A method of manufacturing a cancer specific cytotoxic T lymphocytes cell line for use in treatment of a cancer patient, wherein said method comprises - identifying cancer antigens from nucleic acid obtained from cancer cells of the cancer patient using the steps of a) obtaining a plurality of mutant sequences from the nucleic acid of cancer cells from a cancer patient, said mutant sequences coding for all or a portion of an expressed gene and wherein the mutant sequences each have a mutant position amino acid which substitutes for a wildtype position amino acid located at the same position in the wildtype sequence of the protein, wherein said mutant sequences are obtained using a parallel sequencing platform, said parallel sequencing platform employing parallel processing of said nucleic acid of cancer cells leading to sequence reads and mapping of the sequence reads to a database with reference gene sequences; and b) selecting mutant sequences from those identified in step a) by identifying an HLA class or supertype of the cancer patient and then selecting an amino acid for said HLA class or supertype as the mutant position amino acid using FIG. 7; wherein cancer antigens are identified in that peptides are synthesized comprising said mutant sequences from step b) and evaluated for activating cytotoxic T lymphocytes cell lines prepared from the cancer patient or from an HLA-matched donor, said CTL cell lines obtained by contacting mononuclear cells from the cancer patient or from the HLA-matched donor with cancer cells from the cancer patient; - contacting T cells from the cancer patient or from a HLA matched donor with the cancer antigens in vitro to stimulate cancer specific cytotoxic T lymphocytes cell line. - identifying cancer antigens from nucleic acid obtained from cancer cells of the cancer patient using the steps of a) obtaining a plurality of mutant sequences from the nucleic acid of cancer cells from a cancer patient, said mutant sequences coding for all or a portion of an expressed gene and wherein the mutant sequences each have a mutant position amino acid which substitutes for a wildtype position amino acid located at the same position in the wildtype sequence of the protein, wherein said mutant sequences are obtained using a parallel sequencing platform, said parallel sequencing platform employing parallel processing of said nucleic acid of cancer cells leading to sequence reads and mapping of the sequence reads to a database with reference gene sequences; and b) selecting mutant sequences from those identified in step a) by identifying an HLA class or supertype of the cancer patient and then selecting an amino acid for said HLA class or supertype as the mutant position amino acid using FIG. 7; wherein cancer antigens are identified in that peptides are synthesized comprising said mutant sequences from step b) and evaluated for activating cytotoxic T lymphocytes cell lines prepared from the cancer patient or from an HLA-matched donor, said CTL cell lines obtained by contacting mononuclear cells from the cancer patient or from the HLA-matched donor with cancer cells from the cancer patient; a) obtaining a plurality of mutant sequences from the nucleic acid of cancer cells from a cancer patient, said mutant sequences coding for all or a portion of an expressed gene and wherein the mutant sequences each have a mutant position amino acid which substitutes for a wildtype position amino acid located at the same position in the wildtype sequence of the protein, wherein said mutant sequences are obtained using a parallel sequencing platform, said parallel sequencing platform employing parallel processing of said nucleic acid of cancer cells leading to sequence reads and mapping of the sequence reads to a database with reference gene sequences; and b) selecting mutant sequences from those identified in step a) by identifying an HLA class or supertype of the cancer patient and then selecting an amino acid for said HLA class or supertype as the mutant position amino acid using FIG. 7; wherein cancer antigens are identified in that peptides are synthesized comprising said mutant sequences from step b) and evaluated for activating cytotoxic T lymphocytes cell lines prepared from the cancer patient or from an HLA-matched donor, said CTL cell lines obtained by contacting mononuclear cells from the cancer patient or from the HLA-matched donor with cancer cells from the cancer patient; - contacting T cells from the cancer patient or from a HLA matched donor with the cancer antigens in vitro to stimulate cancer specific cytotoxic T lymphocytes cell line.