Generation of novel killer hybridomas derived from proliferation-suppressed somatic cell hybrids between YACUT T cell lymphoma and normal lymphocytes activated in secondary mixed lymphocyte cultures

• Published in: Journal of immunological methods 1996-06, Vol.192 (1), p.137-147
• Main Authors: Kubota, Koichi, Nakazato, Kazumi, Tamauchi, Hidekazu, Sasahara, Takeshi, Katoh, Hideki
• Format: Article
• Language: English
• Subjects: Animals; Antibody-Dependent Cell Cytotoxicity; Antigens, CD - analysis; Base Sequence; Cell Division - immunology; Cell Fusion; Cell Line, Transformed; Cytotoxicity, Immunologic; Differentiation; Functional T cell hybridoma; Hybrid Cells - classification; Hybrid Cells - cytology; Hybrid Cells - immunology; Hybridomas - immunology; Immunophenotyping; Intratypic T cell hybrid; Killer Cells, Natural - immunology; Leukemia L5178 - immunology; Leukemia L5178 - metabolism; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Lymphoma, T-Cell - immunology; Lymphoma, T-Cell - metabolism; Mice; Mice, Inbred BALB C; Mice, Inbred C3H; Mice, Inbred DBA; Molecular Sequence Data; Receptors, Antigen, T-Cell - biosynthesis; Receptors, IgG - genetics; Somatic cell genetics; Tumor Cells, Cultured
• ISSN: 0022-1759; 1872-7905
• DOI: 10.1016/0022-1759(96)00030-0

Somatic cell hybridization between the YACUT T cell lymphoma cell line with normal lymphocytes activated in secondary mixed lymphocyte cultures (MLCs) consistently yielded IL-2-dependent CD4 −CD8 α + β − FcγRIII + hybrids with cytotoxic function. The hybrids expressed T cell receptors other than that of YACUT origin, and fusion of the YACUT with a CD8 α + β − FcγRIII − T cell line also yielded hybrids with an unexpected CD8 α + β − FcγRIII + phenotype, which two observations strongly suggested that CD8 + T cells became the parental cell of the hybrids. Prolonged growth of the hybrids with IL-2 resulted in the generation of autonomously growing hybrids (hybridomas) without abrogating the cytotoxic function. The hybridomas exhibited MHC-unrestricted cytotoxicity in a Ca 2+-dependent manner without prior stimulation and also mediated antibody-dependent cellular cytotoxicity. These results indicate that novel killer hybridomas can be produced following cell transformation of proliferation-suppressed cytotoxic YACUT × MLC cell hybrids. The killer hybridomas may be of value for analyzing recognition mechanisms and molecules involved in MHC-unrestricted cell-mediated cytotoxicity.