Introduction: HIV, the human immunodeficiency virus, is the etiological agent of acquired immunodeficiency syndrome (AIDS). The aim of this study was to assess the evolution of the viral load in patients under treatme...Introduction: HIV, the human immunodeficiency virus, is the etiological agent of acquired immunodeficiency syndrome (AIDS). The aim of this study was to assess the evolution of the viral load in patients under treatment. Methodology: This was a study carried out from July 2017 to June 2022 at the Point G University Hospital laboratory. The determination of the viral load of patients was carried out by PCR on the ABOTT M2000sp/rt platform. Results: A total of 129 patients infected with HIV-1, aged 19 to 72 years with a mean age of 40.05 years ± 10.71;all on antiretroviral chemotherapy. The female gender predominated among our patients. The most common treatment regimen was 2INTI + 1INNTI with 72.9% followed by 2INTI + 1INI with 13.2%. As for the combinations of molecules, the combination TDF + 3TC + EFV and TDF + 3TC + DTG predominated, respectively 65.1% and 13.2%. 89.9% of our patients had undetectable viremia after 12 months of treatment (p < 0.005) with an average viral load which had evolved from 681315.65 copies/ml ± 1616908.484 to M0 at 5742.36 copies /ml ± 35756.883 at M12 (p Conclusion: Generally speaking, antiretroviral treatment had contributed to controlling viral loads, however the therapeutic combination TDF + 3TC + DTG had made it possible to obtain more patients with undetectable viremia instead.展开更多
Combinations of DNA and recombinant-viral-vector based vaccines are promising AIDS vaccine methods because of their potential for inducing cellular immune responses. It was found that Gag-specific cytotoxic lymphocyte...Combinations of DNA and recombinant-viral-vector based vaccines are promising AIDS vaccine methods because of their potential for inducing cellular immune responses. It was found that Gag-specific cytotoxic lymphocyte (CTL) responses were associated with lowering viremia in an untreated HIV-1 infected cohort. The main objectives of our studies were the construction of DNA and recombinant Sendai virus vector (rSeV) vaccines containing a gag gene from the prevalent Thailand subtype B strain in China and trying to use these vaccines for therapeutic and prophylactic vaccines. The candidate plasmid DNA vaccine pcDNA3.1(+)-gag and recombinant Sendai virus vaccine (rSeV-gag) were constructed separately. It was verified by Western blotting analysis that both DNA and rSeV-gag vaccines expressed the HIV-1 Gag protein correctly and efficiently. Balb/c mice were immunized with these two vaccines in different administration schemes. HIV-1 Gag-specific CTL responses and antibody levels were detected by intracellular cytokine staining assay and enzyme-linked immunosorbant assay (ELISA) respectively. Combined vaccines in a DNA prime/rSeV-gag boost vaccination regimen induced the strongest and most long-lasting Gag-specific CTL and antibody responses. It maintained relatively high levels even 9 weeks post immunization. This data indicated that the prime-boost regimen with DNA and rSeV-gag vaccines may offer promising HIV vaccine regimens.展开更多
To investigate the prevalence of drug-resistance mutations,resistance to antiretroviral drugs,and the subsequent virological response to therapy in treatment-naive and antiretroviral-treated patients infected with HIV...To investigate the prevalence of drug-resistance mutations,resistance to antiretroviral drugs,and the subsequent virological response to therapy in treatment-naive and antiretroviral-treated patients infected with HIV/AIDS in Henan,China,a total of 431 plasma samples were collected in Queshan county between 2003 and 2004,from patients undergoing the antiretroviral regimen Zidovudine + Didanosine + Nevirapine(Azt+Ddi+Nvp).Personal information was collected by face to face interview.Viral load and genotypic drug resistance were tested.Drug resistance mutation data were obtained by analyzing patient-derived sequences through the HIVdb Program(http://hivdb.stanford.edu).Overall,38.5% of treatment-naive patients had undetectable plasma viral load(VL),the rate significantly increased to 61.9% in 0 to 6 months treatment patients(mean 3 months)(P<0.005)but again significantly decrease to 38.6% in 6 to 12 months treatment patients(mean 9 months)(P<0.001)and 40.0% in patients receiving more than 12 months treatment(mean 16 months)(P<0.005).The prevalence of drug resistance in patients who had a detectable VL and available sequences were 7.0%,48.6%,70.8%,72.3% in treatment-na?ve,0 to 6 months treatment,6 to 12 months treatment,and treatment for greater than 12 months patients,respectively.No mutation associated with resistance to Protease inhibitor(PI)was detected in this study.Nucleoside RT inhibitor(NRTI)mutations always emerged after non-nucleoside RT inhibitor(NNRTI)mutations,and were only found in patients treated for more than 6 months,with a frequency less than 5%,with the exception of mutation T215Y(12.8%,6/47)which occurred in patients treated for more than 12 months.NNRTI mutations emerged quickly after therapy begun,and increased significantly in patients treated for more than 6 months(P<0.005),and the most frequent mutations were K103N,V106A,Y181C,G190A.There had been optimal viral suppression in patients undergoing treatment for less than 6 months in Queshan,Henan.The drug resistance strains were highly prevalent in antiretroviral-treated patients,and increased with the continuation of therapy,with many patients encountering virological failure after 6 months therapy.展开更多
AIM: To determine the frequency of various hepatitis C virus (HCV) genotypes present in patients from north eastern Algeria. METHODS: This is a retrospective cross-sectional study of 435 HCV infected patients from nor...AIM: To determine the frequency of various hepatitis C virus (HCV) genotypes present in patients from north eastern Algeria. METHODS: This is a retrospective cross-sectional study of 435 HCV infected patients from northeast Algeria, detected in the Sadelaoud laboratory and diagnosed between January 2010 and December 2012. The patients were diagnosed with HCV infection in their local hospitals and referred to be assessed for HCV genotype before the antiviral treatment. Demographic information (sex, age and address), genotype, subtype and viral load were retrieved from the patient medical records. The serum samples were tested by the type-specific genotyping assay.RESULTS: The majority of the patients (82.5%) were from the central part of the examined region (P = 0.002). The mean age of the patients studied was 53.6 ± 11.5 years. HCV genotype 1 was the most frequent (88.7%), followed by genotypes 2 (8.5%), 4 (1.1%), 3 (0.9%) and 5 (0.2%). Genotype 6 was not detected in these patients. Mixed infection across the HCV subtypes was detected in twenty patients (4.6%). The genotype distribution was related to age and region. Genotype 1 was significantly less frequent in the ≥ 60 age group than in the younger age group (OR = 0.2; 95%CI: 0.1-0.5, P < 0.001). Furthermore, genotype 1 was more frequent in the central part of the examined region than elsewhere (P < 0.01). CONCLUSION: The HCV genotype (type 1b was dominant) distribution in Algeria is different from those in other northern countries of Africa.展开更多
To prepare HIV-1 Vif and hAPOBEC3G and to produce their antibodies, the full length gene fragment of HIV-1 vif was amplified by PCR from a plasmid of HIV-1 NL4.3 cDNA, and the APOBEC3G gene was obtained by RT-PCR from...To prepare HIV-1 Vif and hAPOBEC3G and to produce their antibodies, the full length gene fragment of HIV-1 vif was amplified by PCR from a plasmid of HIV-1 NL4.3 cDNA, and the APOBEC3G gene was obtained by RT-PCR from the total RNA of H9 cells. The resulting DNA construct was cloned into a prokaryotic expression vector (pET-32a). Recombinant pET-vif and pET-APOBEC3G were expressed respectively in Eserichia coli BL21 (DE3) as an insoluble protein. The vector also contained a six-histidine tag at the C-terminus for convenient purification and detection. To express and purify the HIV-1 Vif and hAPOBEC3G in E.coli cells, the accuracy of inserted gene and specificity of proteins were detected by the two enzyme digestion method, SDS-PAGE, and Western blotting. Rabbits were then immunized by Vif or APOBEC3G protein and serum samples were tested by indirect ELISA to determine the level of antibodies. Immunoenzyme and immunofluorescence assays were performed to identify the specificity of polyclonal antibodies. The titer of the anti-Vif antibodies was 1:204800, and that of the anti-APOBEC3G antibodies was 1:102400. Thus the antibodies could detect the antigen expression in the cells, demonstrating that fusion proteins with high purity and their corresponding polyclonal antibodies with high titer and specificity were achieved.展开更多
Contexte:?The fight against HIV/AIDS epidemics is one of the greatest challenges of this century. The epidemic affects generally under-developed countries, and Sub-Saharan Africa are the most concerned. The combined m...Contexte:?The fight against HIV/AIDS epidemics is one of the greatest challenges of this century. The epidemic affects generally under-developed countries, and Sub-Saharan Africa are the most concerned. The combined marketed form known as Triomune was used as first-line treatment in several sub-Saharan African Countries (60% of VIH infected people), including Chad. However, no evaluation has been done for that treatment in the country. Objective: To evaluate the efficacy and safety immuno-virological of Triomune at the General Hospital in N’Djamena/Chad. Methods: 48 HIV-1 positive patients eligible for ARV treatment were enrolled in our study, and they have been then followed for 8 months. We have measured in these patients the CD4 cell count before treatment and at the 8th month of treatment. After 8 months of treatment, we have also evaluated the Lymphocyte T CD4 and the plasma viral load (VL). Comparisons of means of CD4 lymphocytes and plasma CV (≥1000 copies/ml) were used to define treatment failure.?Results:?48 patients were under Triomune regime. The average CD4 count was decreased from 462 ± 179.22 [56 - 981] cells/mm3?before treatment to 327.23 ± 153.77 [10 - 1008] cells/mm3?at the 8th month of treatment. The mean plasma viral load for patients was 66008.62 copies/ml. The failure rate to Triomune was 43.75% (21/48).?Conclusion:?Aside from the side effects already described for Triomune, our study reveals a high treatment failure rate. Hence, there is the need of regular revisions of therapeutic regime administer in the first intention.展开更多
To analyze the variability and phenotype of envelope glycoprotein （Env） of human immunodeficiency virus type 1 （HIV-1） prevalent in Heilongjiang province, cloning of the full-length env gene from the peripheral bl...To analyze the variability and phenotype of envelope glycoprotein （Env） of human immunodeficiency virus type 1 （HIV-1） prevalent in Heilongjiang province, cloning of the full-length env gene from the peripheral blood mononuclear cells （PBMCs） of an HIV-1 positive individual in Heilongjiang province in China was performed by using conserved region primers. The amplified PCR products were cloned into a plasmid vector and sequenced. Phylogenetic analysis was done upon the full-length Env amino acid sequence. Subsequently, an HIV-1 pseudotyped virus bearing the envelope protein was constructed and the infectivity was examined using U87 cell lines expressing CD4 with either CCR5 or CXCR4. As the result, two functional env clones named as CHNHLJ03009c34 （GenBank Accession No： AY905493 ） and CHNHLJ03009c33 were obtained. It was found that the homology between CHNHLJ03009c34 and an HIV-1 subtype B＇ strain, RIA-2, isolated from Yunnan province, was 91.52% through comparing and analyzing full-length Env amino acid sequence of HIV-1 isolated from either China or abroad. Phylogenetic analysis indicated that CHNHLJ03009c34 has the closest molecular relation with strain RIA2 based on analyzing the full-length of the Env, while it became an independent branch upon analyzing the sequences of C2-V3 region of the Env. The secondary structure analysis of the envelope protein showed that the antigenicity and hydrophobicity of the strain demonstrated have no definite difference from that of RL42. Examination of infectivity showed that pseudovirus CHNHLI03009c34 could only infect U87. CD4. CCR5 cells, indicating that it was a RS-tropic HIV-1. In the conclusion, two HIV-1 env clones from an infected individual in Heilongjiang province have been identified as subtype B＇ and RS-tropic HIV-1. This is the first report on the analysis of primary isolates in Heilongjiang province.展开更多
Mpox is a zoonotic infectious disease caused by the mpox virus(MPXV).Historically,the majority of mpox cases have been documented in Central Africa.However,since May 2022,there has been a notable rise in reported case...Mpox is a zoonotic infectious disease caused by the mpox virus(MPXV).Historically,the majority of mpox cases have been documented in Central Africa.However,since May 2022,there has been a notable rise in reported cases from regions beyond Africa.Currently,over 110 countries spanning Europe,North America,South America,Asia,and other territories have reported mpox infections.This report details a case involving a patient who identifies as a man who has sex with men(MSM)and is concurrently infected with MPXV,human immunodeficiency virus type 1(HIV-1),Pneumocystis jiroveci,as well as extensively drug-resistant tuberculosis(XDR-TB).This patient had also received a vaccination for smallpox in the past.Additionally,we provide photographic documentation charting the progression of dermatological manifestations associated with mpox.This case highlights the significance of sexual intercourse as a crucial mode of transmission for mpox.The rapid and widespread dissemination of the MPXV across various regions,especially among MSM communities,underscores the importance of enhancing preventive education efforts targeted at high-risk populations.展开更多
Background:Total human immunodeficiency virus(HIV)DNA and integrated HIV DNA are widely used markers of HIV persistence.Droplet digital polymerase chain reaction(ddPCR)can be used for absolute quantification without n...Background:Total human immunodeficiency virus(HIV)DNA and integrated HIV DNA are widely used markers of HIV persistence.Droplet digital polymerase chain reaction(ddPCR)can be used for absolute quantification without needing a standard curve.Here,we developed duplex ddPCR assays to detect and quantify total HIV DNA and integrated HIV DNA.Methods:The limit of detection,dynamic ranges,sensitivity,and reproducibility were evaluated by plasmid constructs containing both the HIV long terminal repeat(LTR)and human CD3 gene(for total HIV DNA)and ACH-2 cells(for integrated HIV DNA).Forty-two cases on stable suppressive antiretroviral therapy(ART)were assayed in total HIV DNA and integrated HIV DNA.Correlation coefficient analysis was performed on the data related to DNA copies and cluster of differentiation 4 positive(CD4^(+))T-cell counts,CD8^(+)T-cell counts and CD4/CD8 T-cell ratio,respectively.The assay linear dynamic range and lower limit of detection(LLOD)were also assessed.Results:The assay could detect the presence of HIV-1 copies 100%at concentrations of 6.3 copies/reaction,and the estimated LLOD of the ddPCR assay was 4.4 HIV DNA copies/reaction(95%confidence intervals[CI]:3.6-6.5 copies/reaction)with linearity over a 5-log_(10)-unit range in total HIV DNA assay.For the integrated HIV DNA assay,the LLOD was 8.0 copies/reaction(95%CI:5.8-16.6 copies/reaction)with linearity over a 3-log 10-unit range.Total HIV DNA in CD4^(+)T cells was positively associated with integrated HIV DNA(r=0.76,P<0.0001).Meanwhile,both total HIV DNA and integrated HIV DNA in CD4^(+)T cells were inversely correlated with the ratio of CD4/CD8 but positively correlated with the CD8^(+)T-cell counts.Conclusions:This ddPCR assay can quantify total HIV DNA and integrated HIV DNA efficiently with robustness and sensitivity.It can be readily adapted for measuring HIV DNA with non-B clades,and it could be beneficial for testing in clinical trials.展开更多
AIM:To investigate if and how programmed death type-1(PD-1)expression affects the natural course of hepatitis B virus(HBV)infection. METHODS:Sixty-four patients in different natural stages of chronic HBV infection wer...AIM:To investigate if and how programmed death type-1(PD-1)expression affects the natural course of hepatitis B virus(HBV)infection. METHODS:Sixty-four patients in different natural stages of chronic HBV infection were enrolled in this study.PD-1 expression in total T cells was detected by flow cytometry.Levels of total CD8+T cell responses and proliferation in relation to PD-1 expression levels were analyzed with intracellular staining and PD-1/ PD-L1 blockage. RESULTS:The PD-1 expression in T cells was dynamically changed during the natural course of chronic HBV infection,did not significantly increase in the immune tolerance phase,and returned to normal in the inactive virus carrier stage.Blockage of the PD-1/PD-L1 pathway could not affect the T-cell response in the immune tolerance and inactive virus carrier stages of chronic HBV infection.However,it could significantly restore the T-cell response in the immune clearance stage of chronic HBV infection.Furthermore,the PD-1 expression level in T cells was associated with the alanine aminotransferase level during the immune clearance stage of chronic HBV infection. CONCLUSION:The PD-l/PD-L1 pathway plays a different role in T-cell response during the natural course of chronic HBV infection.展开更多
The epidemic of HIV/AIDS is sweeping across the world. It is of great importance to figure out new ways to curb this disease. Epitope-based vaccine is one of these solutions. In this study, a chimeric gene was obtaine...The epidemic of HIV/AIDS is sweeping across the world. It is of great importance to figure out new ways to curb this disease. Epitope-based vaccine is one of these solutions. In this study, a chimeric gene was obtained by combination of a designed HIV-1 multi-epitope gene (MEG) and HIV-1 p24 gene. A re- combinant plasmid pUTA2-MEGp24 was then constructed by inserting MEGp24 gene into the down- stream of the promoter (ATI-P7.5×20) of fowlpox virus (FPV) transfer vector pUTA2. The recombinant plasmid and wild-type FPV 282E4 strain were then co-transfected into CEF cells and homologous re- combination occurred. A recombinant virus expressing HIV-1 protein MEGp24 was screened by ge- nome PCR and Western blot assay. Large scale preparation and purification of the recombinant fowl- pox virus (rFPV) were then carried out. BALB/c mice were immunized intramuscularly with the rFPV for three times on day 0, 14 and 42. Mice were executed and sampled one week after the third inoculation. Anti-HIV-1 antibody in serum and Th1 cytokines in the supernatant of cultured spleen cells were as- sayed by ELISA. The count of T lymphocyte subsets and the CTL activity of spleen lymphocytes were analyzed by flow cytometry and lactate dehydrogenase (LDH) release assay, respectively. The results showed that HIV-1 specific antibody in serum and increased T lymphocyte subsets (CD4+ T, CD8+ T) were detected in the immunization group. CTL target-killing activity and higher secretion of Th1 cyto- kines (IFN-γ and IL-2) of spleen lymphocytes stimulated by H-2d-restricted CTL peptide were observed in immunized mice. We concluded that the rFPV may induce HIV-1 specific immunity especially cellular immunity in mice.展开更多
文摘Introduction: HIV, the human immunodeficiency virus, is the etiological agent of acquired immunodeficiency syndrome (AIDS). The aim of this study was to assess the evolution of the viral load in patients under treatment. Methodology: This was a study carried out from July 2017 to June 2022 at the Point G University Hospital laboratory. The determination of the viral load of patients was carried out by PCR on the ABOTT M2000sp/rt platform. Results: A total of 129 patients infected with HIV-1, aged 19 to 72 years with a mean age of 40.05 years ± 10.71;all on antiretroviral chemotherapy. The female gender predominated among our patients. The most common treatment regimen was 2INTI + 1INNTI with 72.9% followed by 2INTI + 1INI with 13.2%. As for the combinations of molecules, the combination TDF + 3TC + EFV and TDF + 3TC + DTG predominated, respectively 65.1% and 13.2%. 89.9% of our patients had undetectable viremia after 12 months of treatment (p < 0.005) with an average viral load which had evolved from 681315.65 copies/ml ± 1616908.484 to M0 at 5742.36 copies /ml ± 35756.883 at M12 (p Conclusion: Generally speaking, antiretroviral treatment had contributed to controlling viral loads, however the therapeutic combination TDF + 3TC + DTG had made it possible to obtain more patients with undetectable viremia instead.
文摘Combinations of DNA and recombinant-viral-vector based vaccines are promising AIDS vaccine methods because of their potential for inducing cellular immune responses. It was found that Gag-specific cytotoxic lymphocyte (CTL) responses were associated with lowering viremia in an untreated HIV-1 infected cohort. The main objectives of our studies were the construction of DNA and recombinant Sendai virus vector (rSeV) vaccines containing a gag gene from the prevalent Thailand subtype B strain in China and trying to use these vaccines for therapeutic and prophylactic vaccines. The candidate plasmid DNA vaccine pcDNA3.1(+)-gag and recombinant Sendai virus vaccine (rSeV-gag) were constructed separately. It was verified by Western blotting analysis that both DNA and rSeV-gag vaccines expressed the HIV-1 Gag protein correctly and efficiently. Balb/c mice were immunized with these two vaccines in different administration schemes. HIV-1 Gag-specific CTL responses and antibody levels were detected by intracellular cytokine staining assay and enzyme-linked immunosorbant assay (ELISA) respectively. Combined vaccines in a DNA prime/rSeV-gag boost vaccination regimen induced the strongest and most long-lasting Gag-specific CTL and antibody responses. It maintained relatively high levels even 9 weeks post immunization. This data indicated that the prime-boost regimen with DNA and rSeV-gag vaccines may offer promising HIV vaccine regimens.
基金Molecular epidemiology research of HIV-1 Drug resistance in China sponsored by the 973 program (2005CB523103) Molecular epidemiology research and new technologies in HIV surveillance in China sponsored by the 863 program (2006AA02Z418).
文摘To investigate the prevalence of drug-resistance mutations,resistance to antiretroviral drugs,and the subsequent virological response to therapy in treatment-naive and antiretroviral-treated patients infected with HIV/AIDS in Henan,China,a total of 431 plasma samples were collected in Queshan county between 2003 and 2004,from patients undergoing the antiretroviral regimen Zidovudine + Didanosine + Nevirapine(Azt+Ddi+Nvp).Personal information was collected by face to face interview.Viral load and genotypic drug resistance were tested.Drug resistance mutation data were obtained by analyzing patient-derived sequences through the HIVdb Program(http://hivdb.stanford.edu).Overall,38.5% of treatment-naive patients had undetectable plasma viral load(VL),the rate significantly increased to 61.9% in 0 to 6 months treatment patients(mean 3 months)(P<0.005)but again significantly decrease to 38.6% in 6 to 12 months treatment patients(mean 9 months)(P<0.001)and 40.0% in patients receiving more than 12 months treatment(mean 16 months)(P<0.005).The prevalence of drug resistance in patients who had a detectable VL and available sequences were 7.0%,48.6%,70.8%,72.3% in treatment-na?ve,0 to 6 months treatment,6 to 12 months treatment,and treatment for greater than 12 months patients,respectively.No mutation associated with resistance to Protease inhibitor(PI)was detected in this study.Nucleoside RT inhibitor(NRTI)mutations always emerged after non-nucleoside RT inhibitor(NNRTI)mutations,and were only found in patients treated for more than 6 months,with a frequency less than 5%,with the exception of mutation T215Y(12.8%,6/47)which occurred in patients treated for more than 12 months.NNRTI mutations emerged quickly after therapy begun,and increased significantly in patients treated for more than 6 months(P<0.005),and the most frequent mutations were K103N,V106A,Y181C,G190A.There had been optimal viral suppression in patients undergoing treatment for less than 6 months in Queshan,Henan.The drug resistance strains were highly prevalent in antiretroviral-treated patients,and increased with the continuation of therapy,with many patients encountering virological failure after 6 months therapy.
文摘AIM: To determine the frequency of various hepatitis C virus (HCV) genotypes present in patients from north eastern Algeria. METHODS: This is a retrospective cross-sectional study of 435 HCV infected patients from northeast Algeria, detected in the Sadelaoud laboratory and diagnosed between January 2010 and December 2012. The patients were diagnosed with HCV infection in their local hospitals and referred to be assessed for HCV genotype before the antiviral treatment. Demographic information (sex, age and address), genotype, subtype and viral load were retrieved from the patient medical records. The serum samples were tested by the type-specific genotyping assay.RESULTS: The majority of the patients (82.5%) were from the central part of the examined region (P = 0.002). The mean age of the patients studied was 53.6 ± 11.5 years. HCV genotype 1 was the most frequent (88.7%), followed by genotypes 2 (8.5%), 4 (1.1%), 3 (0.9%) and 5 (0.2%). Genotype 6 was not detected in these patients. Mixed infection across the HCV subtypes was detected in twenty patients (4.6%). The genotype distribution was related to age and region. Genotype 1 was significantly less frequent in the ≥ 60 age group than in the younger age group (OR = 0.2; 95%CI: 0.1-0.5, P < 0.001). Furthermore, genotype 1 was more frequent in the central part of the examined region than elsewhere (P < 0.01). CONCLUSION: The HCV genotype (type 1b was dominant) distribution in Algeria is different from those in other northern countries of Africa.
基金National Natural Science Foundation ofChina(30400368)The Natural Science foundation ofBeijing(5072003)Beijing Natural Science foundationProgram and Scientific Research Key Program of BeijingMunicipal commission of Education(KZ20051005001).
文摘To prepare HIV-1 Vif and hAPOBEC3G and to produce their antibodies, the full length gene fragment of HIV-1 vif was amplified by PCR from a plasmid of HIV-1 NL4.3 cDNA, and the APOBEC3G gene was obtained by RT-PCR from the total RNA of H9 cells. The resulting DNA construct was cloned into a prokaryotic expression vector (pET-32a). Recombinant pET-vif and pET-APOBEC3G were expressed respectively in Eserichia coli BL21 (DE3) as an insoluble protein. The vector also contained a six-histidine tag at the C-terminus for convenient purification and detection. To express and purify the HIV-1 Vif and hAPOBEC3G in E.coli cells, the accuracy of inserted gene and specificity of proteins were detected by the two enzyme digestion method, SDS-PAGE, and Western blotting. Rabbits were then immunized by Vif or APOBEC3G protein and serum samples were tested by indirect ELISA to determine the level of antibodies. Immunoenzyme and immunofluorescence assays were performed to identify the specificity of polyclonal antibodies. The titer of the anti-Vif antibodies was 1:204800, and that of the anti-APOBEC3G antibodies was 1:102400. Thus the antibodies could detect the antigen expression in the cells, demonstrating that fusion proteins with high purity and their corresponding polyclonal antibodies with high titer and specificity were achieved.
文摘Contexte:?The fight against HIV/AIDS epidemics is one of the greatest challenges of this century. The epidemic affects generally under-developed countries, and Sub-Saharan Africa are the most concerned. The combined marketed form known as Triomune was used as first-line treatment in several sub-Saharan African Countries (60% of VIH infected people), including Chad. However, no evaluation has been done for that treatment in the country. Objective: To evaluate the efficacy and safety immuno-virological of Triomune at the General Hospital in N’Djamena/Chad. Methods: 48 HIV-1 positive patients eligible for ARV treatment were enrolled in our study, and they have been then followed for 8 months. We have measured in these patients the CD4 cell count before treatment and at the 8th month of treatment. After 8 months of treatment, we have also evaluated the Lymphocyte T CD4 and the plasma viral load (VL). Comparisons of means of CD4 lymphocytes and plasma CV (≥1000 copies/ml) were used to define treatment failure.?Results:?48 patients were under Triomune regime. The average CD4 count was decreased from 462 ± 179.22 [56 - 981] cells/mm3?before treatment to 327.23 ± 153.77 [10 - 1008] cells/mm3?at the 8th month of treatment. The mean plasma viral load for patients was 66008.62 copies/ml. The failure rate to Triomune was 43.75% (21/48).?Conclusion:?Aside from the side effects already described for Triomune, our study reveals a high treatment failure rate. Hence, there is the need of regular revisions of therapeutic regime administer in the first intention.
文摘To analyze the variability and phenotype of envelope glycoprotein （Env） of human immunodeficiency virus type 1 （HIV-1） prevalent in Heilongjiang province, cloning of the full-length env gene from the peripheral blood mononuclear cells （PBMCs） of an HIV-1 positive individual in Heilongjiang province in China was performed by using conserved region primers. The amplified PCR products were cloned into a plasmid vector and sequenced. Phylogenetic analysis was done upon the full-length Env amino acid sequence. Subsequently, an HIV-1 pseudotyped virus bearing the envelope protein was constructed and the infectivity was examined using U87 cell lines expressing CD4 with either CCR5 or CXCR4. As the result, two functional env clones named as CHNHLJ03009c34 （GenBank Accession No： AY905493 ） and CHNHLJ03009c33 were obtained. It was found that the homology between CHNHLJ03009c34 and an HIV-1 subtype B＇ strain, RIA-2, isolated from Yunnan province, was 91.52% through comparing and analyzing full-length Env amino acid sequence of HIV-1 isolated from either China or abroad. Phylogenetic analysis indicated that CHNHLJ03009c34 has the closest molecular relation with strain RIA2 based on analyzing the full-length of the Env, while it became an independent branch upon analyzing the sequences of C2-V3 region of the Env. The secondary structure analysis of the envelope protein showed that the antigenicity and hydrophobicity of the strain demonstrated have no definite difference from that of RL42. Examination of infectivity showed that pseudovirus CHNHLI03009c34 could only infect U87. CD4. CCR5 cells, indicating that it was a RS-tropic HIV-1. In the conclusion, two HIV-1 env clones from an infected individual in Heilongjiang province have been identified as subtype B＇ and RS-tropic HIV-1. This is the first report on the analysis of primary isolates in Heilongjiang province.
基金the National Natural Science Foundation of China(Nos.82241068 and 82241072).
文摘Mpox is a zoonotic infectious disease caused by the mpox virus(MPXV).Historically,the majority of mpox cases have been documented in Central Africa.However,since May 2022,there has been a notable rise in reported cases from regions beyond Africa.Currently,over 110 countries spanning Europe,North America,South America,Asia,and other territories have reported mpox infections.This report details a case involving a patient who identifies as a man who has sex with men(MSM)and is concurrently infected with MPXV,human immunodeficiency virus type 1(HIV-1),Pneumocystis jiroveci,as well as extensively drug-resistant tuberculosis(XDR-TB).This patient had also received a vaccination for smallpox in the past.Additionally,we provide photographic documentation charting the progression of dermatological manifestations associated with mpox.This case highlights the significance of sexual intercourse as a crucial mode of transmission for mpox.The rapid and widespread dissemination of the MPXV across various regions,especially among MSM communities,underscores the importance of enhancing preventive education efforts targeted at high-risk populations.
基金supported by the National Key R&D Program of China(Nos.2021YFC2301900 and 2021YFC2301905)the National 13th Five-Year Grand Program on Key Infectious Disease Control(Nos.2018ZX10301-101 and 2018ZX10301101-001-001)+3 种基金the National Natural Science Foundation of China(Nos.82241072,82072271,and 82272319)the High-Level Public Health Specialized Talents Project of Beijing Municipal Health Commission(Nos.2022-2-018 and 2022-1-007)the Climbing the peak(Dengfeng)Talent Training Program of Beijing Hospitals Authority(No.DFL20191701)Beijing Key Laboratory for HIV/AIDS Research(No.BZ0089).
文摘Background:Total human immunodeficiency virus(HIV)DNA and integrated HIV DNA are widely used markers of HIV persistence.Droplet digital polymerase chain reaction(ddPCR)can be used for absolute quantification without needing a standard curve.Here,we developed duplex ddPCR assays to detect and quantify total HIV DNA and integrated HIV DNA.Methods:The limit of detection,dynamic ranges,sensitivity,and reproducibility were evaluated by plasmid constructs containing both the HIV long terminal repeat(LTR)and human CD3 gene(for total HIV DNA)and ACH-2 cells(for integrated HIV DNA).Forty-two cases on stable suppressive antiretroviral therapy(ART)were assayed in total HIV DNA and integrated HIV DNA.Correlation coefficient analysis was performed on the data related to DNA copies and cluster of differentiation 4 positive(CD4^(+))T-cell counts,CD8^(+)T-cell counts and CD4/CD8 T-cell ratio,respectively.The assay linear dynamic range and lower limit of detection(LLOD)were also assessed.Results:The assay could detect the presence of HIV-1 copies 100%at concentrations of 6.3 copies/reaction,and the estimated LLOD of the ddPCR assay was 4.4 HIV DNA copies/reaction(95%confidence intervals[CI]:3.6-6.5 copies/reaction)with linearity over a 5-log_(10)-unit range in total HIV DNA assay.For the integrated HIV DNA assay,the LLOD was 8.0 copies/reaction(95%CI:5.8-16.6 copies/reaction)with linearity over a 3-log 10-unit range.Total HIV DNA in CD4^(+)T cells was positively associated with integrated HIV DNA(r=0.76,P<0.0001).Meanwhile,both total HIV DNA and integrated HIV DNA in CD4^(+)T cells were inversely correlated with the ratio of CD4/CD8 but positively correlated with the CD8^(+)T-cell counts.Conclusions:This ddPCR assay can quantify total HIV DNA and integrated HIV DNA efficiently with robustness and sensitivity.It can be readily adapted for measuring HIV DNA with non-B clades,and it could be beneficial for testing in clinical trials.
基金Supported by Grants from the"Yucai"Research Program of Changhai Hospital
文摘AIM:To investigate if and how programmed death type-1(PD-1)expression affects the natural course of hepatitis B virus(HBV)infection. METHODS:Sixty-four patients in different natural stages of chronic HBV infection were enrolled in this study.PD-1 expression in total T cells was detected by flow cytometry.Levels of total CD8+T cell responses and proliferation in relation to PD-1 expression levels were analyzed with intracellular staining and PD-1/ PD-L1 blockage. RESULTS:The PD-1 expression in T cells was dynamically changed during the natural course of chronic HBV infection,did not significantly increase in the immune tolerance phase,and returned to normal in the inactive virus carrier stage.Blockage of the PD-1/PD-L1 pathway could not affect the T-cell response in the immune tolerance and inactive virus carrier stages of chronic HBV infection.However,it could significantly restore the T-cell response in the immune clearance stage of chronic HBV infection.Furthermore,the PD-1 expression level in T cells was associated with the alanine aminotransferase level during the immune clearance stage of chronic HBV infection. CONCLUSION:The PD-l/PD-L1 pathway plays a different role in T-cell response during the natural course of chronic HBV infection.
文摘The epidemic of HIV/AIDS is sweeping across the world. It is of great importance to figure out new ways to curb this disease. Epitope-based vaccine is one of these solutions. In this study, a chimeric gene was obtained by combination of a designed HIV-1 multi-epitope gene (MEG) and HIV-1 p24 gene. A re- combinant plasmid pUTA2-MEGp24 was then constructed by inserting MEGp24 gene into the down- stream of the promoter (ATI-P7.5×20) of fowlpox virus (FPV) transfer vector pUTA2. The recombinant plasmid and wild-type FPV 282E4 strain were then co-transfected into CEF cells and homologous re- combination occurred. A recombinant virus expressing HIV-1 protein MEGp24 was screened by ge- nome PCR and Western blot assay. Large scale preparation and purification of the recombinant fowl- pox virus (rFPV) were then carried out. BALB/c mice were immunized intramuscularly with the rFPV for three times on day 0, 14 and 42. Mice were executed and sampled one week after the third inoculation. Anti-HIV-1 antibody in serum and Th1 cytokines in the supernatant of cultured spleen cells were as- sayed by ELISA. The count of T lymphocyte subsets and the CTL activity of spleen lymphocytes were analyzed by flow cytometry and lactate dehydrogenase (LDH) release assay, respectively. The results showed that HIV-1 specific antibody in serum and increased T lymphocyte subsets (CD4+ T, CD8+ T) were detected in the immunization group. CTL target-killing activity and higher secretion of Th1 cyto- kines (IFN-γ and IL-2) of spleen lymphocytes stimulated by H-2d-restricted CTL peptide were observed in immunized mice. We concluded that the rFPV may induce HIV-1 specific immunity especially cellular immunity in mice.
