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Hsp70 release from peripheral blood mononuclear cells.
Claire Hunter-Lavin1, Emma L Davies, Maria M F V G Bacelar
1Chester Centre for Stress Research, Biological Sciences, University College Chester, Parkgate Road, Chester CH1 4BJ, United Kingdom.
Biochemical and Biophysical Research Communications
|October 12, 2004
Summary
Heat shock proteins (Hsps) like Hsp70 are actively secreted from peripheral blood mononuclear cells (PBMCs) into serum. This release occurs through a non-classical pathway, not due to cell damage.
Area of Science:
- Cellular Biology
- Immunology
- Biochemistry
Background:
- Heat shock proteins (Hsps) are increasingly detected in human serum.
- The mechanisms of Hsp release into circulation require elucidation.
Purpose of the Study:
- To investigate the release of Heat Shock Protein 70 (Hsp70) from peripheral blood mononuclear cells (PBMCs).
- To determine if Hsp70 release is due to cellular damage or active secretion.
Main Methods:
- Culturing intact whole blood and purified PBMCs under normal conditions.
- Measuring Hsp70 release rates over 24 hours.
- Assessing cellular damage using viable cell counts and lactate dehydrogenase (LDH) release.
- Evaluating the effect of inhibitors (monensin, methyl-beta-cyclodextrin, methylamine, brefeldin A) on Hsp70 release.
Main Results:
- Intact Hsp70 was actively released from PBMCs in culture and whole blood.
- Hsp70 release was rapid initially (0.1 ng/10^6 cells/h) and sustained at a lower rate.
- No significant cellular damage correlated with Hsp70 release.
- Release was inhibited by monensin, methyl-beta-cyclodextrin, and methylamine, but not brefeldin A.
Conclusions:
- Hsp70 is actively secreted from PBMCs via a non-classical pathway.
- The pathway may involve lysosomal lipid rafts.
- Hsp70 release is independent of cellular damage.