Involvement of MIP-2 and CXCR2 in neutrophil infiltration following injection of late apoptotic cells into the peritoneal cavity.
Apoptotic cells are cleared by phagocytes, such as macrophages, as soon as they appear in vivo. If apoptosis occurs acutely, however, macrophages may be outnumbered by apoptotic cells, which causes late apoptosis. We previously showed that injection of late apoptotic cells into the peritoneal cavity led to transient infiltration of neutrophils. In this study, we examined the involvement of MIP-2 and CXCR2 in the neutrophil infiltration. We first produced a recombinant MIP-2 protein, and a fusion protein between CXCR2 and GST in E. coli, and then generated anti-MIP-2 antibodies and anti-CXCR2 antibodies in rabbits. We then confirmed their specificity by Western blotting analysis and flow cytometry. Injection of late apoptotic cells, such as P388 cells treated with etoposide for 24 hours and CTLL-2 cells cultured in IL-2-free medium for 28 hours, induced neutrophil infiltration into the peritoneal cavity, as expected. The antibodies, but not control antibodies against GST, suppressed the neutrophil infiltration to the level caused by injection of normal (viable) cells, suggesting that MIP-2 and CXCR2 are mainly involved in the neutrophil infiltration caused by late apoptotic cells.
Iyoda
T
T
Division of Molecular Medicine, Department of Biomolecular Science, Faculty of Science, Toho University, Funabashi, Chiba 274-8510, Japan.
Kobayashi
Y
Y
D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000494:pharmacology; D000918:Antibody Specificity / Q000276:immunology; D017209:Apoptosis; D015153:Blotting, Western; D002460:Cell Line; D045744:Cell Line, Tumor; D017690:Cell Transplantation / Q000379:methods; D054426:Chemokine CXCL2; D018925:Chemokines / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D005047:Etoposide / Q000494:pharmacology; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007114:Immunization; D007274:Injections, Intraperitoneal; D007376:Interleukin-2 / Q000172:deficiency; D007958:Leukocyte Count; D008264:Macrophages / Q000166:cytology / Q000187:drug effects / Q000637:transplantation; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D020556:Neutrophil Infiltration / Q000187:drug effects / Q000502:physiology*; D009504:Neutrophils / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D010529:Peritoneal Cavity / Q000166:cytology / Q000502:physiology*; D010538:Peritonitis / Q000139:chemically induced / Q000503:physiopathology; D011817:Rabbits; D023063:Receptors, Interleukin-8B / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D013601:T-Lymphocytes / Q000637:transplantation; D013864:Thioglycolates / Q000494:pharmacology
D000906:Antibodies; D054426:Chemokine CXCL2; D018925:Chemokines; C404915:Cxcl2 protein, mouse; D007376:Interleukin-2; D023063:Receptors, Interleukin-8B; D011993:Recombinant Fusion Proteins; D013864:Thioglycolates; D005047:Etoposide