Suppressive biological activity of a synthetic pentapeptide on highly enriched human and murine marrow hematopoietic progenitors: synergism with recombinant human tumor necrosis factor-alpha and interferon-gamma.
The synthetic pentapeptide pGlu-Glu-Asp-Cys-Lys (SPI) was evaluated in vitro alone and in combination with recombinant human tumor necrosis factor-alpha (TNF-alpha) and/or interferon-gamma (IFN-gamma) for effects on colony formation by hematopoietic progenitor cells (HPC) present in low-density (LD), nonadherent low-density T-lymphocyte-depleted (NALT-), and highly enriched sorted progenitor cells from normal human bone marrow. Progenitor cells in NALT- fractions were further enriched by cell sorting using two-color fluorescence on a Coulter Epics 753 flow cytometry apparatus with My10 and HLA-DR monoclonal antibodies. The sorted My10 DR+ progenitor cell population had a cloning efficiency of up to 38% for granulocyte-macrophage colony-forming units (CFU-GM), erythroid burst-forming units (BFU-E), and multipotential colony-forming units (CFU-GEMM). SP1 inhibited, by up to 86%, each of the colony-forming cells in a dose-dependent fashion. The sensitivities of the different progenitor cells to inhibition by the pentapeptide were the same when My10 DR+ marrow cells were used, and the progenitor cells in the My10 DR+ fraction were more sensitive than the cells in the LD or NALT- fraction to inhibition by SP1. The suppressive activity of SP1 on purified HPC was confirmed when 10(-3) M SP1 completely inhibited colony and cluster formation from a population of mouse bone marrow cells in which one of two cells was a CFU-GM. The effects of SP1 were not absolutely cell-cycle-specific for human HPC, but the non-S-phase cells were less sensitive than the S-phase cells to the suppressive effects of SP1. SP1 synergized with TNF-alpha and/or IFN-gamma to inhibit proliferation of progenitor cells using both LD or My10 DR+ human marrow cells stimulated by recombinant human interleukin 3 (IL-3). These studies suggest that the suppressive effect of SP1 occurs in the absence of certain accessory cells (e.g., monocytes and T-lymphocytes), that this effect may be mediated directly at the level of the HPC, and that this pentapeptide can be considered a candidate modulatory molecule for HPC proliferation.
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Department of Medicine (Hematology/Oncology), Indiana University School of Medicine, Indianapolis 46223.
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Chillemi
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Shen
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Carow
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Broxmeyer
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D000818:Animals; D002455:Cell Division / Q000187:drug effects; D003114:Colony-Forming Units Assay; D004357:Drug Synergism; D005434:Flow Cytometry; D006131:Growth Inhibitors; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D066298:In Vitro Techniques; D007371:Interferon-gamma / Q000494:pharmacology*; D051379:Mice; D009842:Oligopeptides / Q000138:chemical synthesis / Q000494:pharmacology*; D011761:Pyrrolidonecarboxylic Acid / Q000031:analogs & derivatives; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D006131:Growth Inhibitors; D009842:Oligopeptides; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma; C040758:hemoregulatory peptide 5b; D011761:Pyrrolidonecarboxylic Acid