Advances in the research of HIV-1 envelope glycoprotein gp120 inhibitors / 药学学报

From the process of human immunodeficiency virus-1 (HIV-1) invading cells, the combination of gp120 and CD4 is the first step for HIV-1 to invade cells. Interfering with this process can prevent HIV from recognizing target cells and inhibit virus replication. Therefore, HIV-1 gp120 is an important part of the HIV-1 life cycle. Fostesavir, a phosphatate prodrug derived from the gp120 inhibitor BMS-626529 modified by the prodrug strategy, was approved for the treatment of adult patients with multidrug resistant HIV-1 infection by the US FDA and the European Medicines Agency in 2020 and 2021, respectively. In this review, we focus on the research progress of small molecule inhibitors targeting the interaction of gp120-CD4 from the perspective of medicinal chemistry, in order to provide reference for the subsequent research of gp120 inhibitors.

Characteristics of treatment-naïve HBV-infected individuals with HIV-1 coinfection: A cross-sectional study from South India

Purpose: Human immunodeficiency virus-1 (HIV-1) and hepatitis B virus (HBV) coinfection has become a major health problem across the globe. The increased life expectancy of HIV-1 patients due to antiretroviral therapy has led to the emergence of liver disease as a major mortality factor among them. The purpose of the study was to examine the baseline characteristics of HBV in treatment-naïve HBV/HIV coinfection from southern India compared to monoinfected individuals. Materials and Methods: The study was cross sectional in design, and samples were examined from 80 HIV-1, 70 HBV and 35 HBV/HIV-coinfected individuals using chemiluminescent microparticle immunoassay, real-time polymerase chain reaction and flow cytometry assays. Results: There was a significant increase in HBV DNA (P = 0.0001), higher hepatitis B e antigen percentage difference (P = 0.027) and lower CD4 counts (P = 0.01) among the HBV/HIV-coinfected individuals, but no difference in the HIV-1 viral load compared to HIV-1-monoinfected individuals. Also, the aspartate aminotransferase levels, prothrombin time and the international normalised ratio were significantly high among coinfected individuals. Conclusion: These findings conclude that HIV-1 coinfection can have serious implications on the outcome of HBV-related liver disease. To the contrary, HBV infection had no consequence on the progression of HIV-1 disease but distinctly lowered CD4+ T-cells.

Research progress in neurotoxicity of HIV-1 viral protein R / 中华实验和临床病毒学杂志

Viral Protein R (Vpr) is an accessory protein of HIV-1 that increases the infectivity of HIV-1, activates latent viruses, and acts with other accessory proteins. A number of studies have demonstrated the important role of Vpr in the pathogenesis of HIV-1 related neurological diseases. Vpr can disrupt the integrity of the blood-brain barrier, induce neuronal apoptosis, and may be an important causative agent of AIDS dementia syndrome. However, the exact mechanism is still unknown. This review elaborates the neurotoxic effects of HIV-1 Vpr.

Progress in research on neurotoxicity of HIV-1 Nef protein / 中华实验和临床病毒学杂志

Human immunodeficiency virus-1(HIV-1)is the causative agent of acquired immunodeficiency syndrome(AIDS), which can invade the central nervous system and cause a series of neurological disorders such as HIV-associated neurocognitive disorders(HAND). Nef protein is one of the early proteins of HIV-1 virus, it is found that the neurotoxicity of HIV-1 mutant lacking nef gene is less neurotoxic than that of wild strain with nef gene, indicating that Nef protein plays an important role in HIV-1 neuropathogenicity.In this paper, neurotoxicity and mechanism of Nef are reviewed from three aspects, including the effects of Nef on the blood-brain barrier permeability and the direct and indirect damage to neurons.

Analysis of genetic diversity and amino acid sequence of HIV-1 Vpr from a patient with AIDS dementia complex / 中华实验和临床病毒学杂志

Objective@#To study the sequence characteristics and variation of HIV-1 Vpr gene in different parts of an AIDS dementia complex (ADC) patient and provide basis for the study of the neurologic pathogenesis of HIV-1-assciatd dementia.@*Methods@#Genomic DNA was extracted from peripheral samples (lymph nodes, spleen, liver) and central nervous system (meninges, frontal lobe, temporal lobe gray matter, frontal white matter, basal ganglia cortex) of an ADC patient, The Vpr gene was amplified with nested polymerase chain reaction (PCR). PCR products were cloned into the pMD19-T vector. After transformation into DH5α competent E. coli, five positive clones were sequenced. The phylogenetic tree was built and genetic distance was calculated through MEGA6, and the values of ds/dn was calculated through SNAP, then the changes of the amino acid sites were analyzed.@*Results@#HIV-1 Vpr genes isolated from different tissues of the ADC patient had variations. Vpr HIV-1 gene sequences from central nervous system and peripheral tissues were intercrossed together in the phylogenetic tree. Central nervous system and peripheral HXB2 Vpr had no significant differences in genetic distance. The ds/dn of all the HIV-1 Vpr gene sequences were 3.3749.@*Conclusions@#The HIV-1 Vpr sequences were different in the ADC patient, and there were different variations in different parts of the peripheral and central regions. Whether these variations are related to the pathogenesis of ADC remains to be further studied.

Effects of human immunodeficiency virus-1 Tat protein on blood brain barrier and its mechanism / 中华实验和临床病毒学杂志

Human immunodeficiency virus-1 (HIV-1) infects the human body and acts on the related cells of blood brain barrier, causing structural and functional abnormalities of blood brain barrier, which will be beneficial to the invasion of the virus into central nervous system. There are many types of the molecular mechanism of damage of blood brain barrier. Tat protein, as HIV-1 trans-activator of transcription, can damage the vascular endothelial cells, pericytes and astrocytes, and reduce the tight junction protein expression. Some chemicals such as methamphetamine can cooperate with HIV-1 Tat protein damage blood brain barrier. In this paper, the effects and its mechanism of HIV-1 Tat protein on blood brain barrier are reviewed.

The combined anti-HIV-1 effect of chloroquine and antiretroviral drugs in vitro / 药学学报

The study is aimed to evaluate the anti-HIV-1 effect of chloroquine in combination with antihuman immunodeficiency virus (HIV) drugs, and inhibition of plasmacytoid dendritic cells (pDC) activation and type I interferon (IFN-I) production by Toll-like receptor 7 (TLR7) agonist stimulation. We investigated the anti-HIV-1ⅢB, HIV-1KM018 activity of chloroquine and chloroquine combined with rategrivir (RAL), enfuvirtide (T-20), indinavir (IDV) and efavirenz (EFV) in vitro by luciferase activity assay system and ELISA method for p24 antigen. We measured the effect of chloroquine on the activation of pDC in combination with RAL and IDV, respectively. Quantitative PCR was used to evaluate the activity of chloroquine in combination with RAL and IDV in the upregulation of interferon (IFN)-α and IFN-β. Chloroquine showed less cytotoxicity to C8166, TZM-bl and PBMC cells, and the 50% cytotoxic concentration values were 85.02 ±0.28, 73.67 ±5.10 and 91.84 ±4.10 μmol·L-1, respectively. The anti-HIV-1ⅢB activity of chloroquine combination with RAL, T-20, IDV and EFV were moderate in synergy, strong in synergy, additive and moderate antagonism, respectively. The anti-HIV-1KM018 activity of chloroquine in combination with RAL, IDV were moderate synergy, minor synergy. There was no significant difference between the chloroquine monotherapy and chloroquine combined with RAL, IDV in the down-regulation of pDC activation and IFN-α, IFN-β expression levels. We have found that chloroquine combined with different anti-HIV drugs represent different degrees of synergism, antagonism or additive anti-HIV-1 effect. Chloroquine in combination with RAL and IDV did not have influence on the inhibitory effect of chloroquine on pDC activation and type I interferon secretion induced by TLR7 agonist. The results suggest that chloroquine may be used to enhance the therapeutic activities of anti-HIV medicines.

Prevalência de infecção pelos vírus linfotrópicos de células T humanas dos tipos 1 e 2 (HTLV-1 e HTLV-2) e vírus da imunodificiência humana do tipo 1 (HIV-1) em população infectada pelos vírus da hepatite B (HBV) e hepatite C (HCV)

Os vírus linfotrópicos de células T humanas dos tipos 1 (HTLV-1) e 2 (HTLV2),assim como o vírus da imunodeficiência humana (HIV) e os vírus dashepatites B (HBV) e C (HCV) compartilham vias de transmissão, portantocoinfecções por estes vírus podem acontecer e alterar o curso das doençasa eles relacionadas. O presente estudo avaliou a prevalência de infecção porHTLV-1/2 em população com hepatite B e C, infectada ou não pelo HIV, e oimpacto das coinfecções na viremia HBV e HCV. O estudo foi realizado em1.244 amostras de plasma/soro enviadas ao Instituto Adolfo Lutz de SãoPaulo para determinação de carga viral (CV) de HBV e HCV: 622 depacientes com HBV (G1, 327 homens e 295 mulheres, média de idade 45,8anos) e 622 de pacientes com HCV (G2, 343 homens e 279 mulheres, médiade idade de 50,8 anos). A triagem de HTLV-1/2 foi realizada por ensaioimunoenzimático (EIA HTLV-I/II, Gold ELISA, REM) e confirmadas porWestern Blot (HTLV BLOT 2.4, MP Biomedicals) e imunoensaio de linha(INNO-LIA HTLV-I/II, Fujirebio). A pesquisa de infecção por HIV foi realizadapor teste imunocromatográfico (kit Rapid Check HIV 1 e 2, NDI, UniversidadeFederal do Espírito Santo, Brasil) seguido do EIA (GS HIV-1/HIV-2 Plus OEIA, Bio-Rad). A infecção por HTLV-1 foi confirmada em 25 amostras (cincono G1 e 20 no G2)...

The human T-cell lymphotropic virus types 1 and 2 (HTLV-1 and HTLV-2) aswell as the human immunodeficiency virus (HIV) and the hepatitis B virus(HBV) and hepatitis C virus (HCV) share routes of virus transmission; thusco-infections with such viruses can occur and alter the course of subsequentdiseases. The present study aimed at evaluating the prevalence of HTLV-1/-2 in blood samples of individuals with hepatitis B and C, infected or not byHIV, and the impact of co-infections on the HBV and HCV viremia. The studywas conducted with 1,244 plasma/serum samples sent to Instituto AdolfoLutz of São Paulo for measuring HCV and HBV viral load (VL): 622 fromHBV-infected patients (G1, 327 male and 295 female, median age 45.8years), and 622 from HCV-infected patients (G2, 343 male and 279 female,median age 50.8 years). HTLV-1/-2 antibodies were screened by enzymeimmunoassay (EIA, HTLV-I/II, Gold ELISA, REM), and confirmed by Westernblot (HTLV BLOT 2.4, MP Biomedicals), and line immunoassay (INNO-LIAHTLV-I/II, Fujirebio). The HIV infection was detected byimmunochromatographic assay (Rapid Check HIV 1 e 2, NDI, UniversidadeFederal do Espírito Santo, Brasil) and by EIA (GS HIV-1/HIV-2 Plus O EIA,Bio-Rad). HTLV-1 was confirmed in 25 samples (5 in G1 and 20 in G2)...

Effect of downregulation of FBI-1 on proliferation of human breast carcinoma cell line and its mechanism / 中国癌症杂志

Background and purpose: Factor that binds to the inducer of short transcripts of human immuno-deficiency virus-1 (FBI-1) in a variety of malignant tumors showed high expression levels, which may be closely related to tumor proliferation and differentiation, angiogenesis, metastasis, but its relationship with breast cancer has not been fully elucidated. The purpose of this study was to investigate the expression of FBI-1 in breast cancer cells, and to study the effect of FBI-1 gene expression on the proliferation of breast cancer cells and its possible mechanism. Methods:Real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot analysis were applied to detect FBI-1 expression in normal human mammary epithelial cell line MCF-10A and breast cancer cell MCF-7. RNA interference method was used to down-regulate FBI-1 expression in MCF-7 cells. The cell proliferation was measured by CCK-8 kit and colony formation assay. RTFQ-PCR and Western blot were used to detect the expression of FBI-1 and NF-κBp65 in MCF-7 cells before and after the interference of FBI-1 expression. Results: The expression of FBI-1 was higher in breast cancer cells than that in normal human mammary epithelial cells (P<0.05). The effects of FBI-1 down-regulation inhibited proliferation in MCF-7 cells (P<0.05). At the same time, after inhibition of FBI-1, the NF-κBp65 mRNA and protein expression levels were significantly decreased (P<0.05). Conclusion: FBI-1 is highly expressed in breast cancer cells. Down-regulated FBI-1 expression can inhibit the proliferation of breast cancer cells,and its mechanism may be related to the inhibition of NF-κB signaling pathway.

Effect of retinoblastoma binding protein 4 (RBBP4)on Sp1-mediated transcription of HIV long terminal repeat in 293 T cells / 中华临床感染病杂志

Objective To investigate the effect of retinoblastoma binding protein 4 (RBBP4)in Sp1 -mediated HIV long terminal repeat(LTR)transcription.Methods RBBP4 expression vector and Sp1 expression vector were respectively co-transfected into 293 T cells with HIV promoter pHIV-LTR-Luc or Sp1 site mutated pHIV-LTR-sp1 -mut by liposome transfection,and the transfected cells were examined by dual luciferase reporter assay system.The effect of RBBP4 on the binding of Sp1 to LTR was further studied by chromatin immunoprecipitation (ChIP)and electrophoretic mobility shift assay (EMSA).Results The relative firefly luciferase activity activated by Sp1 was decreased from 62.5 to 16 at the dose of 500 ng of RBBP4 expression vector (t =14.52,P <0.01 ).When the Sp1 binding sites were mutated,the effects of 100,300 or 500 ng of RBBP4 expression vector on the firefly luciferase activity of HIV LTR were not statistically significance (t =1 .897,2.357 and 3.162,all P <0.05).ChIP results showed that when the binding of RBBP4 on HIV LTR increased,the binding of Sp1 on HIV LTR increased significantly (t =11 .93,P <0.01 ),while the reduced binding of RBBP4 on HIV LTR significantly attenuated the binding of Sp1 onto LTR(t =11 .38,P <0.01 ).The effect of RBBP4 on the binding of Sp1 to DNA in ChIP assays was further verified by EMSA assays.Conclusion RBBP4 can inhibit the Sp1 -mediated HIV LTR transcription in 293 T cells.

Expression, purification, characterization and immunogenicity of human immunodeficiency virus-1 glycoprotein gp120 derived from insect cells / 中华微生物学和免疫学杂志

Objective To establish an efficient baculovirus-insect cell expression system for the production of human immunodeficiency virus-1 ( HIV-1 ) envelope glycoprotein gp120 and to evaluate the physiochemical properties, antigenicity and immunogenicity of the recombinant protein. Methods The gene encoding HIV-1 NL4-3 gp120 was cloned into the downstream of pH promoter of the baculovirus transfer vec-tor pAcgp67B to construct the recombinant transfer vector pAc-gp120. Expression of the protein of interest was induced in baculovirus-infected High FiveTM insect cells. ELISA, analytical ultracentrifugation and size-exclusion chromatography were carried out to characterize physicochemical properties of the expressed gp120 protein. Immunogenicity of the recombinant gp120 protein was analyzed by HIV neutralization assay after im-munizing BALB/c mice with it. Results The recombinant HIV-1 gp120 protein was successfully obtained from the established insect cell expression system with a purity of more than 90％ and a mean yield of 13 mg/L in four batches. That recombinant HIV-1 gp120 protein was characterized by homogeneity in solution and possessed a good immunoreactivity to neutralizing antibodies and antisera against HIV. Immunogenicity analysis in BALB/c mice demonstrated that the recombinant gp120 protein could induce effective immune re-sponses against HIV-1 NL4-3. Conclusion A simple and scalable approach to obtain homogeneous and im-munogenic HIV-1 gp120 antigen is successfully established, which will promote further investigation of HIV vaccine candidates.

Expression, purification, characterization and immunogenicity of human immunodeficiency virus-1 glycoprotein gp120 derived from insect cells / 中华微生物学和免疫学杂志

Objective To establish an efficient baculovirus-insect cell expression system for the production of human immunodeficiency virus-1 ( HIV-1 ) envelope glycoprotein gp120 and to evaluate the physiochemical properties, antigenicity and immunogenicity of the recombinant protein. Methods The gene encoding HIV-1 NL4-3 gp120 was cloned into the downstream of pH promoter of the baculovirus transfer vec-tor pAcgp67B to construct the recombinant transfer vector pAc-gp120. Expression of the protein of interest was induced in baculovirus-infected High FiveTM insect cells. ELISA, analytical ultracentrifugation and size-exclusion chromatography were carried out to characterize physicochemical properties of the expressed gp120 protein. Immunogenicity of the recombinant gp120 protein was analyzed by HIV neutralization assay after im-munizing BALB/c mice with it. Results The recombinant HIV-1 gp120 protein was successfully obtained from the established insect cell expression system with a purity of more than 90％ and a mean yield of 13 mg/L in four batches. That recombinant HIV-1 gp120 protein was characterized by homogeneity in solution and possessed a good immunoreactivity to neutralizing antibodies and antisera against HIV. Immunogenicity analysis in BALB/c mice demonstrated that the recombinant gp120 protein could induce effective immune re-sponses against HIV-1 NL4-3. Conclusion A simple and scalable approach to obtain homogeneous and im-munogenic HIV-1 gp120 antigen is successfully established, which will promote further investigation of HIV vaccine candidates.

Cytotoxic role of γδT cells to latency cells in patients with early human immunodeficiency virus-1 infection / 基础医学与临床

Objective To investigate the cytotoxicity of γδ T cells to HIV-1 latency cells in patients with early HIV-1 infection.Methods Sixteen early HIV-1-infected patients were enrolled in this study.Peripheral blood mononuclear cells (PBMCs) of patients were isolated and γδ T cells were expanded using zoledronate (5 μmol/L) and interleukin (IL)-2 (1 000 IU/mL) ex vivo.Lactic dehydrogenase (LDH) was used to detect the cytotoxic role of γδ T cells to HIV-1 latency cells(J-Lat Full Length Clonel0.6).The phenotype of γδ T cells before and after expansion and the intensity of GFP in HIV-1 latency cells were detected by flow cytometry.Results Zoledronate plus IL-2 stimulated rapid and large γδ T cells proliferation ex vivo (P＜0.001).γδ T cells showed high cytotoxici ty to latency cells,and the intensity of GFP in latency cells was decreased significantly (P＜0.05).Moreover,expanded γδ T cells displayed cytotoxic NK-like phenotype,the frequency of CD56+ Vδ2 T cells in patients with early HIV-1 infection was significantly higher than that of healthy controls.Conclusions γδ T cell has an ability to eradicate HIV-1 latency,and γδ T cell-based autologous or xenogenous adoptive immunotherapy will have promise prospects to cure HIV-1 infection.

Expression of cytokine‑mRNA in peripheral blood mononuclear cell of human immunodeficiency virus‑1 subtype C infected individuals with opportunistic viral infections from India (South).

Human immunodeficiency virus (HIV) disease progression is associated with a marked change in the level of plasma cytokines. The study reported here investigated the level of mRNA expression of different cytokines: Tumour necrosis factor‑alpha (TNF‑α), interferon (INF)‑gamma, interleukin‑10 (IL‑10) and IL‑21 in the peripheral blood mononuclear cell among the antiretroviral therapy naive subtype C HIV‑1 infected individuals and normal healthy controls by real time polymerase chain reaction. The mRNA expressions of all the 4 cytokines in HIV‑1 infected individuals were significantly higher compared to healthy controls (P value range 0.0004–0.01). The mean level of IL‑10, INF‑gamma and TNF‑α were higher in HIV infected individuals with low CD4 counts (<300 cells/μl). The IL‑10 expression showed a significant negative correlation with CD4 counts (r = −0.25, P = 0.04) while IL‑21 showed a positive correlation with CD4 counts (r = 0.26, P = 0.03). There was a significant negative correlation between the cytomegalovirus (CMV) viral load and IL‑21 expression. Cytokine levels by mRNA detection avoids the inherent problem of measuring plasma level and this study also provide information on the cytokine levels and CD4+ T cell level among HIV‑1 subtype C infected individuals with opportunistic viral infections like CMV.

Gene mutations of human immunodeficiency virus drug resistance from men who have sex with men in Shanghai,2013 / 中华传染病杂志

Objective To investigate the gene mutations of human immunodeficiency virus (HIV)‐1 drug resistance among anti‐retrovirus (ARV ) treated‐naive men who have sex with men (MSM ) in Shanghai to provide evidence‐based data for optimized treatment .Methods All 669 treatment‐nave cases of HIV‐1 infection identified among MSM in 2013 were recruited and their plasma was collected .RNA was extracted and amplified by nest reverse transcription‐polymerase chain reaction ,and DNA was sequenced and then phylogenetically analyzed .Finally ,subtypes were identified and drug resistance was analyzed in comparison with International HIV Drug Resistance Database .Results The pol gene fragments of 645 cases were obtained .Primary drug‐resistance rate was 2 .48% (16/645) ,including mutations conferring resistance to protease inhibitor (PI) (0 .31% ,2/645) ,nucleoside reverse transcriptase inhibitors (NRTI) (0 .16% ,1/645) ,non‐nucleoside reverse transcriptase inhibitors (NNRTI) (1 .70% ,11/645) and both NRTI and NNRTI (0 .31% ,2/645) ,respectively .Mutations conferring resistance to CRF01_AE were 12 cases (2 .99% ) ,while mutations conferring resistance to CRF07_BC and CRF_01B were 0 .61%(1/163) and 4 .65% (2/43 ) including 1 case of CRF52_01B and unidentified CRF_01B , respectively . Resistance to NNRTI in B subtype were 2 .70% (1/37) .Conclusion The prevalence of HIV‐1 drug resistance‐associated mutations among MSM in Shanghai ,2013 is still low ,but resistance to NNRTI is relatively high .CRF01_AE is the major subtype of drug resistance .It is necessary to strengthen the HIV drug resistance surveillance in MSM group in Shanghai .

Analysis of genetic variations of hepatitis C virus 5′untranslated regions gene in human immunodeficiency ;virus-1/hepatitis C virus co-infected patients in Xinj iang / 中华传染病杂志

Objective To investigate the genetic dynamics of hepatitis C virus (HCV)5′untranslated regions (UTR)gene in human immunodeficiency virus (HIV-1)/HCV co-infected patients in Xinjiang.Methods Anti-HCV was tested using enzyme linked immunosorbent assay in 212 HIV-1 infected patients in Xinjiang. RNA was extracted from HIV-1/HCV co-infected samples.Then HCV 5′UTR gene was amplified after reverse transcription using PrimeScriptTMⅡ 1st Strand cDNA Synthesis kit.The sequences were then compared with HCV international standard strains to determine the HCV gene types.The gene distance of 4 sampling virus strains was calculated and the phylogenetic analysis was performed to establish the cladogram.Results One hundred and fifty-seven among the 212 HIV-infected patients were HCV antibody positive.Of them,2 patients were infected through heterosexual transmission and the remaining 155 patients were injection drug users (IDU). HCV 5′UTR was amlipicated and geno-typed successfully in 131 samples.Five HCV genotypes were identified, including 3a (40/131,30.53%),3b (23/131,17.56%),1a (11/131,8.40%),1b (56/131,42.75%),and 6a (1/131,0.76%).The general homology was 87.5%-99.3% in 38 patients,and there were differences in homology in different subtypes.The homology of the 4 sampling sequences from the same sample was 99.3%-100.0%,with the variation of 0 -0.7%.The intragroup gene deviation and deviation with the first sample tended to increase with time.The mean evolutionary rate of HCV 5′UTR in HIV-1 infected patients in Xinjiang was 1.62 × 10 -3 (95%CI :1.52 × 10 -3 -5.38 × 10 -3 ).The similarity and homology of several sequences obtained from the one participant were the highest.Conclusions The prevalence of HCV co-infection among HIV-1 infected patients in Xinjiang is high,especially among IDU.Subtype 1b of HCV is prevalent strain.

Desafios no diagnóstico da infecção pelos Vírus Linfotrópicos de Células T Humanas do tipo 1 e tipo 2 (HTLV-1 e HTLV-2) em pacientes infectados com o HIV-1 / Challenges in the diagnosis of Human T-Cell Lymphotropic Virus type 1 and type 2 (HTLV-1 and HTLV-2) infection in patients infected with HIV-1

Desde a década de 90 o Instituto Adolfo Lutz de São Paulo (IAL) tem realizado o diagnóstico da infecção por Vírus Linfotrópicos de Células T Humanas dos tipos 1 e 2 (HTLV-1 e HTLV-2) e, desde então, têm sido reportadas as dificuldades principalmente no diagnóstico de HTLV-2, em especial em pacientes infectados pelo HIV-1. O presente trabalho teve como objetivo avaliar várias técnicas de diagnóstico disponíveis no momento atual (kits comerciais e testes in house) e estabelecer o melhor algoritmo para ser empregado no diagnóstico de pacientes infectados pelo HIV-1. A população analisada foi composta por dois grupos provenientes de Serviços de Assistência Especializados em HIV/AIDS de São Paulo: um pioneiro [Grupo 1 (G1), n=1.608] e outro com histórico mais recente [Grupo 2 (G2), n=1.383]. Ambos os grupos foram formados, na maioria, por indivíduos do sexo masculino... (AU).

Since the 90 decade, the Instituto Adolfo Lutz (IAL) has performed the diagnosis of Human T-cell Lymphotropic Virus type 1 and type 2 (HTLV-1 and HTLV-2), and thenceforth the difficulties in diagnosing HTLV-2 have been reported, mostly in HIV-infected patients. The present study aimed at evaluating the several diagnostic techniques currently available (commercial kits and in-house assays), and to establish the best algorithm to be employed for diagnosing HTLV-1/-2 in patients infected with HIV-1. The study population was composed by two patient groups attended at HIV/AIDS specialized services care in São Paulo: the pioneer one [Group 1 (G1), n=1,608], and the other with the most recent historical health setting [Group 2 (G2), n=1,383. The majority of the both groups were composed by male patients...(AU).

Investigation of the HIV-1 drug resistance in Guangxi during 2009 to 2012 / 中华微生物学和免疫学杂志

Objective To investigate the HIV-1 drug resistance in Guangxi during 2009 to 2012 and to analyze the correlations between drug resistance and HIV-1 subtypes.Methods Patients with human immunodeficiency virus infection or acquired immune deficiency syndrome ( HIV/AIDS) were randomly re-cruited from different areas in Guangxi.HIV-1 RNA was extracted from blood samples of the subjects and converted into complementary DNA ( cDNA) by using reverse transcription.The pol gene was amplified and sequenced.Subtyping analysis was performed by using the online analysis tool of Genotyping in combination with the MEGA 5.03 software.The HIV resistance mutations were determined and scored with the use of Stanford HIV Drug Resistance Database.Results A total of 196 pol gene sequences were obtained from 103 antiretroviral therapy (ART)-treated subjects (52.55%) and 93 ART-na?ve subjects (47.45%).The 196 pol gene sequences were classified into four subtypes including CRF01_AE, CRF08_BC, CRF07_BC and B, accounting for 48.47%, 44.90%, 6.12%and 0.51%, respectively.The HIV drug resistance rates in sub-jects with and without ART were 10.68% and 7.53%, respectively.Among the 196 subjects, 14 cases showed low level of drug resistance, 3 cases showed moderate level of drug resistance and 4 cases showed high level of resistance.Only one case was resistant to both nucleoside reverse transcriptase inhibitors ( NR-TIs) and non-nucleoside reverse transcriptase inhibitors ( NNRTIs) .The resistance rates of the 196 cases to protease inhibitor (PIs), NRTIs, NNRTIs, and integrase inhibitors (INs) were 6.63%, 3.06%, 11.22%and 8.67%, respectively.The frequencies of PIs-related mutations in subtypes CRF01_AE, CRF07_BC and CRF08_BC were 6.32%, 41.67% and 2.27%, respectively.Most of the PI-related A71V/T mutations were identified in strains belonging to subtype CRF07_BC, accounting for 75% of all A71V/T mutations found in the 196 strains.The NNRTI-related E138A mutations only appeared in strains belonging to subtype CRF08_BC.Conclusion The drug resistance rate among patients with HIV-1/AIDS in Guangxi was higher than the average level in China.The drug resistance rates varied with the subtypes of HIV-1 strains.

Construction of Tre enzyme eukaryotic expression vector and identification of its function in loxLTR sequence-specific recognition / 医学研究生学报

Objective Clearing HIV provirus is the key to cure AIDS .The study was to construct the Tre enzyme eukaryotic expression vector and identify its function in specific recognition of loxLTR sequence in HIV provirus . Methods Tre gene was in-serted into eukaryotic expression vector pcDNA 3.1 gene recombination manipulation by genetic recombination techniques including gene synthesis , PCR, restriction enzyme digestion and ligation .EGFPpA-LoxLTR sequence was inserted into pmCherry-N1 vector and was tested by restriction enzyme digestion , PCR and sequencing .Constructed vectors were electroporated into HeLa cells , then using fluorescence microscopy to observe fluorescence intensity changes . Results PCR, restriction enzyme digestion , electrophoresis and sequencing confirmed that Tre enzyme eukaryotic expression vector had been constructed successfully , and it could specifically recog-nize and cut loxLTR sequence after being transfected into Hela cells . Conclusion Constructed Tre enzyme eukaryotic expression vector can be expressed in Hela cells and specifically recognize loxLTR sequence , which has prepared the experimental ground for fur-ther studies of clearing HIV provirus .

Comportamiento de marcadores serológicos en donantes de sangre del territorio de Colón, 1998-2007 / Behavior of the serologic markers in blood donors in the territory of Colon, 1998-2007

En el laboratorio de Sistema Ultra-Micro-Analítico, del banco de sangre del Hospital Territorial Universitario Dr Mario Muñoz Monroy, del municipio Colón, provincia Matanzas, se realizó un estudio descriptivo prospectivo longitudinal, sobre el comportamiento de marcadores serológicos, donde se determinó la incidencia y prevalencia del antígeno de superficie del virus de la hepatitis B (VHB, HBsAg) y de los anticuerpos contra los virus de la hepatitis C (VHC, anti-VHC) y de la inmunodeficiencia humana 1 y 2 (VIH 1 y 2, anti-VIH 1+2), en donantes de sangre del territorio, y el estimado de infección potencial no detectada transmisible por sangre o riesgo residual (RR) en la sangre donada, en el tiempo comprendido del 1 de enero de 1998 al 31 de diciembre de 2007. La investigación se realizó con todo el universo de donantes útiles y sus respectivas donaciones de sangre, y quedó constituido por 49 749 donantes y 84 932 bolsas de sangre. Los índices de prevalencia (x 100 000 donantes), incidencia (x 100 000 donantes), y estimado de riesgo residual (x 1 000 000 de unidades de sangre donada) en el citado período de tiempo fueron: para el VHB 0,81; 0,17 y 0,20; para el VHC 0,55; 0,12 y 0,23; y para los VIH 1y2 0,005; 0,01x10-2 y 0,02 x10-3, respectivamente, índices bajos según la clasificación internacional; pero no para Cuba con respecto al HBsAg.

We carried out a prospective descriptive longitudinal study on the behavior of the serologic markers in the Ultra-Micro-Analytic System laboratory, of the blood bank of the territorial university hospital Dr Mario Muñoz Monroy, of the municipality of Colon, province of Matanzas. We determined the incidence and prevalence of the surface Hepatitis B virus antigen (HBV, HBsAg) and of the antibodies against the Hepatitis C (HCV, anti HCV) and the human immunodeficiency virus 1 and 2 (HIV 1 and 2, anti-HIV 1+2) in blood donors of the territory, and the estimate of non-detected potential infection transmissible by blood or residual risk (RR) in the donated blood, in the period from January 1st 1998 to December 31st 2007. The research was made with all the universe of utile donors and their respective blood donations, and was formed by 49 749 donors and 84 932 blood bags. The prevalence rates (x 100 000 donors), incidence (x 100 000 donors), and estimated residual risk (x 1 000 000 units of donated blood) in the quoted time period were: for the HBV 0,81; 0,17 and 0,20; for the HCV 0,55; 0,12 and 0,23, and for the HIV 1 and 2 0,005; 0,01x10-2 and 0,02x10-3 respectively, low rates according to the international classification, but not for Cuba with respect of the HBsAg.