Rapid generation of human T cell hybridomas

Using the concept of a selectable surface marker we have generated screenable human T cell hybridomas within 4-7 days of fusion. OKT4+ T cell blasts were fused with MOLT4, an OKT4- T cell line, and OKT4+ fusion products were obtained by indirect rosetting technique. Supernatants of this heterogeneous population of hybrids were screened and those with activities of interest were subjected to cloning on soft agar and re-screening to dissect out various activities. Fusion efficiency, by staining, ranges from 6 to 80% depending on the state of activation of the T blasts and the growth phase of the MOLT4 line. This method allows for immortalization of T cell subpopulations, clones and malignant cells to study factor production and potentially messenger RNA for these factors. Additionally selectable surface markers may be used in human B cell and rodent:human fusions, avoiding the innate toxicity of various selection media.