Mitogen-activated axolotl (Ambystoma mexicanum) splenocytes produce a cytokine that promotes growth of homologous lymphoblasts.
Culture supernatants (PHA-SNs) from axolotl splenocytes cultured with phytohemagglutinin-P (PHA) in medium supplemented with bovine serum albumin (BSA) were collected after 1, 2, and 3 days, pooled, treated to remove residual PHA, precipitated with saturated ammonium sulfate, dialyzed, aliquoted, and stored at -20 degrees C. PHA-SNs stimulated proliferation of homologous lymphoblasts, but not resting splenocytes. SDS-PAGE of metabolically labeled PHA-SNs revealed a band between 14 and 21 kDa. This corresponds to the M(r) of the gel fractions with biological stimulatory activity eluted from PHA-SNs. Blasts absorbed significant bioactivity of PHA-SNs whereas freshly harvested splenocytes did not. Although splenocytes cultured in medium supplemented with 1% fetal bovine serum (FBS) did not proliferate in response to PHA, they did secrete a cytokine with lymphoblast growth-promoting activity. Furthermore, PHA-induced lymphoblasts, initially cultured in medium supplemented with 0.25% BSA, could proliferate in response to PHA-SNs in 1% FBS-supplemented medium.
Koniski
A
A
Department of Microbiology and Immunology, University of Rochester School of Medicine and Dentistry, NY 14642.
Cohen
N
N
D000558:Ambystoma mexicanum / Q000276:immunology*; D000818:Animals; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D007376:Interleukin-2 / Q000096:biosynthesis*; D008213:Lymphocyte Activation / Q000276:immunology*; D010835:Phytohemagglutinins / Q000494:pharmacology; D013154:Spleen / Q000166:cytology / Q000276:immunology*; D013950:Thymus Gland / Q000166:cytology
D017077:Culture Media, Conditioned; D007376:Interleukin-2; D010835:Phytohemagglutinins