Object: To investigate the relationship between chlamydiatrachomatis (CT) and urogenital infection. Method Positive rate of CT in patients with inflammationof urogenital tract was significantly higher than those witho...Object: To investigate the relationship between chlamydiatrachomatis (CT) and urogenital infection. Method Positive rate of CT in patients with inflammationof urogenital tract was significantly higher than those withoutinflammation(P<0.05). Result: There was statistical difference in the males nomatter they were patients with inflammation of urogenitaltract or not (P>H0.05), while there was no statistical differencein females (P>0.05). The incidence of the infection was highamong those aging from 21-50 years old. Conclusion: The clinical manifestations of CT infectionwere obscure, so we should examine CT in patients who haveno symptoms, especially in females and those of high-riskpopulation.展开更多
Objectives: To find out the level and functions of Chlamydia trachomatis heat shock protein (C-hsp60) antibody, anti-spermantibody(ASAb), interleukin 1(IL-1), interleukin 6 (IL-6),interleukin 8 (IL-8), Tumor necrosis ...Objectives: To find out the level and functions of Chlamydia trachomatis heat shock protein (C-hsp60) antibody, anti-spermantibody(ASAb), interleukin 1(IL-1), interleukin 6 (IL-6),interleukin 8 (IL-8), Tumor necrosis factor alpha (TNF-α)and γ-interferon (IFN-γ) in patients with CT-related infertility. Methods: CT-DNA of cervical secretions was detectedthrough polymerase chain reaction (PCR) and migrationinhibiting factor (MIF) was employed to measure IgG titre ofCT MOMP antibody. Western blot was used to determinepresence of C-hsp60 antibody and enzyme-linkedimmunoadsorbent assay (ELISA) measured ASAb of IgG typein blood serum and determine the content of IL-1, IL-6. IL-8.TNF-α. IFN-γ in uterine tube fluid. Results: 68 patients had positive CT-DNA, among which 57(83.8%) had C-hsp60 antibody. Among the 172 patients withnegative CT-DNA, 64 patients (37.2%) also had C-hsp60Antibody. There was a significant difference (P<0.01) betweeninfertile patients and control group patients in the presence ofAAb. Infertile patients with positive CT-DNA had higher levels of IL-1、IL-6. IL-8. TNF-α. IFN-γ in uterine tube fluidcompared to control group patients (P<0.01). Conclusion: Firstly, those patients with negative CT testingfrom cervical secretions cannot be ruled out for CT infectionin deep parts of the body (such as oviduct, pelvic kidney).Detection of C-hsp60 Antibody may help to diagnose suchcases of CT. Secondly, CT infection of the oviduct can raiselevels of IL-1、IL-6、 IL-8、 TNF-α. IFN-γ. The pathogenesis ofinfertility caused by CT infection in the reproductive tractmay be related to cytokine production and inflammatoryresponses mediated by C-hsp60 Antibody, IL-1, IL-6, IL-8,TNF-α, and IFN-γ.展开更多
AIM:To evaluate the direct binding of two main chlamydial biovars(C.trachomatis and C.pneumoniae) to plasma lipoproteins and its effect on chlamydial infection rate in human hepatoma cell line(HepG2 cells). METHODS:Mu...AIM:To evaluate the direct binding of two main chlamydial biovars(C.trachomatis and C.pneumoniae) to plasma lipoproteins and its effect on chlamydial infection rate in human hepatoma cell line(HepG2 cells). METHODS:Murine plasma lipoproteins were fractionated and isolated using fast-performance liquid chromatography(FPLC),spotted on nitrocellulose membrane and incubated with chlamydial suspensions. Direct binding of chlamydial particles to lipoprotein fractions has been studied using lipopolysaccharide-specific antibodies in immuno-dot blot binding assay and immunoprecipitation analysis.Immunostaining protocol as well as flow cytometry analysis have been employed to study the infectivity rate of chlamydial species in HepG2 cells. RESULTS:Elementary bodies of both C.trachomatis and C.pneumoniae bind ApoB-containing fractions of plasma lipoproteins.That binding becomes stronger when heat-denatured FPLC fractions are used, suggesting a primary role of apolipoproteins in interaction between chlamydial particle and lipoprotein. Both chlamydial biovars efficiently propagate in human hepatoma cell line-HepG2 cells even in serum free conditions forming late-stage inclusion bodies and releasing extracellular elementary bodies.Preincubation of C.trachomatis and C.pneumoniae with native ApoB-containing lipoproteins enhances the rate of chlamydial infection in HepG2 cells.CONCLUSION:A productive infection caused by C. trachomatis and C.pneumoniae may take place in human-derived hepatocytes revealing hepatic cells as possible target in chlamydial infection.Obtained results may suggest the participation of lipoprotein receptors in the mechanism of attachment and/or entry of chlamydial particles into target cells.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
文摘Object: To investigate the relationship between chlamydiatrachomatis (CT) and urogenital infection. Method Positive rate of CT in patients with inflammationof urogenital tract was significantly higher than those withoutinflammation(P<0.05). Result: There was statistical difference in the males nomatter they were patients with inflammation of urogenitaltract or not (P>H0.05), while there was no statistical differencein females (P>0.05). The incidence of the infection was highamong those aging from 21-50 years old. Conclusion: The clinical manifestations of CT infectionwere obscure, so we should examine CT in patients who haveno symptoms, especially in females and those of high-riskpopulation.
基金The Project was funded by the Department of Education,Jangsu Province(Fund Number: 00KJD310003).
文摘Objectives: To find out the level and functions of Chlamydia trachomatis heat shock protein (C-hsp60) antibody, anti-spermantibody(ASAb), interleukin 1(IL-1), interleukin 6 (IL-6),interleukin 8 (IL-8), Tumor necrosis factor alpha (TNF-α)and γ-interferon (IFN-γ) in patients with CT-related infertility. Methods: CT-DNA of cervical secretions was detectedthrough polymerase chain reaction (PCR) and migrationinhibiting factor (MIF) was employed to measure IgG titre ofCT MOMP antibody. Western blot was used to determinepresence of C-hsp60 antibody and enzyme-linkedimmunoadsorbent assay (ELISA) measured ASAb of IgG typein blood serum and determine the content of IL-1, IL-6. IL-8.TNF-α. IFN-γ in uterine tube fluid. Results: 68 patients had positive CT-DNA, among which 57(83.8%) had C-hsp60 antibody. Among the 172 patients withnegative CT-DNA, 64 patients (37.2%) also had C-hsp60Antibody. There was a significant difference (P<0.01) betweeninfertile patients and control group patients in the presence ofAAb. Infertile patients with positive CT-DNA had higher levels of IL-1、IL-6. IL-8. TNF-α. IFN-γ in uterine tube fluidcompared to control group patients (P<0.01). Conclusion: Firstly, those patients with negative CT testingfrom cervical secretions cannot be ruled out for CT infectionin deep parts of the body (such as oviduct, pelvic kidney).Detection of C-hsp60 Antibody may help to diagnose suchcases of CT. Secondly, CT infection of the oviduct can raiselevels of IL-1、IL-6、 IL-8、 TNF-α. IFN-γ. The pathogenesis ofinfertility caused by CT infection in the reproductive tractmay be related to cytokine production and inflammatoryresponses mediated by C-hsp60 Antibody, IL-1, IL-6, IL-8,TNF-α, and IFN-γ.
文摘AIM:To evaluate the direct binding of two main chlamydial biovars(C.trachomatis and C.pneumoniae) to plasma lipoproteins and its effect on chlamydial infection rate in human hepatoma cell line(HepG2 cells). METHODS:Murine plasma lipoproteins were fractionated and isolated using fast-performance liquid chromatography(FPLC),spotted on nitrocellulose membrane and incubated with chlamydial suspensions. Direct binding of chlamydial particles to lipoprotein fractions has been studied using lipopolysaccharide-specific antibodies in immuno-dot blot binding assay and immunoprecipitation analysis.Immunostaining protocol as well as flow cytometry analysis have been employed to study the infectivity rate of chlamydial species in HepG2 cells. RESULTS:Elementary bodies of both C.trachomatis and C.pneumoniae bind ApoB-containing fractions of plasma lipoproteins.That binding becomes stronger when heat-denatured FPLC fractions are used, suggesting a primary role of apolipoproteins in interaction between chlamydial particle and lipoprotein. Both chlamydial biovars efficiently propagate in human hepatoma cell line-HepG2 cells even in serum free conditions forming late-stage inclusion bodies and releasing extracellular elementary bodies.Preincubation of C.trachomatis and C.pneumoniae with native ApoB-containing lipoproteins enhances the rate of chlamydial infection in HepG2 cells.CONCLUSION:A productive infection caused by C. trachomatis and C.pneumoniae may take place in human-derived hepatocytes revealing hepatic cells as possible target in chlamydial infection.Obtained results may suggest the participation of lipoprotein receptors in the mechanism of attachment and/or entry of chlamydial particles into target cells.
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.
