Removal of PHA from supernatants containing T-cell growth factor.
Supernatants of human PBL cultures stimulated with PHA contain a lymphokine named T-cell growth factor (TCGF). To remove the contaminant PHA from these supernatants, we have designed a simple and effective method based on the binding properties of this lectin to chicken red blood cells. This procedure completely removes the functional activities of a PHA solution, both in lectin-induced cellular cytotoxicity against erythroid targets and in DNA synthesis of fresh PBL. The TCGF activity remaining in the supernatants after absorption with CRBC is dependent on the cellular concentration employed in the preparation of the supernatants. The absorption procedure does not remove significant TCGF activity in the supernatants prepared at high cellular concentrations (5 X 10(6) cells/ml), whereas in those prepared at low cellular concentrations (1 X 10(6) cells/ml) a partial loss of TCGF activity is detected.
D000818:Animals; D002478:Cells, Cultured; D002645:Chickens / Q000097:blood; D003470:Culture Media / Q000032:analysis*; D006801:Humans; D007158:Immunologic Techniques; D008222:Lymphokines / Q000302:isolation & purification; D010835:Phytohemagglutinins / Q000302:isolation & purification*; D013601:T-Lymphocytes / Q000166:cytology
D003470:Culture Media; D008222:Lymphokines; D010835:Phytohemagglutinins