HSV-mediated transfer of interleukin-10 reduces inflammatory pain through modulation of membrane tumor necrosis factor alpha in spinal cord microglia.
To dissect the molecular basis of the neuroimmune response associated with the genesis of inflammatory (nociceptive) pain, we constructed a herpes simplex virus-based gene transfer vector to express the antiinflammatory cytokine interleukin-10 (IL-10), and used it to examine the effect of IL-10 expression in activated microglial cells in vitro, and in inflammatory pain in vivo. IL-10 reduced the phosphorylation of p38 mitogen-activated protein kinase (MAPK) and decreased the expression of full-length membrane spanning tumor necrosis factor-alpha (mTNFalpha) following lipopolysaccharide stimulation of microglia in vitro. IL-10 also reduced intracellular cleavage of mTNFalpha and release of the soluble cleavage product sTNFalpha. Similar effects on TNFalpha expression were observed when the cells were pretreated with a p38 MAPK inhibitor. In animals, injection of a dilute solution of formalin in the skin resulted in an increase in mTNFalpha in spinal dorsal horn, without detectable sTNFalpha. Local release of IL-10 achieved by gene transfer reduced the number of spontaneous flinches in the early and delayed phases of the formalin test of inflammatory pain. The effect of IL-10 on nocisponsive behavior correlated with a block in phosphorylation of p38 and reduced expression of 26 kDa mTNFalpha in spinal microglia. The results emphasize the key role played by membrane TNFalpha in the spinal neuroimmune response in pain caused by peripheral inflammation.
Zhou
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Z
Department of Neurology, University of Michigan Medical School and GRECC and Neurology Service, VA Ann Arbor Healthcare System, Ann Arbor, MI 48109-0316, USA.
Peng
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Hao
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Fink
D J
DJ
Mata
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D000818:Animals; D002478:Cells, Cultured; D015536:Down-Regulation; D005557:Formaldehyde; D015316:Genetic Therapy / Q000379:methods*; D016753:Interleukin-10 / Q000235:genetics*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D017628:Microglia / Q000276:immunology*; D023421:Models, Animal; D009187:Myelitis / Q000473:pathology / Q000628:therapy; D059408:Pain Management; D010766:Phosphorylation; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D018139:Simplexvirus / Q000235:genetics*; D013116:Spinal Cord / Q000276:immunology; D014161:Transduction, Genetic / Q000379:methods; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism
D008070:Lipopolysaccharides; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10; D005557:Formaldehyde; D048051:p38 Mitogen-Activated Protein Kinases