Development of a molecular platform for HTLV confirmatory diagnosis: importance of the internal amplification control

Finnancial Support: CTC/INCTC, FAPESP, FUNDHERP, FINEP

Influence of INF-gamma gene polymorphism in HTLV-1 proviral load

Financial Support FAPESP, CNPq, CTC/FUNDHERP and INCTC

Upregulation of hsa-miR-125b in HTLV-1 asymptomatic carriers and HTLV-1-associated myelopathy/tropical spastic paraparesis patients

The retrovirus human T lymphotropic virus type 1 (HTLV-1) promotes spastic paraparesis, adult T cell leukaemia and other diseases. Recently, some human microRNAs (miRNAs) have been described as important factors in host-virus interactions. This study compared miRNA expression in control individuals, asymptomatic HTLV-1 carriers and HTLV-1 associated myelopathy (HAM)/tropical spastic paraparesis patients. The proviral load and Tax protein expression were measured in order to characterize the patients. hsa-miR-125b expression was significantly higher in patients than in controls (p = 0.0285) or in the HAM group (p = 0.0312). Therefore, our findings suggest that miR-125b expression can be used to elucidate the mechanisms of viral replication and pathogenic processes.FAPESP [06/59388-4]FAPESPCNPqCNPqFUNDHERPFUNDHER

Avaliação sorológica e molecular do parvovírus B19 (B19V) em doadores de sangue da Fundação Hemocentro de Brasília, Brasil : foco em mulheres em idade fértil

O parvovírus B19 (B19V) pode ser transmitido por via respiratória, verticalmente - da mãe para o feto - e via transfusão de sangue e transplante de órgãos. A infecção por transfusão de sangue ou hemoderivados ocorre devido à resistência do B19V aos métodos de inativação viral. Nosso estudo avaliou a presença do ácido desoxirribonucleico (DNA) B19V e a prevalência de imunoglobulina da classe G (IgG) anti-B19V em mulheres em idade fértil, doadoras de sangue do Distrito Federal, Brasil. Nossos resultados demonstraram a ausência de DNA de B19V nesses doadores. No entanto, foi observada a soroprevalência de IgG anti-B19V em 60,7% dessa população. Este estudo fornece dados importantes da circulação do B19V no Centro-Oeste do Brasil.Parvovirus B19 (B19V) can be transmitted by the respiratory route, vertically - from the mother to the fetus - and via blood transfusion or organ transplantation. Infection by transfusion of blood or blood products occurs due to the resistance of B19V to viral inactivation methods. Our study evaluated the presence of B19V deoxyribonucleic acid (DNA) and the prevalence of anti-B19V class G immunoglobulin (IgG) in women of childbearing age blood donors of the Federal District, Brazil. Our results demonstrated the absence of B19V DNA in these blood donors. However, the seroprevalence for anti-B19V IgG was observed in 60.7% of this population. This study provides important data of B19V circulation in the Center-West of Brazil

Inhibition of Expression of HTLV-1 Structural Genes Mediated by Short Hairpin RNA In Vitro

Background: Human T-lymphotropic virus 1 (HTLV-1) is associated with the T-cell malignancy known as adult T-cell leukemia! lymphoma (ATLL) and with a disorder called HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). Currently, the treatment of these diseases is based on symptom relief. RNA interference (RNAi) technology has been described as an efficient mechanism for development of new therapeutic methods. Thus, the aim of this study was to evaluate the inhibition of HTLV-1 structural proteins using short hairpin RNAs (shRNAs) expressed by non-viral vectors. Materials and Methods: Reporter plasmids that express enhanced green fluorescent protein-Gag (EGFP-Gag) and EGFP-Env fusion proteins and vectors that express shRNAs corresponding to the HTLV-1 gag and env genes were constructed. shRNA vectors and reporter plasmids were simultaneously transfected into HEK 293 cells. Results: Fluorescence microscopy, flow cytometry and real-time PCR showed that shRNAs were effective in inhibiting the fusion proteins. Conclusion: These shRNAs are effective against the expression of structural genes and may provide an approach to the development of new therapeutic agents.Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Centro de Terapia Celular/Fundacao Hemocentro de Ribeirao Preto (CTC/FUNDHERP)Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq

Molecular and phylogenetic analyses of human Parvovirus B19 isolated from Brazilian patients with sickle cell disease and beta-thalassemia major and healthy blood donors

Human Parvovirus B19 (B19V) is a recognized cause of life-threatening conditions among patients with hemoglobinopathies. This study investigates B19V infection in patients with sickle cell disease and beta-thalassemia using different experimental approaches. A total of 183 individuals (144 with sickle cell disease and 39 with beta-thalassemia major) and 100 healthy blood donors were examined for B19V using anti-B19V IgG enzyme immunoassay, quantitative PCR, DNA sequencing, and phylogenetic analysis. Viremia was documented in 18.6% of patients and 1% of donors, and was generally characterized by low viral load (VL); however, acute infections were also observed. Anti-B19V IgG was detected in 65.9% of patients with sickle cell disease and in 60% of donors, whereas the patients with thalassemia exhibited relatively low seroreactivity. The seroprevalence varied among the different age groups. In patients, it progressively increased with age, whereas in donors it reached a plateau. Based on partial NS1 fragments, all isolates detected were classified as subgenotype 1A with a tendency to elicit genetically complex infections. Interestingly, quasispecies occurred in the plasma of not only patients but also donors with even higher heterogeneity. The partial NS1 sequence examined did not exhibit positive selection. Quantitation of B19V with a conservative probe is a technically and practically useful approach. The extensive spread of B19V subgenotype 1A in patients and donors and its recent introduction into the countryside of the Sao Paulo State, Brazil were demonstrated; however, it is difficult to establish a relationship between viral sequences and the clinical outcomes of the infection. J. Med. Virol. 84:16521665, 2012. (c) 2012 Wiley Periodicals, Inc.Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)Instituto Nacional de Ciencia e Tecnologia em Celulas Tronco e Terapia Celular (INCTC)Instituto Nacional de Ciencia e Tecnologia em Celulas Tronco e Terapia Celular (INCTC