JP2014118361A - Msi-specific frameshift peptide (fsp) for prevention and treatment of cancer - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an improved vaccine for higher immune reactions to tumor antigens.SOLUTION: Provided is a vaccine comprising an MSI-specific frameshift peptide (FSP). In particular, FSP comprising amino acid sequences such as NTQIKALNRGLKKKTILKKAGIGMCVKVSSIFFINKQKP(TAF1B(-1)), EIFLPKGRSNSKKKGRRNRIPAVLRTEGEPLHTPSVGMRETTGLGC(HT001(-1)), HSTIKVIKAKKKHREVKRTNSSQLV(AIM2(-1)), or ASPKCIMKEKKSLVRLSSCVPVALMSAMTTSSSQKNITPAILTCC(TGFBR2(-1)) or the like are exemplified.

Description

  The present invention provides a vaccine for the prevention and treatment of cancer characterized by microsatellite instability (MSI). The vaccine contains an MSI-specific frameshift peptide (FSP) or a nucleic acid encoding the FSP that produces a humoral and cellular response to tumor cells.

  Human tumors arise through two major pathways of genomic instability, chromosomal instability and microsatellite instability (MSI) resulting from deficiencies in the DNA mismatch repair system. MSI occurs in a variety of extracolonic malignancies that exhibit a defective DNA mismatch repair system, including 15% of colorectal cancers, as well as endometrial, gastric, small intestine and other organ tumors. MSI cancer can occur sporadically or in association with hereditary tumor syndrome, hereditary nonpolyposis colorectal cancer (HNPCC) or Lynch syndrome.

  MSI colorectal cancer is a large part of the development of MSI tumors as a direct result of mismatch repair deficiencies that result in altered translational reading frames when microsatellite in the gene coding region is affected by mutations. It is characterized by high immunogenicity resulting from the production of the frameshift peptide (FSP) of (Fig. 1).

  FSP is a very promising target in immunotherapy due to the abundance of predictable MSI-specific FSP antigens and the fact that these antigens are directly attributable to the malignant transformation process. The human immune system is a means of eradicating tumor cells, and it is possible to develop effective treatments when immune system components are properly stimulated to recognize and eliminate cancer cells. It is considered possible. For this reason, immunotherapy (which directly or indirectly activates the body's immune system, including compositions and methods for reducing or eradicating cancer) includes the conventional cancer therapy. Has been studied for many years as an adjunct.

  It is generally accepted that tumor growth and metastasis are highly dependent on the ability of the tumor to evade host immune surveillance. Most tumors express antigens that the host immune system can recognize to varying degrees, but in many cases the immune response is inadequate. The inability to induce strong activation of effector T cells may be due to weak immunogenicity of tumor antigens or inappropriate or deleted expression of costimulatory molecules by tumor cells. In most T cells, IL-2 production and proliferation requires a costimulatory signal that coincides with TCR engagement, otherwise the T cell enters a functionally unresponsive state known as clonal anergy. there is a possibility.

  Despite the length of time these therapies have been studied, there remains a need for improved strategies that enhance the immune response to tumor antigens.

  There is also a need in the art for safe and effective compositions that can stimulate the immune system as cancer immunotherapy.

  According to the present invention, safe and effective stimulation of the immune system as cancer immunotherapy is achieved by the subject matter defined in the claims. In vitro data showed that FSP is highly immunogenic and can elicit significant FSP-specific T cell responses in vitro (Linnebacher et al. 2001, Ripberger et al. 2003, Schwitalle et al. 2004). In a further study examining peripheral blood collected from patients with MSI colon cancer, a high frequency of FSP-specific T cell responses were demonstrated (Schwitalle et al. 2008). Despite the large number of FSP-specific T cells in the tumor and peripheral blood, these patients show no signs of autoimmunity, indicating that the FSP vaccination approach has no side effects from an autoimmune perspective Suggested that is expected.

  Immunological analysis in individuals with germline mutations predisposing to hereditary nonpolyposis colorectal cancer (HNPCC) in DNA mismatch repair genes, even in the absence of clinically detectable tumors It was also found to show a cellular immune response against FSP. This suggests that FSP-specific immune responses may be protective in HNPCC individuals, so that FSP vaccination is also used in prophylactic situations as the first specific prophylactic approach in hereditary cancer conditions It is suggested that you can.

In summary,
(A) Frameshift peptide (FSP) is MSI specific and directly results from the pathogenesis of MSI tumors,
(B) Expected to have no clinically relevant side effects,
(C) FSP combinations are expected to target all tumors with MSI,
(D) FSP vaccination is designed for the treatment of 15% of colon cancer and tumors of the endometrium, stomach, small intestine and other organs;
(E) Molecular tumor analysis can identify patients who can benefit from FSP vaccination (targeted therapy),
(F) FSP vaccination can be used as preventive vaccination in high risk groups.

Schematic representation of coding microsatellite instability due to DNA mismatch repair deficiency (Kloor et al., 2010). If the coding microsatellite mutation results in a translation reading frame change, a truncated protein is generated that includes the FSP sequence (red) (eg, TGFBR2 protein). FIG. 5 shows an exemplary T cell response to newly designed FSP in peripheral blood from three patients with MSI colon cancer. It is a figure which shows the humoral immune response with respect to FSP induced | guided | derived from AIM2 (-1), HT001 (-1), TAF1B (-1), and TGFBR2 (-1). ELISA reveals FSP-specific antibody responses directed against neopeptides derived from AIM2 (-1), HT001 (-1), TAF1B (-1), and TGFBR2 (-1) became. The specificity of the peptide was demonstrated by preabsorption of each serum antibody as previously described (Reuschenbach et al., 2008). FIG. 5 shows a cytotoxic response determined by surface expression of CD107a. After stimulating T cells with antigen for 4 weeks, significant FSP-specific responses were observed in various healthy donors. The response occurred only in the presence of antigen presenting B cells and FSP antigen. A typical response is shown as a bar graph. FIG. 5 shows a cytotoxic response determined by surface expression of CD107a. Shown is a representative FACS analysis of the CD107a assay for T cells incubated with B cells as antigen presenting cells in the absence (left panel) or presence (right panel) of FSP antigen HT001 (-1).

  Thus, the present invention provides an MSI tumor specific frameshift peptide (FSP) such as TAF1B (accession number L39061), HT001 (accession number AF113539), AIM2 (accession number AF024714), or TGFBR2 (accession number). A vaccine comprising an FSP derived from NM_003242) or a nucleic acid encoding the FSP, wherein the FSP is capable of inducing an immune response against cancer exhibiting MSI.

In a preferred embodiment, the vaccine of the present invention comprises
(A) FSP comprising or consisting of the following amino acid sequences:
NTQIKALNRGLKKKK TILKKAGIGMCVKVSSIFFINQQP (TAF1B (-1)),
EIFLPKGRSNSKKKK GRNRNRIPAVLRTEGEPLHTSVGMRETTGLGC (HT001 (-1)),
HSTIKVIKAKKK HREVKRTNSSQLV (AIM2 (-1)), or ASPKCIMKEKK SLVRLSSCVPVALMSAMTTSSSQKNITPAILTCC (TGFBR2 (-1)),
(B) a functional equivalent of the FSP of (a), or (c) a combination of the FSP of (a) and / or (b),
Containing.

  The term “functional equivalent” as used herein refers to a variant of FSP that is still capable of eliciting an immune response, eg, against cancer, ie, still useful as an effective vaccine or Regarding fragments. An immune response is defined as a condition that meets at least one of the following criteria: Cytotoxicity assay, or IFN-γ secretion, or perforin expression, or granzyme B, measurable as above background by ELISpot, or intracellular cytokine staining, or cytokine ELISA, or equivalent method Induction of CD8 positive T cells detectable by expression or other cytokines that can be produced by CD8 positive T cells. 2. Induction of CD4 positive T cells detectable by cytokine secretion, measurable as exceeding background by ELISpot, or intracellular cytokine staining, or cytokine ELISA, or equivalent method. Cytokines are IFN-α, IFN-γ, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, IL-13, IL-17, TNF-α, TGF-β, Or other cytokines that can be produced by CD4 positive T cells. 3. Induction of antibodies detectable by Western blot, ELISA and equivalent or related methods. 4. Induction of any type of cellular immune response that is not mediated by the CD8 + T cells or CD4 + T cells according to 4.1 and 2.

  Variants are characterized by amino acid deletions, substitutions, and / or additions. Preferably, the amino acid difference is due to one or more conservative amino acid substitutions. The term “conservative amino acid substitution” refers to the replacement of the aliphatic or hydrophobic amino acids Ala, Val, Leu and Ile, the replacement of hydroxyl residues Ser and Thr, the replacement of acidic residues Asp and Glu, the amide residue Asn. And Gln replacement, replacement of basic residues Lys, Arg and His, replacement of aromatic residues Phe, Tyr and Trp, and replacement of small size amino acids Ala, Ser, Thr, Met and Gly.

  For the generation of peptides exhibiting a certain degree of identity to FSP, amino acid changes are introduced at specific positions in the cloned DNA sequence, for example using genetic manipulation, in order to identify regions important for peptide function be able to. For example, site-directed mutagenesis or alanine scanning mutagenesis (introduction of a single alanine mutation at every residue in the molecule) can be used (Cunningham and Wells, 1989). The resulting mutant molecules can then be tested for immunogenicity using the assay of Example 1.

  Preferably, the variant is characterized by no more than 8 aa substitutions, deletions and / or additions, more preferably no more than 6 aa, and even more preferably no more than 4 aa.

  In FSP fragments, at least 5 consecutive aa, preferably at least 10 consecutive aa, more preferably at least 15 consecutive aa, and even more preferably at least 20 consecutive, of a particular amino acid sequence aa remains. The fragment can still elicit an immune response.

  In a more preferred embodiment, the vaccine of the invention further comprises an adjuvant and / or an immunostimulatory cytokine or chemokine.

  Suitable adjuvants include aluminum salts such as aluminum hydroxide gel (alum) or aluminum phosphate, but may be calcium, iron or zinc salts or insoluble suspensions of acylated tyrosine or acylated sugars, It may be a cationically or anionically derivatized polysaccharide or polyphosphazene. Other known adjuvants include CpG-containing oligonucleotides. This oligonucleotide is characterized in that the CpG dinucleotide is not methylated. Such oligonucleotides are known and are described, for example, in WO 96/02555.

  The use of immunostimulatory cytokines has become an increasingly promising approach in cancer immunotherapy. Its main purpose is to protect patients from tumor-specific T lymphocyte activation, or disease recurrence, which can reject tumor cells from patients with low tumor tissue mass. Strategies that provide high levels of immunostimulatory cytokines locally at the site of the antigen have proven preclinical and clinical efficacy. Preferred immunostimulatory cytokines include IL-2, IL-4, IL-7, IL-12, IFN, GM-CSF and TNF-α.

  Chemokines are small (7 kD to 16 kD) secreted, structurally related soluble proteins involved in leukocyte and dendritic cell chemotaxis, PMN degranulation, and angiogenesis. Chemokines are produced during the early stages of host response to injury, allergens, antigens, or invading microorganisms. Chemokines selectively attract leukocytes to inflammatory foci, inducing both cell migration and activation. Chemokines can enhance innate or specific host immunity against tumors and thus may be useful in combination with FSP.

  The vaccine of the present invention may contain a nucleic acid encoding FSP for DNA immunization, a technique used to effectively stimulate humoral and cellular immune responses against protein antigens. Good. By direct injection of genetic material into a living host, a small amount of the cells produce a transgene product. This inappropriate gene expression in the host has important immunological significance and results in the activation of the host specific immunity against the gene delivery antigen. Direct injection of naked plasmid DNA induces a strong immune response against the antigen encoded by the genetic vaccine. Upon injection of the plasmid DNA construct, the host cell takes up foreign DNA, expresses the viral gene, and FSP is produced intracellularly. This form of antigen presentation and processing induces cellular and humoral immune responses that are restricted by both MHC class I and MHC class II. DNA vaccines usually have two units: a promoter / enhancer sequence followed by an antigen expression unit composed of an antigen (FSP) coding sequence and a polyadenylation sequence, and a production unit composed of sequences necessary for vector amplification and selection. Consists of a vector containing one unit. Construction of vectors containing vaccine inserts is accomplished using recombinant DNA technology, and those skilled in the art are aware of vectors that can be used in this approach. The efficiency of DNA immunization can be improved by stabilizing the DNA against degradation and increasing the efficiency of delivery of DNA to antigen-presenting cells. This has been demonstrated by coating DNA on biodegradable cationic microparticles (eg, poly (lactide-co-glycolide) formulated with cetyltrimethylammonium bromide). Such DNA-coated microparticles can be as effective as recombinant vaccinia virus in inducing CTLs, particularly when mixed with alum. Particles with a diameter of 300 nm are considered most effective for uptake by antigen presenting cells.

  Various expression vectors such as plasmids or viral vectors can be used to contain and express the nucleic acid sequence encoding the FSP of the invention.

  Preferred viral vectors are poxviruses, adenoviruses, retroviruses, herpesviruses, or adeno-associated virus (AAV) vectors. Particularly preferred poxviruses are vaccinia virus, NYVAC, avipox virus, canarypox virus, ALVAC, ALVAC (2), fowlpox virus or TROVAC.

  Recombinant alphavirus-based vectors have also been used to improve the efficiency of DNA vaccination. The gene encoding FSP is inserted into the alphavirus replicon to replace the structural gene, but leave the nonstructural replicase gene intact. Sindbis virus and Semliki Forest virus have been used to construct recombinant alphavirus replicons. However, unlike conventional DNA vaccination, alphavirus vectors are only expressed transiently. Alphavirus replicons elicit immune responses, replicon-induced cytokine responses, or replicon-induced apoptosis due to high levels of protein expressed by this vector, resulting in improved antigen uptake by dendritic cells .

  In a more preferred embodiment, the FSP contains a Tag sequence, preferably at the C-terminus, which may be useful for the purification of recombinantly produced FSP. A preferred Tag sequence is His-Tag. A particularly preferred His-Tag consists of 6 His residues.

  The vaccine of the present invention is administered in an amount suitable for immunization of an individual, and preferably further contains one or more common auxiliaries. The term “amount suitable for immunization of an individual” as used includes any amount of FSP that can immunize an individual. The amount will vary depending on whether the immunization is for prophylactic treatment or therapeutic treatment. In addition, the age, sex and weight of the individual also influence the determination of the amount. Thus, an amount suitable for immunization of an individual represents the amount of active ingredient that is sufficient to affect the course and severity of the tumor, resulting in a reduction or amelioration of such pathology. A “suitable amount for immunization of an individual” can be determined using methods known to those skilled in the art (see, eg, Fingl et al., 1975). The term “individual” as used herein includes any species of individual that may be affected by cancer. Examples of such individuals are humans and animals, and their cells.

  Administration of the vaccine by injection can be performed intramuscularly, subcutaneously, intradermally or in any other application form at various sites in the individual. It may also be convenient to perform approximately equal amounts of one or more “booster boosters”.

  The term “general aid” as used includes any aid suitable for conferring immunity to an individual with a vaccine. Such auxiliaries are, for example, buffered saline solutions, water, emulsions such as oil / water emulsions, wetting agents, sterile solutions and the like.

  The FSP, nucleic acid sequence or vector of the invention may be present in the vaccine itself or in combination with a carrier. Conveniently, the carrier within the individual is not immunogenic. Such a carrier can be an individual's own protein or a foreign protein, or a fragment thereof. Carriers such as serum albumin, fibrinogen or transferrin, or fragments thereof are preferred.

  The vaccine of the present invention may be therapeutic, i.e. administered to treat cancer in which the compound is present or to prevent recurrence of cancer, or prophylactic I.e. the compound is administered to reduce or delay the development of cancer. When the composition is used therapeutically, the composition is administered to a cancer patient to induce an immune response and stabilize the tumor by suppressing or slowing the growth of existing cancer. Or designed to reduce the spread of metastases, reduce tumor size, prevent recurrence of treated cancer, or eliminate cancer cells that are not killed by previous treatments . Vaccines used as prophylactic treatment are designed to be administered to individuals who do not have cancer, elicit an immune response, and target potential cancer cells.

  The invention also relates to the use of FSP or functional equivalents, nucleic acid sequences, or vectors as defined above for the prevention of cancer, for example the production of vaccines for high-risk group prophylaxis or for the treatment of cancer. Related. For example, they can be colorectal cancer, preferably hereditary nonpolyposis colorectal cancer (HNPCC), endometrial cancer, gastric cancer or small intestinal cancer.

In the present invention, it is possible to immunize individuals, particularly humans and animals. Immunization is performed both by antibody induction and CD8 ⁺ T cell stimulation. This makes it possible to take preventive and therapeutic measures against cancer.

  The following examples illustrate the invention in more detail.

Example 1
Detection of FSP-specific T cells in peripheral blood from patients with MSI colon cancer and healthy HNPCC mutation carriers (A) Method (ELISpot assay)
The frequency of FSP-specific peripheral T cells (pTc) was determined using ELISpot analysis to determine the number of IFN-γ secreting Tc specific for newly designed FSPs derived from 3cMS-containing candidate genes. Quantified by determining. The ELISpot assay was coated with mouse anti-human IFN-γ monoclonal antibody (mAb) (Mabtech, Nacka, Sweden) overnight and blocked with serum-containing medium (Multiscreen; Millipore, Bedford, Mass.) It carried out using. pTc (Day 0, 1 × 10 ⁵ / well) was converted to antigen-presenting cells in 200 μl IMDM with 10% human AB serum, and autologous CD40 activated B cells (4 × 10 ⁴ / well, TiBc or pBc, respectively). ) And 6-fold plating. Peptides were added to a final concentration of 10 μg / mL. As a positive control, pTc was treated with 20 nmol / L phorbol-12-myristate-13-acetate in combination with 350 nmol / L ionomycin. After 24 hours incubation at 37 ° C., the plates were washed thoroughly, incubated with biotinylated rabbit anti-human IFN-γ mAb for 4 hours, washed again and incubated with streptavidin-alkaline phosphatase for 2 hours, and finally The washing process was performed. Spots were detected by incubation with NBT / BCIP (Sigma-Aldrich) for 1 hour, the reaction was stopped with water, and the spots were counted by microscope after drying. The method is described in detail in Schwitalle et al., 2008.

(B) Results An ELISpot analysis was performed to determine whether FSP-specific T cell responses were detectable in the peripheral blood of MSI-H CRC patients. Self-pBc showing high expression of MHC class I and II, costimulatory factors (CD40, CD80 and CD86) and B cell specific antigens (CD19 and CD23) were used as antigen presenting cells.

Significant reactivity was observed against newly designed FSPs derived from AIM2 (-1), HT001 (-1), TAF1B (-1) and TGFBR2 (-1):
TAF1B (-1) NTQIKALNRGLKKKK TILKKAGIGMCVKVSSIFFINQQP
HT001 (-1) EIFLPKGRSNSKKK GRRRNRIPAVLRTEGEPLHTSGVGRETTGGLGC
AIM2 (-1) HSTIKVIKAKKKK HREVKRTNSSQLV
TGFBR2 (-1) ASPKCIMKEKK SLVRLSSCVPVALMSAMTTSSSQKNITPILTCCC
(Neopeptide sequences are underlined)

  The results obtained from patients (n = 8) are summarized in Table 1. A typical ELISpot result is shown in FIG.

  The average number of spots from repeated analysis is given for each peptide and test individual. Patients with MSI01-MSI07-MSI-H CRC, MC01-MC06-Healthy HNPCC germline mutation carriers.

Example 2
Detection of FSP-specific humoral immune response in peripheral blood from patients with MSI colon cancer and healthy HNPCC mutation carriers (A) Method (ELISA)
For enzyme-linked immunosorbent assay (ELISA), peptides were coated overnight in PBS at a concentration of 40 μg / ml at 4 ° C. in 96 well polystyrene microtiter plates “Maxisorp” (Nunc, Roskilde, Denmark). After coating, the plates were washed 4 times with PBS (0.05% Tween) and blocked with 0.5% casein in PBS for 1 hour. Peptide binding to microtiter plates and optimal saturated peptide concentration were evaluated using an alkaline phosphatase-peptide competition assay. A control peptide derived from the p16 ^INK4a protein (p16_76-105) was used to monitor the individual background reactivity of each serum, but antibody reactivity to this was not seen in a large cohort. (Reuschenbach et al., 2008). Each serum was diluted 1: 100 with blocking buffer (0.5% casein in PBS) and tested in duplicate for the presence of antibodies against all FSP and control peptides. As a reference for dispersion between plates, one control serum per plate was included and the peptide specific OD of the control serum was used for normalization. Diluted serum (50 μl / well) was incubated for 1 hour, and after the washing step, the plate was incubated with HRP-labeled rabbit anti-human IgG antibody (Jackson Immunoresearch, West Grove, PA; 1: 10000 in blocking buffer) for 1 hour. After washing, 50 μl / well of TMB substrate (Sigma, Deisenhofen, Germany) was added, and 30 minutes later, the enzyme reaction was stopped by adding 50 μl / well of 1N H ₂ SO ₄ . Absorbance was measured at 450 nm (reference wavelength 595 nm). Serum antibody preabsorption for specificity control was performed according to the method described in detail in Reuschenbach et al., 2008.

(B) Results An ELISA analysis was performed to determine whether FSP-specific antibodies were detectable in the peripheral blood of MSI-H CRC patients, healthy Lynch syndrome mutation carriers, and healthy controls. did. Significant reactivity was observed against newly designed FSPs derived from AIM2 (-1), HT001 (-1), TAF1B (-1) and TGFBR2 (-1). The results of the ELISA are shown in FIG.

Example 3
Detection of FSP-specific cytotoxic T cell responses. Surface expression of CD107a on effector T cells upon stimulation with clinical FSP antigen was measured. The CD107a assay is used to demonstrate the secretion of cytotoxic particles containing perforin / granzyme B from effector cells. CD107a molecules are expressed on the surface of cytotoxic particles and become detectable on the cell surface when the particles are released in the context of a cytotoxic T cell response.

  To determine the possibility that the FSP peptide induces a cytotoxic cellular immune response, blood was collected from healthy donors and T cells were stimulated with FSP using dendritic cells as antigen presenting cells. Stimulation was repeated once a week for 4 weeks. After 4 weeks, T cells were harvested, co-incubated with target cells and FSP, and analyzed for peptide-specific induction of cytotoxic T cell responses using the CD107a assay.

  A cytotoxic response was observed in T cells co-incubated with antigen presenting B cells in the presence of antigenic FSP, as determined by surface expression of CD107a (FIG. 4A). Significant responses were observed in various healthy donors after 4 weeks of stimulation of T cells with antigen. A typical response is shown in the bar graph. FIG. 4B shows a representative FACS analysis of the CD107a assay for T cells incubated with B cells that are antigen presenting cells in the absence (left panel) or presence (right panel) of FSP antigen HT001 (-1). .

References
Cunningham and Wells, Science 244 (1989), 1081-1085.

Fingl et al., The Pharmocological Basis of Therapeutics, Goodman and Gilman, eds.Macmillan Publishing Co., New York, pp. 1-46 (1975).

Kloor M, Michel S, von Knebel Doeberitz M.
Immune evasion of microsatellite unstable colorectal cancers.
Int J Cancer. 2010 Mar 2. [Epub ahead of print]

Linnebacher M, Gebert J, Rudy W, Woerner S, Yuan YP, Bork P, von Knebel Doeberitz M. Frameshift peptide-derived T-cell epitopes: a source of novel tumor-specific antigens. Int J Cancer. 2001 Jul 1; 93 (1): 6-11.

Reuschenbach M, Waterboer T, Wallin KL, Einenkel J, Dillner J, Hamsikova E, Eschenbach D, Zimmer H, Heilig B, Kopitz J, Pawlita M, von Knebel Doeberitz M, Wentzensen N.
Characterization of humoral immune responses against p16, p53, HPV16 E6 and HPV16 E7 in patients with HPV-associated cancers.
Int J Cancer. 2008 Dec 1; 123 (11): 2626-31.

Reuschenbach M, Kloor M, Morak M, Wentzensen N, Germann A, Garbe Y, Tariverdian M, Findeisen P, Neumaier M, Holinski-Feder E, von Knebel Doeberitz M.
Serum antibodies against frameshift peptides in microsatellite unstable colorectal cancer patients with Lynch syndrome.
Fam Cancer. 2009 Dec 2. [Epub ahead of print]

Ripberger E, Linnebacher M, Schwitalle Y, Gebert J, von Knebel Doeberitz M. Identification of an HLA-A0201-restricted CTL epitope generated by a tumor-specific frameshift mutation in a coding microsatellite of the OGT gene.J Clin Immunol. 2003 Sep ; 23 (5): 415-23.

Schwitalle Y, Kloor M, Eiermann S, Linnebacher M, Kienle P, Knaebel HP, Tariverdian M, Benner A, von Knebel Doeberitz M. Immune response against frameshift-induced neopeptides in HNPCC patients and healthy HNPCC mutation carriers.Gastroenterology. 2008 Apr; 134 (4): 988-97.

Schwitalle Y, Linnebacher M, Ripberger E, Gebert J, von Knebel Doeberitz M. Immunogenic peptides generated by frameshift mutations in DNA mismatch repair-deficient cancer cells.Cancer Immun. 2004 Nov 25; 4: 14.
Sequence listing

Claims (15)

  A vaccine comprising an MSI-specific frameshift peptide (FSP), wherein the FSP is capable of inducing an immune response against a tumor exhibiting MSI. below:
(A) FSP comprising the following amino acid sequence:
NTQIKALNRGLKKKK TILKKAGIGMCVKVSSIFFINQQP (TAF1B (-1)),
EIFLPKGRSNSKKKK GRNRNRIPAVLRTEGEPLHTSVGMRETTGLGC (HT001 (-1)),
HSTIKVIKAKKHK HREVKRTNSSSQLV (AIM2 (-1)), or ASPKCIMKEKKSLVRLSSCVPVALMSAMMTTSSSQKNITPAILTCC (TGFBR2 (-1)), or (b) capable of eliciting an immune response against cancer that still exhibits MSI, SP (a) An equivalent, or a combination of (c) (a) and / or (b) FSP,
The vaccine according to claim 1, comprising:   The vaccine according to claim 1 or 2, wherein the FSP further comprises a Tag sequence.   The vaccine containing the nucleic acid sequence which codes FSP as described in any one of Claims 1-3, or the vector containing this nucleic acid sequence.   The vaccine according to claim 4, wherein the vector is a plasmid or a viral vector.   6. The vaccine of claim 5, wherein the viral vector is a poxvirus, adenovirus, retrovirus, herpes virus, alphavirus-based vector or adeno-associated virus (AAV) vector.   The vaccine according to claim 6, wherein the poxvirus is vaccinia virus, NYVAC, avipox virus, canarypox virus, ALVAC, ALVAC (2), fowlpox virus or TROVAC.   The vaccine according to any one of claims 1 to 7, further comprising an adjuvant and / or an immunostimulatory cytokine or chemokine.   The vaccine according to any one of claims 1 to 8, wherein the active compound is present in an oil-in-water or water-in-oil emulsion vehicle.   10. A vaccine according to any one of claims 1 to 9, further comprising one or more other antigens.   The vaccine according to any one of claims 1 to 10, which is used in a method for preventing or treating a tumor.   The MSI tumor-specific frameshift peptide (FSP) according to any one of claims 1 to 3, the nucleic acid according to claim 4 or the claim 4 for the preparation of a vaccine for the prevention or treatment of tumors. Use of the vector according to any one of -7.   Use according to claim 11 or 12 for preventive vaccination of high risk groups.   The use according to any one of claims 11 to 13, wherein the tumor is colorectal cancer, endometrial cancer, gastric cancer or small intestine cancer.   15. Use according to claim 14, wherein the colorectal cancer is hereditary non-polyposis colorectal cancer (HNPCC).

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