Partial purification and characterization of the human lymphocyte colony enhancing factor (LCEF).
The cloning potential of PHA-treated T cells is significantly enhanced when lymphocyte culture fluid (LCF) from mitogen-treated lymphocytes is added to the soft agar culture system. The mitogens seem to stimulate the release of a lymphocyte colony enhancing factor (LCEF) into the culture medium. A study of the physico-chemical properties of the LCEF revealed that it is a nondialyzable, heat-labile molecule which migrates in the haptoglobin (2--2) post-transferrin region in acrylamide electrophoresis. It is stable to RNase and DNase but labile to papain and trypsin. The LCEF was partially purified from the crude LCF using a sequence of techniques--ammonium sulphate precipitation, DEAE-cellulose and Bio-Gel P-150 chromatography and disc electrophoresis. The mol. wt of the purified LCEF, determined from gel filtration chromatography, was 90,000--110,000.
D002999:Clone Cells; D003115:Colony-Stimulating Factors / Q000037:antagonists & inhibitors / Q000302:isolation & purification; D003470:Culture Media; D003956:Dialysis; D004591:Electrophoresis, Polyacrylamide Gel; D006358:Hot Temperature; D006801:Humans; D008214:Lymphocytes / Q000276:immunology*; D008222:Lymphokines / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D008970:Molecular Weight
D003115:Colony-Stimulating Factors; D003470:Culture Media; D008222:Lymphokines