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Quantitation, in Vitro Propagation, and Characterization of Preleukemic Cells Induced by Radiation Leukemia Virus¹

Lautenberg Center for General and Tumor Immunology [E. Y., S. E.] and Department of Molecular Genetics [S. E., M. K.], Hebrew University-Hadassah Medical School, Jerusalem 91 010, Israel

Intrathymic (i.t.) inoculation of radiation leukemia virus into C57BL/6 mice induces a population of preleukemic (PL) cells that can progress into mature thymic lymphomas upon transfer into syngeneic recipients. A minimum of 10³ PL thymic cells are required to induce lymphomas in the recipient. Most of the individual lymphomas developed in mice which were inoculated with cells of a single PL thymus, derived from different T-cell precursors. PL thymic cells could be grown in vitro on a feeder layer consisting of splenic stromal cells. Growth medium was supplemented with supernatant harvested from an established radiation leukemia virus-induced lymphoma cell line (SR4). The in vitro-grown PL cells were characterized as Thy-1⁺, CD4⁺, CD8^- T-cells, most of which expressed radiation leukemia virus antigens. Cultured PL cells were found to be nontumorigenic, based on their inability to form s.c. tumors. However, these cells could develop into thymic lymphomas if inoculated i.t. into syngeneic recipients. A culture of PL cells, maintained for 2 mo, showed clonal T-cell receptor arrangement. Lymphomas which developed in several recipient mice upon injection with these PL cells were found to possess the same T-cell receptor arrangement. These results indicate that PL cells can be adapted for in vitro growth while maintaining their preleukemic character.

¹ Supported by Grant 88-00095 of the US-Israel Binational Science Foundation and by Concern Foundation, Los Angeles, CA.

² To whom requests for reprints should be addressed.

Received 3/ 8/90. Accepted 2/ 1/91.

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