Characterization and growth factor requirements of SJL lymphomas. II. Interleukin 5 dependence of the in vitro cell line, cRCS-X, and influence of other cytokines

Abstract

Follicular B cell lymphomas of SJL mice [reticulum cell sarcoma (RCS)] are dependent on syngeneic Ly-1+,2- T cells for their growth. These T cells produce a number of lymphokines in response to stimulation with gamma-irradiated RCS cells including interleukin (IL) 2, interferon-gamma (IFN-gamma), IL4 and IL5, some of which may be required for growth of the tumor. Previous studies have shown that an RCS cell line, cRCS-X, can be maintained in vitro indefinitely, if the cultures are supplemented with gamma-irradiated lymph node (LN) cells or with a preparation of human B cell growth factor (BCGF). In the present studies, the growth requirements of this cell line were analyzed in more detail in short-term assays of both [3H]thymidine incorporation and colony formation in agarose. Recombinant murine IL5 cause dose-dependent proliferation of cRCS-X cells similar to that induced with BCGF. The level of colony formation by cRCS-X cells induced by optimal concentrations of BCGF was not increased further by the addition of IL5, suggesting that the two factors act via a common mechanism. IL1 and IFN-gamma each enhanced cRCS-X proliferation induced by BCGF or IL5 in both assays. The effects of IL1 plus BCGF, IFN-gamma plus BCGF, and IL1 plus IL5 were clearly synergistic. Preincubation of cRCS-X cells with IL1 enhanced their ability to proliferate in response to BCGF or IL5 in [3H]thymidine incorporation assays, but the reverse sequence of cytokine addition showed no effect of IL1. No such effect was seen with IFN-gamma. Indeed, IL1 and IFN-gamma appeared to affect BCGF-induced cRCS-X growth by different mechanisms and their combined effects were greater than that of IL1 or IFN-gamma added separately. None of the other cytokines studied, including IL2, IL3, IL4, IL6, tumor necrosis factor-alpha, granulocyte monocyte-colony-stimulating factor or transforming growth factor-beta, had any detectable effect on cRCS-X cells, either alone or in combination with BCGF or IL5. Like IL5, SJL gamma-irradiated LN cells induced cRCS-X colony-forming units (CFU) in a dose-dependent manner. IL1, or the combination of IL1 plus IFN-gamma, clearly synergized with LN cells in the induction of cRCS-X CFU, suggesting that LN cells contribute IL5. The level of CFU induced by an optimal dose of BCGF was enhanced further in the presence of LN cells and IL1.(ABSTRACT TRUNCATED AT 400 WORDS)