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Membrane-bound proteinase 3 and PAR2 mediate phagocytosis of non-opsonized bacteria in human neutrophils

Cited 8 time in Web of Science Cited 8 time in Scopus
Authors
Kim, Yong C.; Shin, Ji E.; Lee, Sung H.; Chung, Woo-Jae; Choi, Bong K.; Choi, Youngnim; Lee, Yoon-Sik
Issue Date
2011-09
Publisher
PERGAMON-ELSEVIER SCIENCE LTD
Citation
MOLECULAR IMMUNOLOGY; Vol.48, No.15-16, pp.1966-1974
Keywords
NeutrophilsBacteriaPhagocytosisPAR2PR3
Abstract
The molecular mechanisms underlying the non-opsonic phagocytosis of bacteria by neutrophils are poorly understood. We previously reported the efficient uptake of Streptococcus sanguinis by human neutrophils in the absence of opsonins. To characterize the phagocytosis receptor, protein lysates from neutrophils and HL-60 cells were subjected to affinity chromatography using epoxy beads coated with S. sanguinis. Denaturing electrophoresis of the eluted proteins and subsequent mass spectrometry revealed that one of the proteins eluted from neutrophils was proteinase 3 (PR3). Enzymatic cleavage of the glycosylphosphatidylinositol linker of NB1, a co-receptor for membrane-bound PR3 (mPR3), significantly reduced the phagocytosis of S. sanguinis. In addition, the neutralization of mPR3 with antibody reduced both binding and phagocytosis of S. sanguinis. Treatment of neutrophils with a serine proteinase inhibitor indicated that protease activity is required for phagocytosis. Thus, we studied whether protease-activated receptor 2 (PAR2) is involved in signal transmission from mPR3 during this process. Indeed, neutralizing antibodies against PAR2 inhibited phagocytosis and S. sanguinis-induced calcium mobilization desensitized PAR2. Furthermore, the phagocytosis of S. sanguinis and the concomitant activation of Rho family GTPases were inhibited by the intracellular calcium chelator, BAPTA-AM. Collectively, mPR3 acts as a non-opsonic phagocytosis receptor for bacteria probably by activating PAR2 in neutrophils. (C) 2011 Elsevier Ltd. All rights reserved.
ISSN
0161-5890
Language
English
URI
https://hdl.handle.net/10371/80416
DOI
https://doi.org/10.1016/j.molimm.2011.05.026
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College of Dentistry/School of Dentistry (치과대학/치의학대학원)Dept. of Dentistry (치의학과)Journal Papers (저널논문_치의학과)
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