Accuri C6 Flow Cytometer System
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Published online before print December 10, 2009
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© by The Society for Leukocyte Biology
Journal of Leukocyte Biology, doi:10.1189/jlb.0709502

Received for publication July 21, 2009.
Revised November 12, 2009.
Accepted for publication November 15, 2009.

Article

Differential heat shock protein localization in chronic lymphocytic leukemia

Nina C. Dempsey *, Francesca Leoni *, H. Elyse Ireland *, Christine Hoyle {dagger}, and John H. H. Williams *@

*Chester Centre for Stress Research, University of Chester, Chester, United Kingdom; and{dagger}Glan Clwyd Hospital, Bodelwyddan, North Wales

@ To whom correspondence should be addressed. E-mail: john.williams{at}.


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Abstract

Mechanisms behind carcinogenesis and resistance of tumor cells to treatment regimes remain elusive. The major stress proteins Hsp72, Hsp90, and Hsp27 are credible candidates to provide this resistance, as their overexpression in many cancer types is well documented. In addition to being present inside tumor cells, where they confer resistance to apoptosis, Hsp72, in particular, is presented externally, embedded in the cell membrane of cancer cells. This study aimed to investigate the localization of Hsp72, Hsp90, and Hsp27 in leukocytes from patients with CLL and age-matched control subjects. CLL patients were found to express significantly higher levels of iHsp90 (CLL=2463 MFI; control=748 MFI) and iHsp27 (CLL=2190 MFI; control=1031 MFI) in lymphocytes than that expressed by lymphocytes from control subjects. Furthermore, expression of iHsp90 was shown to be related to stage of disease, and expression of iHsp27 correlated with levels of active caspase-3. Patients were found to express very high levels or very low levels of sHsp72 and iHsp72 in CD5+/CD19+ cells, although surface and intracellular datasets did not correlate. Levels of extracellular Hsp72 circulating in the serum were found to correlate with internal levels of Hsp72 and were also found to be significantly lower in patients receiving corticosteroid treatment than in patients not receiving corticosteroid treatment. Finally, analysis of the number of circulating Tregs revealed significantly elevated numbers in CLL patients compared with control subjects.

Key Words: Hsp72 • Hsp90 • Hsp27 • CLL • Tregs