title: Lipopolysaccharide could be internalized into human peripheral blood mononuclear cells and elicit TNF-alpha release, but not via the pathway of toll-like receptor 4 on the cell surface.
abstract: Lipopolysaccharide (LPS), the principal component of the outer membrane of Gram-negative bacteria, stimulates various cell types to release numerous proinflammatory mediators such as TNF-alpha, IL-6 and IL-12, which may damage cells and lead to organ injury, even sepsis and septic shock. Toll-like receptor 4 (TLR4) has been identified as the receptor involved in the recognition of LPS, but the role of LPS uptake in activating signal transduction remains controversial. In the present study, TNF-alpha was used as a marker of macrophages/ monocytes activated by LPS, and CQ was used as an inhibitor of endosome mature in order to definitude what stage of the signal transduction elicited by LPS was interrupted. We found that there indeed existed internalization of LPS and internalization partially participated in LPS signaling since CQ inhibited cytokine release, and decreased accumulation of FITC-LPS in hPBMCs. In contrast, anti-hTLR4 antibody could decrease cytokine release, but had no inhibition on accumulation of FITC-LPS. This result revealed that inhibition of cytokine release was related to reduction of FITC-LPS accumulation in the cells. But TLR4 on the cell surface couldn't participate in internalization of LPS. Thus, LPS signaling and internalization couldn't be viewed as mutually independent processes.
authors.affiliation: Department of Pharmacology, College of Medicine, Third Military Medical University, Chongqing 400038, China. zhouh64@mail.tmmu.com.cn
authors.forename: Hong
authors.initials: H
authors.lastname: Zhou
authors.forename: Guofu
authors.initials: G
authors.lastname: Ding
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.forename: Liangxi
authors.initials: L
authors.lastname: Wang
authors.forename: Yongling
authors.initials: Y
authors.lastname: Lu
authors.forename: Hongwei
authors.initials: H
authors.lastname: Cao
authors.forename: Jiang
authors.initials: J
authors.lastname: Zheng
mesh_terms: D000911:Antibodies, Monoclonal / Q000494:pharmacology; D018501:Antirheumatic Agents / Q000494:pharmacology; D002478:Cells, Cultured; D002738:Chloroquine / Q000494:pharmacology; D004705:Endocytosis / Q000187:drug effects / Q000276:immunology; D006090:Gram-Negative Bacteria / Q000276:immunology*; D006801:Humans; D018664:Interleukin-12 / Q000276:immunology / Q000378:metabolism; D015850:Interleukin-6 / Q000276:immunology / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D008070:Lipopolysaccharides / Q000276:immunology / Q000494:pharmacology*; D012772:Shock, Septic / Q000276:immunology; D051197:Toll-Like Receptor 4 / Q000276:immunology*; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D018501:Antirheumatic Agents; D015850:Interleukin-6; D008070:Lipopolysaccharides; D051197:Toll-Like Receptor 4; D014409:Tumor Necrosis Factor-alpha; D018664:Interleukin-12; D002738:Chloroquine