Evaluation of HIV-1 antiretroviral drug resistance profiles in the peripheral blood reservoir of successfully treated persons using massive sequencing and viral full genome characterization.

BACKGROUND: Antiretroviral therapy has revolutionized HIV treatment, increasing quality and life expectancy of people living with HIV (PLWH). However, the expansion of treatment has resulted in an increase in antiretroviral-resistant viruses, which can be an obstacle to maintenance of successful ART. OBJECTIVES: This study analysed the genetic composition of the HIV near full-length genome (NFLG) from archived proviruses of PLWH under successful ART, and determined the presence/frequency of drug resistance mutations (DRMs) and viral subtype. PATIENTS AND METHODS: Forty-six PLWH from Rio de Janeiro (RJ) and 40 from Rio Grande (RS) had proviral HIV NFLG PCR-amplified and ultradeep sequenced. The presence/frequency of DRMs were analysed in Geneious. Phylogenetic analyses were performed using PhyML and SimPlot. RESULTS: All samples included in the study were sequenced and 69 (80.2%) had the HIV NFLG determined. RJ and RS showed a predominance of HIV subtypes B (78.3%) and C (67.5%), respectively. Overall, 168 DRMs were found in 63 (73.3%) samples, and 105 (62.5%) of them were minority variants. Among DRMs, 41 (39.0%) minority variants and 33 (52.4%) variants with frequency above 20.0% in the viral population were able to confer some degree of resistance to at least one drug in use by respective patients, yet no one showed signs of therapeutic failure. CONCLUSIONS: Our study contributes to the understanding of the impact of DRMs on successful therapy and supports the sustainability of combinatorial ART, because all patients maintained their successful treatment despite the high prevalence of DRMs at low (62.5%) or high (37.5%) frequency.

High prevalence and autochtonous transmission of human pegivirus (HPgV-1) in blood donors in the extreme southern of Brazil.

Recent studies have suggested that human pegivirus 1 (HPgV-1) may have some pathogenic potential. In the southernmost region of Brazil, studies on HPgV-1 are scarce, and circulating genotypes have not yet been identified. The current study aimed to evaluate the prevalence of HPgV-1 among blood donors from the southernmost region of Brazil and identify the genotypes involved with associated factors. A cross-sectional study was conducted with 281 blood donors, who had their plasma subjected to RNA extraction, complementary DNA synthesis, HPgV-1 detection by nested polymerase chain reaction, and subsequent genotyping. The observed prevalence of HPgV-1-RNA was 21.7%. The only variable that was significantly associated with virus infection was the relationship status of the donor. Single or no fixed partner blood donors were twice as likely to have HPgV-1 (95% CI, 1.12 to 4.56; P = 0.02). Genotype 2-subtypes 2b (69%) and 2a (29%)-was the most prevalent. In the absence of risk factors for parenteral transmission, it is likely that sexual transmission was the route of infection in the individuals studied. Further work will be needed to determine whether this virus is inert in the population, or if there are potential deleterious effects in infected individuals.

Human papillomavirus type distribution and HPV16 intratype diversity in southern Brazil in women with and without cervical lesions.

BACKGROUND: Increasing evidence suggests that human papillomavirus (HPV) intratype variants (specific lineages and sublineages) are associated with pathogenesis and progression from HPV infection to persistence and the development of cervical cancer. OBJECTIVES: This study aimed to verify the prevalence of HPV infection and distribution of HPV types and HPV16 variants in southern Brazil in women with normal cytology or intraepithelial lesions. METHODS: HPV typing was determined by L1 gene sequencing. To identify HPV16 variants, the LCR and E6 regions were sequenced, and characteristic single nucleotide variants were identified. FINDINGS: A total of 445 samples were studied, with 355 from cervical scrapes and 90 from cervical biopsies. HPV was detected in 24% and 91% of these samples, respectively. The most prevalent HPV types observed were 16 (cervical, 24%; biopsies, 57%) and 58 (cervical, 12%; biopsies, 12%). Seventy-five percent of the HPV16-positive samples were classified into lineages, with 88% defined as lineage A, 10% as lineage D, and 2% as lineage B. MAIN CONCLUSIONS: This study identified a high frequency of European and North American HPV16 lineages, consistent with the genetic background of the human population in southern Brazil.

Prevalence of human pegivirus (HPgV) infection in patients carrying HIV-1C or non-C in southern Brazil.

Previous studies have demonstrated that coinfection with HPgV is a protective factor for human immunodeficiency virus (HIV)-infected patients, leading to slower disease progression, and longer survival after established disease. The present study sought to estimate the prevalence of HPgV infection and associated risk factors in patients harboring C or non-C HIV-1 subtypes followed-up at HU-FURG, southern Brazil. Samples from 347 HIV-1-infected subjects were subjected to plasma RNA extraction, cDNA synthesis, HPgV RNA detection, and HIV-1 genotyping. The overall prevalence of HPgV RNA was 34%. Individuals aged 18-30 years had higher chances of infection compared with those 50 years or older (95%CI 1.18-52.36, P = 0.03). The number of sexual partner between one and three was a risk factor for HPgV infection (95%CI 1.54-10.23; P < 0.01), as well as the time since diagnosis of HIV-1 ≥ 11 years (95%CI 1.01-2.89; P = 0.04). Patients infected with HIV non-C subtypes had six times more chance of being HPgV-infected when compared to subtype C-infected subjects (95%CI 2.28-14.78; P < 0.01). This was the first study conducted in southern Brazil to find the circulation of HPgV. HIV/HPgV coinfection was associated with a longer survival among HIV+ patients. Of novelty, individuals infected by HIV non-C subtypes were more susceptible to HPgV infection. However, additional studies are needed to correlate the HIV-1 subtypes with HPgV infection and to clarify cellular and molecular pathways through which such associations are ruled. J. Med. Virol 88:2106-2114, 2016. © 2016 Wiley Periodicals, Inc.

Estimating HIV-1 Genetic Diversity in Brazil Through Next-Generation Sequencing.

Approximately 36.7 million people were living with the human immunodeficiency virus (HIV) at the end of 2016 according to UNAIDS, representing a global prevalence rate of 0.8%. In Brazil, an HIV prevalence of 0.24% has been estimated, which represents approximately 830,000 individuals living with the virus. As a touristic and commercial hub in Latin America, Brazil harbors an elevated HIV genetic variability, further contributed by the selective pressure exerted by the host immune system and by antiretroviral treatment. Through the progress of the next-generation sequencing (NGS) techniques, it has been possible to expand the study of HIV genetic diversity, evolutionary, and epidemic processes, allowing the generation of HIV complete or near full-length genomes (NFLG) and improving the characterization of intra- and interhost diversity of viral populations. Greater sensitivity in the detection of viral recombinant forms represents one of the major improvements associated with this development. It is possible to identify unique or circulating recombinant forms using the near full-length viral genomes with increasing accuracy. It also permits the characterization of multiple viral infections within individual hosts. Previous Brazilian studies using NGS to analyze HIV diversity were able to identify several distinct unique and circulating recombinant forms and evidenced dual infections. These data unveiled unprecedented high rates of viral recombination and highlighted that novel recombinants are continually arising in the Brazilian epidemic. In the pooled analysis depicted in this report, HIV subtypes have been determined from HIV-positive patients in five states of Brazil with some of the highest HIV prevalence, three in the Southeast (Rio de Janeiro, São Paulo, and Minas Gerais), one in the Northeast (Pernambuco) and one in the South (Rio Grande do Sul). Combined data analysis showed a significant prevalence of recombinant forms (29%; 101/350), and a similar 26% when only NFLGs were considered. Moreover, the analysis was able to evidence the occurrence of multiple infections in some individuals. Our data highlight the great HIV genetic diversity found in Brazil and unveils a more accurate scenario of the HIV evolutionary dynamics in the region.

Prevalence of herpes simplex virus types 1 and 2 at maternal and fetal sides of the placenta in asymptomatic pregnant women.

PROBLEM: Herpes simplex virus 1 (HSV-1) commonly causes orolabial infection, but can also infect the genital mucosa. In contrast, HSV-2 is usually genital. Genital herpes can transmit the virus vertically to the fetus during pregnancy. We sought to estimate the prevalence of HSV-1/2 on the maternal and fetal sides of the placenta. METHOD OF STUDY: Placental tissues were collected from pregnant women seen at the Rio Grande University Hospital. HSV-1 and HSV-2 were detected by nested PCR. RESULTS: The prevalence of HSV-1 and HSV-2 was, respectively, 28% and 12.6% (maternal side) and 29.9% and 8.3% (fetal side). All HSV-positive women were asymptomatic. Sexual behavior, vaginal delivery, and presence of HSVs on one side of the placenta were risk factors associated with HSV infection. CONCLUSION: The occurrence of HSVs in placental tissue was high, especially for HSV-1. Novel strategies need to be implemented for the management of asymptomatic women who might transmit HSV to their newborns.

Prevalence and vertical transmission of human pegivirus among pregnant women infected with HIV.

OBJECTIVE: To determine the prevalence of human pegivirus (HPgV) and factors associated with vertical transmission among pregnant women infected with HIV. METHOD: A retrospective cross-sectional study was conducted among pregnant women treated at an HIV reference service in Rio Grande, Brazil, between January 1, 2010, and January 1, 2015. The polymerase chain reaction was used to diagnose HPgV infection among the women and their neonates. Clinical, obstetric, and neonatal data were obtained from medical records. RESULTS: Infection with HPgV was detected among 16 (25%) of 63 women and 5 (8%) of 63 newborns, corresponding to a vertical transmission rate of 31%. Multivariate analysis demonstrated that the absence of prenatal care was the only risk factor for vertical transmission of HPgV (prevalence ratio 19.61, 95% confidence interval 1.29-297.48; P=0.032). CONCLUSION: Prenatal care could protect against vertical transmission of HPgV among women infected with HIV; however, studies among HIV-negative individuals are still required to verify this correlation.

Human papillomavirus type distribution and HPV16 intratype diversity in southern Brazil in women with and without cervical lesions

BACKGROUND Increasing evidence suggests that human papillomavirus (HPV) intratype variants (specific lineages and sublineages) are associated with pathogenesis and progression from HPV infection to persistence and the development of cervical cancer. OBJECTIVES This study aimed to verify the prevalence of HPV infection and distribution of HPV types and HPV16 variants in southern Brazil in women with normal cytology or intraepithelial lesions. METHODS HPV typing was determined by L1 gene sequencing. To identify HPV16 variants, the LCR and E6 regions were sequenced, and characteristic single nucleotide variants were identified. FINDINGS A total of 445 samples were studied, with 355 from cervical scrapes and 90 from cervical biopsies. HPV was detected in 24% and 91% of these samples, respectively. The most prevalent HPV types observed were 16 (cervical, 24%; biopsies, 57%) and 58 (cervical, 12%; biopsies, 12%). Seventy-five percent of the HPV16-positive samples were classified into lineages, with 88% defined as lineage A, 10% as lineage D, and 2% as lineage B. MAIN CONCLUSIONS This study identified a high frequency of European and North American HPV16 lineages, consistent with the genetic background of the human population in southern Brazil.

Non-toxic derivatives of LT as potent adjuvants.

The heat-labile enterotoxin of Escherichia coli (LT) consists of an enzymatically active A subunit (LTA) and a pentameric B subunit (LTB). LT has been extensively studied as a potent modulator of immune responses but wild-type LT is toxic and therefore unsuitable for clinical use. Approaches pursued to avoid the toxicity associated with the use of the native toxin while retaining its adjuvant properties have included isolation of subunit B (LTB) and construction of non-toxic LT AB complex mutants, such as LTK63 mutant. Here we review the immunomodulatory characteristics of LTB and LTK63 and their potential as mucosal and parenteral vaccine adjuvants.