Lower High-Density Lipoproteins Levels During Human Immunodeficiency Virus Type 1 Infection Are Associated With Increased Inflammatory Markers and Disease Progression.

INTRODUCTION: High-density lipoproteins (HDL) are responsible for the efflux and transport of cholesterol from peripheral tissues to the liver. In addition, HDL can modulate various immunological mechanisms, including the inflammatory response. Inflammasomes are multiprotein complexes that have been reported to be activated during human immunodeficiency virus type 1 (HIV-1) infection, thus contributing to immune hyperactivation, which is the main pathogenic mechanism of HIV-1 progression. However, the relationship between HDL and inflammasomes in the context of HIV-1 infection is unclear. Therefore, this research aims to explore the association between HDL and the components of the inflammatory response during HIV-1 infection. METHODOLOGY: A cross-sectional study, including 36 HIV-1-infected individuals without antiretroviral treatment and 36 healthy controls matched by sex and age, was conducted. Viral load, CD4+ T-cell counts, serum HDL, and C-reactive protein (CRP) were quantified. Serum cytokine levels, including IL-1ß, IL-6, and IL-18, were assessed by ELISA. The inflammasome-related genes in peripheral blood mononuclear cells were determined by quantitative real-time PCR. RESULTS: HIV-1-infected individuals showed a significant decrease in HDL levels, particularly those subjects with higher viral load and lower CD4+ T-cell counts. Moreover, upregulation of inflammasome-related genes (NLRP3, AIM2, ASC, IL-1ß, and IL-18) was observed, notably in those HIV-1-infected individuals with higher viral loads (above 5,000 copies/mL). Serum levels of IL-6 and CRP were also elevated in HIV-1-infected individuals. Significant negative correlations between HDL and the mRNA of NLRP3, AIM2, ASC, IL-1ß, and IL-18, as well as viral load and CRP were observed in HIV-1-infected individuals. Likewise, a significant positive correlation between HDL and CD4+ T-cell counts was found. CONCLUSION: In summary, our results indicate that HDL might modulate the expression of several key components of the inflammasomes during HIV-1 infection, suggesting a novel role of HDL in modifying the inflammatory state and consequently, the progression of HIV-1 infection.

Discovery of N-arylsulfonyl-3-acylindole benzoyl hydrazone derivatives as anti-HIV-1 agents

The discovery and development of novel inhibitors with activity against variants of human immunodeficiency virus type 1 (HIV-1) is pivotal for overcoming treatment failure. As our ongoing work on research of anti-HIV-1 inhibitors, 32 N-arylsulfonyl-3-acylindole benzoyl hydrazone derivatives were prepared by introduction of the hydrazone fragments on the N-arylsulfonyl-3-acylindolyl skeleton and preliminarily screened in vitro as HIV-1 inhibitors for the first time. Among of all the reported analogues, eight compounds exhibited significant anti-HIV-1 activity, especially N-(3-nitro)phenylsulfonyl-3-acetylindole benzoyl hydrazone (18) and N-(3-nitro)phenylsulfonyl-3-acetyl-6-methylindole benzoyl hydrazone (23) displayed the most potent anti-HIV-1 activity with EC50 values of 0.26 and 0.31 µg/mL, and TI values of >769.23 and >645.16, respectively. It is noteworthy that introduction of R3 as the methyl group and R2 as the hydrogen group could result in more potent compounds. This suggested that introduction of R3 as the methyl group could be taken into account for further preparation of these kinds of compounds as anti-HIV-1 agents

Synthesis and in vitro anti-HIV-1 evaluation of some N-arylsulfonyl-3-formylindoles

As our ongoing work on research of anti-HIV-1 inhibitors, fifteen N-arylsulfonyl-3-formylindoles (3a-o) were designed and prepared through two step synthetic route. Firstly, 3-formylindoles (2a-c) were synthesized via the Vilsmeier-Haack reaction. Subsequently, treatment of 2a-c with the appropriate arylsulfonyl chlorides led to the corresponding target compounds in excellent yields. All analogues were also preliminary evaluated in vitro for their inhibitory activity against HIV-1 replication. Among of all the reported analogues, three compounds 3c, 3g and 3i displayed significant anti-HIV-1 activity, with EC50 values of 9.57, 11.04 and 5.02 µM, and TI values of 31.89, 13.79 and 81.69, respectively. N-m-nitrophenylsulfonyl-3-formylindole (3c) and N-m-nitrophenylsulfonyl-6-methyl-3-formylindole (3i) especially exhibited the best promising anti-HIV-1 activity. In addition, it demonstrated that insertion of a methyl group at the C-6 position of the indolyl ring and a nitro group at the meta position of the arylsulfonyl ring, as in compound 3i, resulted in both low cytotoxicity (CC50= 410.41 µM) and good antiviral activity

Frequent detection of CXCR4-using viruses among Brazilian blood donors with HIV-1 long-standing infection and unknown clinical stage: Analysis of massive parallel sequencing data.

The determination of viral tropism is critically important and highly recommended to guide therapy with the CCR5 antagonist, which does not inhibit the effect of X4-tropic viruses. Here, we report the prevalence of HIV-1×4 HIV strains in 84 proviral DNA massively parallel sequencing "MPS" data from well-defined non-recently infected first-time Brazilian blood donors. The MPS data covering the entire V3 region of the env gene was extracted from our recently generated HIV-1 genomes sequenced by a paired-end protocol (Illumina). Of the 84 MPS data samples, 63 (75%) were derived from donors with long-standing infection and 21 (25%) were lacking stage information. HIV-1 tropism was inferred using Geno2pheno (g2p) [454] algorithm (FPR=1%, 2.5%, and 3.75%). Among the 84 data samples for which tropism was defined by g2p2.5%, 13 (15.5%) participants had detectable CXCR4-using viruses in their MPS reads. Mixed infections with R5 and X4 were observed in 11.9% of the study subjects and minority X4 viruses were detected in 7 (8.3%) of participants. Nine of the 63 (14.3%) subjects with LS infection were predicted by g2p 2.5% to harbor proviral CXCR4-using viruses. Our findings of a high proportion of blood donors (15.5%) harboring CXCR4-using viruses in PBMCs may indicate that this phenomenon is common. These findings may have implications for clinical and therapeutic aspects and may benefit individuals who plan to receive CCR5 antagonists.

Identification of innate immune antiretroviral factors during in vivo and in vitro exposure to HIV-1.

Defensins, RNases and cytokines are present at mucosal barriers, main ports of HIV entry, and are potential mediators of the resistant phenotype exhibited by HIV-1-exposed seronegative individuals (HESN) during sexual exposure. We aimed to determine the role of soluble factors in natural resistance to HIV-1 infection. Vaginal/endocervical/oral mucosal samples were taken from 60 HESN, 60 seropositive (SP) and 61 healthy controls (HC). Human neutrophil peptide 1 (hNP-1), human beta defensin (hBD) 2 and 3, RNases, MIP-1ß and RANTES mRNA transcripts were quantified by qPCR and in vitro single-round, recombinant-based viral infectivity assay was used to evaluate the anti-HIV-1 activity of hBDs and RNases. HESN expressed significantly higher levels of hNP-1, hBDs mRNA in oral mucosa compared to HC (P < 0.05). In genital mucosa, significantly higher mRNA levels of MIP-1ß, RANTES and RNases were found in HESN compared to HC (P < 0.05). HBDs and RNases inhibit HIV-1 replication, particularly R5 at entry, reverse transcription and nuclear import of the viral life cycle. hNP-1, hBDs, MIP-1ß, RANTES and RNases, collectively could contribute to HIV-1 resistance during sexual exposure. Moreover, the inhibition of HIV-1 infection in vitro by hBDs and RNases suggests that they may be exploited as potential antiretrovirals.

In vitro anti-HIV-1 activity of the enzymatic extract enriched with laccase produced by the fungi Ganoderma sp. and Lentinus sp / Actividad anti-VIH-1 in vitro del extracto enzimático enriquecido con lacasa producido por los hongos Ganoderma sp. y Lentinus sp

Background: Natural compounds are a good source for the development of antiretroviral drugs with low cytotoxicity. The laccase enzyme, produced by fungi of the genera Ganoderma sp. and Lentinus sp., inhibits the reverse transcriptase (RT) of the human immunodeficiency virus 1 (HIV-1), in cell-free models in vitro. Objetives: In this study we evaluated the anti-HIV-1 activity of the enzymatic extracts (EE) enriched with laccase, produced by two native species of fungi of the same genera in an in vitro cell culture model. Methods: The inhibition of viral replication was performed using the U373-MAGI cell line infected with recombinant viruses in the presence/absence of the EE and 48 hpi, the percentage of infected cells was evaluated by flow cytometry for green fluorescent protein ­GFP­ and ELISA for p24. The inhibition of the RT was determined by quantification of early and late products of reverse transcription using quantitative PCR. Results: The EEs from Ganoderma sp. and Lentinus sp. inhibited the replication of HIV-1 between 80 and 90% and decreased the production of early and late transcripts between 55,5%-91,3% and 82,1%-93,6% respectively. The EE from Lentinus sp. had the best selectivity index (SI: 8.3). Conclusions: These results suggest the potential anti-HIV-1 activity of the EE for the exploration of an alternative therapy against HIV-1 infection.

Antecedentes: Los compuestos naturales son una buena fuente para el desarrollo de fármacos antirretrovirales con baja citotoxicidad. La enzima lacasa, producida por hongos del género Ganoderma sp. y Lentinus sp., inhibe la transcriptasa reversa (TR) del virus de la inmunodeficiencia humana tipo 1 (VIH-1), en modelos in vitro, libres de células. Objetivos: En este estudio se evaluó la actividad anti-VIH-1 del extracto enzimático (EE) enriquecido con lacasa, producida por dos especies nativas de hongos de los mismos géneros en un modelo in vitro de cultivo celular. Métodos: La inhibición de la replicación viral se realizó usando la línea celular U373-MAGI infectada con virus recombinantes en la presencia/ ausencia del EE y 48 hpi, el porcentaje de células infectadas se evaluó mediante citometría de flujo para GFP y ELISA para p24. La inhibición de la TR se determinó mediante la cuantificación de los productos tempranos y tardíos de la transcripción reversa utilizando una PCR cuantitativa. Resultados: El EE de Ganoderma sp. y Lentinus sp. inhibió la replicación del VIH-1 entre el 80 y 90% y disminuyó la producción de transcriptos tempranos y tardíos entre el 55,5% -91,3% y 82,1% -93,6%, respectivamente. El EE de Lentinus sp. mostró el mejor índice de selectividad (IS: 8.3). Conclusiones: Estos resultados sugieren el potencial anti-VIH-1 del EE para la exploración de una terapia alternativa contra la infección por el VIH-1.

Avaliação dos fatores associados à transmissão vertical de HIV-1 / Evaluation of factors associated with vertical HIV-1 transmission

Resumo Objetivo Comparar a prevalência e os fatores associados à transmissão vertical de HIV-1 entre grávidas tratadas de 1998-2004 e de 2005-2011 em um serviço de referência de cuidado de pacientes com HIV no sul do Brasil. Métodos Estudo descritivo e analítico que usou as bases de dados de laboratórios da Rede Nacional de Laboratórios de CD4 e Carga Viral de DST/Aids do Ministério da Saúde. As grávidas com HIV-1 foram selecionadas em uma pesquisa ativa de informações clínicas e dados obstétricos e neonatais em seus prontuários médicos entre 1998-2011. Resultados Foram analisadas 102 grávidas entre 1998 e 2004 e 251 entre 2005-2011, no total 353 crianças nascidas de grávidas com HIV-1. Observou-se que a transmissão vertical foi de 11,8% entre 1998 e 2004 e de 3,2% entre 2005-2011 (p < 0,001). O maior uso de medicamentos antirretrovirais (p = 0,02), a redução na carga viral (p < 0,001) e o tempo de ruptura de membranas menor do que quatro horas (p < 0,001) foram associados à redução nos fatores de transmissão vertical quando os dois períodos são comparados. Conclusão Observou-se uma redução na taxa de transmissão vertical nos últimos anos. De acordo com as variáveis estudadas, sugere-se que os fatores de risco de transmissão vertical de HIV-1 foram ausência de terapia antirretroviral, alta carga viral das grávidas e tempo de ruptura maior do que quatro horas.

Abstract Objective To compare the prevalence and factors associated with vertical transmission of human immunodeficiency virus 1 (HIV-1) among pregnant women treated in the periods of 1998-2004 and 2005-2011 in a reference service for the care of HIV-infected patients in southern Brazil. Methods This was a descriptive and analytical study that used the databases of laboratories from the CD4 and STDs/AIDS Viral Load National Laboratory Network of the Brazilian Ministry of Health. HIV-1-infected pregnant women were selected after an active search for clinical information and obstetric and neonatal data from their medical records between the years of 1998 and 2011. Results 102 pregnant women were analyzed between 1998 and 2004 and 251 in the period between 2005 and 2011, totaling 353 children born to pregnant women with HIV-1. It was observed that the vertical transmission rate was 11.8% between 1998 and 2004 and 3.2% between 2005 and 2011 (p < 0.001). The increased use of antiretroviral drugs (p = 0.02), the decrease in viral load (p < 0.001), and time of membrane rupture lower than 4 h (p < 0.001) were associated with the decrease of vertical transmission factors when comparing the two periods. Conclusion It was observed a decrease in the rate of vertical transmission in recent years. According to the studied variables, is suggested that the risk factors for vertical transmission of HIV-1 were absence of antiretroviral therapy, high viral load in the pregnant women, and membrane rupture time >4 h.

Evaluation of factors associated with vertical HIV-1 transmission.

OBJECTIVE: To compare the prevalence and factors associated with vertical transmission of human immunodeficiency virus 1 (HIV-1) among pregnant women treated in the periods of 1998-2004 and 2005-2011 in a reference service for the care of HIV-infected patients in southern Brazil. METHODS: This was a descriptive and analytical study that used the databases of laboratories from the CD4 and STDs/AIDS Viral Load National Laboratory Network of the Brazilian Ministry of Health. HIV-1-infected pregnant women were selected after an active search for clinical information and obstetric and neonatal data from their medical records between the years of 1998 and 2011. RESULTS: 102 pregnant women were analyzed between 1998 and 2004 and 251 in the period between 2005 and 2011, totaling 353 children born to pregnant women with HIV-1. It was observed that the vertical transmission rate was 11.8% between 1998 and 2004 and 3.2% between 2005 and 2011 (p<0.001). The increased use of antiretroviral drugs (p=0.02), the decrease in viral load (p<0.001), and time of membrane rupture lower than 4h (p<0.001) were associated with the decrease of vertical transmission factors when comparing the two periods. CONCLUSION: It was observed a decrease in the rate of vertical transmission in recent years. According to the studied variables, is suggested that the risk factors for vertical transmission of HIV-1 were absence of antiretroviral therapy, high viral load in the pregnant women, and membrane rupture time >4h.

Participación de la vitamina D en la patogénesis de la infección por el virus de la inmunodeficiencia humana tipo 1 / Vitamin D involvement in the pathogenesis of human immunodeficiency virus type 1

La vitamina D (VitD), además de su papel en el metabolismo mineral, tiene funciones inmunomoduladoras y podría participar activamente en la fisiopatogénesis de la infección por el VIH-1; sin embargo, la evidencia científica en este campo es limitada y controvertida. La VitD tiene propiedades antiinflamatorias que podrían disminuir la hiperactivación inmunológica, reduciendo el daño asociado a este fenómeno; además, promueve la expresión de péptidos con actividad anti-VIH-1, sustentando su papel protector. En contraste, la VitD activa el promotor del VIH-1 y podría potenciar la replicación del virus; adicionalmente, algunas variantes alélicas en el gen del receptor de la VitD, que aumentan su función, se han asociado con susceptibilidad al VIH-1. Esta revisión presenta evidencia científica sobre el efecto de la vía de la VitD en la patogénesis de la infección por el VIH-1, dada las implicaciones de este tópico en la identificación de nuevos blancos terapéuticos en esta infección.

Beyond its role in mineral metabolism, vitamin D (VitD) has immunomodulatory functions and can actively participate in the physiopathogenesis of HIV-1 infection; however, scientific evidence in this field is limited and controversial. VitD has anti-inflammatory properties that can reduce immune hyperactivation, decreasing the damage associated with this phenomenon. It also promotes the expression of antimicrobial peptides with anti-HIV-1 activity, supporting its protective role. In contrast, VitD activates the HIV-1 promoter and can increase viral replication. Furthermore, a number of allelic variants in the vitamin D receptor gene, which increase its function, have been associated with susceptibility to HIV-1 infection. Given the implications of this topic for the identification of new therapeutic targets in HIV infection, this review presents scientific evidence on the effect of the VitD pathway in HIV-1 pathogenesis.

Activation of Toll-like receptor 2 increases macrophage resistance to HIV-1 infection.

Patients infected with HIV-1, the etiological agent of AIDS, have increased intestinal permeability, which allows for the passage of microbial products, including Toll-like receptor (TLR) ligands, into circulation. The exposure of HIV-1-infected cells to certain TLR agonists affects viral replication, but studies associating viral production with the activation of TLR2 in HIV-1-infected cells are rare and controversial. Here, we report that the TLR2 ligands Zymosan and Pam3CSK4 potently inhibit HIV-1 replication in acutely infected monocyte-derived macrophages and the exposure to TLR2 ligands prior to infection renders macrophages refractory to HIV-1 production. Macrophage treatment with Pam3CSK4 did not change the cellular expression of the HIV-1 entry receptors CD4 and CCR5. Both TLR2 ligands increased the macrophage production of ß-chemokines and IL-10, and the blockage of these soluble factors prevented the inhibitory effect of TLR2 activation on HIV-1 replication. Our findings show that the direct engagement of TLR2 in HIV-1-infected macrophages increase cellular resistance to HIV-1 infection, and that controlling HIV-1 replication with agonists for TLR2 might have implications for the development of antiretroviral therapies.

Transmitted drug-resistance in human immunodeficiency virus-infected adult population in El Salvador, Central America.

El Salvador harbours one of the largest Central American human immunodeficiency virus (HIV) epidemics, but few studies have analysed it in depth. Here, we describe the presence of transmitted drug resistance (TDR) and HIV variants in the HIV-infected adult population in El Salvador. Dried blood spots from 119 HIV-infected antiretroviral-naive adults attended in El Salvador were collected in 2011. The TDR was assessed according to the list recommended by the WHO. HIV-1 variants were described using phylogeny. Pol sequences could be amplified in 88 patients (50.6% men), with a mean age of 35 years. Almost all (96.7%) were infected with HIV through sexual practice and 58.7% were recently diagnosed. The mean CD4(+) count was 474 cells/mm(3) and 43.1% and 15.5% of patients showed moderate (<500 CD4 cells) or severe (<200) immune suppression, respectively. HIV-1 viral load was >100 000 copies/mL in 24.7% of patients and <2000 copies/mL in 9.1%. Five samples (5.7%) harboured any TDR mutation: 2.3% for nucleoside reverse transcriptase inhibitor (NRTI) and non-nucleoside reverse transcriptase inhibitor (NNRTI), and 1.4% for protease inhibitor (PI). All showed only one TDR single-class resistance mutation: M184I (two cases) for NRTI, K101E and K103N for NNRTI and L23I for PI. All viruses excepting one (URF_BG) belonged to subtype B. No phylogenetic TDR networks were found. In conclusion, we report a TDR prevalence of 5.7% in El Salvador, lower than in other Central American studies. Periodical studies are essential to monitor and prevent TDR emergence in low-income and middle-income regions. Also, more efforts are needed to promote early diagnosis and prevention of infection in El Salvador.

Genetic and Immunological Factors Involved in Natural Resistance to HIV-1 Infection.

Infection with Human immunodeficiency virus type-1 (HIV-1) induces severe alterations of the immune system leading to an increased susceptibility to opportunistic infections and malignancies. However, exposure to the virus does not always results in infection. Indeed, there exist individuals who have been repeatedly exposed to HIV-1 but do not exhibit clinical or serological evidence of infection, known as exposed seronegative individuals. Many studies have focused on the different mechanisms involved in natural resistance to HIV-1 infection, and have reported several factors associated with this phenomenon, including the presence of genetic polymorphisms in the viral coreceptors, innate and adaptive immune cells with particular phenotypic and functional features, and molecules such as antibodies and soluble factors that play an important role in defense against infection by HIV-1. The study of these factors could be the key for controlling this viral infection. This review summarizes the main mechanisms involved in resistance to HIV-1 infection.

Validación del método de titulación del virus de la inmunodeficiencia humana tipo 1 / Validation of the titration method for the human immunodeficiency virus type 1

INTRODUCCIÓN: en la validación de los procesos de producción de productos biológicos, la cuantificación viral es esencial para demostrar su seguridad. OBJETIVO: en este trabajo se evaluó el método de titulación del virus de la inmunodeficiencia humana tipo 1, para ello se determinó la exactitud, precisión, linealidad y límite de detección. MÉTODOS: la titulación se realizó por el método del cálculo de la dilución punto final, mediante la visualización del efecto citopático y la determinación de la producción de antígeno p24 por medio de un ensayo inmunoenzimático, la dosis infectiva media en cultivo de células por mililitro se calculó según el método de Reed y Muench. RESULTADOS: la exactitud del método coincidió con el título de referencia, los valores del coeficiente de variación de la precisión (repetibilidad y precisión intermedia) fueron menores que 5 y 10 por ciento, respectivamente; la linealidad mostró un adecuado coeficiente de correlación y el límite de detección se determinó en la dilución 10-12. CONCLUSIONES: el método evaluado demostró un adecuado desempeño para la cuantificación viral.

BACKGROUND: viral quantification is essential to demonstrate biological product safety during the validation of the production process. OBJECTIVE: to evaluate the accuracy, linearity and detection limit of a titration method for the human immunodeficiency virus type 1. METHODS: the titration was performed by calculating the end-point dilution method, through the cytopathic effect assay and the determination of Ag p24 production by an enzyme immunoassay. Mean infective dose in cultured cells per milimeter was estimated by Reed and Muench´s method. RESULTS: the accuracy of the method was similar to that of the reference titration, the variation coefficient figures for accuracy (repeatability and intermediate accuracy) were lower than 5 percent and 10 percent, respectively, the linearity showed adequate correlation coefficient and the detection limit was determined in the 10-12 dilution CONCLUSIONS: the evaluated method was suitable for viral quantification.

Prevalência, fatores de risco e caracterização genética dos vírus linfotrópico de células T humana tipo 1 e 2 em pacientes infectados pelo vírus da imunodeficiência humana tipo 1 nas Cidades de Ribeirão Preto e São Paulo / Prevalence, risk factors and genetic characterization of human T-cell lymphotropic virus types 1 and 2 in patients infected with human immunodeficiency virus type 1 in the cities of Ribeirão Preto and São Paulo

O objetivo deste estudo foi definir a prevalência dos vírus linfotrópico de células T humana tipo 1 e 2 em pacientes positivos para o vírus da imunodeficiência humana tipo 1 no Estado de São Paulo, Brasil. Avaliamos 319 indivíduos atendidos em clínicas de Ribeirão Preto e Capital. Os pacientes foram entrevistados e testados sorologicamente. Foram seqüenciadas as regiões tax e long terminal repeat para diferenciação e determinação do subtipo. A soroprevalência geral foi de 7,5 por cento (24/319) e esteve associada somente com uso de drogas injetáveis e ao vírus da hepatite tipo C (p<0, 001). O genoma viral foi detectado em 13 das 24 amostras, sendo 12 caracterizadas como HTLV-2 subtipo 2c e uma como 1a. Nossos dados mostraram que o uso de drogas injetáveis é um importante fator de risco para a transmissão de HTLV-2 em populações infectadas pelo vírus da imunodeficiência humana tipo 1.

The aim of this study was to define the prevalence of human T cell lymphotropic virus types 1 and 2 in patients who were positive for human immunodeficiency virus type 1 in the State of São Paulo, Brazil. We evaluated 319 individuals infected with HIV type 1 who were attended at specialized clinics in two cities (Ribeirão Preto and São Paulo). The patients were interviewed and tested for antibodies against HTLV types 1 and 2 (Orthoâ HTLV-1/HTLV-2 Ab-Capture enzyme immunoassay). Direct DNA sequencing of polymerase chain reaction products from the tax region of HTLV type 2 and the long terminal repeat region of HTLV types 1 and 2 were performed to differentiate and determine the subtypes. The overall prevalence of anti-HTLV type 1 and 2 antibodies was 7.5 percent (24/319; 95 percent CI: 5.2-11.5). HTLV type 1 and 2 infection was associated with a history of injected drug use and with antibodies for hepatitis C virus (p < 0.001), but not with age (p = 0.2), sex (p = 0.9), sexual behavior or serological markers for sexually transmitted diseases (anti-Treponema pallidum, anti-human herpesvirus type 8 or anti-hepatitis B virus antibodies) (p > 0.05). HTLV DNA was detected in 13 out of 24 samples, of which 12 were characterized as HTLV subtype 2c and one as HTLV subtype 1a. Among the 12 HTLV type 2 samples, seven were from injected drug users, thus indicating that this route is an important risk factor for HTLV type 2 transmission among our population infected with HIV type 1.

Prevalence of HIV type 1 drug resistance mutations in treatment-naïve and experienced patients from resource-limited settings with universal access to antiretroviral therapy: a survey in two small Brazilian cities

Concerns have been raised that universal availability of antiretroviral agents in resource-limited settings might lead to the emergence and spread of resistant strains. We present the largest survey on human immunodeficiency virus type 1 (HIV-1) resistance among treatment-naïve and experienced patients followed in small, relatively underprivileged cities in Brazil with universal availability to standard of care antiretroviral combinations. Samples were collected between 2004 and 2006 from 95 patients followed in the cities of Saquarema and Santo Antonio de Pádua, state of Rio de Janeiro. A proviral fragment encompassing protease and reverse transcriptase (RT) regions was generated and drug susceptibility level was inferred. Among 50 strains from drug-naïve subjects, one (2 percent) had intermediate-level resistance to RT inhibitors. Among 38 patients on therapy as of sampling, 28 (73.7 percent) had plasma viral load (PVL) below detection limit (26 of whom without evidence of resistance mutations) and 11 (28.9 percent) harbored strains with reduced susceptibility. Only two strains harbored both protease and RT inhibitor mutations. Among seven patients who were off-treatment as of sampling, two (28.5 percent) harbored strains with reduced susceptibility to RT inhibitors. The relatively high frequency of undetectable PVL among patients on treatment and the overall low prevalence of resistance-associated mutations are reassuring. Continued surveillance, however, is necessary.

Enfermedad por citomegalovirus en niños chilenos infectados por el virus de la inmunodeficiencia humana-1 / Cytomegalovinas disease in HIV-1 infected Chilean children

Citomegalovirus (CMV) es una infección oportunista frecuente en niños infectados con el virus de la inmunodeficiencia humana tipo 1 (VIH 1) y produce altas tasas de morbilidad y mortalidad. El objetivo de este estudio fue determinar la frecuencia y trascendencia de la enfermedad por CMV en la cohorte de niños chilenos infectados con VIH-1. Se encontró enfermedad por CMV en 28 de 222 niños infectados con VIH-1 (12,6 por cientoo); 92 por cientoo fue clasificado en etapa C y 61 por cientoo en categoría 3 (CDC 1994). La enfermedad pulmonar fue la manifestación más común (25). Las muestras clínicas se obtuvieron de tracto respiratorio, sangre, orina y biopsia. Para la detección de CMV se utilizó, preferentemente, el cultivo acelerado (shell vial), (20/28). Todos fueron tratados con ganciclovir ev. El tiempo promedio de sobrevida fue 42 meses hasta el momento de esta evaluación. Conclusión: La enfermedad por CMV causó morbilidad y mortalidad en los niños chilenos infectados por VIH-1. El diagnóstico y tratamiento precoces son la base para un resultado exitoso.

Cytomegalovirus (CMV) is a frequent opportunistic infection in human inmunodeficiency virus type 1 (HIV-1) infected children associated with significant morbidity and mortality. The aim of this study was to determine the frequency and impact of CMV disease in a prospective ly followed cohort of HIV-1 infected Chilean children. CMV disease was diagnosed in 28 out of 222 HIV infected children (12.6 percent); 92 percent of them were classified in category C and 61 percent in category 3 (CDC, 1994). Lung disease was the most common manifestation (25 children). Samples were obtained from the respiratory tract, blood, urine and tissue biopsies. Shell vial for CMV early antigen detection was the most commonly used diagnostic technique (20/ 28). All patients were treated with iv.ganciclovir and two children died during the CMV episode. The mean survival time for the remaining children is currently 42 months. Conclusion: CMV disease was frequent and caused mortality in HIV-1 infected Chilean children. Early diagnosis and treatment are key for clinical success.

Antiretroviral resistance in individuals presenting therapeutic failure and subtypes of the human immunodeficiency virus type 1 in the Northeast Region of Brazil

This study aimed to analyze human immunodeficiency virus (HIV) mutation profiles related to antiretroviral resistance following therapeutic failure, and the distribution of hiv subtypes in the Northeast Region of Brazil. A total of 576 blood samples from AIDS patients presenting therapeutic failure between 2002 and 2004 were analyzed. The genotyping kit viroSeq® was used to perform viral amplification in order to identify mutations related to hiv pol gene resistance. An index of 91.1 percent of the patients presented mutations for nucleoside reverse transcriptase inhibitors (nrti), 58.7 percent for non-nucleoside reverse transcriptase inhibitors (nnrti), and 94.8 percent for protease inhibitors (pi). The most prevalent mutations were 184V and 215E for nrti, 103N and 190A for nnrti. Most mutations associated with PIs were secondary, but significant frequencies were observed in codons 90 (25.2 percent), 82 (21.1 percent), and 30 (16.2 percent). The resistance index to one class of antiretrovirals was 14 percent, to two classes of antiretrovirals 61 percent, and to three classes 18.9 percent. Subtype B was the most prevalent (82.4 percent) followed by subtype F (11.8 percent). The prevalence of mutations related to nrti and nnrti was the same in the two subtypes, but codon analysis related to PI showed a higher frequency of mutations in codon 63 in subtype B and in codon 36 in subtype F. The present study showed that there was a high frequency of primary mutations, which offered resistance to nrti and nnrti. Monitoring patients with treatment failure is an important tool for aiding physicians in rescue therapy.

Autochthonous horizontal transmission of a CRF02_AG strain revealed by a human immunodeficiency virus type 1 diversity survey in a small city in inner state of Rio de Janeiro, Southeast Brazil

As part of an ongoing study on the features of AIDS spread towards small cities and rural areas, we present a molecular survey of human immunodeficiency virus type 1 (HIV-1) polymerase sequences recovered between 2004 and 2006 from 71 patients receiving care in the city of Saquarema, inner state of Rio de Janeiro. Phylogenetic reconstructions found the two prevalent lineages in the state (subtypes B [59 strains, 83.1 percent], F1 [6 strains; 8.4 percent], and BF1 recombinants [four strains; 5.6 percent]), as well as two (2.8 percent) CRF02_AG strains, which seems to be an emerging lineage in the capital. These CRF02_AG sequences were recovered from a married heterosexual couple who never traveled abroad, thus providing the first molecular evidence of autochthonous horizontal transmission of this lineage of major global importance. Also, three phylogenetic clusters of strains recovered from a total of 18.3 percent of the cohort were uncovered. Their close genetic relatedness suggests they were recovered from patients who probably took part in the same chain of viral spread. In conjunction with our previous surveys from inner Rio de Janeiro, these results suggest that although small cities harbor unique molecular features of HIV-1 infection, they also clearly reflect and may rapidly absorb the diversity recorded in large urban centers.

Comparison of the epidemiology, profile of mutations, and clinical response to antiretrovirals among subtypes B and F of the human immunodeficiency virus type 1

The authors compared demographic aspects and profile of mutations in 80 patients with subtypes B and F of human immunodeficiency type 1 (HIV-1). Genotyping of the pol region of the reverse transcriptase was performed using the ViroSeqTM Genotyping System. A total of 61 (76.2 percent) patients had subtype B and 19 (23.8 percent) subtype F of the HIV-1. Subtype F tended to be more frequent in heterosexuals and women with a low educational level, but without statistical significance. The frequency of mutations related to nucleoside reverse transcriptase inhibitors and protease inhibitors (PI) was the same in the two subtypes, but mutations related to PI at the codons 63, 77, and 71 were more frequent in subtype B, while mutations at the codons 36 and 20 predominated in subtype F. Sixty-two of the 80 patients infected with subtypes B and F were submitted to antiretroviral therapy for an average of 18-22 months. Undetectable viral loads at the end of follow-up were similar in the two groups, representing 63.8 percent of subtype B and 73.3 percent of subtype F (p = 0.715). CD4 lymphocyte counts before and after treatment were similar in the two groups. This study, despite pointing to possible epidemiological and genetic differences among subtypes B and F of HIV-1, suggests that the use of highly active antiretroviral therapy is equally effective against these subtypes.

Molecular evidence that human immunodeficiency virus type 1 dissemination in a small Brazilian city was already taking place in the early 1990s

We recently performed a molecular epidemiology survey of human immunodeficiency virus type 1 (HIV-1) infection in Miracema, a small city in Southeast Brazil, and found multiple monophyletic clusters, consistent with independent introductions and spread of different viral lineages in the city. Here we apply Bayesian coalescent-based methods to the two largest subtype B clusters and estimate that the most recent common ancestors that gave rise to these two transmission chains were in circulation around 1991-1992. The finding that HIV-1 spread in this Brazilian small city was already taking place at a time Aids was considered a problem restricted to large urban centers may have important public health implications.