Infection with Chlamydia trachomatis induces inflammatory pathologies in the urogenital tract that can lead to infertility and ectopic pregnancy. Pathogenesis of infection has been mostly attributed to excessive cytok...Infection with Chlamydia trachomatis induces inflammatory pathologies in the urogenital tract that can lead to infertility and ectopic pregnancy. Pathogenesis of infection has been mostly attributed to excessive cytokine production. However, precise mechanisms on how C. trachomatis triggers this production, and which protein（s） stimulate inflammatory cytokines remains unknown. In the present study, the C. trachomatis pORF5 protein induced tumor necrosis factor alpha （TNF-a）, interleukin-1 beta （IL-1β） and interleukin-8 （IL-8） in dose and time-dependent manners in the THP-1 human monocyte cell line. We found that intracellular p38/mitogen-activated protein kinase （MAPK） and extracellular signal-regulated kinase （ERK）/MAPK signaling pathways were required for the induction of TNF- a, IL-1β and IL-8. Blockade of toll-like receptor 2 （TLR2） signaling reduced induction levels of TNF-a, IL-8 and IL-1β. We concluded that the C. trachomatis pORF5 protein might contribute to the inflammatory processes associated with chlamydial infections.展开更多
The inclusion membrane proteins play potentially important roles in chlamydial biology and pathogenesis.Here we localized and characterized the hypothetical protein CT440 in Chlamydia trachomatis-infected cells.The op...The inclusion membrane proteins play potentially important roles in chlamydial biology and pathogenesis.Here we localized and characterized the hypothetical protein CT440 in Chlamydia trachomatis-infected cells.The open reading frame(ORF) encoding the CT440 protein from the C.trachomatis serovar D genome was cloned into the prokaryotic expression vector pGEX-6p and expressed as a glutathione-S-transferase(GST) fusion protein in E.coli XL1-Blue.The CT440 fusion protein was used to immunize mice to raise antigen-specific antibody.After verification by Western blot and immunofluorescence assay(IFA),the specific antibody was used to localize the endogenous CT440 protein and to detect its expression pattern in Chlamydia-infected cells.Cytosolic expression of CT440 in HeLa cells was also carried out to evaluate the effect of the CT440 protein on the subsequent chlamydial infection.The results showed that the hypothetical protein CT440 was localized in the C.trachomatis inclusion membrane,and was detectable 12 h after chlamydial infection.Expression of CT440 in the cytoplasm did not inhibit the subsequent chlamydial infection.In summary,we have identified a new inclusion membrane protein that may be an important candidate for understanding C.trachomatis pathogenesis.展开更多
基金supported by the National Natural Science Foundation of China(30970165,81102230)Team Project for the Technology Innovation of Higher Education of Hunan Province,China,2010
文摘Infection with Chlamydia trachomatis induces inflammatory pathologies in the urogenital tract that can lead to infertility and ectopic pregnancy. Pathogenesis of infection has been mostly attributed to excessive cytokine production. However, precise mechanisms on how C. trachomatis triggers this production, and which protein（s） stimulate inflammatory cytokines remains unknown. In the present study, the C. trachomatis pORF5 protein induced tumor necrosis factor alpha （TNF-a）, interleukin-1 beta （IL-1β） and interleukin-8 （IL-8） in dose and time-dependent manners in the THP-1 human monocyte cell line. We found that intracellular p38/mitogen-activated protein kinase （MAPK） and extracellular signal-regulated kinase （ERK）/MAPK signaling pathways were required for the induction of TNF- a, IL-1β and IL-8. Blockade of toll-like receptor 2 （TLR2） signaling reduced induction levels of TNF-a, IL-8 and IL-1β. We concluded that the C. trachomatis pORF5 protein might contribute to the inflammatory processes associated with chlamydial infections.
基金supported by the National Natural Science Foundation of China (Grant Nos. 30970165 and 81102230)the Hunan Provincial Natu-ral Science Foundation of China (Grant No. 09JJ3059)the Team Project for the Technology Innovation of Higher Education of Hunan Province,China
文摘The inclusion membrane proteins play potentially important roles in chlamydial biology and pathogenesis.Here we localized and characterized the hypothetical protein CT440 in Chlamydia trachomatis-infected cells.The open reading frame(ORF) encoding the CT440 protein from the C.trachomatis serovar D genome was cloned into the prokaryotic expression vector pGEX-6p and expressed as a glutathione-S-transferase(GST) fusion protein in E.coli XL1-Blue.The CT440 fusion protein was used to immunize mice to raise antigen-specific antibody.After verification by Western blot and immunofluorescence assay(IFA),the specific antibody was used to localize the endogenous CT440 protein and to detect its expression pattern in Chlamydia-infected cells.Cytosolic expression of CT440 in HeLa cells was also carried out to evaluate the effect of the CT440 protein on the subsequent chlamydial infection.The results showed that the hypothetical protein CT440 was localized in the C.trachomatis inclusion membrane,and was detectable 12 h after chlamydial infection.Expression of CT440 in the cytoplasm did not inhibit the subsequent chlamydial infection.In summary,we have identified a new inclusion membrane protein that may be an important candidate for understanding C.trachomatis pathogenesis.
