Accuri C6 Flow Cytometer System
Originally published online as doi:10.1189/jlb.1105693 on March 24, 2006

Published online before print March 24, 2006
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(Journal of Leukocyte Biology. 2006;79:1357-1368.)
© 2006 by Society for Leukocyte Biology

RGS1 and RGS13 mRNA silencing in a human B lymphoma line enhances responsiveness to chemoattractants and impairs desensitization

Jang-Il Han, Ning-Na Huang, Dong-Uk Kim and John H. Kehrl1

Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland

1Correspondence: Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bldg. 10, Room 11B08, 10 Center Dr., MSC 1876, Bethesda, MD 20892. E-mail: jkehrl{at}niaid.nih.gov

Chemokines bind receptors that are members of the G-protein-coupled receptor family. Chemokine receptors transduce intracellular signals by activating heterotrimeric G-proteins. Acting to limit and modulate heterotrimeric G-protein signaling is a family of proteins, termed regulator of G-protein signaling (RGS). Two of these proteins, RGS1 and RGS13, are well-expressed in germinal center B cells and many Burkitt’s lymphoma cell lines. Reducing RGS13 and to a lesser extent RGS1 expression in a Burkitt’s lymphoma cell line enhances responsiveness to two chemokines, CXC chemokine ligand 12 (CXCL12) and CXCL13, and reducing both mRNAs augments the responses more dramatically. The double knock-down (KD) cells respond better to restimulation with CXCL12 or CXCL13 after a primary stimulation with CXCL12 than do the control cells. The double-KD cells also exhibit a greater propensity to polarize and to develop multiple small lamellipodia. These results indicate that RGS1 and RGS13 act together to regulate chemokine receptor signaling in human germinal center B lymphocytes and provide evidence that they contribute significantly to the rapid desensitization of the signaling pathway.

Key Words: B lymphocyte • calcium • chemokine




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