Preparation of T cell growth factor free from interferon and factors stimulating hemopoietic cells and mast cells.
A simple two-step method involving ammonium sulfate precipitation followed by hydrophobic chromatography is described for the separation of T cell growth factor (TCGF) from a number of other factors contained in medium conditioned by concanavalin A-stimulated spleen cells. Thus, granulocyte-macrophage colony-stimulating factor, P cell-stimulating activity, pluripotential stem cell-supporting activity and interferon activity were not detected in TCGF partially purified by these steps. T cell-replacing factor co-purified with TCGF. Macrophage activity factor (MAF) co-purified with TCGF, but the ratio of MAF to TCGF activities was reduced more than 20-fold relative to that in crude conditioned medium. All of the factors were present in the 50-80% saturated ammonium sulfate fraction, however, levels of concanavalin A were reduced by 98% in this step. TCGF, separated in this way from these other regulatory factors will be useful in experiments analyzing the actions of TCGF on mixed populations of cells.
D000818:Animals; D002454:Cell Differentiation; D002845:Chromatography; D003115:Colony-Stimulating Factors / Q000302:isolation & purification / Q000494:pharmacology; D003208:Concanavalin A / Q000302:isolation & purification / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D007372:Interferons / Q000302:isolation & purification / Q000494:pharmacology; D007376:Interleukin-2 / Q000302:isolation & purification* / Q000494:pharmacology / Q000592:standards; D015848:Interleukin-5; D008213:Lymphocyte Activation; D008222:Lymphokines / Q000302:isolation & purification* / Q000494:pharmacology; D016215:Macrophage-Activating Factors; D008297:Male; D008407:Mast Cells / Q000166:cytology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008819:Mice, Nude
D003115:Colony-Stimulating Factors; D007376:Interleukin-2; D015848:Interleukin-5; D008222:Lymphokines; D016215:Macrophage-Activating Factors; D003208:Concanavalin A; D007372:Interferons