Possible role of Toll-like receptor-2 in the intracellular survival of Staphylococcus aureus in murine peritoneal macrophages: involvement of cytokines and anti-oxidant enzymes.
Effects of blocking toll-like receptor-2 (TLR-2) on the survival of Staphylococcus aureus (S. aureus) and cytokine production in peritoneal macrophages of Swiss albino mice were analysed. Macrophages were infected with S. aureus in the presence and absence of anti-TLR-2 antibody. Tumour necrosis factor-α (TNF-α) interleukin-6 (IL-6), interferon-gamma (IFN-γ), interleukin-1β (IL-1β), interleukin-12 (IL-12) and interleukin-10 (IL-10) concentrations were measured. Expressions of TLR-2, NF-κB, MyD 88 were analysed by Western Blot. Expression of TLR-2 was increased in S. aureus-infected macrophages with respect to control and was MyD 88 independent. TLR2 blocking significantly reduced TNF-α, IL-6, IL-1β and IL-10 and increased IFN-γ and IL-12 production. Decreased catalase activity and increased superoxide dismutase (SOD) by S. aureus with concomitant increase in H2 O2 and nitric oxide (NO) were observed in the case of prior TLR-2 blocking. To understand whether catalase contributing in the intracellular survival, was of bacterial origin or not, 3-amino, 1, 2, 4-triazole (ATZ) was used to inhibit specifically macrophage-derived catalase. Catalase enzyme activity from the whole staphylococcal cells in the presence of ATZ suggested that the released catalase were of extracellular origin. From the intracellular survival assay, it was evident that pretreatment of macrophages with ATZ reduces the bacterial burden in macrophages when infected with the recovered bacteria only from the anti-TLR-2 antibody-treated macrophages after phagocytosis. Catalase protein expression from the whole staphylococcal cells recovered after phagocytosis also indicated the catalase release from S. aureus. Capturing of S. aureus via TLR-2 induces inflammatory reactions through activation of NF-κB-signalling pathways which was MyD88-independent.
Department of Physiology, Immunology Laboratory, University of Calcutta, University Colleges of Science and Technology, Calcutta, West Bengal, India.
B
B
Bishayi
D
D
Bandyopadhyay
A
A
Majhi
R
R
Adhikary
D000640:Amitrole / Q000494:pharmacology; D000818:Animals; D000906:Antibodies / Q000276:immunology; D002374:Catalase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D016207:Cytokines / Q000096:biosynthesis / Q000276:immunology; D006861:Hydrogen Peroxide / Q000378:metabolism; D017737:Macrophages, Peritoneal / Q000201:enzymology / Q000276:immunology* / Q000382:microbiology; D008297:Male; D051379:Mice; D053594:Myeloid Differentiation Factor 88 / Q000096:biosynthesis; D052002:NF-kappa B p50 Subunit / Q000096:biosynthesis; D052003:NF-kappa B p52 Subunit / Q000096:biosynthesis; D009569:Nitric Oxide / Q000096:biosynthesis; D010587:Phagocytosis / Q000276:immunology; D013203:Staphylococcal Infections / Q000276:immunology*; D013211:Staphylococcus aureus / Q000201:enzymology* / Q000276:immunology*; D013482:Superoxide Dismutase / Q000096:biosynthesis; D051195:Toll-Like Receptor 2 / Q000276:immunology*; D051996:Transcription Factor RelA / Q000096:biosynthesis / Q000276:immunology
D000906:Antibodies; D016207:Cytokines; C507780:Myd88 protein, mouse; D053594:Myeloid Differentiation Factor 88; D052002:NF-kappa B p50 Subunit; D052003:NF-kappa B p52 Subunit; C498144:Rela protein, mouse; C493486:Tlr2 protein, mouse; D051195:Toll-Like Receptor 2; D051996:Transcription Factor RelA; D009569:Nitric Oxide; D006861:Hydrogen Peroxide; D002374:Catalase; D013482:Superoxide Dismutase; D000640:Amitrole