Lipoarabinomannan induced cytotoxic effects in human mononuclear cells.
Mycobacteria produce large quantities of lipoarabinomannan, a cell wall associated glycolipid, which confers virulence in many of the disease causing members of the genus. We studied the lipoarabinomannan induced altered signaling mechanism in human peripheral mononuclear cells. Lipoarabinomannan isolated from Mycobacterium smegmatis (a non-pathogenic species) at concentrations of 2, 5 and 10 microg ml(-1) was used for the study. It was found that protein kinase C activity of human mononuclear cells was significantly inhibited by lipoarabinomannan at these concentrations in a dose dependent manner. Calcium, phosphatidyl serine and diglyceride dependent phosphorylation of endogenous proteins (mainly 90-, 80-, 66-, 38-, 36- and 34-kDa proteins) were also inhibited. Potentially cytotoxic superoxide anions generated by peripheral blood mononuclear cells were scavenged by lipoarabinomannan. It was also observed that incubation of peripheral blood mononuclear cells with lipoarabinomannan at concentrations of 5 and 10 microg ml(-1) for 6 h directed the cells towards apoptotic cell death, confirmed by DNA degradation analysis, microscopic observation of Wright-Giemsa as well as DAPI stained nuclei. These results clearly demonstrate that lipoarabinomannan from M. smegmatis may exert its cytotoxic activity via inhibition of protein kinase C, a key signaling molecule inside the mononuclear cells, which ultimately affects the phosphorylation of various proteins imperative for cellular defence and survival.
Department of Microbiology, Bose Institute, Calcutta, India.
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Ghosh
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Pal
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Das
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Dasgupta
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Majumdar
D000328:Adult; D017209:Apoptosis; D002118:Calcium / Q000378:metabolism; D004075:Diglycerides / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D005260:Female; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008070:Lipopolysaccharides / Q000633:toxicity*; D008297:Male; D009161:Mycobacterium; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D010718:Phosphatidylserines / Q000378:metabolism; D010750:Phosphoproteins / Q000097:blood; D010766:Phosphorylation; D011493:Protein Kinase C / Q000037:antagonists & inhibitors* / Q000097:blood; D015398:Signal Transduction / Q000187:drug effects; D013481:Superoxides / Q000097:blood
D004075:Diglycerides; D008070:Lipopolysaccharides; D010718:Phosphatidylserines; D010750:Phosphoproteins; C050016:lipoarabinomannan; D013481:Superoxides; D009240:N-Formylmethionine Leucyl-Phenylalanine; D011493:Protein Kinase C; D002118:Calcium