Emergence of HIV‑1 drug‑resistant variants in women following antiretroviral prophylaxis for the prevention of mother to child transmission.

Purpose: Emergence of drug resistance following HIV prophylaxis has an important impact on ART program. Objective: To investigate the emergence of drug resistance in HIV‑1 infected pregnant women. Materials and Methods: Fifty‑three HIV‑1 infected pregnant women who had received 4‑12 weeks of antenatal AZT followed by Nevirapine during delivery and Combivir [AZT + 3TC] for 1 week postpartum (group‑1, n = 48) or who come at the time of delivery and received Nevirapine followed by Combivir for 1 week (group‑2, n = 5) were recruited. Samples were collected prior to the start of the prophylaxis and 5‑8 weeks postpartum. In addition, a third sample was collected between 26‑65 weeks postpartum from 7 women. Amplification of HIV‑1 pol gene and drug resistance analysis was done. Result: Two (3.8%) women in group‑1 showed transmitted drug resistance and they continued to show this even at 6 weeks postpartum. One (2%) woman from group‑1 showed a mutation after 6‑8 weeks of prophylaxis. Among the samples collected between 26‑65 weeks postpartum, 3/7 (43%) showed mutations and all these women belong to group‑1. Women belonging to group‑2 didn’t show mutation prior to or following prophylaxis. Conclusion: In contrast to the available data among pregnant women with ART prophylaxis, our data showed reduced frequency of mutations following 5‑8 weeks of postpartum but an emergence of mutation later (26‑65 weeks). The addition of Combivir with the single dose Nevirapine during delivery and the early stage of the disease with higher CD4 counts could be the reasons for this.

Generation of HIV-1 based bi-cistronic lentiviral vectors for stable gene expression and live cell imaging.

The study of protein-protein interactions, protein localization, protein organization into higher order structures and organelle dynamics in live cells, has greatly enhanced the understanding of various cellular processes. Live cell imaging experiments employ plasmid or viral vectors to express the protein/proteins of interest fused to a fluorescent protein. Unlike plasmid vectors, lentiviral vectors can be introduced into both dividing and non dividing cells, can be pseudotyped to infect a broad or narrow range of cells, and can be used to generate transgenic animals. However, the currently available lentiviral vectors are limited by the choice of fluorescent protein tag, choice of restriction enzyme sites in the Multiple Cloning Sites (MCS) and promoter choice for gene expression. In this report, HIV-1 based bi-cistronic lentiviral vectors have been generated that drive the expression of multiple fluorescent tags (EGFP, mCherry, ECFP, EYFP and dsRed), using two different promoters. The presence of a unique MCS with multiple restriction sites allows the generation of fusion proteins with the fluorescent tag of choice, allowing analysis of multiple fusion proteins in live cell imaging experiments. These novel lentiviral vectors are improved delivery vehicles for gene transfer applications and are important tools for live cell imaging in vivo.

Modificación en la información sobre la infección por VIH/SIDA del personal estomatológico en 10 años / Modification in the information about the HIV/AIDS infection of the stomatologic personnel in 10 years

Fundamento: En 1995 la autora demostró un déficit en la información mínima indispensable sobre la infección por VIH/SIDA, y cómo proceder seguro en los tratamientos a las personas viviendo con esta enfermedad. Con posterioridad, se capacitó a gran parte del personal de post grado y se incluyó en los programas de pre grado. Diez años después, conocimos la situación y la comparamos. Objetivo: Evaluar y comparar la evolución de la información que sobre la infección de VIH/SIDA y aspectos de bioseguridad tenía el personal de estomatología en la provincia Camagüey, diez años después de realizarse las primeras intervenciones educativas para modificar esta información. Método: Se realizó un estudio descriptivo, se aplicó la misma encuesta que al inicio del estudio, a una muestra conformada por doscientos noventa y tres personas entre profesionales, técnicos de atención estomatológica y estudiantes de 4to y 5to año de la carrera de estomatología. Los datos se procesaron en una computadora Pentium a través del paquete estadístico SPSS para Windows. Resultados: Se comprobó que hubo una modificación favorable después de los diez años de un 63,5 por ciento en el nivel de información general que sobre la infección por el VIH/SIDA tenía el personal de estomatología. Conclusión: En relación al estudio anterior, diez años después hubo una modificación favorable en el nivel de información general de la infección por VIH/SIDA en el personal de estomatología.

Background: In 1995 the author demonstrated a deficit in the indispensable minimal information about the HIV/AIDS infection, and how to proceed in the treatments to people living with this disease. Subsequently, a great part of the postgraduate personnel was capacitated and it was included in the pre-graduate programs. Ten years later, we knew the situation and we compare it. Objective: To evaluate and to compare the evolution of the information about the HIV/AIDS infection and the biosecurity aspects had the stomatological personnel in Camagüey province, ten years after of being carried out the first educational interventions to modify this information. Method: A descriptive study was conducted, and it was also applied the same survey of the beginning of the study, to a sample conformed by 293 people among professionals, technicians of stomatological care and students of 4th and 5th year of the stomatological career. Data were processed in a Pentium computer through the SPSS statistical package for Windows. Results: It was verified that there was a favorable modification after the ten years of 63, 5 percent in the level of the general information about the HIV/AIDS infection had the stomatological personnel. Conclusión: In relation to the previous study, in 10 years later had a favorable modification in the level of general information of the HIV/AIDS infection in the stomatologic staff.

The association between HLA-A antigen carrying Bw4 epitope and HIV-1 viral loads and CD+4 T cell counts / 中华检验医学杂志

Objective To investigate the impact of HLA-A antigens carrying Bw4 epitopes on disease course of HIV-1 infection.Methods Three hundred and forty subjects chronically infected with HIV-I were recruited and their HLA-A and B alleles were genotyped with sequence-based high resolution typing assay.HLA-Bw genotypes of these HIV-1 infected subjects were determined and their association with CD+4 T cell counts and viral load were analysed.Results When compared with subjects canting no Bw4 epitopes in HLA-A and HLA-B loci (OBw4) (median of CD+4 T cell counts:294/μ1;plasma viral load median 6.29×104 copies/ml),CD+4 T cell counts in subjects with genotypes of 1Bw4-A and 2Bw4-AA were comparable (307 and 308/μ1,respectively),but higher viral load (1.53×105 and 2.68×105 copies/ml,respectively) was observed.In subjects with Bw4 epitopes in HLA-B alleles but no in HLA-A,significantly higher CD+4 T cell counts (417/μ1,P=0.013) and lower viral load (2.10×104 copies/ml,P=0.007) were observed compared with those without Bw4 in HLA-A and HLA-B.Conclusion HLA-B antigens carrying Bw4 epitope were protective in HIV-1 infection by maintaining higher CD+4 T cell counts and lower viral load,but such protective effect was not observed in HLA-A antigens earring Bw4 epitope.

Studies of interaction between KIR alleles and the progression of HIV-1 infection in Chinese population / 中华微生物学和免疫学杂志

Objective To investigate the killer cell lg-like receptors (KIR) gene frequency of HIV-1 infected slow progressors(SP) and typical progressors(TP), and to analyze the interaction between KIR alleles and the progression of HIV-1 infection in Chinese population. Methods Eighty-one HIV-1 posi-tive individuals including 43 SPs and 38 TPs were recruited. Carriage of KIR genes was assessed using poly-merase chain reaction sequence-specific primers (PCR-SSP) assays. Results KIR2DS3 gene frequency was significantly lower in SP group (3.6%) than that in TP group (14.2%), P =0. 018 ,OR =0. 210,95% CI =0.053-0.833. The number of activating KIR genes was less in SP group than that in TP group, but was not significant (P = 0. 208). Conclusion Lower KIR2DS3 gene frequency may potentially be associated with slower progression to AIDS in Chinese population.

A molecular epidemioiogical survey of 60 human immunodeficiency viruses-1 infected individuals from Honghe district in Yunnan Province / 中华传染病杂志

Objective To investigate the molecular epidemiology of human immunodefieiency viruses (HIV)-1 infected individuals in Honghe district,Yunnan Province and provide the evidence of molecular biology features of HIV-1 infection.Methods HIV-1 pol gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR).Then sequencing and phylogenetic tree analysis were employed tO determine HIV-I subgenotype.The sequence alignment was performed in the database of international drug resistance tO identify resistance-associated mutations.Results The samples from 60 HIV-1 infected individuals were investigated:39 were male,21 were female,with average age 35.5 years old.Thirty-four cases were infected with HIV-I through intravenous drug abuse,12 by sexual contacts,2 were contaminated blood/blood products transfusion and 12 with unknown transmission routes.Phylogenetic analysis revealed that 53 cases (88.3%) were subtype 08-BC,6 (10.0%) were subtype 07-BC and 1 (1.7%) was subtype 01_AE.The total rate of drug resistance associated mutations was 33.3%.The major mutations in protease (PR) and reverse transcriptase (RT) regions accounted for 5.0% and 3 1.7%,respectively.The major mutations in PR region were I541M,V82VFIL,M46MI,which were found in 1 case,respectively.The mutations in RT region were as follows:4 cases were T69D,6 were A62V,1 was D67DE,1 was E44D,3 were V179D,1 was V179E.1 was K238KN,1 was L234T+P236S and 1 was V106E.Conclusions The major transmission route of HIV-I infection in Honghe district,Yunnan Province is through drug injection.The major HIV-1 subtype of HIV-infeeted individuals is 08_BC.PR inhibitor and RT inhibitor drug resistance associated mutations in HlV-1 gene have already existed.

Prokaryotic Expression and Purification of HIV-1 Vif and hAPOBEC3G, Preparation of Polyclonal Antibodies / 中国病毒学·英文版

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.

Caracterización biológica de aislamientos de VIH-1 en pacientes con una evolución clínica rápida

Se estudiaron las características biológicas de 11 cepas de VIH-1 aisladas de pacientes con rápida evolución clínica al sida. Los aislados virales se clasificaron según su cinética de replicación y tropimo celular, con estos criterios se observó que 8 de las cepas aisladas (72,7 %) resultaron de crecimiento rápido alto o lento bajo 3 y con tropismo preferencial a la estirpe linfocítica, como corresponde a los pacientes con sida, mientras 3 (27,3 %) tuvieron características de lenta baja 1. La citopatogenicidad de las cepas se estudió en la línea celular MT4 y se observó que la mayoría (72,7 %) resultó inductora de sincicios (IS), por lo que se comprobó relación in vivo e in vitro de las propiedades biológicas, no ocurrió así en 3 de los cultivos (27,3 %) que se comportaron como no inductores de sincicios.

The biological characteristics of 11 HIV-1 strains isolated from patients with a fast clinical evolution to AIDS were studied. The viral isolates were classified according to their replication kinetics and cell tropism. Taking into account these criteria, it was observed that 8 of the isolated strains (72,7 %) were of rapid high growth (RH) or slow low 3 (SL3) with preferential tropism to the lymphocytic stock, as it corresponds to AIDS patients. 3 (27,3 %) had characteristics of slow low 1 (SL1). The cytopatogenecity of the strains was studied in the MT4 celular line, and it was observed that most of them (72,7 %) were syncytium-inducing strains (SI), which allowed to prove the in vivo and in vitro relation of the biological properties. It was not so in 3 of the cultures (27,3 %) that behaved as non-syncytium inducers.