Signaling molecules involved in production and regulation of IL-1beta by murine peritoneal macrophages in vitro on treatment with concanavalin A.
In the present study we report the activation of murine peritoneal macrophages in vitro on treatment with Concanavalin A (ConA). ConA (10 microg/ml) treatment of macrophages resulted in the transcription of IL-1beta gene at 16 h and maximum production of IL-1beta at 24 h. To investigate the signaling molecules involved in the production of IL-1beta different pharmacological inhibitors were used. It was observed that genestein, wortmannin, H-7, TMB-8, PD98059, SB202190, and tyrophostin (AG490) down regulated the expression of IL-1beta. These observations suggested the involvement of tyrosine kinase, PI3 kinase, protein kinase C, p42/44, p38, Ca(++) and JAK2 signaling molecules in ConA induced production of IL-1beta by macrophages. Maximum protein tyrosine kinase activity and expression of PI3K in macrophages was seen at 5 min, PKC activity and Ca(++) release was found at 10 min after ConA treatment. Maximum expression of phospho-JAK2 at 2.5-5 min, phospho-p42/44 at 5-60 min, phospho-p38 at 15-30 min, phospho-IkappaB and phospho-Stat1 at 30-60 min and phospho-ELK1, c-Fos, phospho-Stat3 at 60 min of ConA treatment was observed. Pharmacological inhibitors were also used to check the cascade of activation of tyrosine kinase, PKC, PI3 kinase, p42/44, p38, JAK kinase and release of Ca(++) from intracellular storage to sort out the signaling pathways involved in the release of IL-1beta by macrophages on treatment with ConA in vitro.
Sodhi
Ajit
A
School of Biotechnology, Faculty of Science, Banaras Hindu University, Varanasi-221005, India. ajit.sodhi@lycos.com
Kesherwani
Varun
V
D000818:Animals; D002118:Calcium / Q000378:metabolism; D003208:Concanavalin A / Q000494:pharmacology*; D005260:Female; D005786:Gene Expression Regulation; D053583:Interleukin-1beta / Q000378:metabolism*; D053614:Janus Kinase 2 / Q000378:metabolism; D008262:Macrophage Activation; D017737:Macrophages, Peritoneal / Q000187:drug effects* / Q000378:metabolism*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D011493:Protein Kinase C / Q000378:metabolism; D015398:Signal Transduction; D014157:Transcription Factors / Q000378:metabolism
D053583:Interleukin-1beta; D014157:Transcription Factors; D003208:Concanavalin A; D019869:Phosphatidylinositol 3-Kinases; C507926:Jak2 protein, mouse; D053614:Janus Kinase 2; D011493:Protein Kinase C; D002118:Calcium