Simultaneous Cellular and Humoral Immune Response against Mutated p53 in a Patient with Lung Cancer

Abstract

We recently identified several Ags recognized by tumor-infiltrating B lymphocyte-derived Ab using SCID mice and a xenografted non-small cell lung cancer system. One of these identified Ags was mutated p53 with a point mutation resulting in the alteration of codon 158 from Arg to Leu. The aim of this study was to ascertain whether cellular immunity against mutated p53 exists in the same patient together with humoral immunity. Two different nona peptides (mutated p53150 and p53155 peptides), including a mutated amino acid derived from p53, were synthesized according to the binding motif of HLA class I of the established cancer cell line A904L from the patient. Mediastinal lymph node lymphocytes of the patient were stimulated weekly with the peptides. The mutated p53155 peptide-stimulated lymphocytes showed specific cytotoxicity against both autologous EBV-transformed B cells pulsed with mutated p53155 peptide and A904L. The mutated p53155 peptide-specific CTL clone in an HLA-Cw*0702 restriction was established and analyzed for its TCR usage. Clonotypic PCR using CDR3-specific primers was applied to the tumor tissue containing the tumor-infiltrating lymphocytes. The specific amplification of PCR was found in the tumor tissue. These results demonstrated that not only B lymphocytes producing specific Ab against the p53 protein, but also CTL against mutated p53, expressed in autologous lung cancer cells exist in the tumor tissue. This approach may allow us to better understand the mechanisms of T and B cell immunity against the same tumor Ag in cancer patients.

Identification of a New Cancer/Germline Gene,KK-LC-1, Encoding an Antigen Recognized by Autologous CTL Induced on Human Lung Adenocarcinoma

“…To generate T-cell clones, limiting dilution was done from the bulk CTL line 7 days after the last stimulation as previously described (14). The cells were seeded at 0.1, 0.3, 1, or 3 per well in 96-well round-bottomed plates and stimulated under the following culture condition: culture medium with irradiated F1121L cells and Rosi EBV-B cells as a feeder in the presence of IL-2 (100 units/mL), IL-4 (5 ng/mL), and IL-7 (5 ng/mL).…”

“…2B). The mutated p53 peptide (TRVLAMAIY), which has been identified as a tumor antigen restricted by HLA-Cw*0702 (14), was used as a negative control.…”

Immunological characterization of missense mutations occurring within cytotoxic T cell‐defined p53 epitopes in HLA‐A*0201+ squamous cell carcinomas of the head and neck

“…33,34 In many instances, anti-p53 antibody responses have been shown to decrease following surgical intervention. Interestingly, the A904L lung cancer cell line was initially characterized as accumulating p53 molecules using antibodies derived from the autologous patient's B cells, 26 which suggests that a Th2-biased anti-p53 immune response might have blocked/ reduced the CTL response against this mutant epitope. Interestingly, unlike most p53 sero-positive patients, the A904 patient was reported to have survived at least 6 years following surgery.…”

“…It was shown to be naturally presented in association with HLA-Cw*0702 molecules using CTL induced/expanded from lymphocytes obtained from the autologous A904 patient. 26 These mutated p53 antigens, which are ''non-self'' and the equivalent of the highly restricted, tumor rejection antigens of murine tumors, such as the Meth A p53 mutant antigen, should induce potent antitumor immunity. 3,27 In general, mice can be readily protected from lethal tumor challenges by prior immunization with irradiated tumor or sub-lethal doses of viable tumor.…”

Immunosuppressive effect of regulatory T lymphocytes in lung cancer, with special reference to their effects on the induction of autologous tumor-specific cytotoxic T lymphocytes

“…Recent clinical studies on immunotherapy indicated favorable therapeutic effects, and suggested that such a strategy might represent an alternative treatment approach for lung cancer (2,3). A large number of tumor-associated antigens have been identified in various human cancers (4)(5)(6). A high level of CD8 T cell infiltration into tumor tissues has been reported to be associated with a better prognosis in colon carcinoma and in ovarian cancer, suggesting that antitumor immunity can be provoked in cancer patients (7,8).…”