Melanoma associated antigenic polypeptide, epitopes thereof and vaccines against melanoma

Abstract

A melanoma associated antigen known as gp100 and peptides derived from the antigen. Gp100 and its peptide derivatives can be used in vaccines for the treatment of melanoma. Another aspect of the invention is host-cells capable of expressing gp100 for the gp100-derived peptides. Furthermore, tumor-infiltrating lymphocytes (TILs) specifically recognizing gp100 are described, as are vaccines with these TILs. Also disclosed are diagnostics for the detection of melanoma and for the monitoring of vaccination.

What is claimed is:  
     
         1 . A peptide consisting of from about 8 to about 12 contiguous amino acids of an amino acid sequence, said amino acid sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:21 and SEQ ID NO:22 in combination with a pharmaceutically acceptable carrier or diluent.  
     
     
         2 . A peptide consisting of from about 8 to about 12 contiguous amino acids of an amino acid sequence, said amino acid sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:21 and SEQ ID NO:22.  
     
     
         3 . The peptide of  claim 2 , wherein the amino acid sequence is that of SEQ ID NO:4.  
     
     
         4 . The peptide of  claim 2 , wherein the amino acid sequence is that of SEQ ID NO:6.  
     
     
         5 . The peptide of  claim 2 , wherein the amino acid sequence is that of SEQ ID NO:21.  
     
     
         6 . The peptide of  claim 2 , wherein the amino acid sequence is that of SEQ ID NO:22.  
     
     
         7 . An immunogenic carrier or marker coupled to the peptide of  claim 2 .  
     
     
         8 . A nucleotide sequence, comprising a nucleotide sequence encoding a melanoma associated antigen, said antigen comprising the amino acid sequence of SEQ ID NO:2.  
     
     
         9 . The nucleotide sequence according to  claim 8 , comprising the nucleotide sequence of SEQ ID NO:1.  
     
     
         10 . A nucleotide sequence, comprising a nucleotide sequence encoding an immunogenic peptide, said peptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22 and SEQ ID NO:23.  
     
     
         11 . The nucleotide sequence according to  claim 10 , comprising a nucleotide sequence selected from the group consisting of the sequences of SEQ ID NOS: 3, 5, 7 and 9.  
     
     
         12 . A test kit for the detection of melanoma cells, comprising at least one primer and a labeled probe directed to the nucleotide sequence according to  claim 8  or its complementary sequence.  
     
     
         13 . A method for the detection of melanoma cells, comprising: 
 a. obtaining a specimen from a patient;    b. subjecting the specimen to methods for the amplification of a nucleotide sequence according to  claim 8  or a part of said sequence coding for an immunogenic fragment;    c. reacting amplified nucleic acid with a complementary labeled probe; and    d. detecting nucleic acid labeled with the probe.    
     
     
         14 . A cloning vehicle comprising the nucleotide sequence of  claim 8 .  
     
     
         15 . A host cell transfected or transformed with the cloning vehicle according to  claim 14 .  
     
     
         16 . The host cell according to  claim 15  selected from the group consisting of murine EL4 cells, murine P8.15 cells and human BLM cells.  
     
     
         17 . The host cell according to  claim 15 , which is an antigen presenting cell.  
     
     
         18 . The host cell according to  claim 15 , which also produces co-stimulating molecules.  
     
     
         19 . A vaccine comprising an antigen or epitope thereof, said antigen comprising the amino acid sequence of SEQ ID NO:2, said vaccine comprising an antigen presenting cell that has been preloaded with a peptide comprising the antigen or epitope.  
     
     
         20 . A vaccine comprising a host cell according to  claim 15 .  
     
     
         21 . A vaccine comprising the nucleotide sequence according to  claim 8 .  
     
     
         22 . A vaccine comprising a T cell receptor against a nucleotide sequence or cells expressing said T cell receptor, wherein said nucleotide sequence comprises a nucleotide sequence encoding a melanoma associated antigen, said antigen comprising the amino acid sequence of SEQ ID NO:2.  
     
     
         23 . A vaccine comprising an antigen or epitope thereof, said antigen comprising the amino aid sequence of SEQ ID NO:2, said vaccine additionally comprising at least one compound selected from the group consisting of an adjuvant, one or more cytokines, antibodies directed against CD2, CD3, CD27, CD28 or other T cell surface antigens, and helper epitopes that stimulate CD4+ or CD8+ T cells.  
     
     
         24 . A method for the generation of antigen reactive tumor infiltrating lymphocytes, comprising: 
 a. taking a sample of a melanoma from a patient;    b. isolating tumor infiltrating lymphocytes from the sample;    c. reacting said lymphocytes with an antigen comprising the amino acid sequence of SEQ ID NO:2; and    d. isolating the lymphocytes binding to said antigen.    
     
     
         25 . Tumor infiltrating lymphocytes capable of binding to a melanoma associated antigen, said antigen comprising the amino acid sequence of SEQ ID NO:2.  
     
     
         26 . Melanoma cells that express an antigen comprising the amino acid sequence of SEQ ID NO:2, said cells transfected with a cloning vehicle having a nucleotide sequence coding for a lymphokine.  
     
     
         27 . A vaccine comprising tumor infiltrating lymphocytes according to  claim 25  and a pharmaceutically acceptable carrier or diluent.  
     
     
         28 . An antibody directed to an immunogenic peptide, said peptide comprising an immunogenic fragment of an antigen comprising the amino acid sequence of SEQ ID NO:2.  
     
     
         29 . A vaccine comprising the antibody according to  claim 28 .  
     
     
         30 . A method for monitoring immunotherapy, comprising detecting the presence of antibodies directed to an immunogenic peptide, said peptide comprising an immunogenic fragment of an antigen comprising the amino acid sequence of SEQ ID NO:2.  
     
     
         31 . A kit for the detection of antibodies according to  claim 28 , comprising a conjugate of a detectable marker and an immunogenic peptide comprising an immunogenic fragment of an antigen comprising the amino acid sequence of SEQ ID NO:2.  
     
     
         32 . A cloning vehicle comprising the nucleotide sequence of  claim 10 .  
     
     
         33 . The host cell of  claim 15  cotransfected with a cloning vehicle comprising a nucleotide sequence coding for an MHC class I allele.  
     
     
         34 . A vaccine comprising the nucleotide sequence according to  claim 10 .  
     
     
         35 . A vaccine comprising a T cell receptor against a peptide or cells expressing said T cell receptor, said peptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22 and SEQ ID NO:23.  
     
     
         36 . A vaccine comprising a peptide and a pharmaceutically acceptable carrier or diluent, said peptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22 and SEQ ID NO:23, said vaccine additionally comprising at least one compound selected from the group consisting of an adjuvant, one or more cytokines, antibodies directed against CD2, CD3, CD27, CD28 or other T cell surface antigens, and helper epitopes that stimulate CD4+ or CD8+ T cells.  
     
     
         37 . The vaccine according to  claim 19  comprising, in addition, at least one compound selected from the group consisting of an adjuvant, one or more cytokines, antibodies directed against CD2, CD3, CD27, CD28 or other T cell surface antigens, and helper epitopes that stimulate CD4+ or CD8+ T cells.  
     
     
         38 . The vaccine according to  claim 20  comprising, in addition, at least one compound selected from the group consisting of an adjuvant, one or more cytokines, antibodies directed against CD2, CD3, CD27, CD28 or other T cell surface antigens, and helper epitopes that stimulate CD4+ or CD8+ T cells.  
     
     
         39 . The vaccine according to  claim 21  comprising, in addition, at least one compound selected from the group consisting of an adjuvant, one or more cytokines, antibodies directed against CD2, CD3, CD27, CD28 or other T cell surface antigens, and helper epitopes that stimulate CD4+ or CD8+ T cells.  
     
     
         40 . The vaccine according to  claim 22  comprising, in addition, at least one compound selected from the group consisting of an adjuvant, one or more cytokines, antibodies directed against CD2, CD3, CD27, CD28 or other T cell surface antigens, and helper epitopes that stimulate CD4+ or CD8+ T cells.  
     
     
         41 . The vaccine according to  claim 34  comprising, in addition, at least one compound selected from the group consisting of an adjuvant, one or more cytokines, antibodies directed against CD2, CD3, CD27 or other T cell surface antigens, and helper epitopes that stimulate CD4+ or CD8+ T cells.  
     
     
         42 . The vaccine according to  claim 35  comprising, in addition, at least one compound selected from the group consisting of an adjuvant, one or more cytokines, antibodies directed against CD2, CD3, CD27, CD28 or other T cell surface antigens, and helper epitopes that stimulate CD4+ or CD8+ T cells.  
     
     
         43 . A vaccine comprising melanoma cells according to  claim 26  and a pharmaceutically acceptable carrier or diluent.  
     
     
         44 . An antibody directed to a peptide, said peptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22 and SEQ ID NO:23.  
     
     
         45 . An immunogenic peptide having 5-20 contiguous amino acids of gp100, said peptide being capable of reacting with T-lymphocytes.  
     
     
         46 . The immunogenic peptide of  claim 45 , wherein said peptide is at least about 9 to 10 amino acids in length.  
     
     
         47 . The immunogenic peptide of  claim 46  having the sequence KTWGQYWQV (SEQ ID NO:22) or KTWGQYWQVL (SEQ ID NO:21).  
     
     
         48 . The immunogenic peptide of  claim 46  wherein the peptide is LLDGTATLRL (SEQ ID NO:4).  
     
     
         49 . The immunogenic peptide of  claim 46 , wherein said peptide contains at least one amino acid modification of said gp100 sequence.  
     
     
         50 . The immunogenic peptide of  claim 47 , wherein said peptide contains at least one amino acid modification of said gp100 sequence.  
     
     
         51 . The immunogenic peptide of  claim 48 , wherein said peptide contains at least one amino acid modification of said gp100 sequence.  
     
     
         52 . The peptide of  claim 5 , wherein said modification includes at least one amino acid substitution in said peptide sequence.  
     
     
         53 . An immunogenic peptide having the formula selected from the group consisting of X 1 X 2 X 3 GQYWQX 4  or X 1 X 2 X 3 PGPVTX 4  wherein: 
 X 1  is any naturally occurring amino acid;    X 2  is any hydrophobic aliphatic amino acid;    X 3  is any naturally occurring amino acid; and    X 4  is any hydrophobic aliphatic amino acid.    
     
     
         54 . The peptide of  claim 53 , wherein the amino acid for X 1  is selected from the group consisting of tyrosine and lysine.  
     
     
         55 . The peptide of  claim 53 , wherein X 2  is selected from the group consisting of leucine and threonine.  
     
     
         56 . The peptide of  claim 53 , wherein X 3  is tyrosine.  
     
     
         57 . The peptide of  claim 53 , wherein X 4  is selected from the group consisting of alanine and valine.  
     
     
         58 . The immunogenic peptide of  claim 45  wherein said peptide is recognized by HLA-A2 restricted tumor infiltrating lymphocyte.  
     
     
         59 . The immunogenic peptide of  claim 53  wherein said peptide is recognized by HLA-A2 restricted tumor infiltrating lymphocyte.  
     
     
         60 . The immunogenic peptide of  claim 45  wherein said peptide is a native, synthetic or recombinant peptide.  
     
     
         61 . The immunogenic peptide of  claim 53  wherein said peptide is a native, synthetic or recombinant peptide.