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Effects of Hematopoietic Suppressor Molecules on the in Vitro Proliferation of Purified Murine Granulocyte-Macrophage Progenitor Cells¹

Departments of Medicine (Hematology/Oncology) [D. E. W., S. C., H. E. B.], Microbiology and Immunology [H. E. B.], and The Walther Medical Research Institute [H. E. B.], Indiana University School of Medicine, Indianapolis, Indiana 46223

Highly purified murine granulocyte-macrophage progenitor cells (CFU-GM) were used as target cells to assess the possible direct effects of purified preparations of recombinant murine -interferon, prostaglandin E, recombinant human heavy chain (acidic) ferritin, and recombinant human tumor necrosis factor (TNF-) on progenitor cells in vitro. Target CFU-GM, with cloning efficiencies of up to 84% and containing 0–3% morphologically recognizable accessory cells at the initiation of the culture period, were plated at a density of 100–150 cells/dish in the presence or absence of pure suppressor molecules. Colony formation was stimulated with either crude pokeweed mitogen-stimulated mouse spleen conditioned medium, pure natural murine macrophage colony-stimulating factor, or pure recombinant murine granulocyte-macrophage colony-stimulating factor. All four suppressor molecules were active in vitro against purified CFU-GM as assessed by their ability to inhibit colony or cluster formation. No apparent difference in the degree of responsiveness to prostaglandin E, -interferon, or human heavy chain (acidic) ferritin was noted in the presence of pokeweed mitogen-stimulated mouse spleen conditioned medium, granulocyte-macrophage colony-stimulating factor, or macrophage colony-stimulating factor. In contrast, TNF- in cultures containing macrophage colony-stimulating factor slightly, but significantly, potentiated colony formation. TNF- also appeared more active at suppressing colony formation at lower concentrations in pokeweed mitogen-stimulated mouse spleen conditioned medium than in granulocyte-macrophage colony-stimulating factor-stimulated cultures of purified CFU-GM. The results suggest that TNF-, human heavy chain (acidic) ferritin, -interferon, and prostaglandin E can act directly at the progenitor cell level.

¹ Supported by USPHS Grants CA-36464 and CA-36740 from the National Cancer Institute to Dr. Broxmeyer. Dr. Williams was supported by NIH Training Program IT 32 AM-07519.

² To whom requests for reprints should be addressed, at Department of Medicine, Room 379 Clinical Building, Indiana University School of Medicine, 541 Clinical Drive, Indianapolis, IN 46223.

Received 9/30/87. Accepted 12/11/87.

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