Characterization of B16 melanoma-specific cytotoxic T lymphocytes.
A B16 melanoma-specific CD8+ T cell line (AB1) was established from the spleen cells of C57BL/6 mice cured of B16 melanoma with interleukin (IL)-12 treatment. The AB1 line exclusively used T cell receptor Vbeta11. The AB1 cells exhibited a cytolytic activity against both syngeneic B16 melanoma and allogeneic P815 mastocytoma, whereas a cold inhibition assay revealed specificity of the AB1 cells against B16 melanoma. Their lostability to kill a class I loss variant of B16 melanoma was restored by the transfection of H-2Kb gene. In addition, their interferon (IFN)-gamma production was significantly suppressed by the addition of anti-H-2Kb monoclonal antibody, and RT-PCR analysis showed that the AB1 line expressed the mRNA encoding IFN-gamma, but not IL-4 or IL-10. The experiment using synthetic peptides of tyrosinase-related protein-2 (TRP-2) revealed that the AB1 cells could recognize TRP-2(181-188) peptide. Moreover, the AB1 cells showed an in vivo antitumor effect against established pulmonary metastases of B16 melanoma. Overall, these results indicate that the Tc1-type Vbeta11+ AB1 cells exert an antitumor activity against syngeneic B16 melanoma through recognition of TRP-2(181-188) peptide in an H-2Kb-restricted manner.
Harada
M
M
Department of Virology, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan. haradam@pop.nci.nih.gov
Tamada
K
K
Abe
K
K
Li
T
T
Onoe
Y
Y
Tada
H
H
Tatsugami
K
K
Ando
T
T
Kimura
G
G
Nomoto
K
K
D000818:Animals; D002460:Cell Line; D016207:Cytokines / Q000378:metabolism; D003602:Cytotoxicity, Immunologic; D005260:Female; D006183:H-2 Antigens / Q000235:genetics / Q000276:immunology; D016219:Immunotherapy, Adoptive; D019746:Intramolecular Oxidoreductases / Q000276:immunology; D008546:Melanoma, Experimental / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D008815:Mice, Inbred Strains; D010446:Peptide Fragments / Q000276:immunology; D010641:Phenotype; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000276:immunology / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013154:Spleen / Q000166:cytology; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D018417:Th1 Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D014407:Tumor Cells, Cultured
D016207:Cytokines; D006183:H-2 Antigens; C032876:H-2Kb protein, mouse; D010446:Peptide Fragments; D016693:Receptors, Antigen, T-Cell, alpha-beta; D019746:Intramolecular Oxidoreductases; C042501:dopachrome isomerase