Detection of synergistic factor and interleukin-3 activity in the serum and ascites fluid of mice bearing the WEHI-3 tumour.
Media conditioned (CM) by WEHI-3 cells (a myelomonocytic leukemia cell line) contains a number of haemopoietic growth factors, including synergistic factor (SF) and interleukin-3 (IL3). We have investigated the production of SF and IL3 in vivo in mice bearing the WEHI-3 tumour. SF and IL3 activity were detected in both the sera and ascites fluids of these mice. SF from the ascites fluid was partially purified by a four-step purification schedule consisting of ammonium sulphate fractionation, DEAE-cellulose, hydroxylapatite, and Sephadex G-100 chromatography. This purification sequence resulted in approximately a 250- and 187-fold purification of SF and IL3 respectively on the initial starting material with a yield of 13 and 9.7% respectively of the initial activity. At each stage of purification, the fractions containing SF co-purified with IL3 activity, further supporting our previous report that SF and IL3 are probably identical molecules. The characteristics of the in-vivo derived (sera and ascites fluid) activities were found to be similar to those of the factors produced in vitro in WEHI-3 cell conditioned media. These results support the conclusion that SF and IL3 are produced in vivo in WEHI-3 tumour bearing mice and are not in vitro artifacts.
D000818:Animals; D001202:Ascitic Fluid; D002460:Cell Line; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D006133:Growth Substances / Q000032:analysis*; D006412:Hematopoietic Stem Cells / Q000187:drug effects; D007377:Interleukin-3; D007942:Leukemia, Experimental / Q000378:metabolism*; D007951:Leukemia, Myeloid / Q000378:metabolism*; D008222:Lymphokines / Q000032:analysis* / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C
D006133:Growth Substances; D007377:Interleukin-3; D008222:Lymphokines