A New Flow Cytometry-Based Single Platform for Universal and Differential Serodiagnosis of HTLV-1/2 Infection.

In the present work, we developed and evaluated the performance of a new flow cytometry-based single platform, referred to as "FC-Duplex IgG1 (HTLV-1/2)", for universal and differential serodiagnosis of HTLV-1/2 infection. The proposed technology employs a system for detection of IgG1 antibodies in a single competitive immunofluorescence platform by flow cytometry using fluorescently labeled MT-2/MoT cell line mix coupled to a highly sensitive development system (Biotin/Streptavidin/Phycoerythrin). The stability of fluorescent labeling and the antigenicity of MT-2 and MoT cell lines were confirmed upon storage at -20°C for 2, 6, and 12 months. The anti-HTLV-1/2 IgG1 reactivity, expressed as percentage of positive fluorescent cells (PPFC), was evaluated for each target antigen along the titration curve of test serum samples (1:32 to 1:4,096). Upon selection of target cell line and serum dilutions with higher segregation score between groups, the performance of "FIX" and "FIX & PERM" protocols was evaluated. The "FIX" protocol presented excellent performance indices (Se = 92%/Sp = 94%/AUC = 0.96; Se = 96%/Sp = 100%/AUC = 0.99) for the universal (HTLV-1/2 vs. NI) and differential (HTLV-1 vs. HTLV-2) diagnosis of HTLV-1 infection, respectively. Optimization of the "FIX" protocol using the principle of synchronous and asynchronous pairwise analysis further improved the performance of "FC-Duplex IgG1 (HTLV-1/2)", using the "FIX" protocol for differential diagnosis of HTLV-1 and HTLV-2 infections (Se = 100%/Sp = 100%/AUC = 1.00). In conclusion, the "FC-Duplex IgG1 (HTLV-1/2)" method represents an innovation in the biotechnology segment with the potential to compose a serological kit for differential diagnosis of HTLV-1/2 infection for reference laboratories and blood centers.

Impact of HIV co-infection on immunological biomarker profile of HTLV-1 infected patients.

The impact of HIV co-infection on the plasma immunological biomarker profile of HTLV-1 infected patients was evaluated. The plasma levels of leukotrienes and chemokines/cytokines were quantified by ELISA and Cytometric Bead Array. A total of 138 volunteers were enrolled and divided into two subgroups ("HTLV-1(+)HIV(-)" and "HTLV-1(+)(HIV(+)"), which were categorized according to the HTLV-1-associated neurological disease (AS, pHAM and HAM). Reference controls were BD and HIV mono-infected patients. HAM(+) exhibited higher CD4+ T-cell counts as compared to HIV+ mono-infected patients and lower HTLV-1 proviral load as compared to mono-infected HAM(-) patients. AS(+) exhibited higher levels of CysLT, CXCL8/IL-8 and lower levels of CCL5/RANTES as compared to AS(-). Increased levels of IL-6 and TNF with reduced levels of CXCL10/IP10 and CCL5/RANTES were observed in co-infected pHAM(+) as compared to mono-infected pHAM(-). HAM(+) patients revealed an increase in CXCL8/IL-8, CCL2/MCP-1, CXCL-10/IP-10, TNF and a decrease in IL-2 as compared to HAM(-) subgroup.

Geographic distribution of human T-lymphotropic virus types 1 and 2 among mothers of newborns tested during neonatal screening, Minas Gerais, Brazil.

OBJECTIVE: To evaluate the geographic distribution of human T-lymphotropic virus types 1 and 2 (HTLV-1/2) in the State of Minas Gerais, Brazil, in puerperal women whose newborns were tested for HTLV-1/2 during neonatal screening, and to overlap seropositivity with social and economic status determinants. METHODS: During September-November 2007, the dry-blood samples taken from newborns on filter paper for routine screening were also tested for maternal IgG anti-HTLV-1/2 antibodies. For reactive samples, the mothers of the newborns had blood drawn to test for these viruses. RESULTS: The study analyzed 55,293 specimens taken from newborns. Of these, 52 (9.4 per 10,000) were reactive and 42 mothers (7.6 per 10,000) were confirmed with HTLV-1/2 infection. HTLV-1/2 geographic distribution was heterogeneous, with a tendency to be higher in the North and North-East parts of Minas Gerais. The highest rates of seropositivity were observed in Vale do Mucuri (55.9 per 10,000) and in Jequitinhonha (16.0 per 10,000), overlapping with the State's worst social and economic indicators. CONCLUSIONS: To our knowledge this was the first time that neonatal screening for HTLV-1/2 was performed in Brazil. This model could be used in other areas with high HTLV-1/2 prevalence rates. The detection of carrier mothers can enable intervention measures, such as providing infant formula to newborns, to be implemented expeditiously to reduce vertical transmission.

Use of an automated pyrosequencing technique for confirmation of sickle cell disease.

BACKGROUND: The diagnosis of sickle cell disease (SCD) is made by hemoglobin assays such as high-performance liquid chromatography (HPLC), isoelectric focusing and cellulose acetate or citrate agar electrophoresis. These assays are easy to perform and used in large-scale newborn screening in many countries. These tests however may not easily differentiate Sß0 thalassemia from SS or identify other hemoglobin variants, and in this case, hemoglobin (HBB) gene sequencing may be necessary. OBJECTIVES: To develop a high throughput DNA based confirmatory assay for SCD and to detect mutations in the HBB gene. METHODS: We developed an automated pyrosequencing technique (PyS) based on QIAGEN technology (Hilden, Germany) to detect homozygous or heterozygous hemoglobin S mutations as well as hemoglobin C mutations. The technique was tested on 2,748 samples from patients enrolled in a multi-center SCD cohort in Brazil. Patients were previously tested using HPLC to diagnose SCD as part of routine clinical care. Any subjects with discrepant results between HPLC and PyS or with heterozygous hemoglobin S detected had Sanger sequencing of the HBB gene. RESULTS: We identified 168 samples with discrepant results between HPLC and PyS and 100 with concordant PyS = heterozygous S and HPLC, which would suggest SB-thalassemia or other heterozygous S variants. The PyS assay correctly identified 1906 (98.7%) of the 1930 HbSS and 628 (98.7%) of the 636 HbSC samples. Of the 179 remaining samples, PyS correctly indicated S heterozygosis in 165 (92.2%). Of the 165 heterozygous S samples confirmed by Sanger as consistent with Sß thalassemia genotype, 84 samples were classified as Sß0 thalassemia and 81 as Sß+ thalassemia. The most frequent beta thalassemia mutations of Sß0 and Sß+ were HBB: c.118C>T (Gln40Stop) and HBB c.92 + 6T> C, respectively. DISCUSSION: The PyS proved to be satisfactory for large-scale confirmatory testing of hemoglobin mutation. Moreover, with this study we were able to describe the most common ß+ and ß0 mutations in SCD patients with Sß-thalassemia in a large multi-institutional SCD cohort in Brazil.

Vestibular-evoked myogenic potential triggered by galvanic vestibular stimulation may reveal subclinical alterations in human T-cell lymphotropic virus type 1-associated myelopathy.

BACKGROUND: Vestibular-evoked myogenic potential triggered by galvanic vestibular stimulation (galvanic-VEMP) evaluates the motor spinal cord and identifies subclinical myelopathies. We used galvanic-VEMP to compare spinal cord function in individuals infected with human T-cell lymphotropic virus type 1 (HTLV-1) from asymptomatic status to HTLV-1-associated myelopathy (HAM). METHODOLOGY/PRINCIPAL FINDINGS: This cross-sectional study with 122 individuals included 26 HTLV-1-asymptomatic carriers, 26 individuals with possible HAM, 25 individuals with HAM, and 45 HTLV-1-seronegative individuals (controls). The groups were similar regarding gender, age, and height. Galvanic stimuli (duration: 400 ms; intensity: 2 mA) were applied bilaterally to the mastoid processes and VEMP was recorded from the gastrocnemius muscle. The electromyographic parameters investigated were the latency and amplitude of the short-latency (SL) and medium-latency (ML) responses. While SL and ML amplitudes were similar between groups, SL and ML latencies were delayed in the HTLV-1 groups compared to the control group (p<0.001). Using neurological examination as the gold standard, ROC curve showed an area under the curve of 0.83 (p<0.001) for SL and 0.86 (p<0.001) for ML to detect spinal cord injury. Sensibility and specificity were, respectively, 76% and 86% for SL and 79% and 85% for ML. Galvanic-VEMP disclosed alterations that were progressive in HTLV-1-neurological disease, ranging from SL delayed latency in HTLV-1-asymptomatic carriers, SL and ML delayed latency in possible HAM group, to absence of VEMP response in HAM group. CONCLUSIONS/SIGNIFICANCE: The worse the galvanic-VEMP response, the more severe the myelopathy. Galvanic-VEMP alteration followed a pattern of alteration and may be a prognostic marker of progression from HTLV-1-asymptomatic carrier to HAM.

[Evaluation of the performance of immunological parameters as indicators for clinical progression of chronic HTLV-1 infection]. / Avaliação do desempenho de parâmetros imunológicos como indicadores de progressão clínica da infecção crônica pelo HTLV-1.

This study evaluated the performance of single and combined laboratory parameters, B-lymphocyte percentages (%LB), T/B cell ratio and %CD8+HLA-DR+/CD8+, to differentiate asymptomatic cases (AS) from HAM/TSP patients (HT) within a population of HTLV-1 seropositive cases. Percentage indices demonstrated that each parameter alone presented moderate performance, with co-negativity of 83 and 91% for %LB and T/B cell ratio, respectively, and co-positivity of 78% for %CD8+HLA-DR+/CD8+. Combined analysis (%CD8+HLA-DR+/CD8+ and T/B cell ratio) did not show any substantial performance enhancement (co-positivity = 75% and co-negativity = 74%). Likelihood ratio analysis using different value ranges for the separate parameters revealed that HTLV-1 seropositive cases with %LB<7%, T/B cell ratio>11 and %CD8+HLA-DR+/CD8+>70% would have, respectively, 11, 19 and 10 times greater chance of belonging to the HT group. Therefore, use of these phenotypic indicators as complementary laboratory methods for monitoring the clinical progression of chronic HTLV-1 infection is recommended.

Reducing the global burden of HTLV-1 infection: An agenda for research and action.

Even though an estimated 10-20 million people worldwide are infected with the oncogenic retrovirus, human T-lymphotropic virus type 1 (HTLV-1), its epidemiology is poorly understood, and little effort has been made to reduce its prevalence. In response to this situation, the Global Virus Network launched a taskforce in 2014 to develop new methods of prevention and treatment of HTLV-1 infection and promote basic research. HTLV-1 is the etiological agent of two life-threatening diseases, adult T-cell leukemia and HTLV-associated myelopathy/tropical spastic paraparesis, for which no effective therapy is currently available. Although the modes of transmission of HTLV-1 resemble those of the more familiar HIV-1, routine diagnostic methods are generally unavailable to support the prevention of new infections. In the present article, the Taskforce proposes a series of actions to expand epidemiological studies; increase research on mechanisms of HTLV-1 persistence, replication and pathogenesis; discover effective treatments; and develop prophylactic and therapeutic vaccines.

Identification of a Novel HIV-1 Circulating Recombinant Form (CRF72_BF1) in Deep Sequencing Data from Blood Donors in Southeastern Brazil.

We report the identification of a novel HIV-1 circulating recombinant form (CRF72_BF1) in deep sequencing data from peripheral blood mononuclear cells (PBMC) of five blood donors in southeastern Brazil. Detection of this circulating recombinant form (CRF) confirms the need for effective surveillance to monitor the prevalence and distribution of HIV variants in a variety of settings in Brazil.

Incidence of human T cell lymphotropic virus type 1-associated myelopathy/tropical spastic paraparesis in a long-term prospective cohort study of initially asymptomatic individuals in Brazil.

The incidence of human T cell lymphotropic virus type 1 (HLTV-1)-associated myelopathy/tropical spastic paraparesis (HAM/TSP) is not well defined in the literature. Several studies have reported different incidence rates, and recent publications suggest a higher incidence and prevalence of HAM/TSP. The interdisciplinary HTLV Research Group (GIPH) is a prospective open cohort study of individuals infected with HTLV-1/2. This study describes the demographic data and HAM/TSP incidence rate observed in 181 HTLV-1-seropositive individuals and compares the results with previous reports in the literature. HAM/TSP was diagnosed on the basis of the World Health Organization diagnostic criteria and De Castro-Costa et al. [Proposal for diagnostic criteria of tropical spastic paraparesis/HTLV-I-associated myelopathy (TSP/HAM). AIDS Res Hum Retroviruses 2006;22:931-935]. Seven HAM/TSP incident cases were observed during the follow-up. The HAM/TSP incidence density was 5.3 cases per 1,000 HTLV-1-seropositive cases per year (95% confidence interval: 2.6-10.9), with a mean follow-up of 7±4 years (range: 1 month to 15 years). HAM/TSP was more frequent in women in their 40s and 50s with probable infection via the sexual route. The HAM/TSP incidence density among HTLV-1-seropositive cases observed in the present study is higher than that in previous studies. HAM/TSP may be underdiagnosed in countries like Brazil where HTLV infection is prevalent. Orientation and prevent transmission of HTLV programs are needed. Currently, preventing HTLV-1 transmission is the most effective way to reduce the impact of HAM/TSP on society.

Strong correlation between tax and HBZ mRNA expression in HAM/TSP patients: distinct markers for the neurologic disease.

BACKGROUND: HTLV-1 proviral load is a risk marker for HAM/TSP, but it is insufficient to determine the disease outcome. HTLV-1 Tax and HBZ proteins have been implicated in HAM/TSP pathogenesis in inducing cell proliferation and cytotoxic T lymphocytes response. OBJECTIVES: To quantify the expression of tax and HBZ mRNA in asymptomatic carriers (AC) and HAM patients, and to investigate their association with HAM/TSP. STUDY DESIGN: We quantified the expression of HTLV-1 tax and HBZ mRNA in 37 AC and 26 HAM patients classified according to proviral load as low (AC(L) and HAM(L): <1% infected cells) or high (AC(H) and HAM(H): >1%). RESULTS: The AC(L) subgroup showed the lowest frequency of individuals expressing tax mRNA in comparison with AC(H), HAM(L) and HAM(H), and tax mRNA load normalized by proviral load was significantly lower in the AC(L). In turn, normalized HBZ mRNA expression was similar in all subgroups. Both tax and HBZ mRNA expression were moderately correlated with proviral load in AC (r=0.6, p<0.001) and were weaker in HAM (r=0.4, p<0.05). In contrast, the correlation between tax and HBZ mRNA load was moderate in AC (r=0.5, p=0.001) and was much stronger in HAM (r=0.8, p<0.001). In addition, HBZ mRNA load, but not tax, was significantly associated with motor disability in HAM patients (p=0.036). CONCLUSIONS: The expression of tax mRNA seems to be best to estimate the risk of HAM/TSP, whereas HBZ mRNA appears to be a surrogate marker to disease progression, indicating that they have important but distinct roles in HAM/TSP pathogenesis.

Immunological profile of HTLV-1-infected patients associated with infectious or autoimmune dermatological disorders.

In the present study, the frequency, the activation and the cytokine and chemokine profile of HTLV-1 carriers with or without dermatological lesions were thoroughly described and compared. The results indicated that HTLV-1-infected patients with dermatological lesions have distinct frequency and activation status when compared to asymptomatic carriers. Alterations in the CD4(+)HLA-DR(+), CD8(+) T cell, macrophage-like and NKT subsets as well as in the serum chemokines CCL5, CXCL8, CXCL9 and CXCL10 were observed in the HTLV-1-infected group with skin lesions. Additionally, HTLV-1 carriers with dermatological skin lesions showed more frequently high proviral load as compared to asymptomatic carriers. The elevated proviral load in HTLV-1 patients with infectious skin lesions correlated significantly with TNF-α/IL-10 ratio, while the same significant correlation was found for the IL-12/IL-10 ratio and the high proviral load in HTLV-1-infected patients with autoimmune skin lesions. All in all, these results suggest a distinct and unique immunological profile in the peripheral blood of HTLV-1-infected patients with skin disorders, and the different nature of skin lesion observed in these patients may be an outcome of a distinct unbalance of the systemic inflammatory response upon HTLV-1 infection.

Blocking vertical transmission of human T cell lymphotropic virus type 1 and 2 through breastfeeding interruption.

BACKGROUND: Human T cell lymphotropic virus type 1 and 2 (HTLV-1/2) causes serious diseases and is endemic in many parts of the world. It is transmitted from mother to child in 15-25% of the cases, primarily through breastfeeding. Proviral load and duration of breastfeeding are thought to play a role in transmission. This study aimed to detect HTLV-seropositive mothers through testing of neonates, to evaluate maternal HTLV proviral load and to measure the rates of transmission blocking when interruption of breastfeeding was implemented. METHODS: Neonates were screened for HTLV-1/2 IgG by enzyme immunoassay using the neonatal screening program of Minas Gerais State, Brazil. Breastfeeding interruption was recommended to those whose mothers were confirmed HTLV-positive. Children were tested by polymerase chain reaction at birth and at 12 months of age. RESULTS: Of 55,293 neonates tested, 42 (0.076%) were positive for HTLV-1 or HTLV-2 IgG. All 42 were polymerase chain reaction-negative at birth and 1 of 37 (2.7%) became antibody-positive after 12 months. His mother had delivered him vaginally and was informed of the positive HTLV-1 polymerase chain reaction after 7 days of breastfeeding. The mother's proviral load was 271 copies/10,000 cells, whereas the average is 109.2 copies/10,000 cells (95% confidence interval: 70.56-147.83). CONCLUSIONS: Maternal HTLV-1 proviral load and the route of delivery may have played a role in the transmission observed. Avoidance of breastfeeding was an effective measure to reduce HTLV transmission. In endemic countries, routine prenatal or neonatal screening combined with formula feeding for mothers confirmed HTLV-positive may be an important strategy to prevent future development of illnesses related to HTLV.

High prevalence of HTLV-1 and 2 viruses in pregnant women in São Luis, state of Maranhão, Brazil.

INTRODUCTION: Human T cell lymphotropic virus type 1 (HTLV-1) is endemic in the Caribbean, Japan, South America and regions of Africa. HTLV-2 is present in Native American populations and associated with IV drug use in Europe and North America. In Brazil, it is estimated that 1.5 million people are infected with HTLV-1/2. The study objective was to determine HTLV-1/2 prevalence in pregnant women in the prenatal care from three public services in São Luis, State of Maranhão, Brazil, and to counsel seropositive women to reduce viral transmission. METHODS: A cross-sectional study was conducted from February to December 2008; women with age of 18 to 45 years, with low risk for sexually transmitted disease (STD) were invited to participate. Blood samples were collected in filter paper, and HTLV-1/2 immunoenzymatic test (ELISA) was performed as a screening test. Women with reactive results were submitted to peripheral venous blood collection for ELISA repetition, followed by Western blot (WB) and real-time PCR to confirm and discriminate the infection between virus types 1 and 2. RESULTS: Of the 2,044 women tested, seven (0.3%) were ELISA reactive and confirmed positive (four were HTLV-1, and three were HTLV-2). All positive women were oriented not to breastfeed their newborns. CONCLUSIONS: This study showed that the virus is present in high prevalence in that population. Further studies covering other segments of the population are necessary to better characterize the presence of HTLV-1/2 in Maranhão and to elicit measures to prevent its spread.

Decline in prevalence and asymmetric distribution of human T cell lymphotropic virus 1 and 2 in blood donors, State of Minas Gerais, Brazil, 1993 to 2007.

INTRODUCTION: Human T cell lymphotropic virus types 1 and 2 (HTLV-1/2) are endemic in Brazil and are screened for in transfusion services since 1993. This study evaluated the evolution of the prevalence of HTLV-1 and 2 in blood donors of the Hemominas Foundation from 1993 to 2007, and its geographical distribution in State of Minas Gerais, Brazil. METHODS: The Hemominas Foundation is a centralized blood center in Minas Gerais, Brazil. The sources of data were the Hemominas Foundation Technical Bulletin and files from the centralized serological laboratory. Donors were tested in the period using enzyme linked immuno sorbent assays (ELISA), followed by Western blot, when repeatedly reactive. The data were analyzed by EPIINFO 6.2 and TABWIN 3.5 softwares. RESULTS: The average seroprevalence in the period 1993-2007 was 0.1%. A steady decline occurred from 0.4% in 1993 to below 0.1% in 2002 and later, with a transient peak of 0.5% in 1994. HTLV reactivity distribution was asymmetrical in the state, with regions of higher prevalence, interspersed with low prevalence areas. Comparison of positive and negative donors verified that increasing age was proportional to virus positivity. Odds ratio for age ranged from 1.43 (30 to 39 years-old) to 3.09 (50 to 65 years-old). Women had a greater chance of being positive (OR-1.64), as previously described. CONCLUSIONS: Possible explanations for HTLV-1/2 prevalence decline are the exclusion of positive donors from the donor pool, an increase in repeat donors and ELISA test improvement, with reduction in the number of false positive results.

HTLV-1 associated myelopathy diagnosed during lepromatous leprosy reaction treatment: a case report.

Leprosy and human T cell lymphotropic virus type 1 infection are prevalent in Brazil. Coinfection by Mycobacterium leprae and HTLV-1 is reviewed and a case is reported. A 59 year-old woman was followed and HTLV-1 associated myelopathy was diagnosed during leprosy treatment. The clinical and neurological aspects of this unusual association were initially reviewed. Immunological markers and the possible prognoses due to the association of the diseases were discussed. The unexpected association of leprosy and HTLV-1 associated myelopathy may occur in endemic areas and causes difficulties in determining the correct diagnosis and adequate management of the neurological manifestations.

Evaluation of the use of real-time PCR for human T cell lymphotropic virus 1 and 2 as a confirmatory test in screening for blood donors.

INTRODUCTION: HTLV-1/2 screening among blood donors commonly utilizes an enzyme-linked immunosorbent assay (EIA), followed by a confirmatory method such as Western blot (WB) if the EIA is positive. However, this algorithm yields a high rate of inconclusive results, and is expensive. METHODS: Two qualitative real-time PCR assays were developed to detect HTLV-1 and 2, and a total of 318 samples were tested (152 blood donors, 108 asymptomatic carriers, 26 HAM/TSP patients and 30 seronegative individuals). RESULTS: The sensitivity and specificity of PCR in comparison with WB results were 99.4% and 98.5%, respectively. PCR tests were more efficient for identifying the virus type, detecting HTLV-2 infection and defining inconclusive cases. CONCLUSIONS: Because real-time PCR is sensitive and practical and costs much less than WB, this technique can be used as a confirmatory test for HTLV in blood banks, as a replacement for WB.

Hepatitis C: geographical diversity and temporal trends of seropositivity in brazilian blood donors from Minas Gerais state, Brazil

Introduction: Hepatitis C virus (HCV) is transmissible through blood products and is a public health problem in Brazil. Monitoring blood donors may help to understand its trends in the general population. The evolution of HCV antibody prevalence in blood donors from Minas Gerais, south-eastern Brazil, and its distribution in first-time donors are shown. Methodology: From 1993 to 2007, 3,249,944 blood donors were screened for HCV-Ab with ELISAs. Results: Of the donors tested, 8,107 were reactive for HCV-Ab (0.25 por cento CI=0.2441-0.2549). The total prevalence decreased from a maximum of 0.91 por cento in 1993 to 0.098 por cento in 2007. Among 1,283,970 first-time donors in the 2003 to 2007 period, there were 1,792 (0.51 por cento) reactive for HCV-Ab. The cities of Sete Lagoas and Passos had the highest prevalence (0.76 por cento), followed by Uberaba (0.62 por cento) and Belo Horizonte (0.60 por cento). Betim, Diamantina and Ituiutaba had the lowest rates (< 0.4 por cento). Discussion: The decrease in the prevalence of HCV-Ab in blood donors may be due to improved tests, better donor selection and deferral of the repeat donors. The asymmetrical distribution may reflect different profiles of exposure to HCV and may help elicit public health measures that, in turn, will help foster a safer donor population...

Introdução: O vírus da hepatite C (HCV) é transmissível por meio de produtos sanguíneos e consiste em um problema de saúde pública no Brasil. O monitoramento de doadores pode contribuir 2013para o entendimento da tendência desta infecção na população geral. Neste estudo, mostramos a evolução da prevalência de HCV em doadores de Minas Gerais, no sudoeste do Brasil e sua distribuição em doadores de primeira vez, que apresentam maior semelhança com a população. Metodologia: De 1993 a 2007, 3.249.944 doadores de sangue foram triados para HCV usando a metodologia de ELISA. Resultados: Dos doadores testados, 8.107 foram reativos para HCV (0,25 por cento CI=0,2441-0,2549). A prevalência total caiu de um máximo de 0,91 por cento in 1993 para 0,098 por cento em 2007. Dentre os 1.283.970 doadores de primeira vez no período de 2003 a 2007 havia 1.792 (0,51 por cento) indivíduos reativos para HCV. As cidades de Sete Lagoas e Passos tiveram a prevalência mais elevada (0,76 por cento), seguidos por Uberaba (0,62 por cento) e Belo Horizonte (0,60 por cento). Betim, Diamantina e Ituiutaba apresentaram as taxas mais baixas (< 0,4 por cento). Discussão: Redução da prevalência de HCV em doadores é significativa para a segurança do estoque de sangue e pode ser devida à melhoria dos testes, à melhor seleção de doadores e a retirada do grupo de doadores daqueles de repetição que ainda não haviam sido testados. A distribuição assimétrica no estado pode refletir os diferentes perfis de exposição ao HCV e podem auxiliar na elaboração de medidas de saúde pública que poderão, por sua vez, contribuir para uma população de doadores mais saudável...

High prevalence of HTLV-1 and 2 viruses in pregnant women in São Luis, state of Maranhão, Brazil / Alta prevalência dos vírus HTLV-1 e 2 em gestantes de São Luis, estado do Maranhão, Brasil

INTRODUCTION: Human T cell lymphotropic virus type 1 (HTLV-1) is endemic in the Caribbean, Japan, South America and regions of Africa. HTLV-2 is present in Native American populations and associated with IV drug use in Europe and North America. In Brazil, it is estimated that 1.5 million people are infected with HTLV-1/2. The study objective was to determine HTLV-1/2 prevalence in pregnant women in the prenatal care from three public services in São Luis, State of Maranhão, Brazil, and to counsel seropositive women to reduce viral transmission. METHODS: A cross-sectional study was conducted from February to December 2008; women with age of 18 to 45 years, with low risk for sexually transmitted disease (STD) were invited to participate. Blood samples were collected in filter paper, and HTLV-1/2 immunoenzymatic test (ELISA) was performed as a screening test. Women with reactive results were submitted to peripheral venous blood collection for ELISA repetition, followed by Western blot (WB) and real-time PCR to confirm and discriminate the infection between virus types 1 and 2. RESULTS: Of the 2,044 women tested, seven (0.3%) were ELISA reactive and confirmed positive (four were HTLV-1, and three were HTLV-2). All positive women were oriented not to breastfeed their newborns. CONCLUSIONS: This study showed that the virus is present in high prevalence in that population. Further studies covering other segments of the population are necessary to better characterize the presence of HTLV-1/2 in Maranhão and to elicit measures to prevent its spread.

INTRODUÇÃO: O vírus linfotrópico de células T humanas tipo 1 (HTLV-1) é endêmico no Caribe, Japão, América do sul e regiões da África. O HTLV-2 está presente em populações indígenas das Américas e usuários de drogas injetáveis na Europa e América do Norte. No Brasil, estimase que 1,5 milhões de pessoas estejam infectadas pelo HTLV-1/2. O objetivo deste estudo foi de determinar a prevalência do HTLV-1/2 em gestantes atendidas no pré-natal de três serviços públicos em São Luis, Maranhão, e orientar as mulheres soropositivas para reduzir a transmissão viral. MÉTODOS: Foi realizado um estudo transversal, de fevereiro a dezembro de 2008, com gestantes de 18 a 45 anos, com baixo risco de doença sexualmente transmissível (DST). Amostras de sangue foram coletadas em papel filtro e submetidas à técnica de ensaio imunoenzimático (ELISA) como teste de triagem para HTLV-1/2. As gestantes com resultado ELISA reativo foram submetidas à coleta de sangue venoso periférico para repetição do ELISA, seguido por WB e PCR em tempo real para confirmar e discriminar a infecção pelos tipos virais. RESULTADOS: Das 2.044 mulheres examinadas, sete (0.3%)tiveram resultados reativos e foram confirmadas soropositivas (quatro para HTLV-1 e três para HTLV-2). Todas as sete gestantes foram orientadas a não dar leite materno aos filhos. CONCLUSÕES: Este estudo mostrou que o HTLV-1/2 está presente em alta prevalência na população das mulheres grávidas estudadas. São necessários outros estudos contemplando diferentes segmentos populacionais para caracterizar a presença do HTLV-1/2 no Maranhão, e permitirmedidas preventivas contra a disseminação viral.

Thrombospondin-1 expression in human T-lymphotropic virus 1 asymptomatic carriers and patients with HTLV-associated myelopathy/tropical spastic paraparesis / Expressão de Trombospondina-1 em indivíduos infectados pelo HTLV-1 assintomáticos e pacientes portadores da mielopatia associada ao HTLV/Paraparesia Espástica Tropical

O Vírus Linfotrópico de células T humanas tipo 1 (HTLV-1) está associado a uma mielopatia (chamada mielopatia associada ao HTLV - HAM/TSP). A trombospondina-1 (TSP-1) é uma proteína da matriz que interfere com a adesão, a motilidade, e a proliferação celular. Níveis deexpressão de RNA mensageiro (mRNA) da trombospondina-1 foram avaliados em indivíduos infectados por HTLV-1: 11 pacientes assintomáticos, 18 com mielopatia ou oligossintomáticos, e 13participantes não-infectados. O RNA de células mononucleares do sangue periférico foi submetido à análise de RT-PCR para trombospondina-1. O número de indivíduos que expressaram esta proteína foi maior no grupo com mielopatia/sintomas (14/18, p igual 0,007). Em geral, a tendência para valores mais elevados de mRNA de trombospondina-1 foi observada no grupo de infectados pelo vírus (p igual 0,062). Os níveis mais elevados de expressão do mRNA foram detectados no início dos sintomas clínicos da HAM/TSP. Estudos adicionais com maior número de amostras são necessários para elucidar melhor o papel desta proteína da matriz na rede inflamatória relacionada à HAM/TSP.