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.

Proprioceptive neuromuscular facilitation in HTLV-I-associated myelopathy/tropical spastic paraparesis

Introduction: Human T cell lymphotropic virus type I-associated myelopathy/tropical spastic paraparesis (HAM/TSP) can impact the independence and motricity of patients. The aims of this study were to estimate the effects of physiotherapy on the functionality of patients with HAM/TSP during the stable phase of the disease using proprioceptive neuromuscular facilitation (PNF) and to compare two methods of treatment delivery. Methods: Fourteen patients with human T cell lymphotropic virus type I (HTLV-I) were randomly allocated into two groups. In group I (seven patients), PNF was applied by the therapist, facilitating the functional activities of rolling, sitting and standing, walking and climbing and descending stairs. In group II (seven patients), PNF was self-administered using an elastic tube, and the same activities were facilitated. Experiments were conducted for 1h twice per week for 12 weeks. Low-back pain, a modified Ashworth scale, the functional independence measure (FIM) and the timed up and go test (TUG) were assessed before and after the interventions. Results: In the within-group evaluation, low-back pain was significantly reduced in both groups, the FIM improved in group II, and the results of the TUG improved in group I. In the inter-group analysis, only the tone was lower in group II than in group I. Conclusions: Both PNF protocols were effective in treating patients with HAM/TSP.

Absence of HTLV-1/2 infection and dermatological diseases in Manaus, State of Amazonas, Brazil

Introduction The prevalence of human T-cell lymphotropic virus types 1 and 2 (HTLV-1/2) infection is heterogeneous across different populations. We tested the hypothesis that HTLV-1/2 infection occurs more often in dermatological patients. Methods A total of 1,091 patients from a tropical dermatology clinic were tested for HTLV-1/2. In parallel, 6865 first-time blood donors from the same geographic area were screened for HTLV-1/2; HTLV-1/2 positive blood donors underwent dermatological examinations. Results The prevalence of HTLV-1/2 in first-time blood donors was 0.14%. No co-occurrence of HTLV-1/2 infection and dermatological conditions was observed. Conclusions Our results challenge the hypothesis that HTLV-1/2 infection occurs more often in dermatological patients.

The use of botulinum toxin type A in the treatment of HTLV-1-associated overactive bladder refractory to conventional therapy

Urinary symptoms occur in 19% of human T-cell lymphotropic virus type 1 (HTLV-1)-infected patients who do not fulfill criteria for HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) and in almost 100% of HAM/TSP patients. Few studies have evaluated therapies for overactive bladder (OAB) caused by HTLV-1 infection. This case report describes the effect of onabotulinum toxin A on the urinary manifestations of three patients with HAM/TSP and OAB symptoms. The patients were intravesically administered 200 units of Botox®. Their incontinence episodes improved, and their OAB symptoms scores (OABSS) reduced significantly. These data indicate that Botox® should be a treatment option for OAB associated with HTLV-1 infection.

Prevalence and genetic characterisation of HTLV-1 and 2 dual infections in patients with pulmonary tuberculosis in Central-West Brazil

Human T-cell lymphotropic virus (HTLV) may impact the clinical course of tuberculosis (TB). Both infections are highly endemic in Brazil. The aim of this study was to assess the prevalence of HTLV-1/2 in TB patients in Central-West Brazil and to perform a genetic characterisation of the respective isolates. Of the 402 patients, six (1.49%) were positive for anti-HTLV and five (1.24%; 95% confidence interval: 0.46-3.05) were infected with HTLV-1/2. Genetic characterisation demonstrated that the four HTLV-1 isolates belonged to the Transcontinental subgroup A of the Cosmopolitan subtype a and that the HTLV-2 isolate belonged to subtype a (HTLV-2a/c). The prevalence of HTLV infection observed in this study is higher than that observed in local blood donors and the HTLV-1 and 2 subtypes identified are consistent with those circulating in Brazil.

Motor behavioral abnormalities and histopathological findings of Wistar rats inoculated with HTLV-1-infected MT2 cells

The objective of the present study was to describe motor behavioral changes in association with histopathological and hematological findings in Wistar rats inoculated intravenously with human T-cell lymphotropic virus type 1 (HTLV-1)-infected MT2 cells. Twenty-five 4-month-old male rats were inoculated with HTLV-1-infected MT2 cells and 13 control rats were inoculated with normal human lymphocytes. The behavior of the rats was observed before and 5, 10, 15, and 20 months after inoculation during a 30-min/rat testing time for 5 consecutive days. During each of 4 periods, a subset of rats was randomly chosen to be sacrificed in order to harvest the spinal cord for histopathological analysis and to obtain blood for serological and molecular studies. Behavioral analyses of the HTLV-1-inoculated rats showed a significant decrease of climbing, walking and freezing, and an increase of scratching, sniffing, biting, licking, and resting/sleeping. Two of the 25 HTLV-1-inoculated rats (8%) developed spastic paraparesis as a major behavioral change. The histopathological changes were few and mild, but in some cases there was diffuse lymphocyte infiltration. The minor and major behavioral changes occurred after 10-20 months of evolution. The long-term observation of Wistar rats inoculated with HTLV-1-infected MT2 cells showed major (spastic paraparesis) and minor motor abnormalities in association with the degree of HTLV-1-induced myelopathy.

Clinical and immunological features of patients with atopy and concomitant HTLV-1 infection

Human T-cell lymphotropic virus type 1 (HTLV-1) induces an exacerbated type 1 immune response characterized by high spontaneous IFN-γ and TNF-α production. Allergic rhinitis and asthma are associated with the type 2 immune response, with elevated secretion of IL-4 and IL-5. The aim of this study was to characterize the immune response in atopic HTLV-1 carriers. The cytokine profile of atopic HTLV-1 carriers (N = 10; all females) was compared with that of non-atopic HTLV-1 carriers (N = 14; 9 females and 5 males). Mean patient age of atopic and non-atopic groups was 45 ± 8 and 38 ± 11 years, respectively. All atopic HTLV-1 carriers had rhinitis with or without asthma and a skin prick test positive for Dermatophagoides pteronyssinus antigen 1 (Derp-1). There was no difference in cytokine levels between the two groups in unstimulated peripheral blood mononuclear cell cultures. In cultures stimulated with Derp-1, IFN-γ levels tended to be higher (P = 0.06) and IL-5 levels were higher (P = 0.02) in atopic HTLV-1 patients than in non-atopic subjects. In contrast, IL-10 was lower (P = 0.004) in atopic than in non-atopic HTLV-1-infected subjects. This study shows that HTLV-1 infection with an exaggerated type 1 immune response does not prevent atopy. In this case, the exacerbated type 1 and type 2 immune responses were due to a lack of IL-10 production, a cytokine that plays an important role in down-modulating type 1 and type 2 immune responses and in preventing the development of chronic inflammatory diseases.

Severity score system for progressive myelopathy: development and validation of a new clinical scale

Progressive myelopathies can be secondary to inborn errors of metabolism (IEM) such as mucopolysaccharidosis, mucolipidosis, and adrenomyeloneuropathy. The available scale, Japanese Orthopaedic Association (JOA) score, was validated only for degenerative vertebral diseases. Our objective is to propose and validate a new scale addressing progressive myelopathies and to present validating data for JOA in these diseases. A new scale, Severity Score System for Progressive Myelopathy (SSPROM), covering motor disability, sphincter dysfunction, spasticity, and sensory losses. Inter- and intra-rater reliabilities were measured. External validation was tested by applying JOA, the Expanded Disability Status Scale (EDSS), the Barthel index, and the Osame Motor Disability Score. Thirty-eight patients, 17 with adrenomyeloneuropathy, 3 with mucopolysaccharidosis I, 3 with mucopolysaccharidosis IV, 2 with mucopolysaccharidosis VI, 2 with mucolipidosis, and 11 with human T-cell lymphotropic virus type-1 (HTLV-1)-associated myelopathy participated in the study. The mean ± SD SSPROM and JOA scores were 74.6 ± 11.4 and 12.4 ± 2.3, respectively. Construct validity for SSPROM (JOA: r = 0.84, P < 0.0001; EDSS: r = -0.83, P < 0.0001; Barthel: r = 0.56, P < 0.002; Osame: r = -0.94, P < 0.0001) and reliability (intra-rater: r = 0.83, P < 0.0001; inter-rater: r = 0.94, P < 0.0001) were demonstrated. The metric properties of JOA were similar to those found in SSPROM. Several clinimetric requirements were met for both SSPROM and JOA scales. Since SSPROM has a wider range, it should be useful for follow-up studies on IEM myelopathies.

Caractérisation des transcrits antisens chez les rétrovirus HTLV et étude comparative des fonctions des protéines traduites à partir de ces transcrits antisens

Le premier membre de la famille des rétrovirus humains HTLV (Virus T-lymphotropique Humain), HTLV-1, a été découvert en 1980 et l’on estime aujourd’hui à plus de 10 millions le nombre d’individus infectés à travers le monde. Après une période de latence d’environ 40 ans, 5% des individus infectés développent des leucémies, des lymphomes adultes de lymphocytes T (ATLL) ou encore une myélopathie associée à HTLV-1/ paraparésie spastique tropicale (HAM/TSP). L’apparition de la maladie serait en grande partie orchestrée par deux protéines virales, soit Tax et HTLV-1 bZIP factor (HBZ). L’expression du génome viral se fait à partir d’un transcrit sens de pleine longueur suite à un épissage alternatif, à l’exception du gène HBZ. HBZ est produite à partir d’un transcrit antisens initié dans la séquence terminale longue répétée (LTR)’3. Elle a été décrite comme étant capable de réguler négativement la transcription virale dépendante de Tax en se dimérisant avec des facteurs de transcription cellulaires tels que CREB-2 et certains membres de la famille Jun. HBZ a aussi un pouvoir prolifératif et bien que nous ne sachions toujours pas le mécanisme moléculaire menant à l’oncogenèse par HBZ, nous savons qu’elle module une multitude de voies de transduction de signaux, dont AP-1. Nous avons récemment mis en évidence un transcrit antisens nommé Antisense Protein of HTLV-2 (APH-2) chez HTLV-2 qui n’est associé qu’à une myélopathie apparentée au HAM/TSP. Ce n’est qu’en 2005 que HTLV-3 et HTLV-4 se sont rajoutés au groupe HTLV. Cependant, aucune corrélation avec le développement d’une quelconque maladie n’a été montrée jusqu’à ce jour. Le premier volet de ce projet de doctorat avait pour objectif de détecter et caractériser les transcrits antisens produits par HTLV-3 et HTLV-4 et d’étudier les protéines traduites à partir de ces transcrits pour ainsi évaluer leurs similitudes et/ou différences avec HBZ et APH-2. Nos études de localisation cellulaire réalisées par microscopie confocale ont montré que APH-3 et APH-4 sont des protéines nucléaires, se retrouvant sous la forme de granules et, dans le cas d’APH-3, partiellement cytoplasmique. Ces granules co-localisent en partie avec HBZ. Les analyses à l’aide d’un gène rapporteur luciférase contenant le LTR 5’ de HTLV-1 ont montré que APH-3 et APH-4 peuvent aussi inhiber la transactivation du LTR 5’ par Tax. Aussi, des études faisant appel au gène rapporteur précédé d’un promoteur de collagénase (site AP-1), ont montré que ces deux protéines, contrairement à HBZ, activent la transcription dépendante de tous les membres des facteurs de transcription de la famille Jun. De plus, les mutants ont montré que le motif fermeture éclair (LZ) atypique de ces protéines est impliqué dans cette régulation. En effet, APH-3 et APH-4 modulent la voie Jun-dépendante en se dimérisant via leur LZ atypique avec la famille Jun et semblent activer la voie par un mécanisme ne faisant pas par d’un domaine activateur autonome. Dans un deuxième volet, nous avions comme objectif d’approfondir nos connaissances sur la localisation nucléolaire de HBZ. Lors de nos analyses, nous avons identifié deux nouveaux partenaires d’interaction, B23 et la nucléoline, qui semblent être associés à sa localisation nucléolaire. En effet, ces interactions sont plus fortes suivant une délétion des domaines AD et bZIP de HBZ qui dans ce cas est localisée strictement au nucléole. De plus, bien que APH-3 et APH-4 puissent se localiser aux nucléoles, HBZ est la seule protéine traduite à partir d’un transcrit antisens pouvant interagir avec B23. Finalement, ces travaux ont clairement mis en évidence que HTLV-3 et HTLV-4 permettent la production de transcrits antisens comme chez d’autres rétrovirus. Les protéines traduites à partir de ces transcrits antisens jouent d’importants rôles dans la réplication rétrovirale mais semblent avoir des fonctions différentes de celles de HBZ au niveau de la régulation de la transcription de la voie Jun. HBZ semble aussi jouer un rôle unique dans le nucléole en ciblant les protéines nucléolaires de la cellule. Ces études démontrent que les protéines produites à partir de transcrits antisens chez les rétrovirus HTLV partagent plusieurs ressemblances, mais démontrent aussi des différences. Ainsi, les APH pourraient, en tant qu’outil comparatif, aider à mieux cibler les mécanismes moléculaires importants utilisés par HBZ pour induire la pathogénèse associée à une infection par HTLV.

Comparison of signal-to-cutoff values in first, second, and third generation enzyme immunoassays for the diagnosis of HTLV-1/2 infection in ""at-risk"" individuals from Sao Paulo, Brazil

Data obtained during routine diagnosis of human T-cell lymphotropic virus type 1 (HTLV-1) and 2 (HTLV-2) in ""at-risk"" individuals from Sao Paulo, Brazil using signal-to-cutoff (S/C) values obtained by first, second, and third generation enzyme immunoassay (EIA) kits, were compared. The highest S/C values were obtained with third generation EIA kits, but no correlation was detected between these values and specific antibody reactivity to HTLV-1, HTLV-2, or untyped HTLV (p = 0.302). In addition, use of these third generation kits resulted in HTLV-1/2 false-positive samples. In contrast, first and second generation EIA kits showed high specificity, and the second generation EIA kits showed the highest efficiency, despite lower S/C values. Using first and second generation EIA kits, significant differences in specific antibody detection of HTLV-1, relative to HTLV-2 (p = 0.019 for first generation and p < 0.001 for second generation EIA kits) and relative to untyped HTLV (p = 0.025 for first generation EIA kits), were observed. These results were explained by the composition and format of the assays. In addition, using receiver operating characteristics (ROC) analysis, a slight adjustment in cutoff values for third generation EIA kits improved their specificities and should be used when HTLV ""at-risk"" populations from this geographic area are to be evaluated. (C) 2009 Elsevier B.V. All rights reserved.

Soroprevalência de HTLV/II, e fatores de risco, em gestantes do município de Botucatu atendidas em unidades básicas de saúde

Pós-graduação em Doenças Tropicais - FMB

Caracterização molecular do HTLV-1/2 em doadores de sangue em Belém, Estado do Pará: primeira descrição do subtipo HTLV-2b na região Amazônica

Este trabalho objetivou a caracterização molecular do vírus linfotrópico de células T humanas infectando doadores de sangue atendidos na Fundação Centro de Hemoterapia e Hematologia do Pará. Amostras de DNA de 79 indivíduos soropositivos para o vírus linfotrópico de células T humanas foram analisadas por meio da reação em cadeia da polimerase para as regiões genômicas pX, env e 5'LTR, de polimorfismos de comprimento de fragmentos de restrição e do seqüenciamento da região 5LTR, com posterior análise filogenética, definindo o tipo e o subtipo do HTLV circulante na população estudada. Observou-se uma maior prevalência de HTLV-1 (71%) em relação ao HTLV-2 (29%). As amostras de HTLV-1 sequenciadas foram classificadas como pertencentes ao subtipo Cosmopolita, subgrupo Transcontinental, sendo as de HTLV-2 identificadas como HTLV-2c. A análise de polimorfismos de comprimento de fragmentos de restrição da região env e do sequenciamento da região 5'LTR, identificou, pela primeira vez na Amazônia Brasileira, uma amostra de HTLV-2b, enfatizando a necessidade de estudos moleculares contínuos na região para melhor entendimento da epidemiologia de transmissão do HTLV na população e permitir a vigilância epidemiológica da emergência de novos tipos e subtipos.

Carga proviral do HTLV-1 e HTLV-2: um método simples através da PCR quantitativa em tempo real

Os vírus linfotrópicos de células T humanas, quando integrados ao genoma da célula hospedeira, provírus, têm como marcador de replicação seu DNA proviral. A carga proviral parece ser um importante fator no desenvolvimento de patologias associadas a estes retrovírus. Neste estudo foi desenvolvida uma metodologia para quantificação absoluta da carga proviral dos HTLV-1 e HTLV-2 através da PCR em tempo real. Cinqüenta e três amostras de doadores de sangue com teste de ELISA reagente foram submetidas à metodologia, que utilizou o sistema TaqMan® para três seqüências alvo: HTLV-1, HTLV-2 e albumina. A quantificação proviral absoluta foi determinada através da proporção relativa entre o genoma do HTLV e o genoma da célula hospedeira, levando em consideração o número de leucócitos. O método apresentado é sensível (215 cópias/mL), prático e simples para quantificação proviral, além de eficiente e adequado para confirmação e discriminação da infecção pelos tipos virais.

Avaliação clínica de doadores de sangue portadores do vírus linfotrópico de células T humanas (HTLV - I/II)

Os vírus linfotrópicos de células T humanas do tipo I e II (HTLV-I/II) são retrovírus que podem ser transmitidos por transfusão de sangue. Estes vírus estão associados com paraparesia espástica tropical (PET), leucemia/linfoma de células T do adulto (L/LTA) e outras doenças sistêmicas imunomediadas. O presente estudo teve como objetivo investigar sinais clínicos de patologias associadas a esses vírus para utilizar na triagem clinica de candidatos à doação de sangue. Usou procedimentos padronizados para avaliação clinica de 30 doadores de sangue soropositivos para HTLV-I/II confirmados pela técnica de reação em cadeia da polimerase (PCR) matriculados no ambulatório do Núcleo de Medicina Tropical da UFPA. Paralelamente, através de interrogatório clínico complementar, estudou grupo controle com 40 candidatos à doação de sangue escolhidos aleatoriamente, que tiveram resultados sorológicos negativos. Dos 30 pacientes examinados, verificou-se que 23 eram portadores de HTLV-I e 07 HTLV-II. Na avaliação clínica, 15 pacientes (50%) não referiram queixas, sendo que 12 pacientes com queixas exclusivamente neurológicas. Observou 05 pacientes com formigamentos; 05 com diminuição da força muscular; 04 com constipação intestinal; 02 com parestesia; 02 com nódulos subcutâneos; 01 com incontinência urinária; 01 com visão borrada; 01 com diminuição do libido. No grupo controle, 05 candidatos (12,5%) referiram queixas. Os resultados indicam que diminuição da força muscular e formigamento devem ser questionados na triagem clínica prévia à doação de sangue para reduzir risco de infecção transfusional.

Prevalência de HTLV-1 e HTLV-2 em portadores de Strongyloides stercoralis, Pará, Brasil

O Strongyloides stercoralis é um nematódeo intestinal de seres humanos causador da estrongiloidíase, doença com distribuição mundial especificamente comum em regiões tropicais e subtropicais. Estudos epidemiológicos têm demonstrado a existência da associação desta parasitose com o Vírus Linfotrópico de Células T Humanas do tipo 1 (HTLV-1). Em regiões onde ambos agentes são endêmicos, a coinfecção pode resultar no desenvolvimento da estrongiloidíase grave, pois o HTLV-1 provoca uma redução na produção dos componentes imunológicos participantes dos mecanismos de defesa contra S. stercoralis. Baseado nessa questão, esse estudo prentedeu contribuir para esclarecer o papel da imunossupressão induzida pelo HTLV-1 e HTLV-2 na persistência e disseminação do Strongyloides stercoralis. Testes sorológicos e moleculares foram utilizados para verificar a frequência da infecção pelo HTLV-1 e HTLV-2 nos portadores de S. stercoralis atendidos no Hospital Universitário João de Barros Barreto em Belém-Pará, no período de Julho de 2009 a Junho de 2011. Nesse estudo observamos a frequência (5,50%) de anticorpos anti-HTLV-1 e HTLV-2 em portadores de Strongyloides stercoralis. A prevalência de HTLV-1 (3,67%) foi superior à de HTLV-2 (0,92%). A análise da amostra estudada mostrou que não houve diferenças estatísticas significativas na frequência do HTLV-1 e HTLV-2 entre homens e mulheres. Quanto à distribuição dos portadores de HTLV-1 e HTLV-2 por faixa etária, observou-se maior frequência do vírus entre os pacientes com idade mais avançada. Baseados nos resultados deste estudo, concluímos que há necessidade de medidas profiláticas que previnam a disseminação do HTLV-1 e HTLV-2 entre portadores de S. stercoralis e como consequência evitar o desenvolvimento de complicações resultantes da associação desses agentes.