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Objective@#To establish a quantitative assay of serum Golgi protein 73 (GP73) using xMAP technology and evaluate its performance.@*Methods@#Monoclonal antibodies against GP73 were prepared and purified, and antibody pair screening was performed by double-antibody sandwich enzyme-linked immunosorbent assay. The screened antibodies were used to construct a Luminex liquid chip detection system, and the analysis performance of the detection system was evaluated. The serum levels of GP73 were detected in 90 clinical samples from healthy controls and patients with chronic hepatitis B infection (CHB) and hepatocellular carcinoma (HCC).@*Results@#Five anti-GP73 monoclonal antibodies were prepared and purified, and 5 antibody pairs were successfully screened. The Luminex liquid chip detection system of GP73 was successfully constructed using 8F10D1 and 10B9F11 antibody pairs. The analytical performance evaluation showed that the sensitivity of this system was 0.25 ng/ml and the dynamic range was 0.25-100 ng/ml. No cross reactivity was observed. The intra- and inter-assay variation for GP73 was <8% and <11%, respectively. The recovery was 83%-92%. The linear regression equation was y=1.141x+ 6.436 (r2=0.998 4, P<0.001). The GP73 concentrations in the serum samples of healthy control, CHB group, and HCC group were 42.8 (38.68, 55.90) ng/ml, 61.49 (43.59, 81) ng/ml, and 122.78 (49.36 liter, 264.55) ng/ml, respectively. The levels of GP73 in HCC group were significantly higher than those in CHB group and healthy controls (P<0.05). Moreover, the levels of GP73 in CHB group were significantly higher than those in healthy controls (P<0.05).@*Conclusions@#A liquid chip detection system of GP73 was successfully constructed. It provides a powerful tool for the clinical application of GP73 in the future.
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Objective:HIV-1 infection was associated with a variety of host genetic factors ,and HLA was one of the factors that contribute to the progression of disease .We analyze the frequency of HLA-A, HLA-B, HLA-C in MSM cohort , and the relationship between HLA gene with disease progression .Methods:PCR-sequence specific primer typing HLA typing was used to detect the allele frequency of HLA gene in 310 patients.HIV Molecular Immunology Database ( HLA Analysis Tools ) to analysis the frequency of HLA.Results:In MSM cohort,HLA-A*1101(34.52%) gene was the highest,followed by HLA-A*0201(31.94%),HLA-C*0102 (27.10%) and HLA-A*2402(26.45%).According to the CD4 cell count of HIV infected patients ,the results showed that there were low levels of HLA-B*4403(1.12%,P=0.0276),HLA-B*1511(2.25%,P=0.0282) and HLA-B*5701(0.37%,P=0.0491) in rapid progressors,while the HLA-C*0304(8.96%,P=0.0319) was higher than that of nonprogressors (4.56%).Conclusion: We obtained the distribution frequency data of HLA-A,HLA-B and HLA-C in MSM cohort.Our data presented here may offer potential cor-relation between dominant effect of HLA alleles and HIV-1 disease progression.And found that the HLA-B*4403,HLA-B*1511, HLA-B*5701 related to slow HIV disease progression and HLA-C*0304 related to accelerate HIV disease progression .
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Objective To analyze the differentiation status of CTL and to evaluate its clinical val-ue in patients with HIV/HCV coinfection .Methods Twenty-eight patients with HIV/HCV coinfection and twelve patients with single HCV infection were enrolled in this study .The technique of Fibro-Scan was used to evaluate liver fibrosis .The viral load of HCV was detected by real-time quantitative PCR .Flow cytometry analysis was performed to measure the differentiation status of CTL .Results Both of the levels of alanine transaminase ( ALT) and alkaline phosphatase ( ALP) in patients with HIV/HCV coinfection were signifi-cantly higher than those in patients with single HCV infection [(53.7464±48.1180) U/L vs (27.4750± 13.9850) U/L, P=0.012;(24.5071±8.1940) g/L vs (16.9667±7.1890) g/L, P=0.009].The liver stiffness of patients with HIV/HCV coinfection was higher than that of patients with single HIV infection [(5.9500, 5.8250) Kpa vs (5.1500, 1.0500) Kpa, P=0.117].Compared with the patients with single HCV infection, the patients with HIV/HCV coinfection showed higher viral loads of HCV [( 6.4768, 5.3434) lg copy/ml vs (2.6815, 1.6990) lg copy/ml, P=0.012], but lower clearance rate of HCV [32.14%vs 75%, P=0.032].Compared with the patients with single HCV infection , the patients with HIV/HCV coinfection showed lower percentages of CD 27+CD28+CTL [(28.265±15.095)%vs (18.068±10.263)%, P=0.017), but higher percentages of CD27+/-CD28+CTL [(62.449 ±14.561)% vs (71.111±12.681)%, P=0.066].A trend toward negative correlation was observed between the percent -age of CD27+CD28+CTL and the degree of liver stiffness (r=-0.310, P=0.058).Conclusion HIV in-fection could accelerate the progression of liver disease in patients with HIV /HCV coinfection by affecting the differentiation of CTL .
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Objective To understand the influence of HIV infection on hepatitis C progress in patients co-infected with HIV and hepatitis C virus (HCV) and related immune mechanism. Methods Twenty eight patients co-infected with HIV/HCV and 12 patients with simplex HCV infection were enrolled. The liver function and hepatic fibrosis progress were evaluated by detecting peripheral blood and with Fibro-Scan. The viral load of HCV was detected by using real time quantitative PCR. And the percentage of Treg/CD4 +T lymphocyte cell was tested by using flow cytometry. Results The levels of ALT and ALP in HIV/HCV co-infection group were (76.16 ± 81.248)U/L,(24.507 1 ± 8.194)g/L respectively,higher than those of simplex HCV infection group [(27.475 0±13.985)U/L,(16.966 7±7.189)g/L],the differences were statistical significant. P value was 0.012 and 0.009 respectively. The liver fibrosis index in HIV/HCV co-infection group was 5.950 0-5.825 0 Kpa,higher than that in simplex HIV infection group(5.150 0-1.050 0 Kpa),and the difference was nearly statistical significant(P=0.077). The HCV viral load in HIV/HCV co-infection group was(6.476 8-5.343 4)lg copy/ml,higher than that in simplex HCV infection group[(1.699 0-2.681 5)lg copy/ml],and the rate of HCV clearance in HIV/HCV co-infection group was 32.14%, lower than that in simplex HCV infection group(75.00%). P value was 0.012 and 0.032 respectively. The percentage of Treg/CD4+T lymphocyte cell in HIV/HCV co-infection group was (7.460 0%-2.287 5%),higher than that in simplex HCV infection group (5.965 0%-2.105 0%),and the difference was significant (P=0.032). The percentage of Treg/CD4 + T lymphocyte cell was significantly related with HCV viral load(ρ=0.350,P=0.027),and HCV viral load was significantly related with the liver fibrosis index(ρ=0.487,P=0.001). Conclusion HIV infection could accelerate the progress of hepatitis C,and Treg cells were involved in this progress.
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Objective To understand the influence of HIV infection on hepatitis C progress in patients co-infected with HIV and hepatitis C virus (HCV) and related immune mechanism. Methods Twenty eight patients co-infected with HIV/HCV and 12 patients with simplex HCV infection were enrolled. The liver function and hepatic fibrosis progress were evaluated by detecting peripheral blood and with Fibro-Scan. The viral load of HCV was detected by using real time quantitative PCR. And the percentage of Treg/CD4 +T lymphocyte cell was tested by using flow cytometry. Results The levels of ALT and ALP in HIV/HCV co-infection group were (76.16 ± 81.248)U/L,(24.507 1 ± 8.194)g/L respectively,higher than those of simplex HCV infection group [(27.475 0±13.985)U/L,(16.966 7±7.189)g/L],the differences were statistical significant. P value was 0.012 and 0.009 respectively. The liver fibrosis index in HIV/HCV co-infection group was 5.950 0-5.825 0 Kpa,higher than that in simplex HIV infection group(5.150 0-1.050 0 Kpa),and the difference was nearly statistical significant(P=0.077). The HCV viral load in HIV/HCV co-infection group was(6.476 8-5.343 4)lg copy/ml,higher than that in simplex HCV infection group[(1.699 0-2.681 5)lg copy/ml],and the rate of HCV clearance in HIV/HCV co-infection group was 32.14%, lower than that in simplex HCV infection group(75.00%). P value was 0.012 and 0.032 respectively. The percentage of Treg/CD4+T lymphocyte cell in HIV/HCV co-infection group was (7.460 0%-2.287 5%),higher than that in simplex HCV infection group (5.965 0%-2.105 0%),and the difference was significant (P=0.032). The percentage of Treg/CD4 + T lymphocyte cell was significantly related with HCV viral load(ρ=0.350,P=0.027),and HCV viral load was significantly related with the liver fibrosis index(ρ=0.487,P=0.001). Conclusion HIV infection could accelerate the progress of hepatitis C,and Treg cells were involved in this progress.
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<p><b>OBJECTIVE</b>To understand the influence of HIV infection on hepatitis C progress in patients co-infected with HIV and hepatitis C virus (HCV) and related immune mechanism.</p><p><b>METHODS</b>Twenty eight patients co-infected with HIV/HCV and 12 patients with simplex HCV infection were enrolled. The liver function and hepatic fibrosis progress were evaluated by detecting peripheral blood and with Fibro-Scan. The viral load of HCV was detected by using real time quantitative PCR. And the percentage of Treg/CD4⁺ T lymphocyte cell was tested by using flow cytometry.</p><p><b>RESULTS</b>The levels of ALT and ALP in HIV/HCV co-infection group were (76.16 ± 81.248) U/L, (24.507 1 ± 8.194) g/L respectively, higher than those of simplex HCV infection group [(27.475 0 ± 13.985) U/L, (16.966 7 ± 7.189) g/L], the differences were statistical significant. P value was 0.012 and 0.009 respectively. The liver fibrosis index in HIV/HCV co-infection group was 5.950 0-5.825 0 Kpa, higher than that in simplex HIV infection group (5.150 0-1.050 0 Kpa), and the difference was nearly statistical significant (P = 0.077). The HCV viral load in HIV/HCV co-infection group was (6.476 8-5.343 4) lg copy/ml, higher than that in simplex HCV infection group [(1.699 0-2.681 5) lg copy/ml], and the rate of HCV clearance in HIV/HCV co-infection group was 32.14%, lower than that in simplex HCV infection group (75.00%). P value was 0.012 and 0.032 respectively. The percentage of Treg/CD4⁺ T lymphocyte cell in HIV/HCV co-infection group was (7.460 0%-2.287 5%), higher than that in simplex HCV infection group (5.965 0%-2.105 0%), and the difference was significant (P = 0.032). The percentage of Treg/CD4⁺ T lymphocyte cell was significantly related with HCV viral load (ρ = 0.350, P = 0.027), and HCV viral load was significantly related with the liver fibrosis index (ρ = 0.487, P = 0.001).</p><p><b>CONCLUSION</b>HIV infection could accelerate the progress of hepatitis C, and Treg cells were involved in this progress.</p>