RESUMO
HIV, HTLV-1/-2, and HCV share routes of transmission, and such virus co-infections could account for worse outcomes of associated diseases. Measuring cytokines/chemokines, CD4 and CD8 T cells, and HIV viral load (VL) in HIV single-infected and co-infected individuals has prognostic value. We analyzed such biomarkers in 129 blood samples of HIV-infected individuals matched for age and sex and divided into six groups (G1 (69 HIV); G2 (9 HIV/HTLV-1); G3 (6 HIV/HTLV-2); G4 (11 HIV/HCV); G5 (19 HIV/HCV/HTLV-1); and G6 (15 HIV/HCV/HTLV-2)). Eight cytokines/chemokines from fifteen analytes could be compared. The highest levels of Th1 and pro-inflammatory cytokines were detected in G2 (IFN-γ) and G6 (IL-6 and IL1-ß) and of chemokines in G1 (MIG, IP10, RANTES), G4 (MCP1), and G6 (MIP1-ß). The highest CD4 cells number and the lowest HIV VL were identified in G3 and the opposite results in G2. Positive correlations between CD4 and CD8 cells counts and IL-6 levels were detected in G2 and G5 and of HIV VL and RANTES in G4. Negative correlations were detected between CD8 and IFN-γ in G4 and HIV VL and RANTES in G6. Despite the small number of the cohort analyzed, and although the cross-sectional study design does not allow firm conclusions, the homogeneity of the characteristics of HIV/HTLV-co-infected individuals regarding age, time and route of HIV acquisition, and criteria for introducing ART enable us to suggest a negative impact of HTLV-1 and a possible protective role of HTLV-2 in HIV infection progression in such patients.
Assuntos
Coinfecção , Infecções por HIV , HIV-1 , Hepatite C , Vírus Linfotrópico T Tipo 1 Humano , Biomarcadores , Quimiocina CCL5 , Quimiocina CXCL10 , Estudos Transversais , HIV-2 , Hepatite C/complicações , Vírus Linfotrópico T Tipo 2 Humano , Humanos , Interleucina-6 , Carga ViralRESUMO
BACKGROUND: A new point-of-care (POC) HIV virus load technology has been recently developed and designed to be utilized in decentralized settings. Alere Technologies GmbH*, Germany, developed the mPIMA HIV-1/2â¯VL plasma test which uses real time PCR technology with 50⯵l and a turnaround time of one hour. OBJECTIVE: Analyze the performance of mPIMA to detect and quantify HIV-1 and HIV-2 and compare with Abbott M2000 assay fooling patients HIV-1 failing ARV therapy. STUDY DESIGN: In this study we evaluate the mPIMA HIV-1/2â¯VL plasma test using 413 specimens from 270 patients failing ARV therapy, and compared its performance with Abbott RealTime HIV-1 Viral Load assay on the m2000 system. In addition, were determined VL in plasma specimens obtained from HIV-2 infected patients. RESULTS: The results strongly indicate that mPIMA HIV-1/2â¯VL plasma test can determine HIV-1 with concordance of 88.9 % (95 % CI 85.4-91.7) the reference test when 1000 HIV-1 VL threshold was used as WHO cutoff to identify therapy failure. The overall correlation between HIV-1 VL was 0928 (Pearson correlation coefficient of Linear regression) and the Bland-Altman showed a mean difference of -0.20 Log cp/mL between the two technology. mPIMA HIV-1/2â¯VL plasma test was also able to measure HIV-2 viral load in 16 specimens from Guinea-Bissau HIV-1/HIV-2 positive samples. CONCLUSIONS: These data support the use of mPIMA HIV-1/2â¯VL plasma test to follow up patients and select patients failing ART, guiding immediate clinical decisions such as adherence counseling or ART regimen switch during the patient consultation.
Assuntos
Infecções por HIV/diagnóstico , Sistemas Automatizados de Assistência Junto ao Leito/normas , RNA Viral/sangue , Carga Viral/instrumentação , Carga Viral/métodos , Fármacos Anti-HIV/uso terapêutico , Alemanha , Infecções por HIV/sangue , Infecções por HIV/tratamento farmacológico , HIV-1/isolamento & purificação , HIV-2/isolamento & purificação , Humanos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Sensibilidade e EspecificidadeAssuntos
Humanos , HIV-1 , HIV-2 , Algoritmos , Ensaio de Imunoadsorção Enzimática/métodos , Estudo de ValidaçãoRESUMO
OBJECTIVE: HIV seroconversion biomarkers are being used in cross-sectional studies for HIV incidence estimation. Bio-Rad Geenius HIV-1/2 Supplemental Assay is an immunochromatographic single-use assay that measures antibodies (Ab) against multiple HIV-1/2 antigens. The objective of this study was to determine whether the Geenius assay could additionally be used for recency estimation. DESIGN: This assay was developed for HIV-1/2 confirmation; however, quantitative data acquired give information on increasing concentration and diversity of antibody responses over time during seroconversion. A quantitative threshold of recent HIV infection was proposed to determine "recent" or "nonrecent" HIV infection; performance using this cutoff was evaluated. METHODS: We tested 2500 highly characterized specimens from research subjects in the United States, Brazil, and Africa with well-defined durations of HIV infection. Regression and frequency estimation were used to estimate assay properties relevant to HIV incidence measurement: mean duration of recent infection (MDRI), false-recent rate, and assay reproducibility and robustness. RESULTS: Using the manufacturer's proposed cutoff index of 1.5 to identify "recent" infection, the assay has an estimated false-recent rate of 4.1% (95% CI: 2.2 to 7.0) and MDRI of 179 days (155 to 201) in specimens from treatment-naive subjects, presenting performance challenges similar to other incidence assays. Lower index cutoffs associated with lower MDRI gave a lower rate of false-recent results. CONCLUSIONS: These data suggest that with additional interpretive analysis of the band intensities using an algorithm and cutoff, the Geenius HIV-1/2 Supplemental Assay can be used to identify recent HIV infection in addition to confirming the presence of HIV-1 and HIV-2 antibodies.
Assuntos
Cromatografia de Afinidade/métodos , Testes Diagnósticos de Rotina/métodos , Anticorpos Anti-HIV/sangue , Infecções por HIV/diagnóstico , Infecções por HIV/epidemiologia , HIV-1/imunologia , HIV-2/imunologia , África , Algoritmos , Brasil , Infecções por HIV/virologia , Incidência , Modelos Teóricos , Estudos Retrospectivos , Estados UnidosRESUMO
Introducción: el estudio de la estabilidad de los componentes y el producto terminado constituye un importante requisito regulatorio en los diagnosticadores. Objetivo: realizar un estudio de estabilidad en tiempo real durante doce meses del sistema inmunoenzimático (ELISA) DAVIH VIH-2. Métodos: se realizó un estudio de estabilidad en tiempo real durante doce meses en tres lotes del diagnosticador DAVIH VIH-2, ELISA indirecto diseñado para la detección de anticuerpos contra el virus de inmunodeficiencia humana tipo 2 en suero o plasma humano. Se controlaron los requisitos de calidad de los componentes de acuerdo a sus especificaciones. Se estudió la normalidad de valores de densidad óptica/valor límite y la homogeneidad de las medias y varianzas mediante las dócimas de Grubbs y Cochran. Se estimó la precisión en los controles positivo y negativo del sistema y en seis muestras con diferente reactividad al virus de inmunodeficiencia humana tipo 2 mediante el cálculo del coeficiente de variación y se confeccionaron las cartas de control de los valores de las medias de densidad óptica respecto al tiempo. Resultados: los requisitos de calidad de cada componente se cumplieron durante 12 meses, excepto las características funcionales del conjugado a partir de los seis meses. Los valores en las dócimas de Grubbs y Cochran fueron menores que los valores críticos tabulados para α del 1 y 5 por ciento por lo que existió homogeneidad en las medias y las varianzas en todo el periodo. El coeficiente de variación se mantuvo inferior al 10 por ciento excepto en las muestras con reactividad media y baja, mientras que en las cartas de control, los valores de densidad óptica se mantuvieron en el límite de la media ±2 desviaciones estándar hasta el noveno mes(AU)
Assuntos
Humanos , Técnicas Imunoenzimáticas/métodos , Reatividade-Estabilidade , Ensaio de Imunoadsorção Enzimática/métodos , HIV-2/imunologiaRESUMO
Post-transcriptional control in both HIV-1 and HIV-2 is a highly regulated process that commences in the nucleus of the host infected cell and finishes by the expression of viral proteins in the cytoplasm. Expression of the unspliced genomic RNA is particularly controlled at the level of RNA splicing, export, and translation. It appears increasingly obvious that all these steps are interconnected and they result in the building of a viral ribonucleoprotein complex (RNP) that must be efficiently translated in the cytosolic compartment. This review summarizes our knowledge about the genesis, localization, and expression of this viral RNP.
Assuntos
Regulação Viral da Expressão Gênica , HIV-1/fisiologia , HIV-2/fisiologia , Interações Hospedeiro-Patógeno , Biossíntese de Proteínas , RNA Viral/metabolismo , Replicação Viral , Núcleo Celular/metabolismo , Citoplasma/metabolismo , HIV-1/genética , HIV-2/genética , Humanos , Ribonucleoproteínas/metabolismo , Proteínas Virais/metabolismoRESUMO
INTRODUCTION: The HIV-2 glycoprotein 36 (gp36) is often used in ELISA. An evaluation of the diagnostic indexes of antigen mixtures with a synthetic peptide of HIV2 gp36 (5) is performed in this study. METHODS: Five mixtures of gp36 (5) and the recombinant proteins of HIV1/2 were prepared. A total of 1306 samples were evaluated with UMELISA HIV1+2 RECOMBINANT used as reference. RESULTS: The variant (V-1) showed very good agreement as regards the reference method. CONCLUSION: The V-1 variant was shown to be highly effective in the immunodiagnosis of HIV 1/2.
Assuntos
Sorodiagnóstico da AIDS/métodos , Ensaio de Imunoadsorção Enzimática/métodos , Antígenos HIV/imunologia , Infecções por HIV/diagnóstico , HIV-1/imunologia , HIV-2/imunologia , Produtos do Gene env do Vírus da Imunodeficiência Humana/imunologia , Especificidade de Anticorpos , Anticorpos Anti-HIV/sangue , Anticorpos Anti-HIV/imunologia , Infecções por HIV/virologia , HIV-1/isolamento & purificação , HIV-2/isolamento & purificação , Humanos , Fragmentos de Peptídeos/síntese química , Fragmentos de Peptídeos/imunologia , Valor Preditivo dos Testes , Proteínas Recombinantes/síntese química , Proteínas Recombinantes/imunologia , Sensibilidade e Especificidade , Produtos do Gene env do Vírus da Imunodeficiência Humana/síntese químicaRESUMO
During the post-transcriptional events of the HIV-2 replication cycle, the full-length unspliced genomic RNA (gRNA) is first used as an mRNA to synthesize Gag and Gag-Pol proteins and then packaged into progeny virions. However, the mechanisms responsible for the coordinate usage of the gRNA during these two mutually exclusive events are poorly understood. Here, we present evidence showing that HIV-2 expression induces stress granule assembly in cultured cells. This contrasts with HIV-1, which interferes with stress granules assembly even upon induced cellular stress. Moreover, we observed that the RNA-binding protein and stress granules assembly factor TIAR associates with the gRNA to form a TIAR-HIV-2 ribonucleoprotein (TH2RNP) complex localizing diffuse in the cytoplasm or aggregated in stress granules. Although the assembly of TH2RNP in stress granules did not require the binding of the Gag protein to the gRNA, we observed that increased levels of Gag promoted both translational arrest and stress granule assembly. Moreover, HIV-2 Gag also localizes to stress granules in the absence of a 'packageable' gRNA. Our results indicate that the HIV-2 gRNA is compartmentalized in stress granules in the absence of active translation prior to being selected for packaging by the Gag polyprotein.
Assuntos
Grânulos Citoplasmáticos/virologia , HIV-2/genética , RNA Viral/metabolismo , Montagem de Vírus , Grânulos Citoplasmáticos/metabolismo , Fator de Iniciação 2 em Eucariotos/metabolismo , Genoma Viral , HIV-2/fisiologia , Células HeLa , Humanos , Biossíntese de Proteínas , RNA Viral/análise , RNA Viral/biossíntese , Proteínas de Ligação a RNA/química , Proteínas de Ligação a RNA/metabolismo , Ribonucleoproteínas/metabolismo , Estresse Fisiológico , Replicação Viral , Produtos do Gene gag do Vírus da Imunodeficiência Humana/biossíntese , Produtos do Gene gag do Vírus da Imunodeficiência Humana/genéticaRESUMO
The presence of infection by human immunodeficiency virus type 2 (HIV-2) in Cuba has been previously documented. However, genetic information on the strains that circulate in the Cuban people is still unknown. The present work constitutes the first study concerning the phylogenetic relationship of HIV-2 Cuban isolates conducted on 13 Cuban patients who were diagnosed with HIV-2. The env sequences were analyzed for the construction of a phylogenetic tree with reference sequences of HIV-2. Phylogenetic analysis of the env gene showed that all the Cuban sequences clustered in group A of HIV-2. The analysis indicated several independent introductions of HIV-2 into Cuba. The results of the study will reinforce the program on the epidemiological surveillance of the infection in Cuba and make possible further molecular evolutionary studies.
Assuntos
Variação Genética , Infecções por HIV/virologia , HIV-2/classificação , HIV-2/genética , Filogenia , Adulto , Idoso , Análise por Conglomerados , Cuba , Feminino , HIV-2/isolamento & purificação , Humanos , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Análise de Sequência de DNA , Adulto Jovem , Produtos do Gene env do Vírus da Imunodeficiência Humana/genéticaRESUMO
O diagnóstico sorológico da infecção pelo HIV-1 e HIV-2 teve início em Cabo Verde em 1987, mas pouco se sabe a respeito da diversidade genética desses vírus nessas ilhas, localizadas na costa Ocidental Africana. Neste estudo, caracterizamos a epidemiologia molecular do HIV-1 e HIV-2 em Cabo Verde, analisamos a origem dos principais clados de HIV introduzidos no país e descrevemos a ocorrência de mutações de resistência aos antirretrovirais (DRM) em indivíduos virgens de tratamento (ARTn) e pacientes em tratamento (ARTexp) oriundos das diferentes ilhas. Amostras de sangue, dados sociodemográfico e clínico-laboratoriais foram obtidos de 221 indivíduos HIV positivos entre 2010-2011. As amostras foram sequenciadas na região da polimerase (1300 pares de bases) e análises filogenéticas e de bootscan foram realizadas para a subtipagem viral. Os algoritmos disponibilizados nos sites Stanford HIV Database e HIV-GRADE e.V. Algorithm Homepage foram utilizados para avaliar a existência de DRM em pacientes positivos para HIV-1 e HIV-2, respectivamente. Os estudos evolutivos e filogeográficos foram realizados através do programa BEASTEntre os 221 pacientes analisados, sendo 169 (76,5 porcento) HIV-1, 43 (19,5 porcento) HIV-2 e 9 de (4,1 porcento) co-infectados pelo HIV-1 e pelo HIV-2, 67 porcento eram do sexo feminino. As medianas de idade foram de 34 (IQR = 1-75) e 47 (IQR = 12-84) para o HIV-1 e HIV-2, respectivamente. A infecção pelo HIV-1 é causada pelo subtipo G (36,6 porcento), CRF02_AG (30,6 porcento), subtipo F1, (9,7 porcento), URFs (10,4 porcento), subtipo B (5,2porcento), CRF05_DF (3,0 porcento), subtipo C (2,2 porcento), CRF06_cpx (0,7 porcento), CRF25_cpx (0,7 porcento) e CRF49_cpx (0,7 porcento), e todas as infecções por HIV-2 pertencem ao grupo A. De acordo com as análises filogeográficas e de origem do HIV, estimase que o HIV-2 foi o primeiro tipo viral introduzido em Cabo Verde e possui relações filogenéticas com sequências referências de Portugal...
HIV-1 and HIV-2 have been detected in Cape Verde since 1987, but little is known regarding the genetic diversity of these viruses in this archipelago, located near the West African coast. In this study, we characterized the molecular epidemiology of HIV-1 and HIV-2, analyzed the origin of the major clades of HIV introduced in CapeVerde and described the occurrence of drug resistance mutations (DRM) among antiretroviral therapy naïve (ARTn) patients and patients under treatment (ARTexp) from different Cape Verde islands. Blood samples, socio-demographic and clinicallaboratorydata were obtained from 221 HIV-positive individuals during 2010-2011.Genetic sequencing of the pol region (1300bp) was obtained and phylogenetic and bootscan analyses were performed for viral subtyping. The evolutionary and phylogeographic studies were performed using the program BEAST. HIV-1 and HIV-2 DRM were evaluated for ARTn and ARTexp patients using the Stanford HIVDatabase and HIV-GRADE e.V. Algorithm Homepage, respectively. Among the 221patients (169 [76.5 percent] HIV-1, 43 [19.5 percent] HIV-2 and 9 [4.1 percent] HIV-1/HIV-2 coinfections), 67 percent were female. The median ages were 34 (IQR=1-75) and 47(IQR=12-84) for HIV-1 and HIV-2, respectively. HIV-1 infections were due to subtypes G (36.6 percent), CRF02_AG (30.6 percent), F1 (9.7 percent), URFs (10.4 percent), B (5.2 percent), CRF05_DF (3.0 percent), C (2.2percent), CRF06_cpx (0.7 percent), CRF25_cpx (0.7 percent) and CRF49_cpx (0.7 percent), whereas all HIV-2 infections belonged to group A. According the HIV phylogeographic analyses, it is estimated that HIV-2 was the first viral type introduced in Cape Verde and has phylogenetic relationships with referral sequencesof Portugal...