betadex and methylamine

There are an increasing number of studies reporting the presence of Hsps in human serum. We have investigated the release of Hsp70 into blood and culture medium from peripheral blood mononuclear cells (PBMCs), and whether this release is due to cell damage or active secretion from the cells. Intact Hsp70 was released from cells within whole blood and from purified PBMCs under normal culture conditions. Hsp70 release was rapid (0.1 ng/10(6) cells/h) over the first 2 h of culture and continued at a reduced rate up to 24 h (<0.025 ng/10(6) cells/h). Using viable cell counts and lactate dehydrogenase release we were able to confirm that the release of Hsp70 was not due to cellular damage. Hsp70 release was inhibited by monensin, methyl-beta-cyclodextrin, and methylamine, but not by brefeldin A. These data suggest that Hsp70 is released from cells via a non-classical pathway, possibly involving lysosomal lipid rafts.

Topics: B-Lymphocytes; beta-Cyclodextrins; Blotting, Western; Brefeldin A; Cell Survival; Culture Media; HSP70 Heat-Shock Proteins; Humans; L-Lactate Dehydrogenase; Leukocytes, Mononuclear; Lysosomes; Membrane Microdomains; Methylamines; Monensin; Protein Synthesis Inhibitors; RNA, Messenger; T-Lymphocytes; Temperature; Time Factors