Production of TNF-alpha, IL-1beta, IL-12 and IFN-gamma in murine peritoneal macrophages on treatment with wheat germ agglutinin in vitro: involvement of tyrosine kinase pathways.
Treatment of macrophages with various doses of wheat germ agglutinin (WGA) for different time intervals resulted in enhanced expression of TNF-alpha, IL-1beta, IL-12 and IFN-gamma. The maximum expressions were observed at 24 h with 100 ng/ml of WGA. Enhanced transcription of cytokines TNF-alpha, IL-1beta, IL-12, and IFN-gamma was observed at 16 h of WGA treatment by RT-PCR. Pharmacological inhibitor of tyrosine kinase, PI3 kinase, protein kinase C, p42/44, p38, JNK and intracellular calcium immobilizing agent down regulated the WGA induced expression of cytokines TNF-alpha, IL-1beta, IL-12 and IFN-gamma. Maximum protein tyrosine kinase activity in macrophages was seen at 5 min of WGA treatment. Maximum cytosolic Ca++ was observed at 10 min of WGA treatment. WGA treated macrophages showed maximum activation of protein kinase C (PKC) and PI3 kinase at 10 min, p42/44, p38 at 15 min and JNK at 30 min. Transcription factor ELK1 was activated at 60 min and IêB, c-Fos and c-Jun at 30 min of WGA treatment. The pharmacological inhibitors were also used to check the cascade of activation of tyrosine kinase, PKC, PI3 kinase, p42/44, p38, JNK and release of calcium from intracellular storage to sort out the signal pathways involved in the release of TNF-alpha, IL-1beta, IL-12, and IFN-gamma by macrophages on treatment with WGA in vitro.
School of Biotechnology, Faculty of Science, Banaras Hindu University, Varanasi 221005, India. ajit.sodhi@lycos.com
Ajit
A
Sodhi
Varun
V
Kesherwani
D000818:Animals; D005260:Female; D005786:Gene Expression Regulation; D007371:Interferon-gamma / Q000096:biosynthesis*; D018664:Interleukin-12 / Q000096:biosynthesis*; D053583:Interleukin-1beta / Q000096:biosynthesis*; D007700:Kinetics; D008264:Macrophages / Q000378:metabolism; D017737:Macrophages, Peritoneal / Q000378:metabolism*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008954:Models, Biological; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis*; D014909:Wheat Germ Agglutinins / Q000378:metabolism*
D053583:Interleukin-1beta; D014409:Tumor Necrosis Factor-alpha; D014909:Wheat Germ Agglutinins; D018664:Interleukin-12; D007371:Interferon-gamma; D011505:Protein-Tyrosine Kinases