In vitro activation of murine peritoneal macrophages by monocyte chemoattractant protein-1: upregulation of CD11b, production of proinflammatory cytokines, and the signal transduction pathway.
The CC chemokine monocyte chemotactic protein-1 (MCP-1) is a major mediator of monocyte/macrophage infiltration at the inflammatory sides under both physiologic and pathologic conditions. We report the ability of MCP-1 to activate murine peritoneal macrophages in vitro for enhanced expression of CD11b, macrophage-mediated cytotoxicity, and production of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 (IL-1). The macrophages treated with MCP-1 in vitro displayed significant cytolytic activity toward TNF-alpha-sensitive L929 cells in a dose-dependent manner. The macrophage-mediated L929 cytotoxicity was blocked in the presence of anti-TNF-alpha antibodies, suggesting the involvement of TNF-alpha. Production of TNF-alpha and IL-1 macrophages on MCP-1 treatment was maximum at 24 h of incubation with 100 ng/ml MCP-1. Enhanced TNF-alpha and IL-1beta mRNA expression was also demonstrated by RT-PCR, which revealed transcription of interferon gamma (IFN-gamma), IL-12, and related T cell-specific chemokine genes, KC and IP-10, in the MCP-1-treated macrophages. The pharmacologic inhibitors pertussis toxin (100 ng/ml), wortmannin (200 ng/ml), H-7 (10 microM), PD98059 (25 microM), and genistein (10 microg/ml) significantly inhibited TNF-alpha and IL-1 production in the MCP1-treated macrophages, suggesting the involvement of G-proteins, phosphoinositol-3-kinase (PI3K), protein kinase C, p42/44 MAPK, and tyrosine kinases in this process.
Biswas
Subhra K
SK
School of Biotechnology, Banaras Hindu University, Varanasi 221005, India.
Sodhi
Ajit
A
D000818:Animals; D039481:CD11b Antigen / Q000378:metabolism*; D018932:Chemokine CCL2 / Q000494:pharmacology*; D054360:Chemokine CXCL1; D054357:Chemokine CXCL10; D018925:Chemokines; D019743:Chemokines, CXC / Q000235:genetics; D016207:Cytokines / Q000096:biosynthesis* / Q000235:genetics; D003602:Cytotoxicity, Immunologic; D005260:Female; D066298:In Vitro Techniques; D018836:Inflammation Mediators / Q000378:metabolism; D007371:Interferon-gamma / Q000235:genetics; D007375:Interleukin-1 / Q000096:biosynthesis / Q000235:genetics; D018664:Interleukin-12 / Q000235:genetics; D008262:Macrophage Activation / Q000187:drug effects*; D017737:Macrophages, Peritoneal / Q000187:drug effects* / Q000276:immunology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000235:genetics; D015854:Up-Regulation / Q000187:drug effects
D039481:CD11b Antigen; D018932:Chemokine CCL2; D054360:Chemokine CXCL1; D054357:Chemokine CXCL10; D018925:Chemokines; D019743:Chemokines, CXC; C475895:Cxcl1 protein, mouse; D016207:Cytokines; D018836:Inflammation Mediators; D007375:Interleukin-1; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; C074302:keratinocyte-derived chemokines; D018664:Interleukin-12; D007371:Interferon-gamma