Expression of human T antigens in interspecies hybridomas

Interspecies hybrids were constructed by fusing normal human male peripheral blood mononuclear cells with BW5147, a HGPRT- thymoma derived from an AKR mouse. Hybrid cells were selected in HAT media in culture dishes containing 1 X 10(7) human red blood cells. Twelve weeks after fusion, hybridomas were diluted to 10-15 cells/well and characterized for their expression of the human immune cell surface antigens HLA-DR, T3, T4, and T8 using fluorescent microscopy and cytographic analysis. More than 70% of the hybrid colonies expressed human T-cell surface antigens. Moreover the specific human repetitive DNA (ALU) bound to DNA sequences isolated from the hybridomas after Southern transfers. However, the same hybrids did not have a statistically significant increase in their chromosome number when compared to the mouse parent cell line. Several of the hybridomas produced a soluble factor capable of stimulating the growth of the IL-2 restricted murine cell line CTLL-2 and supported DNA synthesis in human peripheral T-cell populations. Panning experiments demonstrated that the IL-2 producing hybridomas could be enriched by selecting for the human T-cell surface antigen T3. The results presented here indicate that mouse X human hybridomas which express a broad range of human lymphocyte markers can be constructed and maintained in continuous culture for extended periods of time. It also appears that the T3-Ti receptor complex mediates the proliferation of T cells through the T3 molecules linkage to the secretion and/or production of IL-2. The usefulness of interspecific T-cell hybrids as an immunogenetic research tool as well as the significance of the mapping data are discussed.