Erythropoietin‐independent erythroid colony formation in vitro by hemopoietic cells of mice infected with friend virus

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Abstract
We have investigated the production of erythroid colonies in plasma culture by bone‐marrow and spleen cells taken form C3Hf/Bi mice previously infected with a polycythemic strain of Friend virus (FV). Inclusion of erythropoietin (Epo) in the medium was found unnecessary for erythroid colony formation in vitro by these cells, although it was essential for the production of erythroid colonies by hemopoietic cells from normal animals. Development of erythroid colonies also proceeded unimpeded when cells from FV‐infected animals were cultivated in medium pretreated with rabbit antiserum that was shown to inactivate Epo. Thus, the hemopoietic tissues of FV‐infected mice contained erythroid colony‐forming units (CFU‐Es) which appeared to be Epoindependent. When spleen cells from FV‐infected mice were exposed to antiserum directed against syngeneic FV‐infected spleen cells and complement, and then cultured with or without Epo, the number of erythroid colonies that developed was drastically reduced, indicating that the CFU‐Es in these animals carried FV‐induced antigen(s), and must themselves have been infected with virus. Electron microscopy of erythroid colonies produced by cells from FV‐infected mice revealed the presence of budding and abundant free type‐C virus particles. The efficiency of erythroid colony formation in vitro either with or without Epo by hemopoietic cells from FV‐infected mice was substantially increased over that of cells from normal mice. The increase in the number of CFU‐Es in these animals was due mainly to an increase in the number of Epo‐independent CFU‐Es. Epo‐independent CFU‐Es were first detected in bone marrow and spleen as early as 3 days after FV infection: thereafter their numbers progressively increased for at least 9 days. Hypertransfusion with red blood cells prior to FV infection reduced, while bleeding greatly increased, the efficiency of erythroid colony formation without Epo by cells from the spleens of the infected mice. The phenomenon of erythroid colony formation in plasma cultures lacking Epo provides a sensitive and reliable means of detecting Epo‐independent CFU‐Es, which appear to play a fundamental part in pathogenesis of the disease resulting from infection with the polycythemic strain of FV.