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Objective@#Leukocyte mediated IL-6 (IL-6) plays an important role in chronic viral hepatitis pathogenesis and related liver diseases, We did a large sample-size case-control study and clinical data analysis to find association between IL-6 single nucleotide polymorphism and HBV susceptibility, and to achieve the detection of the body on the expression of IL-6 for the prevention and treatment of HBV infection.@*Methods@#Totally 848 HBV patients and 894 healthy controls in Shenzhen were selected and rs1800796 genotypes were determined by TaqMan assays.@*Results@#The result showed that rs1800796 polymorphism was associated with susceptibility to HBV infection (P=0.0003, odds ratio (OR)=1.43, the difference was statistically significant.@*Conclusions@#The single nucleotide polymorphism of IL-6rs1800796 locus was associated with the susceptibility of HBV infection in Chinese population, and the rs1800796 G allele is a protective gene for HBV infection.
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Objective To investigate the cellular immunologic response of TH 17/Treg cells in the peripheral blood of pelvic tuberculosis patients and explore their roles in the pathogenesis of pelvic tuberculosis.Methods The intracellular flow cytometry was performed to evaluate the expressions of TH 17 and Treg cells in 46 pelvic tuberculosis patients and 25 healthy controls in childbearing age.Twenty-eight of the 46 pelvic tuberculosis patients were followed up to monitor the variation of the TH17/Treg cells after 3 months and 6 months of anti-tuberculosis treatment.Results The percentage of TH 17 cells in the peripheral blood of pelvic tuberculosis patients was (3.26 ± 1.30) ％ which was significantly lower than that of healthy controls [(4.92 ± 1.71) ％,P ＜ 0.01].The percentage of Treg cells in the patients was (5.18 ± 1.53) ％ which was significantly higher than that of healthy controls [(3.26 ± 1.10) ％,P ＜ 0.01].The percentage of TH17 cells in the pelvic tuberculosis patients after 6 months of treatment was (4.67 ± 1.75) ％ which was significantly higher than that in the patients before treatment and after 3 months treatment [(3.26 ± 1.30) ％,P ＜ 0.01 and (3.70 ± 1.06) ％,P ＜0.01,respectively].The percentage of Treg cells in pelvic tuberculosis patient after 6 months of treatment was (3.93 ±0.94)％ which was significantly lower than that in the patients before treatment and after 3 months of treatment [(5.18 ± 1.53)％,P ＜0.01 and (4.94 ± 1.51) ％,P ＜ 0.01,respectively].The percentage of Treg cells in the patients after 6 months of treatment was still significantly higher than that of controls (P ＜ 0.05).The TH 17/Treg ratio before treatment was significantly lower than that of healthy controls (P ＜ 0.01),and the TH 17/Treg ratio was increased after 3 months of treatment but it did not show significant difference compared with that before treatment.The TH 17/Treg ratio after 6 months of treatment (1.18 ± 0.34) ％ was significantly increased in contrast to those after 3 months of treatment and before treatment [(0.77 ± 0.21) ％,P ＜ 0.01 and (0.55 ± 0.13) ％,P ＜ 0.01,respectively].The TH 17/Treg ratio could not rise to the normal level even after 6 months of treatment.Conclusion Both the TH 17 and Treg cells may involve in the immunologic responses of pelvic tuberculosis patients and the imbalance of TH1T/Treg cells may remain persistently.
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Objective To evaluate the diagnostic value of two tuberculosis-specific IFN-γ release assays in latent tuberculosis infection among HIV-infected individuals. Methods The levels of tuberculosis antigen-specific IFN-γin 102 HIV patients from AIDS Outpatient Clinic of Shenzhen Third People's Hospital were detected by in-house tuberculosis-specific IFN-γ ELISpot assay and commercial T-SPOT TB kit, and tuberculin skin test (TST) were done at the same time. There were 66 males and 36 females,and the average age was 35. Results Seventeen HIV infected patients were positive in both IFN-γ ELISpot and T-SPOT TB methods, the sensitivity, specificity positive predictive value(PPV), negative predictive value(NPV) and compliance rates of ELISpot were 94. 4% ,94. 0% ,77. 3% ,98. 8% and 94. 1% ,respectively. Three patients were positive in both IFN-γELISpot and T-SPOT TB methods, the sensitivity, specificity, PPV, NPV and compliance rates of TST were 16. 7%, 98. 8%, 75.0%, 84. 7% and 84. 3%, respectively. The average number of spots using three kinds of antigen ESAT-6, Pool A,Pool B obtained were 26. 89 ±5. 77,18. 96 ±4. 75 and 14. 51 ± 3.77, respectively. Only ESAT-6 and Pool B have a statistically significant difference (H=7.557,P = 0.022 9), no significant difference was shown between other groups. There was no significant difference between the positive rate and the CD4+ T cellls number(x2 =0. 860 8 ,P =0. 650 2) ,as the same as the T-SPOT TB (x2 = 1. 396 4, P = 0. 497 5 ). Conclusions The performance of this in-house tuberculosis-specific IFN-γ ELISPot assay was comparable to T-SPOT assay in diagnosis of latent tuberculosis infection, and the sensitivity and specificity of both these two assays were all much higher than TST. They canbe recommended in diagnosing latent tuberculosis infection in HIV infected patients.
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ObjectiveTo evaluate the IL-17 expression in HIV/tuberculosis-coinfected patients and its role in the pathogenesis of this coinfection.MethodsFifty-four HIV infected patients were divided into three groups:simple HIV infected group,HIV with latent tuberculosis infection (HIV+ LTBI) group and HIV coinfected with active tuberculosis (HIV+ ATB) group.The whole blood intracellular cytokine staining was performed and samples were then detected by BD FACSCanto.The expressions of CD4+ IL-17+ T cells and CD4+ IFNγ+ T cells were analyzed using FACSDiva software.Comparison between groups was done by independent sample t test.ResultsThe CD4+ T cell count and viral load among these three groups were comparable.There were no significant difference of the expression of CD4+ IL-17+ T cells between simple HIV infected group and HIV+ LTBI group (1.40 ± 1.01) ％ vs (1.29±0.86) ％,(t=0.336,P＞0.05),but both of these two groups were much higher than HIV+ATB group (t=3.680,t=2.516,P＜0.05).There were no significant differences of the expression of CD4+ IFNγ+ T cells among these three groups [(32.8±24.0)％ vs (40.3±1 21.9) ％ vs (46.1±31.2)％,(t=-0.939,t=-1.602,t=-0.646,P＞0.05)].ConclusionThe Th17 response is down-regulated in HIV/tuberculosis-coinfected patients,which may play an important antitubercular role in the pathogenesis of coinfection.
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T-6 specific IFN-γ ELISPOT has higher specificity, sensitivity, the positive and negative predicative value. Therefore, the ELISPOT warrant for further improvement and clinical application.
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Objective To develop an ELISA(Enzyme-Linked Immunosorbent Assay)diagnostic kit for early rapid detection of sarum anti-EV71 antibody and evaluate its clinical application value.Methods Recombinant protein VP1 of EV71 were prepared and purified as an immobilized antigen for establishment of an indirect ELISA for detection of serum anti-EV71 IgM and anti-EV71 IgG.Compared with RT-PCR.isolation of EV71 and micro-neutralizing assay.the clinical application value of anti-EV71 IgM and anti-EV71 ISG in the diagnosis of EV71 disease was evaluated.Results In comparison with RT-PCR.the sensitivity,specificity,positive predictive value and negative predictive value of anti-EV71 IgM antibody were 83%,85%,81%and 87%,respectively.The sensitivity,specificity,positive predictive value and negative predictive value of anti-EV71 IgG antibody were 72%,74%,68%and 77%.respectively.Compared with viral isolation assay.the sensitivity and specificity of anti-EV71 IgM antibody were 85%and 97%,respectively.The sensitivity and specificity of anti-EV71 IsG antibody were 75%and 77%,respectively.In addition.the titers of anti-EV71 IgG antibody were significantly correlated with the titers of neutralizing antibody to EV71 by linear regression analysis(r=0.72,P＜0.05).Finally,the serum titers of anti-IgG from patients with EV71 associated hand food and mouth disease at convalescent stage exhibited significantly higher than that of the same patients at acute stage(P＜0.01),but the titers of anti-IgM had no significant difference(P＞0.05).Conclusions With VP1 recombinant protein used as an immobilized antigen,an indirect ELISA diagnostic kit was successfully develooed for detection of serum anti-human EV71 IgM and anti-human EV71 IgG antibodies.
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Objective To assess the validity of a newly developed in-house ELISPOT IFN-γ release assay (IGRA) for the detection of latent tuberculosis infection among HIV infected individuals. Methods In-house ELISPOT assay were performed, together with a tuberculin skin test in 205 health controls and 110 HIV infected individuals , who had no signs of active tuberculosis at time of enrolment . Results Using the ELISPOT assay, positivity rates for the 205 health controls, 110 HIV infected individuals and 47 AIDS patients on highly active antiretrovial therapy (HAART) were 7. 3% , 24.5% , 29. 8% , respectively. These results indicated that the positive rates obtained from HIV infected individuals (include patient on HAART) was significantly higher than health controls( P ＜ 0.001). We found no significant correlation between the CD4 cell count and positivity of ELISPOT assay (P ＞0.05 ). The proportion of subjects with a positive response to ELISPOT assay were higher than the proportion of tuberculin skin test(TST) responders(P＜0.0001) in HIV infected individuals. Conclusion Our study indicates that IGRA using M. tuberculosis specific antigens are likely to retain their validity for the diagnosis of LTBI among HIV positive individuals.