RESUMEN
Previous serological studies on the Arara do Laranjal Indian group revealed extensive HTLV-2 infections. A collection of 97 new samples from the Arara were found repeatedly negative using three different commercial enzyme immunoassays. Eight samples that exhibited optical density readings close to the cut-off value were re-evaluated using Western blot (GeneLab 2.4, Singapore) assay. One sample was found to be non-reactive, five exhibited indeterminate patterns, one was classified as HTLV, and one was confirmed as HTLV-2. Peripheral blood mononuclear cell DNA of the eight samples were subjected to nested PCR and restriction fragment length polymorphism (RFLP) analysis of the pX and env regions, and nucleotide sequencing of the 5'-LTR region. All produced amplification products of pX, but env could be amplified in only one sample with the commonly used primers. RFLP analysis of the pX region using TaqI confirmed HTLV-2 infection. Nucleotide sequencing of the 5'-LTR region was performed in three samples (HTLV-2, HTLV and indeterminate based on Western blot pattern). Phylogenetic analysis of a 449-nt fragment using the Neighbour-Joining method clearly demonstrated that the three samples clustered within the HTLV-2c molecular subtype. The present study confirms the wide dissemination of the HTLV-2c subtype among linguistically and culturally distinct Amazonian Indian groups, and emphasizes the unique occurrence of infection by this subtype in Brazil. Moreover, it emphasizes the limitation of employing the present serological screening assays in blood banks, epidemiological studies, and the importance of molecular assays in the confirmatory procedures for the primary detection of HTLV-2 infections.
Asunto(s)
Infecciones por HTLV-I/inmunología , Anticuerpos Anti-HTLV-II/sangre , Virus Linfotrópico T Tipo 2 Humano/aislamiento & purificación , Filogenia , Brasil/epidemiología , Técnicas para Inmunoenzimas , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Virus ARN/aislamiento & purificación , Sensibilidad y EspecificidadRESUMEN
Blood samples from native Indians in the Kararao village (Kayapo), were analysed using serological and molecular methods to characterize infection and analyse transmission of HTLV-II. Specific reactivity was observed in 3/26 individuals, of which two samples were from a mother and child. RFLP analysis of the pX and env regions confirmed HTLV-II infection. Nucleotide sequence of the 5' LTR segment and phylogenetic analysis showed a high similarity (98%) between the three samples and prototype HTLV-IIa (Mot), and confirmed the occurrence of the HTLV-IIc subtype. There was a high genetic similarity (99.9%) between the mother and child samples and the only difference was a deletion of two nucleotides (TC) in the mother sequence. Previous epidemiological studies among native Indians from Brazil have provided evidence of intrafamilial and vertical transmission of HTLV-IIc. The present study now provides molecular evidence of mother-to-child transmission of HTLV-IIc, a mechanism that is in large part responsible for the endemicity of HTLV in these relatively closed populations. Although the actual route of transmission is unknown, breast feeding would appear to be most likely.
Asunto(s)
Infecciones por HTLV-II/sangre , Infecciones por HTLV-II/transmisión , Virus Linfotrópico T Tipo 2 Humano/genética , Indígenas Sudamericanos , Transmisión Vertical de Enfermedad Infecciosa/estadística & datos numéricos , ARN Viral/sangre , Secuencia de Bases , Brasil , Humanos , Datos de Secuencia Molecular , Filogenia , Salud RuralRESUMEN
Analysis of human T-lymphotropic virus type II (HTLV-II) isolates from North America and Europe have demonstrated the existence of two molecular subtypes of the virus, HTLV-IIa and HTLV-IIb. Recently, studies on HTLV-II infections in Brazil have revealed isolates that are related phylogenetically to the HTLV-IIa subtype but have a HTLV-IIb phenotype with respect to the transactivating protein, tax. To more clearly define this relationship, HTLV-II was isolated from peripheral blood of an IVDA from Sao Paulo, Brazil (SP-WV), and the complete provirus was cloned and sequenced. Comparison of HTLV-II(SP-WV) nucleotide sequences to other available complete HTLV-II proviral sequences revealed that HTLV-II(SP-WV) is most closely related to HTLV-II(Mo), the prototypic HTLV-IIa subtype sequence. Phylogenetic analysis of LTR, env, and tax regions unequivocally demonstrated that HTLV-II(SP-WV) and all other Brazilian sequences examined are members of the IIa subtype. The predicted amino acid sequences of the major coding regions of HTLV-II(SP-WV) are also most closely related to HTLV-II(Mo), with the important exception of tax. The tax protein encoded by HTLV-II(SP-WV) is 96-99% identical to the tax of IIb isolates and is similar in that it has an additional 25 amino acids at the carboxy-terminus compared to the HTLV-II(Mo) tax with which it shares 91% identity. Analysis of tax stop codon usage of a number of HTLV-IIa isolates from North American, Europe, and Brazil demonstrated that isolates from the last region appear to be unique in their extended tax phenotype. It could be demonstrated that the extended tax proteins in the HTLV-IIb and Brazilian isolates had equivalent ability to transactivate the viral LTR, and studies with deletion mutants indicated that the extended C-terminus is not essential for transactivation. In contrast, the HTLV-IIa tax was found to have a greatly diminished ability to transactivate the viral LTR, which appeared to be a consequence of reduced expression of the protein. The studies show that although the Brazilian strains do not represent an entirely new subtype based on nucleotide sequence analysis they are a phenotypically unique molecular variant within the HTLV-IIa subtype.
Asunto(s)
Virus Linfotrópico T Tipo 2 Humano/clasificación , Virus Linfotrópico T Tipo 2 Humano/genética , Brasil , Clonación Molecular , ADN Viral/química , Productos del Gen env/genética , Productos del Gen tax/genética , Genoma Viral , Humanos , Datos de Secuencia Molecular , Fenotipo , Filogenia , Secuencias Repetidas Terminales/genética , Activación TranscripcionalRESUMEN
The occurrence of HTLV-I/II and HIV-1 coinfections have been shown to be frequent, probably in consequence of their similar modes of transmission. This paper presents the prevalence of coinfection of HTLV among HIV-1 infected and AIDS patients in Belém, State of Pará, Brazil. A group of 149 patients attending the AIDS Reference Unit of the State Department of Health was tested for the presence of antibodies to HTLV-I/II using an enzyme immunoassay and the positive reactions were confirmed with a Western blot that discriminates between HTLV-I and HTLV-II infections. Four patients (2.7%) were positive to HTLV-I, seven (4.7%) to HTLV-II and one (0.7%) showed an indeterminate pattern of reaction. The present results show for the first time in Belém not only the occurrence of HTLV-II/HIV-1 coinfections but also a higher prevalence of HTLV-II in relation to HTLV-I. Furthermore, it also enlarges the geographical limits of the endemic area for HTLV-II in the Amazon region of Brazil.
Asunto(s)
Seropositividad para VIH/complicaciones , Infecciones por HTLV-I/complicaciones , Infecciones por HTLV-I/epidemiología , Infecciones por HTLV-II/complicaciones , Infecciones por HTLV-II/epidemiología , Adolescente , Adulto , Anciano , Brasil/epidemiología , Femenino , Seropositividad para VIH/epidemiología , Infecciones por HTLV-I/diagnóstico , Infecciones por HTLV-II/diagnóstico , Humanos , Masculino , Persona de Mediana Edad , Prevalencia , Factores de RiesgoRESUMEN
The human lymphotropic viruses type I (HTLV-I) and type II (HTLV-II) are members of a group of mammalian retroviruses with similar biological properties, and blood transfusion is an important route of transmission. HTLV-I is endemic in a number of different geographical areas and is associated with several clinical disorders. HTLV-II is endemic in several Indian groups of the Americas and intravenous drug abusers in North and South America, Europe and Southeast Asia. During the year of 1995, all blood donors tested positive to HTLV-I/II in the State Blood Bank (HEMOPA), were directed to a physician and to the Virus Laboratory at the Universidade Federal do Pará for counselling and laboratory diagnosis confirmation. Thirty-five sera were tested by an enzyme immune assay, and a Western blot that discriminates HTLV-I and HTLV-II infection. Two HTLV-II positive samples were submitted to PCR analysis of pX and env genomic region, and confirmed to be of subtype IIa. This is the first detection in Belém of the presence of HTLV-IIa infection among blood donors. This result emphasizes that HTLV-II is also present in urban areas of the Amazon region of Brazil and highlights the need to include screening tests that are capable to detect antibodies for both types of HTLV.
Asunto(s)
Donantes de Sangre , Infecciones por HTLV-II/diagnóstico , Salud Urbana/estadística & datos numéricos , Adulto , Brasil/epidemiología , Niño , Femenino , Infecciones por HTLV-II/epidemiología , Humanos , Masculino , Persona de Mediana EdadRESUMEN
The occurence of HTLV-I/II and HIV-1 coinfections have been shown to be frequent, probably in consequence of their similar modes of transmission. This paper presents the prevalence of coinfection of HTLV among HIV-1 infected and AIDS patients in Belém, State of Pará, Brazil. A group of 149 patients attending the AIDS Reference Unit of the State Department of Health was tested for the presence of antibodies to HTLV-I/II using an enzyme immunoassay and the positive reactions were confirmed with a Western blot that discriminates between HTLV-I and HTLV-II infections. Four patients (2.7 per cent) were positive to HTLV-I, seven (4.7 per cent) to HTLV-II and one (0.7 per cent) showed an indeterminate pattern of reation. The present results show for the first time in Belém not only the occurrence of HTLV-II/HIV-1 coinfections, but also a higher prevalence of HTLV-II in relation to HTLV-I. Furthermore. it also enlarges the geographical limits of the endemic area for HTLV-II in the Amazon region of Brazil.
Asunto(s)
Humanos , Seropositividad para VIH , VIH-1 , Anticuerpos Anti-HTLV-I , Anticuerpos Anti-HTLV-II , Síndrome de Inmunodeficiencia Adquirida , BrasilRESUMEN
Os vírus linfotrópicos de células T humanas tipo I (HTLV-I) e tipo II (HTLV-II) são membros de um grupo de retrovírus de mamíferos com propriedades biológicas similares que apresentam como uma das principais rotas de transmissão a transfusão sangüínea. O HTLV-I é endêmico em diferentes áreas geográficas e está associado a vários distúrbios clínicos. O HTLV-II é endêmico em vários grupos indígenas das Américas e em usuários de drogas intravenosas na América do Norte e do Sul, Europa e Sudeste da Ásia. Durante o ano de 1995, todos os doadores de sangue positivos para HTLV-I/II no Banco de Sangue do Estado (HEMOPA), foram direcionados a um médico e ao Laboratório de Virologia na Universidade Federal do Pará, para consulta, aconselhamento e confirmação do diagnóstico laboratorial. Trinta e cinco soros foram testados por um ensaio imunoenzimático e confirmados por um Western blot que discrimina as infecções por HTLV-I e HTLV-II. Amostras soropositivas para HTLV-II foram submetidas à reação em cadeia da polimerase (PCR) para as regiões genômicas env e pX e confirmaram ser do subtipo IIa. Esta é a primeira detecção, em Belém, da presença da infecção pelo HTLV-IIa em doadores de sangue. Estes resultados enfatizam que o HTLV-II está presente em áreas urbanas da região Amazônica e a necessidade de incluir testes de triagem capazes de detectar anticorpos para ambos os tipos de HTLV.
The human lymphotropic viruses type I (HTLV-I) and type II (HTLV-II) are members of a group of mammalian retroviruses with similar biological properties, and blood transfusion is an important route of transmission. HTLV-I is endemic in a number of different geographical areas and is associated with several clinical disorders. HTLV-II is endemic in several Indian groups of the Americas and intravenous drug abusers in North and South America, Europe and Southeast Asia. During the year of 1995, all blood donors tested positive to HTLV-I/II in the State Blood Bank (HEMOPA), were directed to a physician and to the Virus Laboratory at the Universidade Federal do Pará for counselling and laboratory diagnosis confirmation. Thirty-five sera were tested by an enzyme immune assay, and a Western blot that discriminates HTLV-I and HTLV-II infection. Two HTLV-II positive samples were submitted to PCR analysis of pX and env genomic region, and confirmed to be of subtype IIa. This is the first detection in Belém of the presence of HTLV-IIa infection among blood donors. This result emphasizes that HTLV-II is also present in urban areas of the Amazon region of Brazil and highlights the need to include screening tests that are capable to detect antibodies for both types of HTLV.
Asunto(s)
Adulto , Niño , Femenino , Humanos , Masculino , Persona de Mediana Edad , Donantes de Sangre , Infecciones por HTLV-II/diagnóstico , Salud Urbana/estadística & datos numéricos , Brasil/epidemiología , Infecciones por HTLV-II/epidemiologíaRESUMEN
Human T cell leukemia virus type II (HTLV-II) is endemic in a number of native American populations and high rates of infection have also been demonstrated in intravenous drug abusers (IVDAs). Studies of virus isolates in the latter population have shown the existence of two closely related subtypes of the virus, HTLV-IIa and HTLV-IIb. To characterize the viruses present in native Americans, we analyzed by nucleotide sequence analysis the proviruses from the Wayu, an aboriginal population residing in Colombia, South America. The results showed HTLV-IIb infection in this population, and also demonstrated remarkable conservation of sequence when compared to the proviruses in IVDAs.