TNF-α modulates TLR2-dependent responses during mycobacterial infection.
Multifunctional roles of tumor necrosis factor-alpha (TNF-α) during the mycobacterial pathogenesis make it an important molecule to understand and to examine the course of infection. Identification and analysis of TNF-α response can largely contribute to determine the potential host mediators for therapeutic intervention against tuberculosis. The current chapter describes several methods to assess the ability of TNF-α signaling to modulate toll-like receptor (TLR)2 signaling, another key player in mycobacterial infection and its responses. Experiments involving neutralizing antibodies, antagonists, pharmacological inhibitors, and siRNA-mediated gene silencing are discussed in this chapter to establish the role of TNF-α signaling. The widely used protein and mRNA analysis readouts like enzyme-linked immunosorbent assay (ELISA), immunoblotting, fluorescence-activated cell sorting (FACS), and quantitative real-time RT-PCR are useful to estimate and confirm the mediators involved in TNF-α and TLR2 signaling.
Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore, Karnataka, 560012, India.
Sahana
S
Holla
Jamma
J
Trinath
Kithiganahalli Narayanaswamy
KN
Balaji
D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology; D017737:Macrophages, Peritoneal / Q000166:cytology* / Q000378:metabolism / Q000382:microbiology; D051379:Mice; D008810:Mice, Inbred C57BL; D009161:Mycobacterium / Q000472:pathogenicity*; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D051195:Toll-Like Receptor 2 / Q000378:metabolism*; D014376:Tuberculosis / Q000276:immunology* / Q000503:physiopathology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D008070:Lipopolysaccharides; D016328:NF-kappa B; D012333:RNA, Messenger; D051195:Toll-Like Receptor 2; D014409:Tumor Necrosis Factor-alpha