PMID- 8132324 OWN - NLM STAT- MEDLINE DCOM- 19940421 LR - 20210526 IS - 0019-9567 (Print) IS - 1098-5522 (Electronic) IS - 0019-9567 (Linking) VI - 62 IP - 4 DP - 1994 Apr TI - Induction of early-response genes KC and JE by mycobacterial lipoarabinomannans: regulation of KC expression in murine macrophages by Lsh/Ity/Bcg (candidate Nramp). PG - 1176-84 AB - The murine chromosome 1 gene Lsh/Ity/Bcg (candidate Nramp) regulates macrophage activation for antimicrobial activity against Salmonella typhimurium, Leishmania donovani, and Mycobacterium spp. To determine early events in the activation pathway, the ability of mycobacterial lipoarabinomannan (LAM) to induce early gene (KC and JE) expression in macrophages from susceptible (S) C57BL/10ScSn (Lshs) and congenic resistant (R) B10.L-Lshr mice was investigated. Stimulation with 1.8 microgram of arabinofuranosyl-terminated LAM (AraLAM) per ml resulted in similar kinetics for KC or JE expression in S and R macrophages. However, whereas JE/glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNA ratios remained equivalent, R macrophages consistently showed enhanced KC/GAPDH ratios within 30 to 40 min of stimulation compared with S macrophages. Significant differences in KC/GAPDH ratios were observed throughout the peak period (0.5 to 6 h) of the KC response and with doses of AraLAM ranging from 0.01 to 2.5 micrograms/ml. Heavily mannosylated LAM from virulent Mycobacterium tuberculosis Erdman, in doses of up to 2.5 micrograms/ml, failed to stimulate KC or JE in S or R macrophages. Gamma interferon alone (25 U/ml) stimulated equivalent JE expression in S and R macrophages and synergized with AraLAM to enhance JE in both. In contrast, AraLAM-induced KC expression was inhibited in the presence of gamma interferon. Agonist/inhibitor studies were undertaken to determine the signal transduction pathways mediating KC expression. The protein kinase C (PKC) inhibitor Calphostin C (200 nM) inhibited AraLAM-induced KC by 34% +/- 4% in S macrophages and 43% +/- 5% in R macrophages; the cyclic AMP-dependent PKA inhibitor KT5720 (2 microM) inhibited AraLAM-induced KC by 33% +/- 4% (S) and 25% +/- 5% (R). A role for Ca2+ was indicated because ionophore alone stimulated KC expression and synergized with AraLAM to give a dramatically enhanced response. Induction of KC was also inhibited by (i) blocking constitutive nitric oxide (NO) production by preincubation of macrophages with NG-monomethyl-L-arginine (400 microM) (48% +/- 8% [S] and 40% +/- 11% [R]) and (ii) incubation of macrophages with the cyclic GMP-dependent kinase inhibitor KT5823 (4 microM) (65% +/- 4% [S] and 72% +/- 6% [R]). The manner in which these PKC-, PKA-, and Ca(2+)-dependent, NO-mediated cyclic GMP-dependent kinase signal transduction pathways may relate to function of the candidate Lsh/Ity/Bcg gene Nramp is discussed. FAU - Roach, T I AU - Roach TI AD - Department of Medicine, University of Cambridge Clinical School, Addenbrooke's Hospital, United Kingdom. FAU - Chatterjee, D AU - Chatterjee D FAU - Blackwell, J M AU - Blackwell JM LA - eng GR - N01-AI-05074/AI/NIAID NIH HHS/United States GR - Wellcome Trust/United Kingdom PT - Journal Article PT - Research Support, Non-U.S. Gov't PT - Research Support, U.S. Gov't, P.H.S. PL - United States TA - Infect Immun JT - Infection and immunity JID - 0246127 RN - 0 (Carrier Proteins) RN - 0 (Cation Transport Proteins) RN - 0 (Chemokine CCL2) RN - 0 (Chemokine CXCL1) RN - 0 (Chemokines) RN - 0 (Chemokines, CXC) RN - 0 (Chemotactic Factors) RN - 0 (Cxcl1 protein, mouse) RN - 0 (Cytokines) RN - 0 (Iron-Binding Proteins) RN - 0 (Lipopolysaccharides) RN - 0 (Membrane Proteins) RN - 0 (lipoarabinomannan) RN - 0 (natural resistance-associated macrophage protein 1) RN - 0 (solute carrier family 11- (proton-coupled divalent metal ion transporters), member 2) RN - 147037-79-4 (keratinocyte-derived chemokines) RN - 31C4KY9ESH (Nitric Oxide) RN - 82115-62-6 (Interferon-gamma) SB - IM GS - Lsh/Ity/Bcg MH - Animals MH - Carrier Proteins/*genetics MH - *Cation Transport Proteins MH - Cells, Cultured MH - Chemokine CCL2 MH - Chemokine CXCL1 MH - Chemokines MH - Chemokines, CXC MH - Chemotactic Factors/*genetics MH - Cytokines/*genetics MH - Dose-Response Relationship, Drug MH - *Gene Expression Regulation MH - Interferon-gamma/pharmacology MH - *Iron-Binding Proteins MH - Lipopolysaccharides/*pharmacology MH - Macrophages/*metabolism MH - Membrane Proteins/*genetics MH - Mice MH - Mice, Inbred C57BL MH - Mycobacterium/pathogenicity MH - Nitric Oxide/physiology MH - Signal Transduction PMC - PMC186252 EDAT- 1994/04/01 00:00 MHDA- 1994/04/01 00:01 PMCR- 1994/04/01 CRDT- 1994/04/01 00:00 PHST- 1994/04/01 00:00 [pubmed] PHST- 1994/04/01 00:01 [medline] PHST- 1994/04/01 00:00 [entrez] PHST- 1994/04/01 00:00 [pmc-release] AID - 10.1128/iai.62.4.1176-1184.1994 [doi] PST - ppublish SO - Infect Immun. 1994 Apr;62(4):1176-84. doi: 10.1128/iai.62.4.1176-1184.1994.