In vitro activation of murine peritoneal macrophages by ultraviolet B radiation: upregulation of CD18, production of NO, proinflammatory cytokines and a signal transduction pathway.
In the present study, we report the activation of murine peritoneal macrophages in vitro on irradiation with sublethal dose of UVB (50 mJ/cm(2)). The activation involves enhanced expression of CD18 molecule and production of nitric oxide (NO), tumor necrosis factor (TNF-alpha) and interleukin-1 (IL-1). Production of NO, TNF-alpha and IL-1 by the macrophages on UVB irradiation was found to peak at 24 h of incubation post UVB irradiation. Increased iNOS, TNF-alpha and IL-1beta mRNAs expression was also observed by reverse transcription and polymerase chain reaction (RT-PCR). The RT-PCR results also showed the increased transcription of IL-6, IL-12, TLR2 and TLR4 genes in UVB-irradiated macrophages. Increased expression of phospho-IkappaB was also observed by immunoblotting in UVB-irradiated macrophages. Investigating the signal transduction pathway responsible for the NO, TNF-alpha and IL-1 production by the UVB-irradiated macrophages, it was observed that the pharmacological inhibitors pertussis toxin, wortmannin, PD98059, SB202190 and genistein blocked NO, TNF-alpha and IL-1 production suggesting the probable involvement of G-proteins, phosphoinositol-3-kinase, p42/44, p38 mitogen activated protein kinases and tyrosine kinases in the above process.
Sethi
Gautam
G
School of Biotechnology, Banaras Hindu University, Varanasi 221005, India.
Sodhi
Ajit
A
D000818:Animals; D018821:CD18 Antigens / Q000096:biosynthesis* / Q000235:genetics; D002478:Cells, Cultured / Q000187:drug effects / Q000528:radiation effects; D016207:Cytokines / Q000096:biosynthesis* / Q000235:genetics; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D005786:Gene Expression Regulation / Q000528:radiation effects; D051550:I-kappa B Kinase; D007378:Interleukins / Q000096:biosynthesis / Q000235:genetics; D007739:L Cells; D008262:Macrophage Activation / Q000528:radiation effects*; D017737:Macrophages, Peritoneal / Q000187:drug effects / Q000528:radiation effects*; D008297:Male; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D009569:Nitric Oxide / Q000096:biosynthesis* / Q000235:genetics; D037342:Pertussis Toxin / Q000494:pharmacology; D010766:Phosphorylation / Q000528:radiation effects; D011499:Protein Processing, Post-Translational / Q000528:radiation effects; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D011956:Receptors, Cell Surface / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects / Q000528:radiation effects; D013047:Specific Pathogen-Free Organisms; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D051193:Toll-Like Receptors; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000235:genetics; D014466:Ultraviolet Rays
D018821:CD18 Antigens; D016207:Cytokines; D004791:Enzyme Inhibitors; D007378:Interleukins; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D051193:Toll-Like Receptors; D014409:Tumor Necrosis Factor-alpha; D009569:Nitric Oxide; D037342:Pertussis Toxin; D017346:Protein Serine-Threonine Kinases; C496564:Chuk protein, mouse; D051550:I-kappa B Kinase; C496562:Ikbkb protein, mouse; C496566:Ikbke protein, mouse