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The release of microparticles by Jurkat leukemia T cells treated with staurosporine and related kinase inhibitors to induce apoptosis

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Abstract

Microparticles (MPs) are small membrane-bound vesicles released from cells undergoing activation or cell death. These particles display potent biological activities that can impact on physiologic and pathologic processes. Previous studies with the Jurkat T leukemia cell line demonstrated that staurosporine (STS) induces the release of MPs as cells undergo apoptosis. To investigate further this process, we tested the effects of STS, its analogue, 7-hydroxystaurosporine (UCN-01), and other protein kinase C (PKC) and cyclin-dependent kinase (CDK) inhibitors. FACS analysis was used to assess MP release. Results of these studies indicate that STS and UCN-01 induce MP release by Jurkat cells; in contrast, other PKC and CDK inhibitors failed to induce comparable release, suggesting that release does not result from simple inhibition of either kinase alone. Time course experiments indicated that STS-induced particle release occurred as early as 2 h after treatment, with the early release MPs displaying low levels of binding of annexin V and propidium iodide (PI). Early-release MPs, however, matured in culture to an annexin V- and PI-positive phenotype. Together, these results indicate that STS and UCN-01 induce MPs that are phenotypically distinct and reflect specific patterns of kinase inhibition during apoptosis.

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Acknowledgments

This research was funded by grants from the VA Merit Review program, Alliance for Lupus Research and NIH grant AI082402.

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Authors and Affiliations

  1. Division of Rheumatology and Immunology, Department of Medicine, Duke University, Durham, NC, USA

    Anirudh J. Ullal & David S. Pisetsky

  2. Medical Research Service, Durham Veterans Administration Medical Center, Durham, NC, USA

    David S. Pisetsky

  3. VA Medical Center, 151G, 508 Fulton Street, Durham, NC, 27705, USA

    Anirudh J. Ullal

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Correspondence to Anirudh J. Ullal.

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Ullal, A.J., Pisetsky, D.S. The release of microparticles by Jurkat leukemia T cells treated with staurosporine and related kinase inhibitors to induce apoptosis. Apoptosis 15, 586–596 (2010). https://doi.org/10.1007/s10495-010-0470-3

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