQ ID NO: 72. The antigenic peptide according to the present invention is more preferably a sequence variant of the D20 fragment (human reference peptide) "SLFLGILSV" (SEQ ID NO: 266), such as an antigenic peptide comprising or consisting of the amino acid sequences represented by SEQ ID NOs: 81-87, 312, and 370-374, for example, an antigenic peptide comprising or consisting of the amino acid sequence represented by SEQ ID NO: 86.

[0093] In one embodiment, the antigenic peptide according to the present invention is a sequence variety of the tumor antigen CD22 (human reference peptide) fragment, 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 present invention is a sequence variety of the tumor antigen CD22 fragment, such as an antigenic peptide having a core sequence consisting of an amino acid sequence represented by any of SEQ ID NOs: 290 to 293. Therefore, the antigenic peptide according to the present invention is a sequence variety of the tumor antigen CD22 fragment containing or consisting of an MHC class I consensus sequence represented by any of SEQ ID NOs: 313 to 316. In a particular embodiment, the antigenic peptide according to the present invention is HLA-A2 represented by any of SEQ ID NOs: 375 to 390. * A specific HLA molecule (HLA-A2) such as an antigenic peptide containing or consisting of the 02 consensus sequence. *02) is a sequence variety of the tumor antigen CD22 fragment that binds to 02). More preferably, the antigenic peptide according to the present invention is a microbiota sequence variety of the tumor antigen CD22 fragment, such as an antigenic peptide containing or consisting of an amino acid sequence represented by any of SEQ ID NOs: 88 to 110. More preferably, the antigenic peptide according to the present invention is a sequence variety of the CD22 fragment (human reference peptide) "FLSNDTVQL" (SEQ ID NO: 267), such as an antigenic peptide containing or consisting of an amino acid sequence represented by SEQ ID NOs: 88 to 96, 313, and 375 to 380. The antigenic peptide according to the present invention is also more preferably a sequence variety of the CD22 fragment (human reference peptide) "HLLGPWLLL" (SEQ ID NO: 268), such as an antigenic peptide containing or consisting of an amino acid sequence represented by SEQ ID NOs: 97 to 101, 314, and 381 to 383. The antigenic peptide according to the present invention is more preferably a sequence variant of the CD22 fragment (human reference peptide) "ILILAICGL" (SEQ ID NO: 269), such as an antigenic peptide containing or consisting of the amino acid sequences represented by SEQ ID NOs: 102-105, 315, and 384-386. The antigenic peptide according to the present invention is more preferably a sequence variant of the CD22 fragment (human reference peptide) "WVFEHPETL" (SEQ ID NO: 270), such as an antigenic peptide containing or consisting of the amino acid sequences represented by SEQ ID NOs: 106-110, 316, 387-390, and 485-488, for example, an antigenic peptide containing or consisting of the amino acid sequence represented by any of SEQ ID NOs: 107, 108, 109, and 110.

[0094] In one embodiment, the antigenic peptide according to the present invention is a sequence variety 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 present invention is a sequence variety of the tumor antigen CD37, such as an antigenic peptide having a core sequence consisting of an amino acid sequence represented by any of SEQ ID NOs: 294 to 299. Therefore, the antigenic peptide according to the present invention is a sequence variety of the tumor antigen CD37, including or consisting of an MHC class I consensus sequence represented by any of SEQ ID NOs: 317 to 322. In a particular embodiment, the antigenic peptide according to the present invention is an HLA-A2 represented by any of SEQ ID NOs: 391 to 425. * A specific HLA molecule (HLA-A2) such as an antigenic peptide containing or consisting of the 02 consensus sequence. *02) is a sequence variant of the tumor antigen CD37 fragment that binds to 02). More preferably, the antigenic peptide according to the present invention is a microbiota sequence variant of the tumor antigen CD37 fragment, such as an antigenic peptide containing or consisting of an amino acid sequence represented by any of SEQ ID NOs: 111 to 162. More preferably, the antigenic peptide according to the present invention is a sequence variant of the CD37 fragment (human reference peptide) "GLAFVPLQI" (SEQ ID NO: 271), such as an antigenic peptide containing or consisting of an amino acid sequence represented by SEQ ID NOs: 111 to 130, 317, 391 to 402, and 489 to 493, for example, an antigenic peptide containing or consisting of an amino acid sequence represented by any of SEQ ID NOs: 114, 117, 119, 120, 491, and 493. The antigenic peptide according to the present invention is more preferably a sequence variant of the CD37 fragment (human reference peptide) "GLYFGMLLL" (SEQ ID NO: 272), such as an antigenic peptide containing or consisting of the amino acid sequences represented by SEQ ID NOs: 131-136, 318, and 403-406. The antigenic peptide according to the present invention is more preferably a sequence variant of the CD37 fragment (human reference peptide) "ILILAICGL" (SEQ ID NO: 273), such as an antigenic peptide containing or consisting of the amino acid sequences represented by SEQ ID NOs: 137-147, 319, and 407-414. The antigenic peptide according to the present invention is more preferably a sequence variant of the CD37 fragment (human reference peptide) "LLLLFATQI" (SEQ ID NO: 274), such as an antigenic peptide containing or consisting of the amino acid sequences represented by SEQ ID NOs: 148-155, 320, and 415-420. The antigenic peptide according to the present invention is more preferably a sequence variant of the CD37 fragment (human reference peptide) "SIVGICLGV" (SEQ ID NO: 275), such as an antigenic peptide containing or consisting of the amino acid sequences represented by SEQ ID NOs: 156-157, 321, and 421. The antigenic peptide according to the present invention is also more preferably a sequence variant of the CD37 fragment (human reference peptide) "SLIKYFLFV" (SEQ ID NO: 276), such as an antigenic peptide containing or consisting of the amino acid sequences represented by SEQ ID NOs: 158-162, 322, and 422-425.

[0095] In one embodiment, the antigenic peptide according to the present invention is a sequence variety 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), or "PLPGLLFGA" (SEQ ID NO: 280). In a preferred embodiment, the antigenic peptide according to the present invention is a sequence variety of the tumor antigen TNFRSF13C, such as an antigenic peptide having a core sequence consisting of an amino acid sequence represented by any of SEQ ID NOs: 300 to 303. Therefore, the antigenic peptide according to the present invention is a sequence variety of the tumor antigen TNFRSF13C, which includes or consists of an MHC class I consensus sequence represented by any of SEQ ID NOs: 323 to 326. In a particular embodiment, the antigenic peptide according to the present invention is an HLA-A2 represented by any of SEQ ID NOs: 426 to 472. * 02 Consensus sequences and other specific HLA molecules (HLA-A2 *02) is a sequence variant of the tumor antigen TNFRSF13C fragment that binds to 02). More preferably, the antigenic peptide according to the present invention is a microbiota sequence variant of the tumor antigen TNFRSF13C fragment, such as an antigenic peptide containing or consisting of an amino acid sequence represented by any of SEQ ID NOs: 163 to 257. More preferably, 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 containing or consisting of an amino acid sequence represented by SEQ ID NOs: 163 to 170, 323, and 426 to 428. The antigenic peptide according to the present invention is also more preferably a sequence variant of the TNFRSF13C fragment (human reference peptide) "GLALVLALV" (SEQ ID NO: 278), such as an antigenic peptide containing or consisting of an amino acid sequence represented by SEQ ID NOs: 171 to 209, 324, and 429 to 446. The antigenic peptide according to the present invention is more preferably a sequence variant of the TNFRSF13C fragment (human reference peptide) "LLFGAPALL" (SEQ ID NO: 279), such as an antigenic peptide comprising or consisting of amino acid sequences represented by SEQ ID NOs: 210-225, 325, 447-455, and 494-500, for example, an antigenic peptide comprising or consisting of amino acid sequences represented by any of SEQ ID NOs: 212, 217, 220, and 224. The antigenic peptide according to the present invention is more preferably a sequence variant of the TNFRSF13C fragment (human reference peptide) "LLFGAPALL" (SEQ ID NO: 279), such as an antigenic peptide comprising or consisting of amino acid sequences represented by SEQ ID NOs: 226-257, 326, and 456-472, for example, an antigenic peptide comprising or consisting of amino acid sequences represented by SEQ ID NO: 227.

[0096] Preferably, the antigenic peptide according to the present invention comprises or consists of an MHC class I consensus sequence represented by any of SEQ ID NOs: 304-312, 315-316, and 325. More preferably, the antigenic peptide according to the present invention comprises an HLA-A sequence represented by any 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. * It includes or consists of the 02 consensus sequence.

[0097] In some embodiments, the antigenic peptide comprises or consists of an amino acid sequence represented by any of SEQ ID NOs: 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 an amino acid sequence represented by SEQ ID NOs: 34-35, 65-70, 476- The antigenic peptide comprises or consists of an amino acid sequence represented by any of 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 an amino acid sequence represented by any of SEQ ID NOs. 65-70, 476-484, 106-110, 485-488, 111-130, 489-493, 210-225, and 494-500.

[0098] In some embodiments, the antigenic peptide comprises or consists of an amino acid sequence represented by any 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 present invention comprises or consists of an amino acid sequence represented by any of SEQ ID NOs: 10, 21, 33, 35, 39, 40, 61, 65, 68, 72, 86, 110, 114, and 220. Even more preferably, the antigenic peptide according to the present invention comprises or consists of an amino acid sequence represented by any of SEQ ID NOs: 10, 21, 33, 35, 39, 40, 110, 114, and 220. Most preferably, the antigenic peptide according to the present invention comprises or consists of an amino acid sequence represented by any of SEQ ID NOs: 10, 110, 114, and 220. More preferably, the antigenic peptide according to the present invention comprises or consists of an amino acid sequence represented by any of SEQ ID NOs: 65, 110, 114, and 220. In some embodiments, the antigenic peptide comprises or consists of an amino acid sequence represented by SEQ ID NOs: 65. In some embodiments, the antigenic peptide comprises or consists of an amino acid sequence represented by SEQ ID NOs: 110. In some embodiments, the antigenic peptide comprises or consists of an amino acid sequence represented by SEQ ID NOs: 114. In some embodiments, the antigenic peptide comprises or consists of an amino acid sequence represented by SEQ ID NOs: 220.

[0099] As illustrated in the examples herein, certain antigenic peptides according to the present invention result in a potent improvement of the immune response to themselves, and most importantly, a potent improvement of the immune response to peptides contained in the tumor antigen that have amino acid similarity to those contained therein, even if the human reference peptide contained in the tumor antigen is tolerogenic.

[0100] Advantageously, the antigenic peptide according to the present invention may be in the form of an immunogenic compound for use in the prevention or treatment of B-cell malignancies.

[0101] Immunogenic compounds containing antigenic peptides according to the present invention In a further embodiment, the present invention also provides immunogenic compounds comprising the antigenic peptide according to the present invention as described above. In particular, preferred embodiments of the antigenic peptide described above also apply to immunogenic compounds according to the present invention. For example, the antigenic peptide contained in the immunogenic compound preferably comprises or comprises an MHC class I consensus sequence represented by any of SEQ ID NOs: 304-326, such as an antigenic peptide comprising or comprising an amino acid sequence represented by any of SEQ ID NOs: 1-247 and 476-500. For example, an antigenic peptide according to the present invention comprising or comprising an amino acid sequence represented by any of SEQ ID NOs: 10, 21, 33, 35, 39, 40, 61, 65, 68, 72, 86, 110, 114, and 220 is even more preferred. For example, an antigenic peptide according to the present invention comprising or comprising an amino acid sequence represented by any of SEQ ID NOs: 10, 21, 33, 35, 39, 40, 110, 114, and 220 is even more preferred. For example, antigenic peptides according to the present invention comprising or consisting of an amino acid sequence represented by any of SEQ ID NOs: 10, 110, 114, and 220 are more preferred. For example, antigenic peptides according to the present invention comprising or consisting of an amino acid sequence represented by any of SEQ ID NOs: 65, 110, 114, and 220 are even more preferred. Furthermore, a combination thereof, i.e., an immunogenic compound comprising different antigenic peptides according to the present invention, is preferred.

[0102] As used herein, the term “immunogenic compound” means a compound that, when administered to a mammal, in particular to a human organism, can induce, increase, prolong, or maintain an immune response, especially a compound that induces, increases, prolongs, or maintains an immune response.

[0103] Generally, the term "immunogenic compound" includes all types of compounds, including the antigenic peptide according to the present invention. For example, the antigenic peptide according to the present invention can be bound to a carrier molecule, or the antigenic peptide according to the present invention can be contained in a polypeptide or protein (these polypeptides or proteins may exist "separately," that is, not bound to any other compound, or the polypeptide or protein containing the antigenic peptide can be bound to a carrier molecule).

[0104] Preferably, the immunogenic compound according to the present invention comprises the antigenic peptide and a carrier molecule, in particular, the antigenic peptide (or polypeptide or protein containing the antigenic peptide) being bound to the carrier molecule. The preferred carrier molecule is a carrier protein or a carrier peptide. According to a preferred embodiment, the antigenic peptide as defined above, or the polypeptide / protein containing the antigenic peptide, is bound to the carrier protein or carrier peptide, for example, by covalent or non-covalent bonds. Alternatively, such carrier protein or carrier peptide as described herein may be co-administered (separately) in the form of an immunoadjuvant (i.e., not as an "immunogenic compound" but as a co-administration / combination therapy as described herein and below).

[0105] The carrier molecule may also be a lipid or lipid-like moiety. In this case, the immunogenic compound may be a lipopeptide. As used herein, the term “lipopeptide” means a molecule containing a lipid or lipid-like moiety covalently bonded to a peptide moiety. Generally, “lipids” are soluble in nonpolar solvents, but typically “lipids” are insoluble in (or not readily soluble in) water. Examples of lipids or lipid-like moieties include, but are not limited to, fatty acids, waxes, sterols, monoglycerides, diglycerides, triglycerides, and phospholipids. The lipid may be a fatty acid, glycerolipid, glycerophospholipid, sphingolipid, sterollipid, prenolipid, glycolipid, or polyketide. Preferably, the lipid is a fatty acid or a derivative thereof (such as monoglycerides, diglycerides, triglycerides, and phospholipids). Fatty acids typically contain a hydrocarbon chain ending in a carboxylic acid group. Fatty acids may be saturated or unsaturated. Fatty acids can be bonded to functional groups, such as oxygen, halogens, nitrogen, or sulfur. Preferred fatty acids include myristic acid (CH3(CH2)), which has 14 carbon atoms. 12 COOH) or palmitic acid (CH3(CH2) having 16 carbon atoms 14 These include saturated or unsaturated long-chain fatty acids such as COOH, and phospholipids such as phosphatidylglycerol (PG).

[0106] Preferably, the antigenic peptides described herein, or polypeptides / proteins containing such antigenic peptides, can be co-administered with proteins / peptides having immunoadjuvant properties (e.g., providing stimulation of CD4+ Th1 cells), or conjugated, for example, by covalent or non-covalent bonds. The antigenic peptides described herein preferably bind to MHC class I, but CD4+ helper epitopes can be further used to provide an efficient immune response. Th1 helper cells secrete interferon-gamma (IFN-γ), tumor necrosis factor alpha (TNF-α), and interleukin-2 (IL-2), thereby enhancing the expression of dendritic cell (DC) and T cell costimulatory signals, and thus maintaining efficient DC activation and specific CTL activation (Galaine et al., Interest of Tumor-Specific CD4 T Helper 1 Cells for Therapeutic Anticancer Vaccine. Vaccines (Basel). 2015 Jun 30;3(3):490-502).

[0107] For example, the adjuvant peptide / protein may preferably be a non-tumor antigen that evokes immunological memory or provides non-specific help, or a specific tumor-derived helper peptide. Several helper peptides for providing non-specific T cell help, such as tetanus helper peptide, keyhole limpet hemocyanin peptide, or PADRE peptide, have been documented in the literature (Adotevi et al., Targeting antitumor CD4 helper T cells with universal tumor-reactive helper peptides derived from telomerase for cancer vaccine. Hum Vaccin Immunother. 2013 May;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. 2011 Sep-Oct;17(5):343-50). Therefore, tetanus helper peptide, keyhole limpet hemocyanin peptide, and PADRE peptide are preferred examples of such adjuvant peptides / proteins. Furthermore, specific tumor-derived helper peptides are preferred. Specific tumor-derived helper peptides are typically represented by MHC class II, particularly HLA-DR, HLA-DP, or HLA-DQ. Specific tumor-derived helper peptides can be fragments of sequences of shared overexpressed tumor antigens such as HER2, NY-ESO-1, hTERT, or IL13RA2. Such fragments preferably have a length of 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. In particular, fragments of shared overexpressed tumor antigens such as HER2, NY-ESO-1, and hTERT having a length of 13 to 24 amino acids are preferred.Preferred fragments bind to MHC class II and can therefore be identified, for example, using the MHC class II binding prediction tool in the IEDB (Immune epitope database and analysis resource; contractually supported by the National Institute of Allergy and Infectious Diseases, part of the Department of Health and Human Services, U.S. National Institutes of Health; URL: http: / / www.iedb.org / ; http: / / tools.iedb.org / mhcii / ). Preferably, the adjuvant peptide / protein may be the HHD-DR3 peptide of 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, particularly helper peptides, include the UCP2 peptide (e.g., described in International Publication WO2013 / 135553A1 or Dosset et al., Clin Cancer Res. 2012 Nov 15;18(22):6284-95) and the BIRC5 peptide (e.g., described in European Patent Application Publication 2119726A1 or Widenmeyer et al., Int J Cancer. 2012 Jul 1;131(1):140-9). The most preferred helper peptide is the UCP2 peptide (amino acid sequence: KSVWSKLQSIGIRQH; SEQ ID NO: 475, e.g., described in International Publication WO2013 / 135553A1 or Dosset et al., Clin Cancer Res. 2012 Nov 15;18(22):6284-95).

[0108] The immunogenic compound according to the present invention is preferably a polypeptide or protein containing the antigenic peptide according to the present invention. Preferably, such protein or polypeptide is a recombinant protein or polypeptide, for example, a fusion protein. The term "recombinant" means that it does not exist in nature.

[0109] In preferred embodiments, the immunogenic compound according to the present invention comprises or consists of a polypeptide of formula (I). PepNt-CORE-PepCt (I) During the ceremony, - "PepNt" consists of polypeptides with a length of 0 to 500 amino acid residues, and is located at the N-terminus of the polypeptide of formula (I). - "CORE" consists of the antigenic peptide according to the present invention as defined above. - "PepCt" consists of a polypeptide with a length of 0 to 500 amino acid residues and is located at the C-terminus of the polypeptide of formula (I).

[0110] For example, the immunogenic compound may include or consist of a polypeptide of formula (Ia) or (Ib). PepNt-CORE (Ia); or CORE-PepCt(Ib) In the formula, "PepNt", "PepCt", and "CORE" are defined as above.

[0111] Preferably, the polypeptide of formula (I), (Ia), or (Ib) is a fusion peptide or fusion protein, in particular a recombinant fusion peptide or protein.

[0112] The polypeptides or immunogenic compounds defined above are also 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, It is preferable to include 9 to 1000 amino acids, such as 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" can be defined as needed.

[0113] Therefore, the "PepNt" and "PepCt" defined above are 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 It can contain 0 to 500 amino acid residues, such as 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.

[0114] The type of carrier molecule used to generate the immunogenic compound of the present invention, such as an immunogenic compound containing or consisting of a polypeptide of formula (I) bound to a carrier molecule, is within the realm of common technical knowledge for those skilled in the art. In particular, the function of the carrier molecule is to provide cytokine help (or T cell help) to enhance the immune response against tumor antigens.

[0115] Preferably, the antigenic peptide is bound to a carrier molecule, particularly a carrier protein, preferably by covalent or non-covalent bonds. The carrier molecule to which the peptide is optionally bound can be selected from a wide variety of known carriers. Examples of carrier molecules for vaccine purposes include human or bovine serum albumin, keyhole limpet hemocyanin (KLH), and proteins such as fatty acids. Other embodiments of the carrier molecule to which the antigenic peptide of formula (I) can be covalently bound include bacterial toxins or toxoids, such as diphtheria, cholera, E. coli thermal instability, or tetanus toxoids, meningococcal outer membrane protein (European Patent Application No. 0372501), synthetic peptides (European Patent Applications No. 0378881 and No. 0427347), heat shock proteins (International Publication No. WO93 / 17712), pertussis protein (International Publication No. WO98 / 58668), H. influenza-derived protein D (International Publication No. WO00 / 56360), and C. difficile-derived toxin A or B (International Publication No. WO00 / 61761).

[0116] More preferably, the carrier protein or carrier peptide is a protein / peptide having immunoadjuvant properties, such as providing stimulation of CD4+ Th1 cells as described herein. A preferred example is a non-tumor antigen that evokes immunological memory or provides nonspecific help, or may be a specific tumor-derived helper peptide such as tetanus helper peptide, keyhole limpet hemocyanin peptide, or PADRE peptide. Another preferred example is a specific tumor-derived helper peptide that can be presented by MHC II, particularly HLA-DR, HLA-DP, or HLA-DQ, such as a fragment of a shared overexpressed tumor antigen such as HER2, NY-ESO-1, or hTERT. In a preferred embodiment, the carrier protein or carrier peptide is a protein / peptide having immunoadjuvant properties and may be the HHD-DR3 carrier peptide MAKTIAYDEEARRGLERGLN (SEQ ID NO: 473). In particular, "PepNt" and / or "PepCt" may correspond to carrier proteins or carrier peptides 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, particularly helper peptides, include the UCP2 peptide (e.g., described in International Publication WO2013 / 135553A1 or Dosset et al., Clin Cancer Res. 2012 Nov 15;18(22):6284-95) and the BIRC5 peptide (e.g., described in European Patent Application Publication 2119726A1 or Widenmeyer et al., Int J Cancer. 2012 Jul 1;131(1):140-9).The most preferred helper peptide is the UCP2 peptide (amino acid sequence: KSVWSKLQSIGIRQH; SEQ ID NO: 475).

[0117] Furthermore, in polypeptides relating to formula (I), (Ia), or (Ib), "PepNt" and / or "PepCt" can preferably correspond to proteins / peptides having immunoadjuvant properties, such as providing stimulation of CD4+Th1 cells as described herein.

[0118] Furthermore, the immunogenic compound may include or consist of a protein / peptide having immunoadjuvant properties, such as providing stimulation of CD4+Th1 cells as described herein, which is covalently bonded to the N-terminus of the antigenic peptide according to the present invention or to the N-terminus of a polypeptide / protein containing the antigenic peptide.

[0119] Preferably, the antigenic peptide (or polypeptide / protein containing the antigenic peptide) according to the present invention is covalently bonded to the carrier molecule via a linker moiety.

[0120] Preferred linker agents include those named GMBS, sulfo-GMBS, SMPB, and sulfo-SMPB.

[0121] In some embodiments of the immunogenic compounds defined above, the linker agent is selected from the group consisting of GMBS (N-[γ-maleimidobutyryl-oxy]succinimide ester), sulfo-GMBS (N-[γ-maleimidobutyryl-oxy]sulfosuccinimide) ester), SMPB (succinimidyl 4-[p-maleimidophenyl]butyrate), and sulfo-SMPB (sulfosuccinimidyl 4-[p-maleimidophenyl]butyrate).

[0122] Methods for conjugating two proteins with a linker agent, more specifically, a linker agent selected from the group consisting of GMBS, sulfo-GMBS, SMPB, and sulfo-SMPB, are well known to those skilled in the art. For example, such protocols are disclosed in leaflets publicly available from the Pierce Company (Illinois, USA). GMBS, sulfo-GMBS, SMPB, and sulfo-SMPB consist of heterobifunctional linkers containing both an N-hydroxysuccinimide (NHS) ester group and a maleimide group. Conjugation using GMBS, sulfo-GMBS, SMPB, or sulfo-SMPB is typically carried out in a two-step procedure. In the first step, an amine-containing protein is reacted with the linker agent in several-fold molar excess at pH 7-9 to form an amide bond, after which the excess unreacted linker agent is usually removed by desalting or dialysis. In the second step, a sulfhydryl-containing molecule (for example, the peptide of formula (I)) is added and reacted with the maleimide group already bound to the first protein at pH 6.5-7.5 to form a stable thioether bond.

[0123] By using SMPB or sulfo-SMPB as a linker agent for covalently bonding the antigenic peptide (or polypeptide / protein containing the antigenic peptide, such as the polypeptide of formula (I)) according to the present invention to an amine-containing carrier protein, the following conjugate of formula (II) can be obtained. [ka] During the ceremony, -R1 consists of one reactive group of the amine-containing carrier protein, and the NH group bound thereto is derived from (i) an alpha-amino group located at the N-terminus of the amine-containing carrier protein or (ii) a side-chain amino group derived from a lysine (K) amino acid residue of the amine-containing carrier protein; and -R2 consists of an antigenic peptide according to the present invention (or a polypeptide / protein containing the antigenic peptide, such as the polypeptide of formula (I)), and the sulfur (S) atom bound thereto originates from the sulfhydryl (SH) group of a cysteine ​​residue located at the N-terminus or C-terminus of the peptide of formula (I). In some embodiments, the sulfhydryl moiety can be a non-natural amino acid or part of any other molecule present at the terminus of the peptide of formula (I).

[0124] By using GMBS or sulfo-GMBS as a linker agent for covalently bonding the antigenic peptide (or polypeptide / protein containing the antigenic peptide, such as the polypeptide of formula (I)) according to the present invention to an amine-containing carrier protein, particularly a CRM197 carrier protein, the following conjugate of formula (III) can be obtained. [ka] During the ceremony, -R1 consists of one reactive group of the amine-containing carrier protein, and the NH group bound thereto is derived from (i) an alpha-amino group located at the N-terminus of the amine-containing carrier protein or (ii) a side-chain amino group derived from a lysine (K) amino acid residue of the amine-containing carrier protein; and -R2 consists of an antigenic peptide according to the present invention (or a polypeptide / protein containing the antigenic peptide, such as the polypeptide of formula (I)), and the sulfur (S) atom bound thereto originates from the sulfhydryl (SH) group of a cysteine ​​residue located at the N-terminus or C-terminus of the peptide of formula (I). In some embodiments, the sulfhydryl moiety can be a non-natural amino acid or part of any other molecule present at the terminus of the peptide of formula (I).

[0125] Peptide-MHC (pMHC) multimers containing antigenic peptides In a further embodiment, the present invention also provides a peptide-MHC (pMHC) multimer comprising an antigenic peptide according to the present invention.

[0126] 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, an “MHC multimer” is an oligomeric form of an MHC molecule. The primary function of an MHC molecule is to bind to an antigen. According to the present invention, the antigen is the antigenic peptide according to the present invention. Therefore, a complex of MHC proteins “loaded” with the antigenic peptide of the present invention usually means that the antigenic peptide of the present invention is bound to one or more MHC proteins. Examples of “peptide-MHC multimers” (pMHC) of the present invention include, but are 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 general term meaning that encompasses the histocompatibility antigen system described in various species, including human leukocyte antigens (HLA). In humans, there are three major distinct gene loci that encode MHC class I molecules, namely HLA-A, HLA-B, and HLA-C. * 01, HLA-A * 02, and HLA-A * 11 are examples of various MHC class I alleles that can be expressed from these loci.

[0127] In one embodiment of the present invention, the pMHC multimer is a peptide / MHC class I multimer. In another specific embodiment, the pMHC multimer is an HLA corresponding to an MHC class I / peptide multimer. Thus, the pMHC multimer can be an HLA-peptide multimer selected from the group consisting of HLA-A-peptide multimer, HLA-B-peptide multimer, HLA-C-peptide multimer, HLA-E-peptide multimer, MICA-peptide multimer, and MICB-peptide multimer.

[0128] Methods for obtaining pHMC multimers are known in the art and are described, for example, in International Publications WO96 / 26962 and 01 / 18053, which are incorporated herein by reference.

[0129] In addition to the MHC molecule and the antigenic peptide of the present invention, the pMHC may include further components such as a multimerizing agent and / or a label (for example, for visualization). Examples of labels include, but are not limited to, fluorescent labels such as fluorescently labeled proteins such as streptavidin. Examples of fluorescent labels include allophycocyanin (APC), phycoerythrin (PE), R-phycoerythrin (R-PE), and fluorescein isothiocyanate (FITC). A preferred label is biotin.

[0130] In one embodiment of the present invention, the pMHC multimer can be used to visualize T cell populations that are specific to HLA corresponding to the MHC class I peptide complex or MHC class I / peptide complex described herein. For example, the pMHC multimer may be a multimer in which the heavy chain of the MHC is biotinylated, which allows for combination as a tetramer with streptavidin. Such a pMHC tetramer can be used to visualize reactive populations by immunofluorescence because it exhibits high avidity to appropriate TCR-carrying T lymphocytes. In another embodiment of the present invention, the pMHC multimer can be used for detection and / or isolation of T cell populations specific to the pMHC complex described herein by screening (flow cytometry or immunomagnetic screening).

[0131] Antigenic peptide-specific cytotoxic T lymphocytes (CTL) In a further embodiment, the present invention also provides antigenic peptide-specific cytotoxic T lymphocytes (CTLs) according to the present invention, and in particular antigenic peptide-specific activated cytotoxic T lymphocytes (CTLs) according to the present invention.

[0132] The present invention further provides a method for producing antigenic peptide-specific cytotoxic T lymphocytes (CTLs), particularly activated antigenic peptide-specific cytotoxic T lymphocytes (CTLs), 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 the antigenic peptide according to the present 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 present invention. The step of contacting the CTLs with an antigen-loaded human class I or II MHC molecule expressed on the surface of the antigen-presenting cell or the artificial construct mimicking the antigen-presenting cell can be performed for a period of time sufficient to specifically activate the CTLs. Preferably, the antigenic peptide is one of the preferred antigenic peptides described above, such as an antigenic peptide comprising or consisting of an amino acid sequence represented by any of SEQ ID NOs. 65, 110, 114, and 220.

[0133] T cells directed towards the antigenic peptide of the present invention (activated) are useful in therapy. In particular, activated T cells produced by the above method selectively recognize cells that abnormally express polypeptides containing the amino acid sequence of SEQ ID NOs. 258-280 (i.e., tumor antigens), such as polypeptides containing the amino acid sequence represented by any of SEQ ID NOs. 264, 270, 271, and 279.

[0134] Preferably, the (activated) cytotoxic T lymphocytes (CTLs) according to the present invention, which are specific to the antigenic peptide of the present invention, may have (present / express) memory markers. Such memory markers are preferably memory markers for intestinal memory cells such as CCR9, CXCR3, CD103, CX3CR1, and α4β7+.

[0135] The (activated) cytotoxic T lymphocytes (CTLs) according to the present invention, which are specific to the antigenic peptide of the present invention, preferably proliferate more in number and more strongly after vaccination with the antigenic peptide of the present invention (derived from a human microbiota sequence) compared to vaccination with peptides not derived from the microbiota sequence, such as the human (reference) sequence and / or synthetic peptides (e.g., including mutations (e.g., artificially introduced mutations)). In other words, vaccination of a target with the antigenic peptide of the present invention preferably increases the number of (activated) cytotoxic T lymphocytes (CTLs) specific to the antigenic peptide of the present invention compared to vaccination with each human peptide or synthetic peptide (not derived from the microbiota) relating to the same reference epitope.

[0136] The (activated) cytotoxic T lymphocytes (CTLs) according to the present invention, which are specific to the antigenic peptide of the present invention, preferably proliferate more / more strongly and / or faster after vaccination in subjects having the peptide in the gut (expressed by the subject's microbiota) (e.g., the peptide can be detected in a stool sample of the subject) compared to subjects not having the peptide in the gut (not expressed by the subject's microbiota) (e.g., subjects for which the peptide is not detected in a stool sample). In particular, subjects having the peptide in the gut (expressed by the subject's microbiota) can respond more quickly (faster T cell proliferation) and / or have T cells derived from the desired type of Tc1.

[0137] Cells loaded with antigenic peptides or immunogenic compounds In a further embodiment, the present invention also provides cells loaded with an antigenic peptide according to the present invention, or an immunogenic compound containing the antigenic peptide according to the present invention as described above. In particular, preferred embodiments of the antigenic peptide described above also apply to cells according to the present invention. For example, the antigenic peptide loaded into the cells, or contained in the immunogenic compound loaded into the cells, preferably comprises or consists of an MHC class I consensus sequence represented by any of SEQ ID NOs: 304-326, such as an antigenic peptide comprising or consisting of an amino acid sequence represented by any of SEQ ID NOs: 1-247 and 476-500. For example, an antigenic peptide according to the present invention comprising or consisting of an amino acid sequence represented by any of SEQ ID NOs: 10, 21, 33, 35, 39, 40, 61, 65, 68, 72, 86, 110, 114, and 220 is even more preferred. For example, an antigenic peptide according to the present invention comprising or consisting of an amino acid sequence represented by any of SEQ ID NOs: 10, 21, 33, 35, 39, 40, 110, 114, and 220 is even more preferred. For example, antigenic peptides according to the present invention that include or consist of an amino acid sequence represented by any of SEQ ID NOs: 10, 110, 114, and 220 are more preferred. For example, antigenic peptides according to the present invention that include or consist of an amino acid sequence represented by any of SEQ ID NOs: 65, 110, 114, and 220 are even more preferred. Furthermore, a combination thereof, i.e., cells loaded with different antigenic peptides (or their respective immunogenic compounds) according to the present invention, is preferred.

[0138] The preferred cells to which the antigenic peptide or immunogenic compound according to the present invention is loaded are antigen-presenting cells (APCs), more preferably dendritic cells (DCs).

[0139] The primary function of APCs is to process antigens and present them on the cell surface to T cells of the immune system, thereby initiating and regulating the in vivo T cell response, which is of particular interest. In the context of the present invention, it is preferable that APCs are loaded with the antigenic peptide and / or immunogenic compound according to the present invention. This can be done by exposing APCs to the antigenic peptide and / or immunogenic compound in vitro (see 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 immunotolerance of tumors: a new perspective for dendritic cell therapy. J Immunotoxicol. 2014 Oct;11(4):311-8).

[0140] The preferred APC according to the present invention is dendritic cells (DCs). In fact, it may be advantageous to combine at least one antigenic peptide or immunogenic compound according to the present invention with DCs, because they are the most potent APCs and have often been reported to be functionally deficient in cancer patients. DCs can be readily obtained by technicians in the art from healthy, compatible donors (i.e., the DCs are HLA-related) or from the patient themselves if they are functional (i.e., the DCs are autologous), for example, by direct isolation from peripheral blood or by induction from peripheral blood cells such as CD14+ monocytes or CD34+ hematopoietic progenitor cells (Figdor CG, de Vries IJ, Lesterhuis WJ, Melief CJ. Dendritic cell immunotherapy: mapping the way. Nat Med. 2004 May;10(5):475-80). In fact, DCs can be distinguished from other cells in peripheral blood by surface markers such as S100, p55, CD83, and / or OX62, and can therefore be isolated and purified based on these markers using cell culture techniques well known in the art.

[0141] Host cells containing nucleic acids and nucleic acids that encode antigenic peptides In further embodiments, the present invention also provides nucleic acids encoding antigenic peptides according to the present invention, polypeptides of formula (I) as defined above, or immunogenic compounds according to the present invention, which are peptides or proteins. In particular, preferred embodiments of the antigenic peptides described above also apply to nucleic acids according to the present invention. For example, antigenic peptides encoded by nucleic acids preferably consist of an MHC class I consensus sequence represented by any of SEQ ID NOs: 304-326, such as antigenic peptides consisting of an amino acid sequence represented by any of SEQ ID NOs: 1-247 and 476-500. For example, antigenic peptides according to the present invention consisting of an amino acid sequence represented by any of SEQ ID NOs: 10, 21, 33, 35, 39, 40, 61, 65, 68, 72, 86, 110, 114, and 220 are even more preferred. For example, antigenic peptides according to the present invention consisting of an amino acid sequence represented by any of SEQ ID NOs: 10, 21, 33, 35, 39, 40, 110, 114, and 220 are even more preferred. For example, antigenic peptides according to the present invention comprising or consisting of an amino acid sequence represented by any of SEQ ID NOs: 10, 110, 114, and 220 are more preferred. For example, antigenic peptides according to the present invention comprising or consisting of an amino acid sequence represented by any of SEQ ID NOs: 65, 110, 114, and 220 are even more preferred. Furthermore, a combination thereof, i.e., nucleic acids encoding different antigenic peptides according to the present invention, is preferred.

[0142] The nucleic acid preferably comprises single-stranded, double-stranded, or partially double-stranded nucleic acids, and is 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 recombinant polynucleotides, vectors, oligonucleotides, RNA molecules such as rRNA, mRNA, or tRNA, or the DNA molecules described above. Therefore, the nucleic acid (molecule) is preferably 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.

[0143] In the fields of therapeutics, diagnostics, reagents, and biological assays, the ability to deliver nucleic acids (e.g., ribonucleic acid (RNA)) into cells, whether in vitro, in vivo, in situ, or ex vivo, to induce intracellular translation of the nucleic acid and production of the encoded peptide of interest, for example, is of great interest. Of particular importance is the delivery and function of unintegrated polynucleotides. Therefore, it is preferable that nucleic acids such as mRNA are not integrated into the host chromosome. Generally, nucleic acids such as mRNA can be optimized for the expression of the antigenic peptide of the present invention by methods known in the art, such as codon optimization. Furthermore, the nucleic acid can be modified, for example, to increase its stability, extend its lifespan, and / or improve the expression of the antigenic peptide of the present invention. Therefore, optimized or modified mRNA (mmRNA) encoding the antigenic peptide according to the present invention is preferred. The mRNAs described herein are distinguished from wild-type mRNA in their functional and / or structural design features for optimal mRNA delivery and / or optimal expression of the antigenic peptide of the present invention (see, for example, International Publications WO2013 / 151672A2, WO2013 / 101690A1, and WO2013 / 052523A, which are incorporated herein by reference). Generally, nucleic acids are delivered "naked" or can be associated with a carrier, such as a cationic carrier. Cationic carriers (positively charged) usually readily associate with negatively charged nucleic acids. The carrier can be any type, including, for example, polymers, proteins, lipids, and nanoparticles. Cationic lipids and nanoparticles (in particular lipid nanoparticles, LNPs) are preferred for nucleic acid delivery. Accordingly, the present invention also provides nucleic acids described herein that associate with a carrier (e.g., lipids, in particular cationic lipids or LNPs).

[0144] In some embodiments, the nucleic acid molecule can be a vector. As used in the context of this invention, the term “vector” means a nucleic acid molecule, preferably an artificial nucleic acid molecule, i.e., a nucleic acid molecule that does not exist in nature. A vector in the context of this invention is suitable for incorporating or accommodating a desired nucleic acid sequence. Such a vector may be a conserved vector, an expression vector, a cloning vector, a transfer vector, and the like. A conserved vector is a vector that allows for the convenient storage of a nucleic acid molecule. Therefore, such a vector may contain a sequence corresponding to, for example, a desired antigenic peptide according to the present invention. An expression vector can be used to produce expression products such as RNA, such as mRNA, or peptides, polypeptides, or proteins. For example, an expression vector may contain a sequence necessary for the transcription of a sequence of the vector, such as a promoter sequence. A cloning vector is a vector that typically contains a cloning site that can be used to incorporate a nucleic acid sequence into the vector. A cloning vector may be, for example, a plasmid vector or a bacteriophage vector. A transfer vector may be a vector suitable for transferring a nucleic acid molecule to a cell or organism, such as a viral vector. A vector in the context of this invention may be, for example, an RNA vector or a DNA vector. Preferably, the vector is a DNA molecule. For example, a vector in the sense of the present application includes a sequence suitable for amplification of the vector, such as a cloning site, a selection marker such as an antibiotic resistance factor, and an origin of replication. Preferably, a vector in the context of the present application is a plasmid vector. Preferably, a vector in the context of the present application is an expression vector. A preferred vector is a vector for expression in bacterial cells. More preferably, the vector is useful for expression in a so-called "live bacterial vaccine vector," in which live bacterial cells (bacteria or bacterial spores, e.g., endospores, exospores, or microbial cysts) can function as a vaccine.A favorable example is described in da Silva et al., Live bacterial vaccine vectors: an overview; Braz J Microbiol. 2015 Mar 4;45(4):1117-29.

[0145] The nucleic acid encoding the antigenic peptide according to the present invention may be in the form of a naked nucleic acid or a nucleic acid cloned into a plasmid or viral vector (Tregoning and Kinnear, Using Plasmids as DNA Vaccines for Infectious Diseases. Microbiol Spectr. 2014 Dec;2(6). doi: 10.1128 / microbiolspec.PLAS-0028-2014), the latter being particularly preferred. Suitable viral vectors according to the present invention include, but 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 skill of those skilled in the art.

[0146] In a further embodiment, the present invention also provides host cells comprising nucleic acids according to the present invention. Preferably, a combination thereof, i.e., host cells comprising different nucleic acids according to the present invention encoding different antigenic peptides according to the present invention, is also preferred.

[0147] Preferably, the nucleic acid contained in the host cell is preferably a vector. Preferably, the host cell is a bacterial cell. Such a host cell can preferably be used for the production of an antigenic peptide or an immunogenic compound according to the present invention. Furthermore, such a host cell can also be the active ingredient of a vaccine.

[0148] Preferably, the host cell is a bacterial cell, and more preferably, an intestinal bacterial cell. The term "intestinal bacterial cell" refers to bacteria present in the (human) intestines.

[0149] Such bacterial host cells can function as “live bacterial vaccine vectors,” and live bacterial cells (bacteria or bacterial spores, e.g., endospores, exospores, or microbial cysts) can function as vaccines. 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.

[0150] Bacterial cells (bacteria or bacterial spores, e.g., endospores, exospores, or microbial cysts), particularly (whole) intestinal bacterial species, may be advantageous because they have the potential to elicit a higher immune response than the (poly)peptides or nucleic acids they contain.

[0151] Alternatively, the bacterial cells according to the present invention, particularly intestinal bacteria, can be in the form of probiotics, i.e., live intestinal bacteria, and therefore can be used as a food additive due to the health benefits they can provide. They can be freeze-dried, for example, as granules, pills, or capsules, or directly mixed with dairy products for consumption.

[0152] Nanoparticles containing antigenic peptides or immunogenic compounds In a further embodiment, the present invention also provides nanoparticles comprising the following, in particular nanoparticles loaded with the following: - At least one antigenic peptide according to the present invention, or - At least one immunogenic compound according to the present invention; and Optional adjuvant.

[0153] In particular, the preferred embodiments of the antigenic peptide described above also apply to nanoparticles according to the present invention. For example, the antigenic peptide loaded onto the nanoparticles, or the antigenic peptide contained in the immunogenic compound loaded onto the nanoparticles, preferably contains or comprises an MHC class I consensus sequence represented by any of SEQ ID NOs: 304 to 326, such as an antigenic peptide containing or comprising an amino acid sequence represented by any of SEQ ID NOs: 1 to 247 and 476 to 500. For example, an antigenic peptide according to the present invention containing or comprising an amino acid sequence represented by any of SEQ ID NOs: 10, 21, 33, 35, 39, 40, 61, 65, 68, 72, 86, 110, 114, and 220 is even more preferred. For example, an antigenic peptide according to the present invention containing or comprising an amino acid sequence represented by any of SEQ ID NOs: 10, 21, 33, 35, 39, 40, 110, 114, and 220 is even more preferred. For example, an antigenic peptide according to the present invention containing or comprising an amino acid sequence represented by any of SEQ ID NOs: 10, 110, 114, and 220 is even more preferred. For example, antigenic peptides according to the present invention comprising or consisting of an amino acid sequence represented by any of SEQ ID NOs. 65, 110, 114, and 220 are more preferred. Furthermore, a combination thereof, i.e., nanoparticles loaded with different antigenic peptides (or their respective immunogenic compounds) according to the present invention, is preferred.

[0154] Nanoparticles, particularly those for use as vaccines, are known in the art and are 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. In particular, the nanoparticles are used for the delivery of the antigenic peptide (or the immunogenic compound / polypeptide / protein / nucleic acid containing the antigenic peptide) and can optionally act as an adjuvant. The antigenic peptide (the immunogenic compound / polypeptide / protein / nucleic acid containing the antigenic peptide) is typically encapsulated within the nanoparticle or linked / bonded (coated) to the surface of the nanoparticle ("coating"). Compared to conventional approaches, nanoparticles can protect the payload (antigen / adjuvant) from the surrounding biological environment, extend its half-life, minimize systemic toxicity, facilitate delivery to APCs, and even directly induce the activation of TAA-specific T cells. Preferably, the nanoparticles have a size (diameter) of 300 nm or less, more preferably 200 nm or less, and most preferably 100 nm or less. Such nanoparticles are adequately protected from uptake by phagocytic cells, exhibit high structural integrity in circulation, have a long circulation time, can accumulate in tumor growth sites, and can penetrate deep into tumor masses.

[0155] Examples of nanoparticles include polymer nanoparticles such as poly(ethylene glycol) (PEG) and poly(D,L-lactic acid-coglycolic acid) (PLGA); inorganic nanoparticles such as gold nanoparticles, iron oxide beads, iron oxide zinc nanoparticles, carbon nanotubes, and mesoporous silica nanoparticles; liposomes such as cationic liposomes; immunostimulatory complexes (ISCOM); virus-like particles (VLP); and self-assembling proteins.

[0156] Polymer nanoparticles are nanoparticles based on / containing polymers such as poly(D,L-lactide-coglycolide) (PLG), poly(D,L-lactic acid-coglycolic acid) (PLGA), poly(γ-glutamic acid) (γ-PGA), poly(ethylene glycol) (PEG), and polystyrene. Polymer nanoparticles can capture antigens (e.g., the antigenic peptide or a (poly)peptide containing the antigenic peptide) or conjugate / conjugate to antigens (e.g., the antigenic peptide or a (poly)peptide containing the antigenic peptide). Because polymer nanoparticles have a slow biodegradation rate, they can be used, for example, for delivery to specific cells or for sustained-release antigen release. For example, γ-PGA nanoparticles can be used to encapsulate hydrophobic antigens. Polystyrene nanoparticles can be surface-modified with various functional groups and can therefore be conjugated to various antigens. Using polymers such as poly(L-lactic acid) (PLA), PLGA, PEG, and natural polymers such as polysaccharides, hydrogel nanoparticles, a type of nano-sized hydrophilic three-dimensional polymer network, can be synthesized. Nanogels possess desirable properties such as a flexible mesh size, a large surface area for multivalent conjugation, high water content, and high antigen loading capacity. Therefore, preferred nanoparticles are nanogels such as chitosan nanogels. Preferred polymer nanoparticles are nanoparticles based on / containing PEG and PLGA.

[0157] Inorganic nanoparticles are nanoparticles based on / containing inorganic substances, and examples of such nanoparticles include gold nanoparticles, iron oxide beads, iron oxide zinc nanoparticles, carbon nanoparticles (e.g., carbon nanotubes), and mesoporous silica nanoparticles. Inorganic nanoparticles offer a robust structure and controllable synthesis. For example, gold nanoparticles can be easily manufactured in various shapes, such as spherical, rod-shaped, and cubic. Inorganic nanoparticles may be surface-modified with, for example, carbohydrates. Carbon nanoparticles offer good biocompatibility and can be manufactured, for example, as nanotubes or (mesoporous) spheres. For example, multiple copies of the antigenic peptide (or (poly)peptide) according to the present invention can be conjugated onto carbon nanoparticles such as 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 functionalities such as cell recognition, absorption of specific biomolecules, improved cell interaction, and enhanced cellular uptake. Mesoporous silica nanoparticles are particularly preferred.

[0158] Liposomes are typically formed from phospholipids such as 1,2-dioleoyl-3-trimethylammonium propane (DOTAP). Cationic liposomes are generally preferred. Liposomes self-assemble with a phospholipid bilayer shell and an aqueous core. Liposomes can be produced as unilamellar vesicles (having a single phospholipid bilayer) or multilamellar vesicles (having several concentric phospholipid shells separated by a layer of water). Thus, antigens can be encapsulated within the core or between different layers / shells. Preferred liposome systems are those approved for human use, such as Inflexal® V and Epaxal®.

[0159] Immunostimulatory complexes (ISCOMs) are cage-like particles with a diameter of approximately 40 nm, colloidal saponin-containing micelles, and are formed from, for example, the saponin adjuvant Quil-A, cholesterol, phospholipids, and the (poly)peptide antigen (e.g., the antigenic peptide or polypeptide containing it). These spherical particles can capture the antigen through nonpolar interactions. Two types of ISCOMs have been reported, both consisting of cholesterol, phospholipids (usually phosphatidylethanolamine or phosphatidylcholine), and saponins (such as Quil-A).

[0160] Virus-like particles (VLPs) are self-assembling nanoparticles formed by the self-assembly of biocompatible capsid proteins. Due to their naturally optimized nanoparticle size and repeating structural order, VLPs can induce a potent immune response. VLPs can originate from a variety of viruses, ranging in size from 20 nm to 800 nm, typically from 20 nm to 150 nm. VLPs can be manipulated to express additional peptides or proteins by fusing these peptides / proteins with the particle or by expressing multiple antigens. Furthermore, bioconjugated VLPs can be generated by chemically conjugating antigens to the viral surface.

[0161] Examples of self-assembling proteins include ferritin and major vault proteins (MVPs). Ferritin is a protein that can self-assemble into a nearly spherical 10 nm structure. 96 units of MVP can self-assemble into barrel-shaped vault nanoparticles approximately 40 nm wide and 70 nm long. Antigens genetically fused with minimal interaction domains can be packaged within the vault nanoparticles by a self-assembly process when mixed with MVPs. Thus, the antigen (such as the antigenic peptide or polypeptide containing the same according to the present invention) can be fused to a self-assembling protein or a fragment / domain thereof (such as the minimal interaction domain of an MVP). Therefore, the present invention also provides a fusion protein comprising a self-assembling protein (or a fragment / domain thereof) and an antigenic peptide according to the present invention.

[0162] Generally, preferred examples of nanoparticles (NPs) include iron oxide beads, polystyrene microspheres, poly(γ-glutamic acid) (γ-PGA) NPs, iron oxide-zinc oxide NPs, cationized gelatin NPs, Pluronic®-stabilized poly(propylene sulfide) (PPS) NPs, PLGA NPs, (cationic) liposomes, (pH-responsive) polymer micelles, PLGA, cancer cell membrane coated PLGA, lipid-calcium phosphate (LCP) NPs, liposome-protamine-hyaluronic acid (LPH) NPs, polystyrene latex beads, magnetic beads, iron dextran particles, and quantum dot nanocrystals.

[0163] Preferably, the nanoparticles further comprise an adjuvant, such as a Toll-like receptor (TLR) agonist. Thereafter, the antigenic peptide (the immunogenic compound / polypeptide / protein / nucleic acid comprising the antigenic peptide) can be delivered together with the adjuvant to antigen-presenting cells (APCs), such as dendritic cells (DCs). The adjuvant may be encapsulated by the nanoparticles or conjugated to the surface of the nanoparticles, preferably in the same way as the antigenic peptide.

[0164] Particularly preferred adjuvants are polyinosinic acid:polycytidylic acid (also known as "Poly-I:C") and / or its derivative, poly-ICLC. Poly-I:C is a mismatched double-stranded RNA, where one strand is a polymer of inosinic acid and the other is a polymer of cytidylic acid. Poly-I:C is an immunostimulant known to interact with Toll-like receptor 3 (TLR3). Poly-I:C is structurally similar to double-stranded RNA, the "natural" stimulant of TLR3. Therefore, poly-I:C can be considered a synthetic analogue of double-stranded RNA. Poly-ICLC is a synthetic complex of carboxymethylcellulose, polyinosinic acid-polycytidylic acid, and poly-L-lysine double-stranded RNA. Like poly-I:C, poly-ICLC is also a ligand for TLR3. Poly-I:C and poly-ICLC typically stimulate the release of cytotoxic cytokines. A preferred example of poly-ICLC is Hiltonol®.

[0165] Pharmaceutical composition In a further embodiment, the present invention also provides a pharmaceutical composition comprising at least one of the following: - Antigenic peptides according to the present invention as described herein, - Immunogenic compounds according to the present invention as described herein, -Naparticles according to the present invention as described herein, - Cells according to the present invention as described herein, - Nucleic acids according to the present invention as described herein, -Host cells according to the present invention as described herein, and / or -The cytotoxic T lymphocytes according to the present invention as described herein, and Optionally, one or more pharmaceutically acceptable excipients or carriers.

[0166] Therefore, the present invention provides a pharmaceutical composition comprising (at least) one antigenic peptide according to the present invention as described herein. Furthermore, the present invention also provides a pharmaceutical composition comprising (at least) one immunogenic compound according to the present invention as described herein. Furthermore, the present invention also provides a pharmaceutical composition comprising (at least) one nanoparticle according to the present invention as described herein. Furthermore, the present invention also provides a pharmaceutical composition comprising (at least) one cell according to the present invention as described herein. Furthermore, the present invention also provides a pharmaceutical composition comprising (at least) one nucleic acid according to the present invention as described herein. Furthermore, the present invention also provides a pharmaceutical composition comprising (at least) one host cell according to the present invention as described herein. Furthermore, the present invention also provides a pharmaceutical composition comprising (at least) one cytotoxic T lymphocyte according to the present invention as described herein.

[0167] In particular, the preferred embodiments of the antigenic peptide described above also apply to the pharmaceutical composition according to the present invention. For example, the antigenic peptide contained in the pharmaceutical composition, or the antigenic peptide contained in any of the immunogenic compound, nanoparticles, cells, nucleic acids, or host cells contained in the pharmaceutical composition, preferably comprises or consists of an amino acid sequence represented by any of SEQ ID NOs: 1 to 247 and 476 to 500. For example, an antigenic peptide according to the present invention comprising or consisting of an amino acid sequence represented by any of SEQ ID NOs: 10, 21, 33, 35, 39, 40, 61, 65, 68, 72, 86, 110, 114, and 220 is even more preferred. For example, an antigenic peptide according to the present invention comprising or consisting of an amino acid sequence represented by any of SEQ ID NOs: 10, 21, 33, 35, 39, 40, 110, 114, and 220 is even more preferred. For example, an antigenic peptide according to the present invention comprising or consisting of an amino acid sequence represented by any of SEQ ID NOs: 10, 110, 114, and 220 is even more preferred. For example, an antigenic peptide according to the present invention comprising or consisting of an amino acid sequence represented by any of SEQ ID NOs. 65, 110, 114, and 220 is even more preferred.

[0168] Furthermore, a combination thereof, i.e., a pharmaceutical composition containing different antigenic peptides according to the present invention, is preferred. For example, the pharmaceutical composition may include the following: (i) at least two different antigenic peptides according to the present invention; (ii) at least two different immunogenic compounds according to the present invention; (iii) at least two different nanoparticles according to the present invention; (iv) At least two different nucleic acids according to the present invention; and / or (v) At least two different cytotoxic T lymphocytes according to the present invention.

[0169] Therefore, the pharmaceutical composition may contain at least "two different components," preferably three or four different components (of the pharmaceutical composition according to the present invention). In general, the expression "different components" as used herein means (1) A first component such as an antigenic peptide according to the present invention as described herein, an immunogenic compound according to the present invention as described herein, nanoparticles according to the present invention as described herein, cells according to the present invention as described herein, nucleic acids according to the present invention as described herein, host cells according to the present invention as described herein, or cytotoxic T lymphocytes according to the present invention as described herein, and (2) Meaning at least one other component (different from the first component; in the case of three or more different components, each component is different from each other), such as the anticancer agent described above, a different antigenic peptide according to the present invention as described herein, a different immunogenic compound according to the present invention as described herein, a different nanoparticle according to the present invention as described herein, a different cell according to the present invention as described herein, a different nucleic acid according to the present invention as described herein, a different host cell according to the present invention as described herein, a different cytotoxic T lymphocyte according to the present invention as described herein, or one or more human tumor antigens (one or more fragments thereof) in any form ("naked" as an immunogenic compound according to the present invention, a nanoparticle according to the present invention, a (host) cell according to the present invention, or a nucleic acid according to the present invention as described herein).

[0170] Therefore, the “different components” are preferably the active components (as described above) in the context of the disease to be prevented and / or treated (B-cell malignancies). In other words, each of the different components may also be useful for the prevention and / or treatment of the cancer, even when administered separately (not in combination as described herein). However, combination administration (i.e., combined administration) usually preferably synergistically enhances their preventive and / or therapeutic effects (such as immune responses).

[0171] 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 relate to different antigenic peptides of the present invention as described herein.

[0172] For example, the at least three or four different active ingredients are preferably of the same type, but each differs (only) in respect of a different antigenic peptide. More preferably, -The first component relates to an antigenic peptide comprising or consisting of a (microbiota) sequence variant of a fragment of the human tumor antigen CD22; -The (different) second component relates to an antigenic peptide comprising or consisting of a (microbiota) sequence variant of a fragment of the human tumor antigen CD37; -The (different) third component relates to an antigenic peptide comprising or consisting of a (microbiota) sequence variant of a fragment of the human tumor antigen TNFRSF13C; -Optionally, the (different) fourth component relates to an antigenic peptide comprising or consisting of a (microbiota) sequence variant of a fragment of the human tumor antigen MS4A1 (CD20).

[0173] Even more comfortable, -The first component relates to an antigenic peptide containing or comprising the (microbiota) sequence variant of Sequence ID No. 270; -The (different) second component is an antigenic peptide containing or consisting of the (microbiota) sequence variant of Sequence ID No. 271; -The (different) third component is an antigenic peptide comprising or consisting of the (microbiota) sequence variant of Sequence ID No. 279; -Optionally, the (different) fourth component relates to an antigenic peptide containing or consisting of the (microbiota) sequence variant of Sequence ID No. 264.

[0174] Even more comfortable, -The first component relates to an antigenic peptide comprising or consisting of an amino acid sequence represented by Sequence ID No. 110; -The (different) second component is an antigenic peptide comprising or consisting of the amino acid sequence represented by Sequence ID No. 114; -The (different) third component is an antigenic peptide comprising or consisting of the amino acid sequence represented by Sequence ID No. 220; -Optionally, the (different) fourth component relates to an antigenic peptide comprising or consisting of the amino acid sequence described in SEQ ID NO: 65.

[0175] Preferably, the pharmaceutical composition comprises at least two different antigenic peptides according to the present invention.

[0176] Preferably, the pharmaceutical composition comprises a first antigenic peptide according to the present invention, comprising or consisting of a (microbiota) sequence variant of a fragment of the human tumor antigen CD22, and a second antigenic peptide according to the present invention, comprising or consisting of a (microbiota) sequence variant of a fragment of the human tumor antigen TNFRSF13C. Preferably, the first antigenic peptide comprises or consists of an antigenic peptide comprising or consisting of an amino acid sequence represented by any of SEQ ID NOs: 106-110, 316, 387-390, and 485-488, for example, an antigenic peptide comprising or consisting of an amino acid sequence represented by any of SEQ ID NOs: 107, 108, 109, and 110, comprising a (microbiota) sequence variant of the CD22 fragment (human reference peptide) "WVFEHPETL" (SEQ ID NO: 270), and the second antigen The antigenic peptide comprises or consists of an antigenic peptide containing or comprising the amino acid sequences represented by SEQ ID NOs: 210-225, 325, 447-455, and 494-500, for example, any of SEQ ID NOs: 212, 217, 220, and 224, in particular an antigenic peptide containing or comprising the amino acid sequences represented by SEQ ID NOs: 220, 325, and 450, and comprises or consists of a (microbiota) sequence variant of the TNFRSF13C fragment (human reference peptide) "LLFGAPALL" (SEQ ID NO: 279). More preferably, the first antigenic peptide comprises or consists of the amino acid sequences represented by SEQ ID NOs: 110, 387, and 390, and the second antigenic peptide comprises or consists of the amino acid sequences represented by SEQ ID NOs: 220 and 450. Even more preferably, the pharmaceutical composition comprises an antigenic peptide containing or comprising SEQ ID NO: 110 and an antigenic peptide containing or comprising SEQ ID NO: 220.

[0177] More preferably, the pharmaceutical composition comprises at least three different antigenic peptides according to the present invention.

[0178] In particular, the pharmaceutical composition may include a first antigenic peptide according to the present invention comprising or comprising a (microbiota) sequence variant of a fragment of the human tumor antigen CD22, a second antigenic peptide according to the present invention comprising or comprising a (microbiota) sequence variant of a fragment of the human tumor antigen TNFRSF13C, and a third antigenic peptide according to the present invention comprising or comprising a (microbiota) sequence variant of a fragment of the human tumor antigen CD37. Preferably, the pharmaceutical composition comprises a first antigenic peptide comprising or consisting of an antigenic peptide comprising or consisting of an amino acid sequence represented by SEQ ID NOs: 106-110, 316, 387-390, and 485-488, for example, an antigenic peptide comprising or consisting of an amino acid sequence represented by any of SEQ ID NOs: 107, 108, 109, and 110, and an antigenic peptide comprising or consisting of an amino acid sequence represented by SEQ ID NOs: 210-225, 325, 447-455, and 494-500, for example, any of SEQ ID NOs: 212, 217, 220, and 224, particularly the amino acid sequence represented by SEQ ID NOs: 220, 325, and 450 The present invention comprises a second antigenic peptide comprising or consisting of a (microbiota) sequence variant of the TNFRSF13C fragment (human reference peptide) "LLFGAPALL" (SEQ ID NO: 279), such as an antigenic peptide comprising or consisting of a sequence; and a third antigenic peptide comprising or consisting of an antigenic peptide comprising or consisting of an amino acid sequence represented by SEQ ID NOs: 111-130, 317, 391-402, and 489-493, for example, any of SEQ ID NOs: 114, 117, 119, 120, 491, and 493, in particular, an antigenic peptide comprising or consisting of an amino acid sequence represented by SEQ ID NOs: 113-116, 324, and 392-393, such as an antigenic peptide comprising or consisting of an amino acid sequence; and a third antigenic peptide comprising or consisting of a (microbiota) sequence variant of the CD37 fragment (human reference peptide) "GLAFVPLQI" (SEQ ID NO: 271), such as an antigenic peptide comprising or consisting of an amino acid sequence represented by SEQ ID NOs: 114, 117, 119, 120, 491, and 493.

[0179] More preferably, the pharmaceutical composition comprises at least four different antigenic peptides according to the present invention.

[0180] In particular, the pharmaceutical composition may include a first antigenic peptide according to the present invention, comprising or comprising a (microbiota) sequence variant of the human tumor antigen CD22; a second antigenic peptide according to the present invention, comprising or comprising a (microbiota) sequence variant of the human tumor antigen TNFRSF13C; a third antigenic peptide according to the present invention, comprising or comprising a (microbiota) sequence variant of the human tumor antigen CD37; and a fourth antigenic peptide according to the present invention, comprising or comprising a (microbiota) sequence variant of the human tumor antigen CD19 or CD20.Preferably, the pharmaceutical composition comprises a first antigenic peptide comprising or consisting of an antigenic peptide comprising or consisting of an amino acid sequence represented by SEQ ID NOs: 106-110, 316, 387-390, and 485-488, for example, an antigenic peptide comprising or consisting of an amino acid sequence represented by any of SEQ ID NOs: 107, 108, 109, and 110, and a first antigenic peptide comprising or consisting of a (microbiota) sequence variant of the CD22 fragment (human reference peptide) "WVFEHPETL" (SEQ ID NO: 270), and SEQ ID NO: 2 A second antigenic peptide comprising or consisting of an antigenic peptide containing or comprising an amino acid sequence represented by 10-225, 325, 447-455, and 494-500, for example, any of SEQ ID NOs. 212, 217, 220, and 224, in particular SEQ ID NOs. 220, 325, and 450, and an antigenic peptide comprising or consisting of a (microbiota) sequence variant of the TNFRSF13C fragment (human reference peptide) "LLFGAPALL" (SEQ ID NO. 279). , antigenic peptides comprising or consisting of amino acid sequences represented by SEQ ID NOs: 111-130, 317, 391-402, and 489-493, for example, any of SEQ ID NOs: 114, 117, 119, 120, 491, and 493, in particular antigenic peptides comprising or consisting of amino acid sequences represented by SEQ ID NOs: 113-116, 324, 392-393, etc., including (microbiota) sequence variants of the CD37 fragment (human reference peptide) "GLAFVPLQI" (SEQ ID NO: 271) or The present invention comprises a third antigenic peptide comprising, and a fourth antigenic peptide comprising or comprising an antigenic peptide containing or comprising an amino acid sequence represented by SEQ ID NOs. 65-70, 310, 361-364, and 476-484, for example, an antigenic peptide containing or comprising an amino acid sequence represented by any of SEQ ID NOs. 65, 68, 70, and 477, which is a (microbiota) sequence variant of the CD20(MS4A1) fragment (human reference peptide) "IMNSLSLFA" (SEQ ID NO: 264).

[0181] Preferably, the pharmaceutical composition is - A first antigenic peptide according to the present invention comprising or consisting of a (microbiota) sequence variant of the human tumor antigen CD22 fragment, - A second antigenic peptide according to the present invention comprising or consisting of a (microbiota) sequence variant of the human tumor antigen TNFRSF13C fragment, - A third antigenic peptide according to the present invention comprising or consisting of a (microbiota) sequence variant of the human tumor antigen CD37 fragment, -Optionally, a fourth antigenic peptide according to the present invention comprising or comprising a (microbiota) sequence variant of the human tumor antigen CD20 fragment.

[0182] More preferably, the pharmaceutical composition - A first antigenic peptide comprising or consisting of an antigenic peptide containing or consisting of the amino acid sequences represented by SEQ ID NOs: 106-110, 316, and 387-390, and the (microbiota) sequence variant of the CD22 fragment (human reference peptide) "WVFEHPETL" (SEQ ID NO: 270), - A second antigenic peptide comprising or consisting of an antigenic peptide containing or consisting of the amino acid sequences represented by SEQ ID NOs: 220, 325, and 450, and an antigenic peptide comprising or consisting of the (microbiota) sequence variant of the TNFRSF13C fragment (human reference peptide) "LLFGAPALL" (SEQ ID NO: 279), - A third antigenic peptide comprising or consisting of an antigenic peptide containing or consisting of the amino acid sequences represented by SEQ ID NOs: 113-116, 324, and 392-393, and the (microbiota) sequence variant of the CD37 fragment (human reference peptide) "GLAFVPLQI" (SEQ ID NO: 271), -Optionally, a fourth antigenic peptide comprising or consisting of an antigenic peptide containing or consisting of an (microbiota) sequence variant of the CD20 fragment (human reference peptide) "IMNSLSLFA" (SEQ ID NO: 264), such as an antigenic peptide containing or consisting of the amino acid sequences represented by SEQ ID NOs: 65-70, 310, and 361-364.

[0183] More preferably, the pharmaceutical composition comprises an antigenic peptide according to the present invention comprising or consisting of an amino acid sequence represented by SEQ ID NOs: 110, 114, 220, and optionally SEQ ID NO: 65.

[0184] In some embodiments, the pharmaceutical composition does not contain any further antigenic peptides (in addition to the antigenic peptide of the present invention described above).

[0185] It is understood that the aforementioned pharmaceutical composition may also include, instead of the preferred combination of antigenic peptides described above, a combination of each of the immunogenic compounds of the present invention, a combination of each of the nanoparticles of the present invention, or a combination of each of the nucleic acids of the present invention.

[0186] Preferably, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients or carriers.

[0187] The pharmaceutical composition of the present invention may be in any form suitable for the purposes of the present invention. For example, the composition may be in a form suitable for parenteral, enteral, or topical administration, such as a liquid suspension, a solid dosage form (granules, pills, capsules, or tablets), or a paste or gel. Selecting an appropriate form of the composition for the intended purpose is within the scope of the skill of those skilled in the art.

[0188] The composition according to the present invention may further contain other activators that can enhance the effects of, for example, antigenic peptides or immunogenic compounds. Alternatively, the composition may not contain other activators (i.e., anything other than antigenic peptides, immunogenic compounds, nanoparticles, cells, nucleic acids, and / or host cells according to the present invention).

[0189] 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 antigenic peptides according to the present invention or by immunogenic compounds according to the present invention contained in the composition. Preferably, the pharmaceutical composition further comprises one or more immunoadjuvant substances. Pharmaceutical compositions, in particular immunogenic compositions, may also be referred to herein as “vaccine compositions.”

[0190] Preferably, the pharmaceutical composition further comprises at least one immunostimulant to increase, enhance, prolong, or maintain the immune response mediated by the antigenic peptide. Preferred immunostimulants according to the present invention include, but are not limited to, immune adjuvants, antigen-presenting cells, and combinations thereof. Preferably, the immunostimulant is an immune adjuvant or antigen-presenting cells (APCs).

[0191] Preferably, the immunostimulator is an immunoadjuvant. Some immunoadjuvants can favorably work to prolong the duration of the interaction between the antigen and the immune system, while others can mobilize and activate innate immune cells to induce an adaptive response. Adjuvants belonging to the former category include, but are not limited to, mineral compounds such as alum, aluminum hydroxide, aluminum phosphate, and calcium hydroxide; and oily emulsions such as paraffin oil, starch oil, Freund's complete / incomplete adjuvants (FCA / FIA), and saponins (e.g., those derived from plants such as quillaja, soybean, and senega). Adjuvants belonging to the latter category include, but are not limited to, bacterial products 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 for Toll-like receptors (TLRs) such as imiquimod, reximod, or MPL; exosomes such as exosomes derived from dendritic cells (DCs) or tumor cells; heat shock proteins (HSPs such as gp96, hsp90, hsp70, calreticulin, hsp110, hsp170), pathogen-associated molecular patterns (PAMPs), trehalose dimycolate (TDM), muramyl dipeptide (MDP), and polysaccharides (PLS) such as polysaccharide-K.

[0192] More preferably, the immunoadjuvant is a protein / peptide ("helper" peptide) having immunoadjuvant properties, such as providing stimulation of CD4+ Th1 cells, as described herein. A preferred example is a non-tumor antigen that may be a specific tumor-derived helper peptide, such as a tetanus helper peptide, keyhole limpet hemocyanin peptide, or PADRE peptide, which evokes immunological memory or provides nonspecific help. Another preferred example is a specific tumor-derived helper peptide, which may be presented by MHC II, particularly HLA-DR, HLA-DP, or HLA-DQ, for example, a fragment of a shared overexpressed tumor antigen (e.g., HER2, NY-ESO-1, hTERT, or IL13RA2 as described above). In particular, the immunoadjuvant may be the HHD-DR3 peptide of sequence MAKTIAYDEEARRGLERGLN (SEQ ID NO: 473). This peptide is another example of a helper peptide preferred (having immunoadjuvant properties) in the context of the present 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 immunoadjuvants, particularly helper peptides, include the UCP2 peptide (e.g., International Publication WO2013 / 135553A1 or Dosset et al., Clin Cancer Res. 2012 Nov 15;18(22):6284-95) and the BIRC5 peptide (e.g., European Patent Application Publication 2119726A1 or Widenmeyer et al., Int J Cancer. 2012 Jul 1;131(1):140-9). The most preferred helper peptide is the UCP2 peptide (amino acid sequence: KSVWSKLQSIGIRQH; SEQ ID NO: 475).

[0193] Preferably, the pharmaceutical composition comprises at least two different antigenic peptides according to the present invention and a helper peptide, preferably a UCP2 peptide (SEQ ID NO: 475).

[0194] In particular, the pharmaceutical composition may include a first antigenic peptide according to the present invention comprising or consisting of a sequence variant of the human tumor antigen CD22 fragment, a second antigenic peptide according to the present invention comprising or consisting of a sequence variant of the human tumor antigen TNFRSF13C fragment, and a helper peptide, preferably a UCP2 peptide (SEQ ID NO: 475). Preferably, the pharmaceutical composition includes a first antigenic peptide comprising or consisting 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 represented by SEQ ID NOs: 106-110, 316, and 387-390, a second antigenic peptide comprising or consisting 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 represented by SEQ ID NOs: 220, 325, and 450, and a helper peptide, preferably a UCP2 peptide (SEQ ID NO: 475). More preferably, the first antigenic peptide comprises or consists of amino acid sequences represented by SEQ ID NOs: 110, 387, and 390, and the second antigenic peptide comprises or consists of amino acid sequences represented by SEQ ID NOs: 220 and 450. Even 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).

[0195] More preferably, the pharmaceutical composition comprises at least three different antigenic peptides according to the present invention and a helper peptide, preferably a UCP2 peptide (SEQ ID NO: 475).

[0196] In particular, the pharmaceutical composition may include a first antigenic peptide according to the present invention comprising or comprising a (microbiota) sequence variant of the human tumor antigen CD22 fragment, a second antigenic peptide according to the present invention comprising or comprising a (microbiota) sequence variant of the human tumor antigen TNFRSF13C fragment, a third antigenic peptide according to the present invention comprising or comprising a (microbiota) sequence variant of the human tumor antigen CD37 fragment, and a helper peptide, preferably a UCP2 peptide (SEQ ID NO: 475). Preferably, the pharmaceutical composition comprises: a first antigenic peptide comprising or consisting of an antigenic peptide containing or consisting of the amino acid sequences represented by SEQ ID NOs: 106-110, 316, and 387-390, which is a (microbiota) sequence variant of the CD22 fragment (human reference peptide) "WVFEHPETL" (SEQ ID NO: 270); a second antigenic peptide comprising or consisting of an antigenic peptide containing or consisting of the amino acid sequences represented by SEQ ID NOs: 220, 325, and 450, which is a (microbiota) sequence variant of the TNFRSF13C fragment (human reference peptide) "LLFGAPALL" (SEQ ID NO: 279); a third antigenic peptide comprising or consisting of an antigenic peptide containing or consisting of the amino acid sequences represented by SEQ ID NOs: 113-116, 324, and 392-393, which is a (microbiota) sequence variant of the CD37 fragment (human reference peptide) "GLAFVPLQI" (SEQ ID NO: 271); and a helper peptide, preferably a UCP2 peptide (SEQ ID NO: 475).

[0197] Most preferably, the pharmaceutical composition comprises at least four different antigenic peptides according to the present invention and a helper peptide, preferably a UCP2 peptide (SEQ ID NO: 475).

[0198] In particular, the pharmaceutical composition may include a first antigenic peptide according to the present invention comprising or comprising a (microbiota) sequence variant of the human tumor antigen CD22 fragment, a second antigenic peptide according to the present invention comprising or comprising a (microbiota) sequence variant of the human tumor antigen TNFRSF13C fragment, a third antigenic peptide according to the present invention comprising or comprising a (microbiota) sequence variant of the human tumor antigen CD37 fragment, a fourth antigenic peptide according to the present invention comprising or comprising a (microbiota) sequence variant of the human tumor antigen CD19 or CD20 fragment, and a helper peptide, preferably a UCP2 peptide (SEQ ID NO: 475).

[0199] Preferably, the pharmaceutical composition comprises a first antigenic peptide according to the present invention comprising or comprising a (microbiota) sequence variant of the human tumor antigen CD22 fragment, a second antigenic peptide according to the present invention comprising or comprising a (microbiota) sequence variant of the human tumor antigen TNFRSF13C fragment, a third antigenic peptide according to the present invention comprising or comprising a (microbiota) sequence variant of the human tumor antigen CD37 fragment, a fourth antigenic peptide according to the present invention (optionally) comprising or comprising a (microbiota) sequence variant of the human tumor antigen CD20 fragment, and a helper peptide, preferably a UCP2 peptide (SEQ ID NO: 475). More preferably, the pharmaceutical composition comprises a first antigenic peptide comprising or consisting of an antigenic peptide containing or comprising the amino acid sequences represented by SEQ ID NOs: 106-110, 316, and 387-390, which is a (microbiota) sequence variant of the CD22 fragment (human reference peptide) "WVFEHPETL" (SEQ ID NO: 270), and a second antigenic peptide comprising or consisting of an antigenic peptide containing or comprising the amino acid sequences represented by SEQ ID NOs: 220, 325, and 450, which is a (microbiota) sequence variant of the TNFRSF13C fragment (human reference peptide) "LLFGAPALL" (SEQ ID NO: 279), and SEQ ID NOs: 113-116, The material comprises a third antigenic peptide containing or comprising a (microbiota) sequence variant of the CD37 fragment (human reference peptide) "GLAFVPLQI" (SEQ ID NO: 271), such as an antigenic peptide containing or comprising the amino acid sequences represented by 324 and 392-393; a fourth antigenic peptide containing or comprising a (microbiota) sequence variant of the CD20 fragment (human reference peptide) "IMNSLSLFA" (SEQ ID NO: 264), such as an antigenic peptide containing or comprising the amino acid sequences represented by SEQ ID NOs: 65-70, 310, and 361-364; and a helper peptide, preferably a UCP2 peptide (SEQ ID NO: 475).

[0200] A particularly preferred immunoadjuvant is polyinosinate:polycytidylic acid (also known as "Poly-I:C") and / or its derivative, poly-ICLC. Poly-I:C is a mismatched double-stranded RNA, where one strand is a polymer of inosinate and the other is a polymer of cytidylic acid. Poly-I:C is an immunostimulant known to interact with Toll-like receptor 3 (TLR3). Poly-I:C is structurally similar to double-stranded RNA, the "natural" stimulant of TLR3. Therefore, poly-I:C can be considered a synthetic analogue of double-stranded RNA. Poly-ICLC is a synthetic complex of carboxymethylcellulose, polyinosinate-polycytidylic acid, and poly-L-lysine double-stranded RNA. Like poly-I:C, poly-ICLC is also a ligand for TLR3. Poly-I:C and poly-ICLC typically stimulate the release of cytotoxic cytokines. A preferred example of poly-ICLC is Hiltonol®.

[0201] Most preferably, the adjuvant is a montandide such as Montanide ISA 51 VG and / or Montanide ISA 720 VG. When these adjuvants are mixed with a water-based antigenic medium, they form a stable water-in-oil emulsion. Montanide ISA 51 VG is based on a blend of mannidomonoleate surfactant and mineral oil, while Montanide ISA 720 VG uses non-mineral oil (Aucouturier J, Dupuis L, Deville S, Ascarateil S, Ganne V. Montanide ISA 720 and 51: a new generation of water in oil emulsions as adjuvants for human vaccines. Expert Rev Vaccines. 2002 Jun;1(1):111-8; Ascarateil S, Puget A, Koziol ME. Safety data of Montanide ISA 51 VG and Montanide ISA 720 VG, 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).

[0202] Furthermore, the immunostimulant is preferably an antigen-presenting cell (APC). APCs are particularly interesting because their main function is to process antigens and present them on the cell surface to T cells of the immune system, thereby initiating and regulating the in vivo T cell response. In this composition, the APC is preferably loaded with the antigenic peptide and / or immunogenic compound according to the present invention, which can be done by exposing the APC to the antigenic peptide and / or immunogenic compound in vitro (Rizzo et al., Methods Mol Biol. 2014;1139:41-4; Rolinski and Hus, J Immunotoxicol. 2014 Oct;11(4):311-8).

[0203] Preferably, the APC is a dendritic cell (DC). DCs are the most potent APCs and are often reported to be functionally deficient in cancer patients. DCs can be readily obtained by technicians in the art from healthy, compatible donors (i.e., the dendritic cells are HLA-associated) or from the patient themselves if they are functional (i.e., the DCs are autologous), for example, by direct isolation from peripheral blood or by induction from peripheral blood cells such as CD14+ monocytes or CD34+ hematopoietic progenitor cells (Emens et al., 2008). In fact, DCs can be distinguished from other cells in peripheral blood by surface markers such as S100, p55, CD83, and / or OX62, and can therefore be isolated and purified based on these markers using cell culture techniques well known in the art.

[0204] In a preferred embodiment, the pharmaceutical composition may further comprise at least one anticancer therapeutic agent. The therapeutic agent is therefore preferably capable of preventing and / or treating the same type of cancer as those for which the antigenic peptide according to the present invention is used. Preferably, the anticancer therapeutic agent is selected from antibodies, CAR-T cells, tumor cell lysates, chemotherapeutic agents, radiotherapeutic agents, immune checkpoint modulators, and combinations thereof.

[0205] Antibodies are particularly advantageous in cancer treatment because they can bind to specific antigens on the surface of cancer cells to direct the treatment to the tumor (i.e., these are called tumor-targeting antibodies) or block immune checkpoints that are dysregulated in cancer (i.e., these are referred to herein as immunomodulatory antibodies). The purpose of the latter type of antibody is to inhibit cancer's immune resistance, which can be particularly observed against tumor antigen-specific T cells. In fact, as is well known in the art, under normal physiological conditions, immune checkpoints are important for maintaining self-tolerance (i.e., preventing autoimmunity), protecting tissues from damage when the immune system responds to pathogenic infection. However, in cancer, the expression of immune checkpoints can become dysregulated as a key mechanism of immune resistance.The aforementioned resistance is prominently observed in melanoma, ovarian cancer, lung cancer, glioblastoma, breast cancer, and pancreatic cancer with respect to the PD-L1 checkpoint (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) T lymphocyte-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 for malignant melanoma. Cancer. 2010 Apr 1;116(7):1757-66).Other examples of immune checkpoints include, but are not limited to, PD-L2, PD-1, CD80, CD86, CTLA-4, B7H3, B7H4, PVR, TIGIT, GAL9, LAG-3, GITR, CD137, TIM3, VISTA, and VISTA-R (Pico de Coana 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 cancer immunotherapy. Nat Rev Cancer. 2012 Mar 22;12(4):252-64).

[0206] Antibodies are typically used for the aforementioned purposes in the form of naked monoclonal antibodies (i.e., unconjugated) or conjugated to another molecule that may be toxic to cells or radioactive.

[0207] Well-known monoclonal tumor target antibodies used in cancer immunotherapy include alemtuzumab (chronic lymphocytic leukemia), bevacizumab (colorectal cancer, glioblastoma multiforme, cervical cancer, lung cancer, renal cancer), brentuximab / vedotin (lymphoma), blinatumab (acute lymphoblastic leukemia), catumakisomab (malignant ascites in EPCAM+ cancer), cetuximab (head and neck cancer, colorectal cancer), denosumab (breast cancer, prostate cancer, and bone cancer), gemtuzumab / ozogamicin (acute myeloid leukemia), iblitzumab Examples include, but are not limited to, bu / tiuxetan (non-Hodgkin 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 lymphoma), siltuximab (multicentric Castleman disease), tocitumomab (non-Hodgkin lymphoma), and trastuzumab (breast cancer, gastric cancer, and gastroesophageal cancer). On the other hand, examples of immunomodulatory antibodies include, but are not limited to, ipilimumab (melanoma), which blocks the CTLA4-dependent immune checkpoint; nivolumab (melanoma, lung cancer) and prembrolizubumab (melanoma), which both block the PDCD1-dependent immune checkpoint; and MPDL3280A, MEDI4736, MEDI0680, and MSB0010718C (Sharma and Allison, The future of immune checkpoint therapy. Science. 2015 Apr 3;348(6230):56-61), which all block the PD-L1-dependent immune checkpoint.

[0208] Other antibodies for cancer immunotherapy are listed in Buque 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. Mol Immunol. 2015 Oct;67(2 Pt A):28-45; Simpson and Caballero, Monoclonal antibodies for the therapy of cancer MC Proc. 2014; 8(Suppl 4):O6; and on the Antibody Society website (website link http: / / www.antibodysociety.org / news / approved_mabs.php, which lists therapeutic monoclonal antibodies approved or under review in the European Union or the United States).

[0209] Adoptive cell immunotherapy using chimeric antigen receptor (CAR) T cells is transforming the treatment of B-cell non-Hodgkin lymphoma (NHL), particularly advanced B-cell lymphoma. For example, CD19-targeted CAR T cells are a new standard of treatment for DLBCL patients who are resistant to at least two existing lines of treatment. Two CAR T cell products, axicabtagene ciloleucel (axi-cel) (KTE-019) (YESCARTA) TM ) and tisagenlecleucel(CTL019)(KYMRIAH TM ) has received approval from the U.S. Food and Drug Administration for the treatment of DLBCL that is resistant after two lines of treatment. A third product, lisocabtagene maraleucel (liso-cel) (JCAR017), is currently in clinical trials. Other CAR T cells include CD20-CAR-T cells.

[0210] Tumor cell lysates can also be used in combination with antigenic peptides according to the present invention. Tumor cells can actually prime an immune response by presenting endogenous peptide-MHC complexes and via host dendritic cells (DCs) that can process and present antigens delivered by the lysates. This broadens 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).

[0211] Standard chemotherapy and radiotherapy agents are described in detail in the literature, particularly by Baskar et al. (*Cancer and radiation therapy: current advances and future directions. Int J Med Sci. 2012;9(3):193-9), 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. (*Review of therapeutic drug monitoring of anticancer drugs part two--targeted therapies. Eur J Cancer. 2014 Aug;50(12):2020-36), and therefore do not need to be described further herein. A list of such drugs and medications is also available on the cancer.gov website (http: / / www.cancer.gov / about-cancer / treatment / drugs).

[0212] Preferably, an immune checkpoint modulator for use with an antigenic peptide as 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.

[0213] More preferably, the immune checkpoint modulator is an activator of a (co)stimulatory checkpoint molecule or an inhibitor of an inhibitory checkpoint molecule, or a combination thereof. Therefore, the immune checkpoint modulator is more preferably (i) an activator of 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.

[0214] More preferably, the immune checkpoint modulator is an inhibitor of an inhibitory checkpoint molecule (but preferably not an inhibitor of an stimulant checkpoint molecule). Therefore, the immune checkpoint modulator is 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 an inhibitor of their ligands.

[0215] Furthermore, the immune checkpoint modulator is preferably an activator of an irritant or co-stimulatory checkpoint molecule (but preferably not an activator of an inhibitory checkpoint molecule). Therefore, the immune checkpoint modulator is more preferably an activator of CD27, CD28, CD40, CD122, CD137, OX40, GITR, and / or ICOS, or an activator of their ligands.

[0216] The immune checkpoint modulator is more preferably a modulator of the CD40 pathway, the IDO pathway, the LAG3 pathway, the CTLA-4 pathway, and / or the PD-1 pathway. In particular, 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.

[0217] Therefore, the checkpoint modulator for use with the antigenic peptide can be selected from known modulators of the CTLA-4 pathway or the PD-1 pathway. Preferably, the checkpoint modulator for use with the antigenic peptide as defined herein can be selected from known modulators of the CTLA-4 pathway or the PD-1 pathway. Particularly preferably, the immune checkpoint modulator is a PD-1 inhibitor. Preferred inhibitors of the CTLA-4 and PD-1 pathways include Yervoy® (Ipilimumab; Bristol Myers Squibb) and Tremelimumab (Pfizer / MedImmune), as well as Opdivo® (Nivolumab; Bristol Myers Squibb), Keytruda® (Pembrolizumab (also known as Lambrolizumab or MK-3475); Merck), Imfinzi® (Durvalumab (also known as MEDI4736); MedImmune / AstraZeneca), Tecentriq® (Atezolizumab (also known as MPDL3280A); Roche / Genentech), Pidilizumab (CT-011; CureTech), MEDI0680 (AMP-514; AstraZeneca), and Bavencio® (Avelumab; Merck) Examples include monoclonal antibodies for KGaA / Pfizer (also known as MSB-0010718C), MIH1 (Affymetrix), LY3300054 (Eli Lilly), and Spartalizumab (also known as PDR001; Novartis).More preferred checkpoint inhibitors include the CTLA-4 inhibitors Yervoy® (Ipilimumab; Bristol Myers Squibb) and Tremelimumab (Pfizer / MedImmune), and the PD-1 inhibitors Opdivo® (Nivolumab; Bristol Myers Squibb), Keytruda® (Pembrolizumab; Merck), Pidilizumab (CT-011; CureTech), MEDI0680 (AMP-514; AstraZeneca), and AMP-224 (PD-L2Fc fusion protein; MedImmune).

[0218] The immune checkpoint modulators for use in combination with antigenic peptides as defined herein are also selected from the group consisting of Pembrolizumab, Ipilimumab, Nivolumab, Atezolizumab, Durvalumab, Tremelimumab, Avelumab, Spartalizumab, LAG525 (anti-LAG-3 monoclonal antibody), Epacadostat (also known as INCB24360; IDO inhibitor), Varlilumab (anti-CD27 monoclonal antibody), Urelumab (anti-CD137 monoclonal antibody), AMP-224, and CM-24 (anti-CEACAM1 monoclonal antibody).

[0219] Selecting an appropriate immunosuppressant anticancer agent for the purposes of the present invention is within the scope of the skills of those skilled in the art. For example, if the prevention or treatment of melanoma is desired, a lysate and / or antibody ipilimumab from melanoma cells may be used, preferably together with an appropriate antigenic peptide. An appropriate antigenic peptide can be selected by (i) selecting an appropriate tumor antigen for a particular type of cancer known in the art and / or listed in Table 1B herein, and (ii) selecting an appropriate antigenic peptide according to the present invention for the selected tumor antigens described above, for example, in Table 1A.

[0220] The aforementioned anticancer therapeutic agent may also be administered in combination with the composition of the present invention, simultaneously, separately, or sequentially. When the composition and the therapeutic agent are administered separately or sequentially, they may be administered in different dosage forms.

[0221] Therefore, in another aspect, the present invention relates to the above-described composition of the present invention and at least one anticancer agent as a combination preparation for simultaneous, separate, or sequential administration. In other words, the present invention proposes the combination of the above-described composition of the present invention and at least one anticancer agent for simultaneous, separate, or sequential administration.

[0222] Kit of Parts In a further embodiment, the present invention also provides a kit of parts (also referred to herein as a “kit”) comprising at least one of the following: - Antigenic peptides according to the present invention as described herein, - Immunogenic compounds according to the present invention as described herein, -Naparticles according to the present invention as described herein, - Cells according to the present invention as described herein, - Nucleic acids according to the present invention as described herein, -Host cells according to the present invention as described herein, -Cytotoxic T lymphocytes according to the present invention as described herein, and / or -The pharmaceutical composition according to the present invention as described herein.

[0223] In particular, the preferred embodiments of the antigenic peptide described above also apply to the kit according to the present invention. For example, the antigenic peptide contained in the kit, or the antigenic peptide contained in any of the immunogenic compound, nanoparticles, cells, nucleic acids, host cells, or pharmaceutical composition contained in the kit, preferably contains or comprises an amino acid sequence represented by any of SEQ ID NOs: 1 to 247 and 476 to 500. For example, an antigenic peptide according to the present invention containing or comprising an amino acid sequence represented by any of SEQ ID NOs: 10, 21, 33, 35, 39, 40, 61, 65, 68, 72, 86, 110, 114, and 220 is even more preferred. For example, an antigenic peptide according to the present invention containing or comprising an amino acid sequence represented by any of SEQ ID NOs: 10, 21, 33, 35, 39, 40, 110, 114, and 220 is even more preferred. For example, an antigenic peptide according to the present invention containing or comprising an amino acid sequence represented by any of SEQ ID NOs: 10, 110, 114, and 220 is even more preferred. For example, an antigenic peptide according to the present invention comprising or consisting of an amino acid sequence represented by any of SEQ ID NOs. 65, 110, 114, and 220 is even more preferred.

[0224] Furthermore, a combination thereof, i.e., a kit containing different antigenic peptides according to the present invention, is preferred. In particular, the kit of parts according to the present invention may contain two or more of the above-mentioned components, for example, 2, 3, 4, 5, 6, 7, 8, 9, or 10 different components. For example, the kit of parts 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 different components included in the kit of parts described above have different antigenic peptides according to the present invention, for example, one component relating to a first antigenic peptide and one component relating 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 present invention. For example, the kit may include at least two different antigenic peptides according to the present invention. For example, the kit may include at least two different nanoparticles according to the present invention. For example, the kit may include at least two different nucleic acids according to the present invention. For example, the kit may include at least two different cytotoxic T lymphocytes according to the present invention.

[0225] The preferred combination of components such as the antigenic peptide according to the present invention contained in the aforementioned kit corresponds to the preferred combination of components such as the antigenic peptide according to the present invention contained in the pharmaceutical composition described above.

[0226] Accordingly, the present invention provides a kit comprising (at least one) antigenic peptide according to the present invention as described herein. Furthermore, the present invention also provides a kit comprising (at least one) immunogenic compound according to the present invention as described herein. Furthermore, the present invention also provides a kit comprising (at least one) nanoparticles according to the present invention as described herein. Furthermore, the present invention also provides a kit comprising (at least one) cells according to the present invention as described herein. Furthermore, the present invention also provides a kit comprising (at least one) nucleic acid according to the present invention as described herein. Furthermore, the present invention also provides a kit comprising (at least one) host cell according to the present invention as described herein.

[0227] The various components of the kit of parts may be packaged in one or more containers. The components may be provided in lyophilized or dry form, or dissolved in a suitable buffer. The kit may also include further reagents, such as preservatives, growth media, and / or buffers and washing solutions for preserving and / or reconstituting the components.

[0228] Accordingly, the present invention provides a kit comprising at least two, preferably three, different antigenic peptides (or immunogenic compounds, nanoparticles, nucleic acids, cells, etc., with different antigenic peptides) as described herein, and optionally a helper peptide such as UCP2 peptide and / or an adjuvant such as MONTANIDE ISA 51. The different antigenic peptides (or immunogenic compounds, nanoparticles, nucleic acids, cells, etc., with different antigenic peptides) may be contained in the same or different containers. For example, the kit may include a (single) container containing a first antigenic peptide and a second antigenic peptide as described herein. The (single) container may further also include a helper peptide such as UCP2. Optionally, the first and second antigenic peptides (and optionally the helper peptide) contained in the (single) container may be combined and formulated, for example, in water for injection and / or dimethyl sulfoxide (DMSO). Furthermore, the kit may include a further container (different from the container containing the antigenic peptides) containing an adjuvant such as MONTANIDE ISA 51.

[0229] Therefore, the kit is (i) a first vial optionally comprising one or more antigenic peptides of the present invention (e.g., at least 200 or 300 μg of each antigenic peptide) formulated in water for injection and dimethyl sulfoxide (DMSO), and optionally a helper peptide such as UCP2 (e.g., at least 200 or 300 μg of the helper peptide); and (ii) Preferably, the mixture includes a second vial containing MONTANIDE ISA 51 (for example, at least 0.4 or 0.5 ml).

[0230] Furthermore, the kit may include one or more (e.g., two or three) syringes, such as silicone-free syringes and rubber-free syringes. The kit may also include connectors such as I-connectors.

[0231] Non-limiting examples of such connectors include the following. The I-connector developed by Green Peptide (Japan), The connector of reference DIDRACDLLFT of Didanorm (France), The I-connector of Promepla (Monaco) (ref: ODG0015ST), and The I-connector of Smiths medical (USA) (ref: MX494).

[0232] The syringe is preferably suitable for MONTANIDE, i.e., silicon-free and rubber-free (i.e., no rubber tip on the plunger), and preferably also latex-free. Non-limiting examples of such syringes include the following. 2 ml INKJET (Ref: 4606701V, B-Braun, Germany), 5 ml INKJET (Ref: 4606710V, B-Braun, Germany), 2 ml Norm-Ject (Ref: 4020.000V0, Henke Sass Wolf GMBH, Germany), and 5 ml Norm-Ject (Ref: 4050.000V0, Henke Sass Wolf GMBH, Germany).

[0233] For example, the kit can 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) formulated in water for injection and dimethyl sulfoxide (DMSO), and optionally at least 300 μg of UCP2, (ii) a second vial containing at least 0.5 ml of MONTANIDE ISA 51, (iii) two silicon-free and rubber-free syringes, and (iv) an I-connector.

[0234] Optionally, the kit can also include vials of water for injection and / or vial adapters. For example, it can also include a sterile needle for vaccinating a patient after obtaining the emulsion. The syringe within the kit can be, for example, a 2 ml syringe.

[0235] Furthermore, the kit of parts according to the invention can optionally include instructions for use. Thus, the kit preferably includes a package insert or instruction leaflet containing instructions for preventing or treating B cell malignancies by 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 acids according to the invention, the host cells according to the invention, or the pharmaceutical composition according to the invention.

[0236] In addition to any of the above-described components, the kit preferably also includes an anti-cancer therapeutic agent as described herein.

[0237] Furthermore, the present invention also provides a vaccination kit for treating, preventing, and / or stabilizing B cell malignancies, which includes the pharmaceutical composition described herein or the vaccine described herein, as well as instructions for using the pharmaceutical composition or the vaccine in the prevention and / or treatment of B cell malignancies.

[0238] Medical Treatments and Uses As described above, the composition of the present invention can be particularly useful for therapeutic purposes (as a drug), particularly for inducing a specific immune response against specific tumor antigens / proteins, for example, for preventing or treating B cell malignancies (such as B cell lymphoma) in patients who need it.

[0239] In view of this, the present invention provides the following for use in medicine, particularly for the prevention and / or treatment of B cell malignancies. - The antigenic peptide according to the invention as described herein, - The immunogenic compound according to the invention as described herein, - The nanoparticles according to the invention as described herein, -The cytotoxic T lymphocytes (CTLs) according to the present invention as described herein, - Cells according to the present invention as described herein, - Nucleic acids according to the present invention as described herein, -Host cells according to the present invention as described herein, -Pharmaceutical compositions according to the present specification, - A kit according to the present invention as described herein, or -The combination of the present invention as described herein.

[0240] In particular, preferred embodiments of the antigenic peptide described above also apply to the use of the present invention for use in the prevention and / or treatment of B-cell malignancies. For example, the antigenic peptide used in the prevention and / or treatment of B-cell malignancies, or the antigenic peptide contained in any of the immunogenic compounds, nanoparticles, cells, nucleic acids, host cells, or pharmaceutical compositions used in the prevention and / or treatment of cancer, preferably comprises or consists of an amino acid sequence represented by any of SEQ ID NOs: 1-257 and 476-500. In some embodiments, the antigenic peptide according to the present invention preferably comprises or consists of an amino acid sequence represented by any of SEQ ID NOs: 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 SEQ ID NOs: 34-35, 65-70 , comprising or consisting of an amino acid sequence represented by any of 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 an amino acid sequence represented by any 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 an amino acid sequence represented by any 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. For example, an antigenic peptide according to the present invention comprising or consisting of an amino acid sequence represented by any of SEQ ID NOs: 10, 21, 33, 35, 39, 40, 61, 65, 68, 72, 86, 110, 114, and 220 is even more preferred. For example, an antigenic peptide according to the present invention comprising or consisting of an amino acid sequence represented by any of SEQ ID NOs: 10, 21, 33, 35, 39, 40, 110, 114, and 220 is even more preferred.For example, an antigenic peptide according to the present invention comprising or consisting of an amino acid sequence represented by any of SEQ ID NOs: 10, 110, 114, and 220 is even more preferred. For example, an antigenic peptide according to the present invention comprising or consisting of an amino acid sequence represented by any of SEQ ID NOs: 65, 110, 114, and 220 is even more preferred.

[0241] Furthermore, a combination thereof, i.e., different antigenic peptides according to the present invention for use in the prevention and / or treatment of B-cell malignancies, is preferred. In particular, two or more 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 different components used for the prevention and / or treatment of B-cell malignancies described above are different antigenic peptides according to the present invention, for example, one component relating to a first antigenic peptide and one component relating to a second antigenic peptide (different from the first antigenic peptide). For example, at least two different immunogenic compounds according to the present 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 present invention can be used for the prevention and / or treatment of B-cell malignancies. For example, at least two different nanoparticles according to the present invention can be used in the prevention and / or treatment of B-cell malignancies. For example, at least two different nucleic acids according to the present invention can be used in the prevention and / or treatment of B-cell malignancies.

[0242] Accordingly, the present invention provides (at least one) antigenic peptide according to the present invention as described herein for use in the prevention and / or treatment of B-cell malignancies. Furthermore, the present invention also provides (at least one) immunogenic compound according to the present invention as described herein for use in the prevention and / or treatment of B-cell malignancies. Furthermore, the present invention also provides (at least one) nanoparticles according to the present invention as described herein for use in the prevention and / or treatment of B-cell malignancies. Furthermore, the present invention also provides (at least one) cell according to the present invention as described herein for use in the prevention and / or treatment of B-cell malignancies. Furthermore, the present invention also provides (at least one) nucleic acid according to the present invention as described herein for use in the prevention and / or treatment of B-cell malignancies. Furthermore, the present invention also provides (at least one) host cell according to the present invention as described herein for use in the prevention and / or treatment of B-cell malignancies. Furthermore, the present invention also provides (at least one) pharmaceutical composition according to the present invention as described herein for use in the prevention and / or treatment of B-cell malignancies. Furthermore, the present invention also provides a kit according to the present invention as described herein for use in the prevention and / or treatment of B-cell malignancies.

[0243] Accordingly, the present invention also provides a method for preventing (reducing the incidence thereof) and / or treating B-cell malignancies or for initiating, enhancing, or prolonging an antitumor response to B-cell malignancies in subjects requiring such treatment, the method comprising administering the following to the subjects: That is, as described herein, - Antigenic peptide according to the present invention, - Immunogenic compounds according to the present invention, -Napnea according to the present invention, - Cells according to the present invention -The cytotoxic T lymphocytes (CTLs) according to the present invention as described herein, - Nucleic acids according to the present invention, - Host cell according to the present invention, - Pharmaceutical composition according to the present invention, - A kit according to the present invention, or - Combined use according to the present invention.

[0244] Preferably, the B-cell malignancies to be treated include leukemias and lymphomas, such as acute myeloid (or myeloid) leukemia (AML), chronic myeloid (or myeloid) leukemia (CML), acute lymphoblastic (or lymphoblastic) leukemia (ALL), B-chronic lymphocytic leukemia (BCLL), chronic lymphocytic leukemia (CLL, Richter), hairy cell leukemia (HCL), lymphoplasmacytic lymphoma (LPC), or Waldenström Examples include chloroglobulinemia, prolymphocytic leukemia (PLL), small lymphocytic lymphoma (SLL), mantle cell lymphoma (MCL), marginal zone lymphoma (MZL), Burkitt lymphoma (BL), Hodgkin lymphoma (HL), non-Hodgkin lymphoma (NHL), anaplastic large cell lymphoma (ALCL), follicular lymphoma (FL), refractory follicular lymphoma, diffuse large B-cell lymphoma (DLBCL), and multiple myeloma (MM). In some embodiments, the B-cell malignancies are 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 low-grade (slowly progressive) NHL, high-grade NHL, diffuse large B-cell lymphoma (DLBCL), NOS (de novo and transformed from indolent) follicular lymphoma (FL), primary mediastinal large B-cell lymphoma (PMBCL), T-cell / histiocyte-rich large B-cell lymphoma (TCHRBCL), Burkitt lymphoma, mantle cell lymphoma (MCL), and / or follicular lymphoma (FL), optionally follicular lymphoma grade 3B (FL3B).

[0245] Furthermore, the present invention relates to the CD8 in the subject. + The present invention provides a method for inducing or improving an immune response to one or more epitopes dependent on cytotoxic T cells, the method comprising administering one of the following to the subject: That is, as described in this specification, - the antigenic peptide according to the present invention, - the immunogenic compound according to the present invention, - the nanoparticles according to the present invention, - the cytotoxic T lymphocytes (CTLs) according to the present invention, - the cells according to the present invention, - the nucleic acids according to the present invention, - the host cells according to the present invention, - the pharmaceutical compositions according to the present invention, - the kits according to the present invention, or - the combinations according to the present invention.

[0246] CD8 + The immune response that depends on the CD8 response can be determined by evaluating pro-inflammatory cytokine responses such as inflammatory responses, IFN-γ, TNF-α, and an increase in the expression of one or more of IL-2 mRNA or protein, relative to the levels before administration of the compounds of the present invention. 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 by HLA peptide multimer staining, ELISPOT assay, and delayed-type hypersensitivity testing. It can also be measured indirectly by an increase in antigen-specific serum antibodies that depend on antigen-specific T helper cells.

[0247] The present invention also provides a method for inducing or improving an immune response against one or more antigens or antigenic epitopes restricted by a plurality of MHC class I molecules in a subject, the method comprising administering to the subject any of the following. That is, as described in this specification, - the antigenic peptide according to the present invention, - the immunogenic compound according to the present invention, - the nanoparticles according to the present invention, - the cytotoxic T lymphocytes (CTLs) according to the present invention, - the cells according to the present invention, - the nucleic acids according to the present invention, - the host cells according to the present invention, - the pharmaceutical compositions according to the present invention, - A kit according to the present invention, or - Combined use according to the present invention.

[0248] Methods for inducing or improving an immune response to multiple epitopes described herein, restricted by multiple MHC class I molecules, in a subject can be determined by evaluating the cytokine response, including an increase in the expression of one or more IFN-γ, TNF-α, and IL-2 mRNA or protein, relative to pre-administration levels of the compound of the present invention, after in vitro stimulation of T cells with individual peptides bound to separate MHC class I molecules on antigen-presenting cells. Restriction to MHC class I molecules can also be verified by using antigen-presenting cells expressing MHC class I molecules or by using an MHC class I blocking antibody. It can also be measured by the increase in the frequency or absolute number of antigen-specific T cells after administration of the compound of the present invention, as measured by HLA peptide multimer staining using multimers assembled with MHC class I molecules.

[0249] Therefore, in another embodiment, the present invention also provides the following: For use as a pharmaceutical agent, as described herein, - Antigenic peptide according to the present invention, - Immunogenic compounds according to the present invention, -Napnea according to the present invention, -Cytotoxic T lymphocytes (CTLs) according to the present invention, - Cells according to the present invention - Nucleic acids according to the present invention, - Host cell according to the present invention, - Pharmaceutical composition according to the present invention, - A kit according to the present invention, or - Combined use according to the present invention.

[0250] The present invention relates, more particularly, to the compositions defined above for use as vaccines for immunotherapy. Furthermore, the following are described herein: - Antigenic peptide according to the present invention, - Immunogenic compounds according to the present invention, -Napnea according to the present invention, -Cytotoxic T lymphocytes (CTLs) according to the present invention, - Cells according to the present invention - Nucleic acids according to the present invention, - Host cell according to the present invention, - Pharmaceutical composition according to the present invention, - A kit according to the present invention, or -The combination according to the present invention can be used as a vaccine, in particular as a vaccine for (cancer) immunotherapy.

[0251] As used in the context of the present invention, the term "vaccine" typically means a (biological) preparation that provides innate and / or adaptive immunity against a particular disease, preferably a B-cell malignancy. Therefore, vaccines, in particular, support the innate and / or adaptive immune responses of the immune system being treated. For example, the antigenic peptides according to the present invention typically induce or support an adaptive immune response in the patient being treated.

[0252] In the context of the present invention, the vaccine (composition) can induce a specific immune response against tumor antigens and is therefore preferably used to prevent or treat B-cell malignancies.

[0253] Accordingly, in preferred embodiments, the present invention relates to the compositions defined above for use in the prevention and / or treatment of cancer in subjects requiring it. More preferably, the present invention relates to the use of the compositions of the present invention for producing agents for the prevention or treatment of cancer in subjects requiring it. In other words, the present invention relates to a method for the prevention or treatment of cancer in subjects requiring it, comprising administering an effective amount of the compositions of the present invention to the subjects.

[0254] Preferably, as described herein. - Antigenic peptide according to the present invention, - Immunogenic compounds according to the present invention, -Napnea according to the present invention, -Cytotoxic T lymphocytes (CTLs) according to the present invention, - Cells according to the present invention - Nucleic acids according to the present invention, - Host cell according to the present invention, - Pharmaceutical composition according to the present invention, - A kit according to the present invention, or - The cancers targeted for prevention and / or treatment by the combination therapy according to the present invention relate to the (reference) tumor antigens of the antigenic peptides described herein. That is, a suitable antigenic peptide can be selected by (i) selecting a suitable tumor antigen for a particular type of cancer known in the art, and (ii) selecting a suitable antigenic peptide according to the present invention for the selected tumor antigens described above, for example, in 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 malignancies targeted for prevention or treatment, and / or based on human genes / human tumor antigens involved in the B-cell malignancies.

[0255] Generally, the antigenic peptide of the present invention can be administered "naked," or in the form of an immunogenic compound according to the present invention, cells according to the present invention loaded with it, 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.

[0256] In preferred embodiments, they can be administered in the form of microorganisms, such as enterobacteriaceae. Whole enterobacteriaceae may also be advantageous because they have the potential to elicit a higher immune response than the (poly)peptides or nucleic acids they contain. Alternatively, the enterobacteriaceae according to the present invention may be in the form of probiotics, i.e., live enterobacteriaceae, and therefore can be used as a food additive due to the health benefits they can provide. They can be freeze-dried, for example, as granules, pills, or capsules, or mixed directly with dairy products for consumption.

[0257] Methods of administration are well known to those skilled in the art. With respect to the compositions of the present invention, they can be administered directly to the subject, to the affected organ (i.e., topically) or systemically (i.e., enterally or parenterally), or ex vivo to cells derived from the subject or human cell lines to be later administered to the subject, or in vitro to select a subpopulation of immune cells derived from the subject and later re-administer to the subject. Enteral administration includes oral and rectal administration, as well as administration via gastric tube, duodenal tube, or gastrostomy. Parenteral administration includes, in particular, subcutaneous, intravenous, intramuscular, intra-arterial, intradermal, intraosseous, intracerebral, and intrathecal injection. The method of administration often depends on the antigenic peptide and / or immunogenic compound present in the composition, as well as the type of cancer being treated and other activators that may be included in the composition. For example, if the immunogenic compound is a nucleic acid as defined above, the administration is preferably intramuscular or intradermal injection, and if the nucleic acid is cloned into a viral vector, oral / nasal administration is particularly preferred. Alternatively, if the antigenic peptide and / or immunogenic compound is a (poly)peptide as defined above or loaded into nanoparticles as described herein, the administration is preferably intramuscular, intradermal, or oral. However, more alternatively, if the antigenic peptide and / or immunogenic compound is delivered in the form of enteric bacteria as defined above, in particular if the enteric bacteria are in the form of probiotics, the administration is preferably oral.

[0258] The antigenic peptides, immunogenic compounds, and nucleic acids according to the present invention can be further encapsulated to facilitate their administration to targets requiring them. For example, they can be encapsulated in peptide nanocarriers (preferably when the immunogenic compound is a nucleic acid or (poly)peptide), viromosomes (preferably when the immunogenic compound is a nucleic acid or (poly)peptide), or lipid-based carrier systems such as liposome-polycation-DNA complexes (preferably when 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., Peptide Vaccine: Progress and Challenges. Vaccines (Basel). 2014 Jul 2;2(3):515-36).

[0259] The composition may also be administered two or more times to achieve the desired effect. In a preferred embodiment, the composition is administered repeatedly, at least two times, preferably three or more times. This can be done over a long period, such as weekly, bi-weekly, monthly, yearly, or for several years, after the initial administration, to ensure that the target is adequately immunized.

[0260] Combination therapy The antigenic peptide, immunogenic compound, nanoparticles, cells, nucleic acids, host cells, and pharmaceutical compositions according to the present invention can be administered, in particular, by method and use according to the present invention, either alone or in combination with adjuvants useful for the treatment and / or prevention of cancer, such as anticancer agents.

[0261] Therefore, the therapeutic agent can preferably prevent and / or treat the same type of cancer in which the antigenic peptide according to the present invention is used. Particularly preferred anticancer therapeutic agents according to the present invention include, but are not limited to, antibodies, CAR-T cells, tumor cell lysates, chemotherapeutic agents, radiotherapeutic agents, immune checkpoint modulators, and combinations thereof.

[0262] Antibodies are particularly advantageous in cancer treatment because they can bind to specific antigens on the surface of cancer cells and direct treatment to the tumor (i.e., these are called tumor-targeting antibodies), or because they can block immune checkpoints that are dysregulated in cancer (i.e., these are referred to herein as immunomodulatory antibodies). The purpose of the latter type of antibody is to inhibit cancer's immune resistance, which is particularly observable against tumor antigen-specific T cells. In fact, as is well known in the art, under normal physiological conditions, immune checkpoints are important for maintaining self-tolerance (i.e., preventing autoimmunity) and protecting tissues from damage when the immune system responds to pathogenic infection. However, in cancer, the expression of immune checkpoints can become dysregulated as an important mechanism of immune resistance. The aforementioned resistance is significantly observed in melanoma, ovarian cancer, lung cancer, glioblastoma, breast cancer, and pancreatic cancer with respect to the PD-L1 checkpoint (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) T lymphocyte-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 for malignant melanoma. Cancer. 2010 Apr 1;116(7):1757-66).Other examples of immune checkpoints include, but are not limited to, PD-L2, PD-1, CD80, CD86, CTLA4, B7H3, B7H4, PVR, TIGIT, GAL9, LAG-3, GITR, CD137, TIM3, VISTA, and VISTA-R (Pico de Coana 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 cancer immunotherapy. Nat Rev Cancer. 2012 Mar 22;12(4):252-64).

[0263] Antibodies are typically used for the aforementioned purposes either as naked monoclonal antibodies (i.e., unconjugated) or in a conjugated form with another molecule that may be toxic to cells or radioactive.

[0264] Well-known monoclonal tumor target antibodies used in cancer immunotherapy include alemtuzumab (chronic lymphocytic leukemia), bevacizumab (colorectal cancer, glioblastoma multiforme, cervical cancer, lung cancer, renal cancer), brentuximab / vedotin (lymphoma), blinatumab (acute lymphoblastic leukemia), catumakisomab (malignant ascites in EPCAM+ cancer), cetuximab (head and neck cancer, colorectal cancer), denosumab (breast cancer, prostate cancer, and bone cancer), gemtuzumab / ozogamicin (acute myeloid leukemia), and ibritumab. Examples include, but are not limited to, tiuxetan (non-Hodgkin 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 lymphoma), siltuximab (multicentric Castleman disease), tocitumomab (non-Hodgkin lymphoma), and trastuzumab (breast cancer, gastric cancer, and gastroesophageal cancer). On the other hand, examples of immunomodulatory antibodies include, but are not limited to, ipilimumab (melanoma), which blocks the CTLA4-dependent immune checkpoint; nivolumab (melanoma, lung cancer) and prembrolizubumab (melanoma), which both block the PDCD1-dependent immune checkpoint; and MPDL3280A, MEDI4736, MEDI0680, and MSB0010718C (Sharma and Allison, The future of immune checkpoint therapy. Science. 2015 Apr 3;348(6230):56-61), which all block the PD-L1-dependent immune checkpoint.

[0265] Other antibodies for cancer immunotherapy are listed in Buque 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. Mol Immunol. 2015 Oct;67(2 Pt A):28-45), and Simpson and Caballero, Monoclonal antibodies for the therapy of cancer MC Proc. 2014; 8(Suppl 4): O6, as well as on the Antibody Society website (website link http: / / www.antibodysociety.org / news / approved_mabs.php, which lists therapeutic monoclonal antibodies approved or under review in the European Union or the United States).

[0266] Adoptive cell immunotherapy using chimeric antigen receptor (CAR) T cells is transforming the treatment of B-cell non-Hodgkin lymphoma (NHL), particularly advanced B-cell lymphoma. For example, CD19-targeted CAR T cells are a new standard of treatment for DLBCL patients who are resistant to at least two existing lines of treatment. Two CAR T cell products, axicabtagene ciloleucel (axi-cel) (KTE-019) (YESCARTA) TM ) and tisagenlecleucel(CTL019)(KYMRIAH TM ) has received approval from the U.S. Food and Drug Administration for the treatment of DLBCL that is resistant after two lines of treatment. A third product, lisocabtagene maraleucel (liso-cel) (JCAR017), is currently in clinical trials. Other CAR T cells include CD20-CAR-T cells.

[0267] Tumor cell lysates can also be used in combination with antigenic peptides according to the present invention. Tumor cells can actually prime an immune response by presenting endogenous peptide-MHC complexes and via host dendritic cells (DCs) that can process and present antigens delivered by the lysates. This broadens 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).

[0268] Standard chemotherapy and radiotherapy agents are described in detail in the literature, particularly by Baskar et al. (*Cancer and radiation therapy: current advances and future directions. Int J Med Sci. 2012;9(3):193-9), 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. (*Review of therapeutic drug monitoring of anticancer drugs part two--targeted therapies. Eur J Cancer. 2014 Aug;50(12):2020-36), and therefore do not need to be described further herein. A list of such drugs and medications is also available on the cancer.gov website (http: / / www.cancer.gov / about-cancer / treatment / drugs).

[0269] Preferably, an immune checkpoint modulator for use with an antigenic peptide as 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.

[0270] More preferably, the immune checkpoint modulator is an activator of a (co)stimulatory checkpoint molecule or an inhibitor of an inhibitory checkpoint molecule, or a combination thereof. Therefore, the immune checkpoint modulator is more preferably (i) an activator of 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.

[0271] More preferably, the immune checkpoint modulator is an inhibitor of an inhibitory checkpoint molecule (but preferably not an inhibitor of an stimulant checkpoint molecule). Therefore, the immune checkpoint modulator is 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 an inhibitor of their ligands.

[0272] Furthermore, the immune checkpoint modulator is preferably an activator of an irritant or co-stimulatory checkpoint molecule (but preferably not an activator of an inhibitory checkpoint molecule). Therefore, the immune checkpoint modulator is more preferably an activator of CD27, CD28, CD40, CD122, CD137, OX40, GITR, and / or ICOS, or an activator of their ligands.

[0273] The immune checkpoint modulator is more preferably a modulator of the CD40 pathway, the IDO pathway, the LAG3 pathway, the CTLA-4 pathway, and / or the PD-1 pathway. In particular, 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.

[0274] Therefore, the checkpoint modulator for use with the antigenic peptide can be selected from known modulators of the CTLA-4 pathway or the PD-1 pathway. Preferably, the checkpoint modulator for use with the antigenic peptide as defined herein can be selected from known modulators of the CTLA-4 pathway or the PD-1 pathway. Particularly preferably, the immune checkpoint modulator is a PD-1 inhibitor. Preferred inhibitors of the CTLA-4 and PD-1 pathways include Yervoy® (Ipilimumab; Bristol Myers Squibb) and Tremelimumab (Pfizer / MedImmune), as well as Opdivo® (Nivolumab; Bristol Myers Squibb), Keytruda® (Pembrolizumab (also known as Lambrolizumab or MK-3475); Merck), Imfinzi® (Durvalumab (also known as MEDI4736); MedImmune / AstraZeneca), Tecentriq® (Atezolizumab (also known as MPDL3280A); Roche / Genentech), Pidilizumab (CT-011; CureTech), MEDI0680 (AMP-514; AstraZeneca), and Bavencio® (Avelumab; Merck) Examples include monoclonal antibodies for KGaA / Pfizer (also known as MSB-0010718C), MIH1 (Affymetrix), LY3300054 (Eli Lilly), and Spartalizumab (also known as PDR001; Novartis).More preferred checkpoint inhibitors include the CTLA-4 inhibitors Yervoy® (Ipilimumab; Bristol Myers Squibb) and Tremelimumab (Pfizer / MedImmune), and the PD-1 inhibitors Opdivo® (Nivolumab; Bristol Myers Squibb), Keytruda® (Pembrolizumab; Merck), Pidilizumab (CT-011; CureTech), MEDI0680 (AMP-514; AstraZeneca), and AMP-224 (PD-L2Fc fusion protein; MedImmune).

[0275] Immunotherapy checkpoint modulators for use in combination with antigenic peptides as defined herein are also selected from the group consisting of Pembrolizumab, Ipilimumab, Nivolumab, Atezolizumab, MEDI4736, Durvalumab, Tremelimumab, Avelumab, Spartalizumab, LAG525 (anti-LAG-3 monoclonal antibody), Epacadostat (formally INCB24360; IDO inhibitor), Varlilumab (anti-CD27 monoclonal antibody), Urelumab (anti-CD137 monoclonal antibody), AMP-224, and CM-24 (anti-CEACAM1 monoclonal antibody).

[0276] Selecting an appropriate immunoanti-cancer agent for the purposes of this invention is within the scope of the skills of those skilled in the art.

[0277] The aforementioned anticancer therapeutic agents can also be administered together with the antigenic peptides, immunogenic compounds, nanoparticles, cells, nucleic acids, host cells, or pharmaceutical compositions according to the present invention, in the same or different pharmaceutical forms, almost simultaneously or sequentially, as described herein. Accordingly, the present invention proposes the combined use of the compositions of the present invention with at least one of the aforementioned anticancer therapeutic agents for simultaneous, separate, or sequential administration as described herein.

[0278] Furthermore, the present invention also relates to the combination of at least two different antigenic peptides according to the present invention for use, for example, in the prevention and / or treatment of B-cell malignancies. Furthermore, the present invention also relates to the combination of at least two different immunogenic compounds according to the present invention for use, for example, in the prevention and / or treatment of B-cell malignancies. Furthermore, the present invention also relates to the combination of at least two different nanoparticles according to the present invention for use in the prevention and / or treatment of B-cell malignancies. Furthermore, the present invention also relates to the combination of at least two different nucleic acids according to the present invention for use in the prevention and / or treatment of B-cell malignancies.

[0279] Therefore, according to a preferred embodiment, at least two antigenic peptides according to the present invention can be administered in combination, for example, in the same pharmaceutical composition. For example, at least three antigenic peptides, at least four antigenic peptides, at least five antigenic peptides, at least six antigenic peptides, at least seven antigenic peptides, at least eight antigenic peptides, at least nine antigenic peptides, at least ten antigenic peptides, at least eleven antigenic peptides, at least twelve antigenic peptides, at least thirteen antigenic peptides, at least fourteen antigenic peptides, at least fifteen antigenic peptides, at least twenty antigenic peptides, at least twenty-five antigenic peptides, at least fifty antigenic peptides, and at least one hundred antigenic peptides can 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 skill of a person skilled in the art.

[0280] In a particularly preferred embodiment, two different antigenic peptides according to the present invention (for example, relating to the same type of B-cell malignancy and / or the same reference antigen) are used in combination. For example, (i) at least two different immunogenic compounds according to the present invention; (ii) at least two different antigenic peptides according to the present invention; (iii) at least two different nanoparticles according to the present invention; or (iv) At least two different nucleic acids according to the present invention can be used in combination.

[0281] For example, the present invention preferably relates to at least two different antigenic peptides according to the present invention for use in the prevention and / or treatment of B-cell malignancies, (i) A first antigenic peptide according to the present invention, (ii) To provide a combination with a second antigenic peptide (different from the first) according to the present invention.

[0282] For example, the present invention preferably relates to at least two different immunogenic compounds according to the present invention for use in the prevention and / or treatment of B-cell malignancies, in particular, (i) an immunogenic compound according to the present invention comprising a first antigenic peptide according to the present invention, (ii) To provide a combination with an immunogenic compound according to the present invention, which comprises a second antigenic peptide (different from the first) according to the present invention.

[0283] For example, the present invention preferably relates to at least two different nanoparticles according to the present invention for use in the prevention and / or treatment of B-cell malignancies, in particular, (i) Nanoparticles according to the present invention comprising a first antigenic peptide according to the present invention, (ii) To provide a combination with nanoparticles according to the present invention that include a second antigenic peptide (different from the first) according to the present invention.

[0284] For example, the present invention preferably relates to at least two different nucleic acids according to the present invention for use in the prevention and / or treatment of B-cell malignancies, (i) A nucleic acid according to the present invention comprising a polypeptide encoding a first antigenic peptide according to the present invention, (ii) To provide a combination with nucleic acid according to the present invention, which comprises a polypeptide encoding a second antigenic peptide (different from the first) according to the present invention.

[0285] For example, the present invention preferably relates to at least two different cytotoxic T lymphocytes according to the present invention for use in the prevention and / or treatment of B-cell malignancies, in particular, (i) Cytotoxic T lymphocytes according to the present invention that are specific to the first antigenic peptide according to the present invention, (ii) To provide a combination with cytotoxic T lymphocytes according to the present invention that are specific to a second antigenic peptide (different from the first) according to the present invention.

[0286] In the combination according to the present invention, such combination of components such as antigenic peptides according to the present invention is preferred, which corresponds to the preferred combination of components such as antigenic peptides according to the present invention contained in the pharmaceutical composition described above.

[0287] Furthermore, the antigenic peptide according to the present invention can also be used in combination with the corresponding (human) tumor antigen epitope (as described above with respect to the peptide "family"). This enables / supports highly efficient selection of T cell clones against tumors. In particular, the antigenic peptide according to the present invention and the corresponding (human) tumor antigen epitope can be co-administered. Such co-administration can be almost simultaneous or sequential, and in sequential administration, it is preferable that the antigenic peptide according to the present invention is administered first, followed by the corresponding (human) tumor antigen epitope. In particular, the antigenic peptide according to the present invention can be administered first, and the corresponding (human) tumor antigen epitope can be used as a (re)boost. For example, the antigenic peptide according to SEQ ID NO: 10 can be used in combination with the reference peptide according to SEQ ID NO: 258. In another example, the antigenic peptide according to SEQ ID NO: 110 can be used in combination with the reference peptide according to SEQ ID NO: 270. In another example, the antigenic peptide according to SEQ ID NO: 114 can be used in combination with the reference peptide according to SEQ ID NO: 271. In another example, the antigenic peptide according to SEQ ID NO: 220 can be used in combination with the reference peptide according to SEQ ID NO: 279.

[0288] (a) an antigenic peptide according to the present invention and a corresponding (human) tumor antigen epitope or (b) a peptide used in combination, such as two different antigenic peptides according to the present invention, -In the same immunogenic compound according to the present invention or in a different immunogenic compound according to the present invention, - Loaded into the same nanoparticles according to the present invention or into different nanoparticles according to the present invention, -Loaded into the same cell or different cells according to the present invention, -The same nucleic acid or different nucleic acids (encoded by) according to the present invention, - Expressed by the same host cell according to the present invention or by different host cells according to the present invention, or -It can be administered (by being included) in the same pharmaceutical composition according to the present invention or in different pharmaceutical compositions according to the present invention.

[0289] In general, for example, the expression “two different components” in the context of combination therapy for use according to the present invention is defined above (in the context of the pharmaceutical composition). In particular, (1) A first component such as an antigenic peptide according to the present invention as described herein, an immunogenic compound according to the present invention as described herein, a nanoparticle according to the present invention as described herein, a cell according to the present invention as described herein, a nucleic acid according to the present invention as described herein, a host cell according to the present invention as described herein, or a pharmaceutical composition according to the present invention as described herein; and (2) Meaning a second component (different from the first component), such as the anticancer therapeutic agent described above, a different antigenic peptide according to the present invention as described herein, a different immunogenic compound according to the present invention as described herein, a different nanoparticle according to the present invention as described herein, a different cell according to the present invention as described herein, a different nucleic acid according to the present invention as described herein, a different host cell according to the present invention as described herein, a different pharmaceutical composition according to the present invention as described herein, or one or more (fragments) of a human tumor antigen ("naked" as an immunogenic compound according to the present invention, a nanoparticle according to the present invention, a (host) cell according to the present invention, a nucleic acid according to the present invention, or a pharmaceutical composition according to the present invention).

[0290] Therefore, the “two different components” referred to herein in the context of combination therapy for use according to the present invention are preferably active components in the context of the disease to be prevented and / or treated (B-cell malignancies). In other words, each of at least two different components may also be useful for preventing and / or treating the cancer even when administered separately (rather than in combination as described herein), although the combination (i.e., combination administration) usually synergistically enhances their prophylactic and / or therapeutic effects (e.g., immune response).

[0291] Accordingly, the present invention also provides a combination of at least two different antigenic peptides according to the present invention as described herein. In this context, the at least two different antigenic peptides can be in any form, e.g., "naked," such as being contained in immunogenic compounds, nanoparticles, (pharmaceutical) compositions, or cells into which they are loaded, or encoded by nucleic acids (e.g., vectors). Thus, the at least two different antigenic peptides can be contained in at least two different components (combined). In a preferred embodiment, the at least two different components of the combination according to the present invention are at least two different antigenic peptides according to the present invention (in any form, e.g., contained in immunogenic compounds, nanoparticles, cells, pharmaceutical compositions, or encoded by nucleic acids, etc.).

[0292] Preferably, at least two different components of the combination for use according to the present invention relate to the same type of cancer, for example, the same or different antigens relating to said cancer and / or the same or different (reference) epitopes within the antigen relating to said cancer. More preferably, the at least two different components relate to the same tumor antigen.

[0293] In certain embodiments, the at least two distinct components for use according to the present invention are contained in the same or different compositions. In certain embodiments, the at least two distinct components for use according to the present invention are administered via the same or different routes of administration. In certain embodiments, the at least two distinct components for use according to the present invention are administered substantially simultaneously (concurrently) or sequentially.

[0294] Preferably, the at least two different components used in combination according to the present invention are administered approximately simultaneously. More generally, the first component is preferably administered approximately simultaneously with the second component, and the at least two different components used in combination according to the present invention are preferably administered in the same form (i.e., as the same type of formulation, e.g., nanoparticles, pharmaceutical composition, etc.).

[0295] As used herein, “approximately simultaneously” means, in particular, simultaneous administration, or administration of the second component immediately after administration of the first component, or administration of the first component immediately after administration of the second component. Those skilled in the art will understand that “immediately after” includes the time required to prepare the second dose, in particular the time required to expose and disinfect the site for the second dose, and the appropriate adjustment of the “administration device” (e.g., syringe, pump, etc.). Simultaneous administration also includes cases where the administration periods of the first and second components overlap, or, for example, one component is administered by infusion or the like over a longer period, such as 30 minutes, 1 hour, 2 hours, or more, and the other component is administered at some point during such a long period. When different routes of administration and / or different administration sites are used, it is particularly preferable to administer the first and second components approximately simultaneously.

[0296] Furthermore, it is preferable that the at least two different components of the combination for use according to the present invention be administered sequentially. More generally, it is preferable that the first component and the second component be administered sequentially, and that the at least two different components of the combination for use according to the present invention be administered in the same form (i.e., as the same type of formulation, e.g., nanoparticles, pharmaceutical composition, etc.).

[0297] This means that the first component is administered before or after the second component. In continuous administration, the time between the administration of the first component and the administration of the second component is preferably within one week, more preferably within three days, even more preferably within two days, and most preferably within 24 hours. It is particularly preferable that the first component and the second component are administered on the same day, and the time between the administration of the first component and the administration of the second component is preferably within six hours, more preferably within three hours, even more preferably within two hours, and most preferably within one hour.

[0298] Preferably, the first and second components are administered via the same route of administration. More generally, it is preferable that the first and second components are administered via the same route of administration, and the at least two different components of the combination for use according to the present invention are preferably administered in the same form (i.e., as the same type of formulation, e.g., nanoparticles, pharmaceutical composition, etc.).

[0299] Furthermore, it is preferable that the at least two different components of the combination for use according to the present invention are administered via separate routes of administration. More generally, it is preferable that the first and second components are administered via separate routes of administration, and that the at least two different components of the combination for use according to the present invention are administered in the same form (i.e., as the same type of formulation, e.g., nanoparticles, pharmaceutical composition, etc.).

[0300] Preferably, the at least two different components for use according to the present invention are contained in the same composition. More generally, it is preferable that the first and second components are contained in the same composition, and the at least two different components for use according to the present invention are preferably administered in the same form (i.e., as the same type of formulation, such as nanoparticles).

[0301] Furthermore, it is preferable that the at least two different components for use according to the present invention are contained in different compositions. More generally, it is preferable that the first component and the second component are contained in different compositions, and the at least two different components for use according to the present invention are preferably administered in the same form (i.e., as the same type of formulation, such as nanoparticles).

[0302] In particular, the present invention provides a pharmaceutical composition comprising a first antigenic peptide according to the present invention, which comprises or consists of a sequence variant of the human tumor antigen fragment.

[0303] In particular, the present invention provides a combination for use in the prevention and / or treatment of B-cell malignancies, for example, comprising a first antigenic peptide according to the present invention comprising or consisting of a microbiota sequence variant of the human tumor antigen CD22 fragment, and a second antigenic peptide according to the present invention comprising or consisting of a sequence variant of the human tumor antigen TNFRSF13C fragment. 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 represented by SEQ ID NOs: 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 represented by SEQ ID NOs: 220, 325, and 450. More preferably, the first antigenic peptide comprises or consists of amino acid sequences represented by SEQ ID NOs: 110, 387, and 390, and the second antigenic peptide comprises or consists of amino acid sequences represented by SEQ ID NOs: 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.

[0304] More preferably, the combination according to the present invention (for example, for use in the prevention and / or treatment of B-cell malignancies) comprises at least three different components as described above, in particular at least three different antigenic peptides according to the present invention. The above description relating to the combination of two different components also applies to the case of three different components.

[0305] Most preferably, the combination according to the present invention (for example, for use in the prevention and / or treatment of B-cell malignancies) comprises at least four different components as described above, in particular at least four different antigenic peptides according to the present invention. The above description relating to the combination of two different components also applies to the case of four different components.

[0306] For example, the combinations for use in the prevention and / or treatment of B-cell malignancies are understood to include, instead of the preferred combinations of the antigenic peptides, combinations of the immunogenic compounds of the present invention, combinations of the nanoparticles of the present invention, or combinations of the nucleic acids of the present invention.

[0307] A brief description of the attached drawings is provided below. These drawings are intended to illustrate the present invention in more detail. However, these drawings are not intended to limit the subject matter of the present invention in any way. [Brief explanation of the drawing]

[0308] [Figure 1] Figure 1 shows the in vitro affinity of the antigenic peptide CD22-B1 compared to the corresponding human CD22 epitope CD22-H1 for Example 1. [Figure 2] Figure 2 shows the in vitro affinity of the antigenic peptide CD37-B1 compared to the corresponding human CD37 epitope CD37-H1 for Example 1. [Figure 3] Figure 3 shows the in vitro affinity of the antigenic peptide CD19-B1 compared to the corresponding human CD19 epitope CD19-H1, with respect to Example 1. [Figure 4] Figure 4 shows the in vitro affinity of the antigenic peptide CD19-B2 compared to the corresponding human CD19 epitope CD19-H2, for Example 1. [Figure 5] Figure 5 shows the in vitro affinity of the antigenic peptide TNFRSF13C-B1 compared to the corresponding human TNFRSF13C epitope TNFRSF13C-H1, with respect to Example 1. [Figure 6] Figure 6 shows the ELISPOT results for cross-reactivity between HHD DR1 HLA-A2 transgenic mice vaccinated with the antigenic peptide CD22-B1 and the corresponding human peptide CD22-H1, as shown in the figure, for Example 2. The data are shown as the number of spots per T cell totaling 1.106. [Figure 7]Figure 7 shows the ELISPOT results for cross-reactivity between HHD DR1 HLA-A2 transgenic mice vaccinated with the antigenic peptide TNFRSF13C-B1 and the corresponding human peptide TNFRSF13C-H1, as shown in the figure, for Example 2. The data are shown as the number of spots per T cell totaling 1.106. [Figure 8] Figure 8 shows the ELISPOT results for cross-reactivity between HHD DR1 HLA-A2 transgenic mice vaccinated with the antigenic peptide CD37-B1 and the corresponding human peptide CD37-H1, as shown in the figure, for Example 2. The data are shown as the number of spots per T cell totaling 1.106. [Figure 9] Figure 9 shows the ELISPOT results for cross-reactivity between HHD DR1 HLA-A2 transgenic mice vaccinated with the antigenic peptide CD19-B2 and the corresponding human peptide CD19-H2, as shown in the figure, for Example 2. The data are shown as the number of spots per T cell totaling 1.106. [Figure 10] Figure 10 shows the ELISPOT results for cross-reactivity between HHD DR1 HLA-A2 transgenic mice vaccinated with the antigenic peptide CD19-B1 and the corresponding human peptide CD19-H1, as shown in the figure, for Example 2. The data are shown as the number of spots per T cell totaling 1.106. [Figure 11] Figure 11 shows the ELISPOT results for cross-reactivity between HHD DR3 HLA-A2 transgenic mice vaccinated with the antigenic peptide CD22-B1 and the corresponding human peptide CD22-H1, as shown in the figure, for Example 2. The data are shown as the number of spots per T cell totaling 1.106. [Figure 12]Figure 12 shows the ELISPOT results for cross-reactivity between HHD DR3 HLA-A2 transgenic mice vaccinated with the antigenic peptide TNFRSF13C-B1 and the corresponding human peptide TNFRSF13C-H1, as shown in the figure, for Example 2. The data are shown as the number of spots per T cell totaling 1.106. [Figure 13] Figure 13 shows the ELISPOT results for cross-reactivity between HHD DR3 HLA-A2 transgenic mice vaccinated with the antigenic peptide CD37-B1 and the corresponding human peptide CD37-H1, as shown in the figure, for Example 2. The data are shown as the number of spots per T cell totaling 1.106. [Figure 14] Figure 14 shows the in vitro affinity of the antigenic peptide MS4A1-B4 compared to the corresponding human MS4A1 epitope MS4A1-H4, with respect to Example 1. [Figure 15] Figure 15 shows the ELISPOT results for cross-reactivity between HHD DR1 HLA-A2 transgenic mice vaccinated with the antigenic peptide MS4A1-B4 and the corresponding human peptide MS4A1-H4, as shown in the figure, for Example 2. The data are shown as the number of spots per T cell totaling 1.106. [Figure 16] Figure 16 shows the ELISPOT results for cross-reactivity between HHD DR3 HLA-A2 transgenic mice vaccinated with the antigenic peptide MS4A1-B4 and the corresponding human peptide MS4A1-H4, as shown in the figure, for Example 2. The data are shown as the number of spots per T cell totaling 1.106. [Figure 17] Figure 17 shows the detection of CD22-B1, CD37B1, TNFRSF13C-B1, and MS4A1-B4 peptide-specific CD8+ T cells in peripheral blood from a healthy donor (HLA-A2 positive) in Example 3. [Figure 18-1]Figure 18 shows the cytotoxic activity of CD22-B1, CD37B1, TNFRSF13C-B1, and MS4A1-B4 peptide-specific human T cell clones grown in vitro by microbiome-derived peptide stimulation, relating to Example 3. CD22-B1, CD37B1, TNFRSF13C-B1, and MS4A1-B4 peptide-specific T cells have the ability to kill T2 cells loaded with bacterial or human peptides. [Figure 18-2] Figure 18 shows the cytotoxic activity of CD22-B1, CD37B1, TNFRSF13C-B1, and MS4A1-B4 peptide-specific human T cell clones grown in vitro by microbiome-derived peptide stimulation, relating to Example 3. CD22-B1, CD37B1, TNFRSF13C-B1, and MS4A1-B4 peptide-specific T cells have the ability to kill T2 cells loaded with bacterial or human peptides. [Figure 19] Figure 19 shows the in vitro affinity of the antigenic peptides CD22-B1, CD22-B12, and CD22-B13 compared to the corresponding human CD22 epitope CD22-H1, for Example 4. [Figure 20] Figure 20 shows the in vitro affinity of the antigenic peptides CD37-B1, CD37-B12, CD37-B13, CD37-B14, and CD37-B15 compared to the corresponding human CD37 epitope CD37-H1, for Example 4. [Figure 21] Figure 21 shows the in vitro affinity of the antigenic peptides TNFRSF13C-B1, TNFRSF13C-B11, TNFRSF13C-B12, and TNFRSF13C-B13 compared to the corresponding human TNFRSF13C epitope TNFRSF13C-H1, in relation to Example 4. [Figure 22] Figure 22 shows the in vitro affinity of the antigenic peptides MS4A1-B4, MS4A1-B42, and MS4A1-B43 compared to the corresponding human MS4A1 epitope MS4A1-H4, with respect to Example 4. [Examples]

[0309] The following are specific examples illustrating various embodiments and aspects of the present invention. However, the present invention is not limited in scope by the specific embodiments described herein. The following preparation examples and embodiments are provided so that those skilled in the art may better understand and implement the present invention. However, the present invention is not limited in scope by the exemplary embodiments, and the embodiments are intended only to illustrate one aspect of the present invention, and functionally equivalent methods are within the scope of the present invention. In fact, in addition to those described herein, various modifications of the present invention will be readily apparent to those skilled in the art from the above description, the accompanying drawings, and the following embodiments. All such modifications are within the scope of the appended claims.

[0310] Example 1: The antigenic peptide is HLA-A * It has superior affinity for the 0201 allele. Next, HLA-A of various selected antigenic peptides and corresponding fragments of human tumor antigens (human reference peptides) *The binding affinity to the 0201 allele was confirmed in vitro. Specifically, the antigenic peptide of SEQ ID NO: 110 ("YIFEHPELLL," also known herein as CD22-B1) was compared with the corresponding reference human peptide derived from CD22 ("WVFEHPETL," SEQ ID NO: 270, also known herein as CD22-H1). Furthermore, the antigenic peptide of SEQ ID NO: 109 ("YVFEHPELL," also known herein as CD22-B11) was compared with the corresponding reference human peptide derived from CD22 ("WVFEHPETL," SEQ ID NO: 270, also known herein as CD22-H1). Furthermore, the antigenic peptide of SEQ ID NO: 114 ("FLAFVPLQL," also known herein as CD37-B1) was compared with the corresponding reference human peptide derived from CD37 ("GLAFVPLQI," SEQ ID NO: 271, also known herein as CD37-H1). Furthermore, the antigenic peptide of SEQ ID NO: 117 ("GMAFVPLLL," also known herein as CD37-B11) was compared with the corresponding reference human peptide derived from CD37 ("GLAFVPLQI," SEQ ID NO: 271, also known herein as CD37-H1). Furthermore, the antigenic peptide of SEQ ID NO: 34 ("LLVGILHLV," also known herein as CD19-B1) was compared with the corresponding reference human peptide derived from CD19 ("SLVGILHLQ," SEQ ID NO: 260, also known herein as CD19-H1). Furthermore, the antigenic peptide of SEQ ID NO: 10 ("TLLFLTPML," also known herein as CD19-B2) was compared with the corresponding reference human peptide derived from CD19 ("FLLFLTPME," SEQ ID NO: 258, also known herein as CD19-H2). Furthermore, the antigenic peptide of SEQ ID NO: 39 ("YLAYLIFEL," also known herein as CD19-B6) was compared with the corresponding reference human peptide derived from CD19 ("TLAYLIFCL," SEQ ID NO: 261, also known herein as CD19-H6). In addition, the antigenic peptide of SEQ ID NO: 40 ("LQMGGFYLL," also known herein as CD19-B7) was compared with the corresponding reference human peptide derived from CD19 ("QQMGGFYLC," SEQ ID NO: 262, also known herein as CD19-H7).Furthermore, the antigenic peptide of SEQ ID NO: 220 ("LMFGAPALV," also known herein as TNFRSF13C-B1) was compared with the corresponding reference human peptide derived from TNFRSF13C ("LLFGAPALL," SEQ ID NO: 279, also known herein as TNFRSF13C-H1). Furthermore, the antigenic peptide of SEQ ID NO: 231 ("ILPGLLFGL," also known herein as TNFRSF13C-B31) was compared with the corresponding reference human peptide derived from TNFRSF13C ("PLPGLLFGA," SEQ ID NO: 280, also known herein as TNFRSF13C-H3). Furthermore, the antigenic peptide of SEQ ID NO: 227 ("FMPGLLFGA," also known herein as TNFRSF13C-B33) was compared with the corresponding reference human peptide derived from TNFRSF13C ("PLPGLLFGA," SEQ ID NO: 280, also known herein as TNFRSF13C-H3). Furthermore, the antigenic peptide of SEQ ID NO: 61 ("YILGGLLMV," also known herein as MS4A1-B12) was compared with the corresponding reference human peptide derived from MS4A1 (also known as CD20) ("IALGGLLMI," SEQ ID NO: 263, also known herein as MS4A1-H1). Furthermore, the antigenic peptide of SEQ ID NO: 72 ("ILIPAGIYL," also known herein as MS4A1-B3) was compared with the corresponding reference human peptide derived from MS4A1 (also known as CD20) ("LMIPAGIYA," SEQ ID NO: 265, also known herein as MS4A1-H3). Furthermore, the antigenic peptide of SEQ ID NO: 65 ("AMNSLSLYI," also known herein as MS4A1-B4) was compared with the corresponding reference human peptide derived from MS4A1 (also known as CD20) ("IMNSLSLFA," SEQ ID NO: 264, also known herein as MS4A1-H4). Furthermore, the antigenic peptide of SEQ ID NO: 86 ("YLFLGILSL," also known herein as MS4A1-B5) was compared with the corresponding reference human peptide derived from MS4A1 (also known as CD20) ("SLFLGILSV," SEQ ID NO: 266, also known herein as MS4A1-H5).

[0311] A. Materials and Methods A1. Measurement of peptide affinity to T2 cell lines The experimental protocol is HLA-A * This is similar to what was verified for the peptides presented by 0201 (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):3411-21). The affinity measurement of each peptide is performed using HLA-A * This is achieved in human tumor cells T2 that express the 0201 molecule but are TAP1 / 2 negative and unable to present endogenous peptides.

[0312] T2 cells (5.10 per well) 4 Cells are incubated in serum-free medium (TexMacs) supplemented with 100 ng / μl of β2-microglobulin, with peptide concentrations decreasing from 100 μM to 0.1 μM (4 points: 100 μM, 10 μM, 1 μM, 0.1 μM) at 37°C for 16 hours. The cells are then washed twice and marked with anti-HLA-A2 antibody (clone BB7.2, BD Pharmagen) conjugated to PE.

[0313] The analysis will be performed using FACS (Macsquant analyzer 10-Miltenyi).

[0314] For each peptide concentration, the geometric mean of the labels associated with the peptide of interest was subtracted from the background noise, and the HLA-A obtained with the reference peptide HIV pol 589-597 at a concentration of 100 μM was obtained. * The geometric mean percentage of the 0202 label is reported. Next, the relative affinity is determined as follows: Relative affinity = HLA-A *Concentrations of each peptide that induce 20% of O201 expression / HLA-A * The concentration of the reference peptide that induces 20% of 0201 expression.

[0315] A2. Solubilization of peptides Each peptide is solubilized considering its amino acid composition. For peptides that do not contain cysteine, methionine, or tryptophan, DMSO may be added up to a maximum of 10% of the total volume. Other peptides are resuspended in water or PBS (pH 7.4).

[0316] B. Results The average relative fluorescence intensity values ​​of T2 cells obtained at various concentrations of each peptide (the data are normalized to the average fluorescence of the HIV peptide; i.e., a value of 100 is equivalent to the best binding observed with the HIV peptide) are shown in Table 2 below. [Table 2]

[0317] Table 3 below summarizes the concentration required to induce 20% of HLA-A2 expression and the in vitro binding affinity for each peptide tested. * (Normalized to the HIV-pol concentration of the peptide that induces 20% of HLA-A2 expression in the same experiment). [Table 3]

[0318] Furthermore, Figures 1 to 5 and Figure 14 show the results of the selected examples, namely, the results of comparing the antigenic peptide CD22-B1 with the corresponding human CD22 fragment CD22-H1 (Figure 1), the results of comparing the antigenic peptide CD37-B1 with the corresponding human CD37 fragment CD37-H1 (Figure 2), the results of comparing the antigenic peptide CD19-B1 with the corresponding human CD19 fragment CD19-H1 (Figure 3), the results of comparing the antigenic peptide CD19-B2 with the corresponding human CD19 fragment CD19-H2 (Figure 4), the results of comparing the antigenic peptide TNFRSF13C-B1 with the corresponding human TNFRSF13C fragment TNFRSF13C-H1 (Figure 5), and the results of comparing the antigenic peptide MS4A1-B4 with the corresponding human MS4A1 fragment MS4A1-B4 (Figure 14).

[0319] In summary, the results indicate that the antigenic peptide according to the present invention is at least similar to the corresponding human tumor antigen fragment in terms of HLA-A * It is shown that the antigenic peptides exhibit binding affinity to 0201. In most cases, the binding affinity observed for the antigenic peptides according to the present invention was stronger than the binding affinity for the corresponding human epitopes. Although not bound by any theory, it is assumed that the strong binding affinity of the antigenic peptides according to the present invention reflects their ability to enhance the immune response (i.e., their immunogenicity).

[0320] Example 2: Immunogenicity of CD22-B1, CD19B1, CD19-B2, CD37B1, TNFRSF13C-B1, and MS4A1-B4 and cross-reactivity with corresponding human peptides in HLA-A2 transgenic mice. A. Materials and Methods A.1 Mouse Model Simply put, HLA-A2 HHD-DR1 humanized mouse (C57BL / 6JB2mtm1UncIAb- / -Tg(HLA-DRA, HLA-DRB1) * 0101)#GjhTg(HLA-A / H2-D / B2M)1Bpe) or HHD-DR3 humanized mouse (C57BL / 6JB2mtm1UncIAb- / -Tg(HLA-DRA, HLA-DRB1 *The mice were randomly assigned to experimental groups (based on sex and age) and immunized with a specific vaccine peptide (vacc-pAg) conjugated to a common helper peptide (h-pAg UCP2; sequence: KSVWSKLQSIGIRQH; SEQ ID NO: 475; for HHD DR1 mice or h-pAg DR3 mice; sequence: MAKTIAYDEEARRGLERGLN; SEQ ID NO: 473; for HHD DR3 mice) (scheduled in Table 4 below).

[0321] Table 4. Composition of the experimental group. h-pAg: "Helper" peptide; vacc-pAg: Vaccine peptide. The number of boost injections is shown in parentheses. [Table 4]

[0322] The peptides were prepared as follows: • vacc-pAg: CD22-B1, CD19-B1, CD19-B2, CD37-B1, TNFRSF13C-B1, and MS4A1-B4; all manufactured and prepared at a concentration of 4 mg / ml (4 mM); • Resuspend h-pAg:DR3 or UCP2 in pure distilled water at a concentration of 10 mg / mL.

[0323] The peptide formulation (emulsion) to be injected was freshly prepared for each group on the day of injection. Mixes for 10 animals were prepared using a 2 mL Luer-lock syringe (4606701V, B BRAUN) and a Luer connector (Cole-Parmer, 45502-22). 500 μL of the peptide mixture in syringe 1 was emulsified with 500 μL of IFA in syringe 2 as quickly as possible until a thick (white foamy) emulsion was formed. Each emulsion was prepared in excess to compensate for the dead volume at the time of injection.

[0324] The animals were immunized with a prime injection on day 0 (d0) and with a boost injection on day 14. 100 μL of oil-based emulsion was subcutaneously injected into the base of the tail of each mouse. This oil-based emulsion contained the following: • 60 nMole of vacc-pAg; 105 nMole of UCP2 helper peptide (in HHD-DR1 mice) or 65 nMole of DR3 helper peptide (in HHD-DR3 mice); • 10 μL of PBS to make a total volume of 50 μL (per mouse); • Incomplete Freund's adjuvant (IFA) was added in a 1:1 (v:v) ratio (50 μL per mouse).

[0325] A.2 Analysis Seven days after the boost injection (i.e., day 21), the animals were euthanized and their spleens were collected. Splenocytes were prepared by mechanically disrupting the organ, followed by 70 μm filtration and Ficoll density gradient purification.

[0326] The cell suspension was further used in the ELISPOT-IFNγ assay (Table 5). The cells were cultured in 200 μL of complete T cell medium. The experimental conditions (two sets) were as follows: total cell count per well was 2 × 10⁵ when cultured in the presence of various pAg (10 μM) or medium alone; total cell count was 2 × 10⁴ when cultured in the presence of CD3 / CD28-loaded bead particles (T cell activation / proliferation kit, 130-093-627, Miltenyi) (bead-to-cell ratio 1:1). The cultures were evaluated for IFNγ secretion ability according to the manufacturer's instructions (incubated for approximately 16-18 hours before assay) (Diaclone Kit Murine IFNγ ELISpot, 862.031-005PC). The peptides used for restimulation are listed in Table 5.

[0327] Table 5. Setup of the ELISPOT-IFNγ assay [Table 5]

[0328] Spots were counted using the CTL ELISpot reader. Data plotting and statistical analysis were performed using Prism-5 software (GraphPad Software Inc.).

[0329] B. Results All mice were 8–13 weeks old at the start of the experiment. Both males and females were used in this study. Animals were housed in groups of up to 6 per cage. At sacrifice, flow cytometry analysis of the spleen T cell population showed that the majority belonged to the CD4+ T cell subset.

[0330] After plating and incubation with appropriate stimulation, IFNγ-producing cells were generated and counted. The data totaled 1.10 6 The number of spots per T cell was shown. Next, the group mean was plotted using the individual mean (taken from 3 sequences). Statistical analysis for comparison (against medium conditions) was performed using the unpaired nonparametric test (Mann-Whitney). ** :p<0.01; * (p<0.05).

[0331] Overall, the antigenic peptides according to the present invention (CD19-B1, CD19-B2, CD22-B1, CD37-B1, TNFRSF13C-B1, and MS4A1-B4) induced a significant T cell response in the ELISPOT-IFNγ assay in HHD DR1 mice (Figures 6-10). The immunogenicity of CD22-B1, CD37-B1, TNFRSF13C-B1, and MS4A1-B4 was confirmed in HHD DR1 mice (Figures 11-13).

[0332] The results described above (Figure 6) demonstrate that immunizing HHD-DR1 mice with CD22-B1 can induce T cells that respond strongly after inoculation with either CD22-B1 or the corresponding human peptide CD22-H1. Therefore, CD22-B2 possesses potent immunogenicity and can elicit an effective immune response to the corresponding human peptide.

[0333] These results were confirmed in HHD DR3 mice that express human HLA-A2 and HLA-DR3 MHC and lack mouse H-2 class I and class II MHC (Figure 11).

[0334] The results described above (Figure 7) demonstrate that immunizing HHD-DR1 mice with TNFRSF13C-B1 can induce T cells that respond strongly to either TNFRSF13C-B1 or the corresponding human peptide TNFRSF13-H1. Therefore, TNFRSF13C-B1 possesses potent immunogenicity and can elicit an effective immune response to the corresponding human peptide.

[0335] These results were confirmed in HHD DR3 mice that express human HLA-A2 and HLA-DR3 MHC and lack mouse H-2 class I and class II MHC (Figure 12).

[0336] The results described above (Figure 8) demonstrate that immunizing HHD-DR1 mice with CD37-B1 can induce T cells that respond strongly after inoculation with either CD37-B1 or the corresponding human peptide CD37-H1. Therefore, CD37-B2 possesses potent immunogenicity and can elicit an effective immune response to the corresponding human peptide.

[0337] These results were confirmed in HHD DR3 mice that express human HLA-A2 and HLA-DR3 MHC and lack mouse H-2 class I and class II MHC (Figure 13).

[0338] The results described above (Figure 9) demonstrate that immunizing HHD-DR1 mice with CD19-B2 can induce T cells that respond strongly to either CD19-B2 or the corresponding human peptide CD19-H2. Therefore, CD19-B2 possesses potent immunogenicity and can elicit an effective immune response to the corresponding human peptide.

[0339] The results described above (Figure 10) demonstrate that immunizing HHD-DR1 mice with CD19-B1 can induce T cells that respond strongly to either CD19-B1 or the corresponding human peptide CD19-H1. Therefore, CD19-B1 possesses potent immunogenicity and can elicit an effective immune response to the corresponding human peptide.

[0340] The results described above (Figure 15) demonstrate that immunizing HHD-DR1 mice with MS4A1-B4 can induce T cells that respond strongly after inoculation with either MS4A1-B4 or the corresponding human peptide MS4A1-H4. Therefore, MS4A1-B4 possesses potent immunogenicity and can elicit an effective immune response to the corresponding human peptide.

[0341] These results were confirmed in HHD DR3 mice that express human HLA-A2 and HLA-DR3 MHC and lack mouse H-2 class I and class II MHC (Figure 16).

[0342] In summary, these 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—induced a potent immune response. Cross-reactivity of T cells generated for the corresponding human peptides—CD19-B1, CD19-B2, CD22-B1, CD37-B1, TNFRSF13C-B1, and MS4A1-B4—was demonstrated in HHD DR3 and HHD DR1 mice.

[0343] Therefore, these results provide experimental evidence that antigen-based immunotherapy can improve the T cell response in vivo, and that the antigenic peptide according to the present invention is particularly effective for that purpose.

[0344] Example 3: Ex vivo cytotoxic effects of CD22-B1, CD37B1, TNFRSF13C-B1, and MS4A1-B4 specific CD8 human T cells. Numerous studies support the concept of a repertoire of T cells specific to microbial peptides. While the number of microbial-specific T cells for peptides is assumed to be small, it is likely sufficient to be reactivated by vaccination.

[0345] To identify and functionally characterize human circulating CD22-B1, CD37B1, TNFRSF13C-B1, and MS4A1-B4 specific T cells, in vitro proliferation protocols have been developed to detect T cells specific to each antigenic peptide and examine their cytotoxic activity.

[0346] 3.1 Identification of antigenic peptide-specific CD8 T cells in humans In vitro growth methods and specific pMHC multimers are 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 respective human equivalents. Several HLA-A * PBMCs were collected from healthy donors (up to 19 donors), enriched after selecting CD137 and CD8 cells, and subjected to multiple in vitro proliferation cycles using T2 cells loaded with EO2463 peptide to increase the number of specific T cell clones. Detection of OMP peptide-specific CD8 T cells using cytometry analysis with fluorescent multimers was performed on the enriched CD8 T cell population.

[0347] Figure 17 shows the results obtained from one healthy HLA-A2 donor. In this donor, cell proliferation 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%).

[0348] In conclusion, these results indicate that healthy HLA-A2 donors have CD8 T cells in their blood that can recognize microbiome-derived peptides, and importantly, human equivalent peptides.

[0349] 3.2 Antigenic peptide-specific CD8 T cell toxicity function Using CD8+ T cells grown according to the above procedure, cytotoxicity assays were performed in the presence of target cells and effector cells in various ratios, and their cytotoxicity was evaluated using flow cytometry readings. Target cells were T2 cell lines loaded with bacterial peptides or equivalent human peptides. Negative controls included unloaded T2 cells and T2 cells loaded with unrelated peptides. As shown in Figure 18, antigenic peptide-specific human T cell clones grown in vitro were able to kill T2 cells loaded with all bacterial peptides, CD22-B1, CD37B1, TNFRSF13C-B1, and MS4A1-B4. More importantly, in vitro-grown 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 when cross-reactivity was observed by staining (Figure 17).

[0350] Overall, these results indicate the existence of T cell clones in healthy volunteers that can recognize microbial peptides and kill targets with microbial peptides and their human equivalents. These data are particularly promising because the T cell clones were obtained from healthy donors, and therefore, it was anticipated that specific T cell clones could be efficiently amplified in patients exposed to immunization with the antigenic peptides of the present invention.

[0351] Example 4: Further antigenic peptides are HLA-A * It has excellent affinity for the 0201 allele. Next, the HLA-A of the further selected antigenic peptides and the corresponding fragments of human tumor antigens (human reference peptides) * The binding affinity to the 0201 allele was confirmed in vitro.

[0352] Specifically, the antigenic peptides of SEQ ID NO: 110 ("YIFEHPELL," also known herein as CD22-B1), SEQ ID NO: 107 ("LIFEHPERV," also known herein as CD22-B12), and SEQ ID NO: 108 ("RVFEHPELV," also known herein as CD22-B13) were compared with the corresponding reference human peptide derived from CD22 ("WVFEHPETL," SEQ ID NO: 270, also known herein as CD22-H1).

[0353] Furthermore, the antigenic peptides of SEQ ID NO: 114 ("FLAFVPLQL," also known herein as CD37-B1), SEQ ID NO: 119 ("ILAFVPLYL," also known herein as CD37-B12), SEQ ID NO: 120 ("IMAFVPLAV," also known herein as CD37-B13), SEQ ID NO: 491 ("FLAFVPLDV," also known herein as CD37-B14), and SEQ ID NO: 493 ("VLAFVPLGV," also known herein as CD37-B15) were compared with the corresponding reference human peptide derived from CD37 ("GLAFVPLQI," SEQ ID NO: 271, also known herein as CD37-H1).

[0354] Furthermore, the antigenic peptides of SEQ ID NO: 220 ("LMFGAPALV," also known herein as TNFRSF13C-B1), SEQ ID NO: 212 ("FLFGAPASA," also known herein as TNFRSF13C-B11), SEQ ID NO: 217 ("LLFGAPAGV," also known herein as TNFRSF13C-B12), and SEQ ID NO: 224 ("VLFGAPAYL," also known herein as TNFRSF13C-B13) were compared with TNFRSF13C ("LLFGAPALL," SEQ ID NO: 279, also known herein as TNFRSF13C-H1).

[0355] Furthermore, the antigenic peptides of sequence number 65 ("AMNSLSLYI," also known herein as MS4A1-B4), sequence number 70 ("YMNSLSLAL," also known herein as MS4A1-B42), and sequence number 477 ("AMNSLSLTV," also known herein as MS4A1-B43) were compared with the corresponding reference human peptide derived from MS4A1 (also known as CD20) ("IMNSLSLFA," sequence number 264, also known herein as MS4A1-H4).

[0356] A. Materials and Methods A1. Measurement of peptide affinity for T2 cell lines The experimental protocol is HLA-A * This is similar to the validation performed on the peptide presented by 0201 (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):3411-21). Peptide affinity measurement is performed using HLA-A * This is achieved in human tumor cells T2 that express the 0201 molecule but are TAP1 / 2 negative and unable to present endogenous peptides.

[0357] T2 cells (5.10 per well) 4 Cells are incubated in serum-free medium (TexMacs) supplemented with 100 ng / μl of β2-microglobulin, with peptide concentrations decreasing from 100 μM to 0.1 μM (4 points: 100 μM, 10 μM, 1 μM, 0.1 μM) at 37°C for 16 hours. The cells are then washed twice and marked with anti-HLA-A2 antibody (clone BB7.2, BD Pharmagen) conjugated to PE.

[0358] The analysis will be performed using FACS (Macsquant analyzer 10-Miltenyi).

[0359] For each peptide concentration, the geometric mean of the labels associated with the peptide of interest was subtracted from the background noise, and the HLA-A obtained with the reference peptide HIV pol 589-597 at a concentration of 100 μM was obtained. * The percentage is reported as the geometric mean of the 0202 marker.

[0360] A2. Solubilization of peptides Each peptide is solubilized considering its amino acid composition. For peptides that do not contain cysteine, methionine, or tryptophan, DMSO may be added up to a maximum of 10% of the total volume. Other peptides are resuspended in water or PBS (pH 7.4).

[0361] B. Results The results are shown in Figures 19 to 22. For each of the tested human reference epitopes CD22-H1 (Figure 19), CD37-H1 (Figure 20), TNFRSF13C-H1 (Figure 21), and MS4A1-H4 (Figure 22), the respective antigenic peptides according to the present invention are HLA-A * It exhibits stronger binding affinity to 0201.

[0362] In summary, the above results indicate that the antigenic peptide according to the present invention is more effective than the corresponding human tumor antigen fragment in terms of HLA-A * This indicates a stronger binding affinity to 0201. As outlined above, although not bound by any theory, such strong binding affinity of the antigenic peptide according to the present invention is thought to reflect its ability to enhance the immune response (i.e., immunogenicity).

Claims

[Claim 1] An antigenic peptide comprising or consisting of an amino acid sequence represented by any of SEQ ID NOs: 10, 34, 114, 110, 65, and 220.

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