RESUMO
HTLV-1 is the virus that causes T cell lymphoma/leukemia in adults and a neurological disorder known as HTLV-associated myelopathy or tropical spastic paraparesis. One of the transmission means is through contaminated blood and its byproducts. Because of the risk of HTLV-associated infections, screening for HTLV was introduced for Brazilian blood donors in 1993. Most of the diagnostic kits used in the national blood banks are bought from foreign companies. Brazil does not have the technology to produce this material and there is a need to produce diagnostic systems with national technology. In this study, we show the expression of gp21/HTLV-1 in Escherichia coli and its reactivity towards monoclonal antibodies and the antibodies of infected patients. Expressing these proteins is the first step towards obtaining diagnostic kits with Brazilian biotechnology.
Assuntos
Clonagem Molecular , Produtos do Gene env/química , Vírus Linfotrópico T Tipo 1 Humano/química , Produtos do Gene env do Vírus da Imunodeficiência Humana/genética , Anticorpos Monoclonais/genética , Anticorpos Monoclonais/imunologia , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Produtos do Gene env/genética , Produtos do Gene env/imunologia , Vetores Genéticos , Anticorpos Anti-HTLV-I/genética , Anticorpos Anti-HTLV-I/imunologia , Vírus Linfotrópico T Tipo 1 Humano/genética , Vírus Linfotrópico T Tipo 1 Humano/imunologia , Humanos , Immunoblotting , Reação em Cadeia da Polimerase , Produtos do Gene env do Vírus da Imunodeficiência Humana/isolamento & purificaçãoRESUMO
O HTLV-1 é o vírus causador da leucemia/linfoma de célula T no adulto e de uma desordem neurológica conhecida por mielopatia associada ao HTLV ou paraparesia espástica tropical. Um dos modos de transmissão é pelo sangue contaminado e seus subprodutos e, devido ao risco de infecções associadas ao HTLV sua pesquisa na triagem de doadores de sangue foi introduzida no Brasil a partir de 1993. Os kits diagnósticos utilizados nos bancos de sangue nacionais são na sua maioria comprados de empresas estrangeiras. O Brasil não detém a tecnologia para produção deste material e há a necessidade de produção de sistemas de diagnóstico com tecnologia nacional. Neste trabalho, mostramos a expressão da gp21/HTLV-1 em Escherichia coli e sua reatividade frente a anticorpos monoclonais e de pacientes infectados. Expressar tais proteínas é o primeiro passo para obtenção de conjuntos diagnósticos com tecnologia brasileira.
HTLV-1 is the virus that causes T cell lymphoma/leukemia in adults and a neurological disorder known as HTLV-associated myelopathy or tropical spastic paraparesis. One of the transmission means is through contaminated blood and its byproducts. Because of the risk of HTLV-associated infections, screening for HTLV was introduced for Brazilian blood donors in 1993. Most of the diagnostic kits used in the national blood banks are bought from foreign companies. Brazil does not have the technology to produce this material and there is a need to produce diagnostic systems with national technology. In this study, we show the expression of gp21/HTLV-1 in Escherichia coli and its reactivity towards monoclonal antibodies and the antibodies of infected patients. Expressing these proteins is the first step towards obtaining diagnostic kits with Brazilian biotechnology.
Assuntos
Humanos , Clonagem Molecular , Produtos do Gene env/química , Vírus Linfotrópico T Tipo 1 Humano/química , Proteínas Oncogênicas de Retroviridae/genética , Anticorpos Monoclonais/genética , Anticorpos Monoclonais/imunologia , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Vetores Genéticos , Produtos do Gene env/genética , Produtos do Gene env/imunologia , Anticorpos Anti-HTLV-I/genética , Anticorpos Anti-HTLV-I/imunologia , Vírus Linfotrópico T Tipo 1 Humano/genética , Vírus Linfotrópico T Tipo 1 Humano/imunologia , Immunoblotting , Reação em Cadeia da Polimerase , Proteínas Oncogênicas de Retroviridae/isolamento & purificaçãoRESUMO
Monomeric and chimeric synthetic peptides were used as coating antigens in four different mixtures in a solid phase immunoassay to select an optimal combination for the detection of antibodies to human T-cell lymphotropic virus (HTLV) in serum samples. The peptides, P-13 (gp21 I), Q5 (gp21 II)-GG-(gp46 II), and Q (gp46 I)-GG-(p19 I), represent immunodominant sequences from transmembrane protein (gp21), envelope protein (gp46), and core protein (p19) of HTLV I/II viruses; they were the most antigenic and specific peptides in previous studies. The sequences of the chimeric peptides were separated by two glycine residues. An indirect UltramicroEnzyme-linked immunosorbent assay (UMELISA) was used to evaluate the antigenicity of these peptide mixtures by using samples from anti-HTLV I/II PRP205(M), (n = 20), HTLV I-infected individuals from Cuba (n = 7), and HTLV I-positive sera from Colombia and Chile (n = 9). The specificity was evaluated with healthy blood donor sera (n = 300), anti-HIV 1-positive samples (n = 10), and other seropositive samples to different infectious agents. The highest sensitivity and specificity was obtained with mixture 1, which could be very useful in the immunodiagnostic of HTLV infection.
Assuntos
Anticorpos Anti-HTLV-I/imunologia , Antígenos HTLV-I/imunologia , Infecções por HTLV-I/diagnóstico , Infecções por HTLV-I/imunologia , Anticorpos Anti-HTLV-II/imunologia , Antígenos HTLV-II/imunologia , Infecções por HTLV-II/imunologia , Peptídeos/imunologia , Ensaio de Imunoadsorção Enzimática/métodos , Anticorpos Anti-HTLV-I/sangue , Antígenos HTLV-I/química , Infecções por HTLV-I/sangue , Anticorpos Anti-HTLV-II/sangue , Antígenos HTLV-II/química , Infecções por HTLV-II/sangue , Humanos , Peptídeos/química , Sensibilidade e EspecificidadeRESUMO
To investigate the significance of serological human T-cell lymphotropic virus type 1 (HLTV-1) Gag indeterminate Western blot (WB) patterns in the Caribbean, a 6-year (1993 to 1998) cross-sectional study was conducted with 37,724 blood donors from Guadeloupe (French West Indies), whose sera were routinely screened by enzyme immunoassay (EIA) for the presence of HTLV-1 and -2 antibodies. By using stringent WB criteria, 77 donors (0.20%) were confirmed HTLV-1 seropositive, whereas 150 (0.40%; P < 0.001) were considered HTLV seroindeterminate. Among them, 41.3% (62) exhibited a typical HTLV-1 Gag indeterminate profile (HGIP). Furthermore 76 (50.7%) out of the 150 HTLV-seroindeterminate subjects were sequentially retested, with a mean duration of follow-up of 18.3 months (range, 1 to 70 months). Of these, 55 (72.4%) were still EIA positive and maintained the same WB profile whereas the others became EIA negative. This follow-up survey included 33 persons with an HGIP. Twenty-three of them (69.7%) had profiles that did not evolve over time. Moreover, no case of HTLV-1 seroconversion could be documented over time by studying such sequential samples. HTLV-1 seroprevalence was characterized by an age-dependent curve, a uniform excess in females, a significant relation with hepatitis B core (HBc) antibodies, and a microcluster distribution along the Atlantic coast of Guadeloupe. In contrast, the persons with an HGIP were significantly younger, had a 1:1 sex ratio, did not present any association with HBc antibodies, and were not clustered along the Atlantic façade. These divergent epidemiological features, together with discordant serological screening test results for subjects with HGIP and with the lack of HTLV-1 proviral sequences detected by PCR in their peripheral blood mononuclear cell DNA, strongly suggest that an HGIP does not reflect true HTLV-1 infection. In regard to these data, healthy blood donors with HGIP should be reassured that they are unlikely to be infected with HTLV-1 or HTLV-2.
Assuntos
Produtos do Gene gag/imunologia , Anticorpos Anti-HTLV-I/sangue , Infecções por HTLV-I/epidemiologia , Vírus Linfotrópico T Tipo 1 Humano , Proteínas Virais/imunologia , Adolescente , Adulto , Idoso , Doadores de Sangue , Western Blotting , Região do Caribe/epidemiologia , Portador Sadio/virologia , Estudos Transversais , DNA Viral/sangue , Feminino , Anticorpos Anti-HTLV-I/imunologia , Infecções por HTLV-I/virologia , Vírus Linfotrópico T Tipo 1 Humano/genética , Vírus Linfotrópico T Tipo 1 Humano/imunologia , Vírus Linfotrópico T Tipo 1 Humano/isolamento & purificação , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da PolimeraseRESUMO
We report eight patients with familial tropical spastic paraparesis belonging to four families. The diagnosis was reached by clinical, radiological and electrophysiological studies. Human lymphotropic virus type I infection was confirmed by ELISA, immunofluorescence essays. Western blot and polymerase chain reaction in DNA of peripheral blood mononuclear cells, using primers for tax and 5'ltr genes. In all these families there was a vertical transmission of the disease from the first to the second generation. All patients improved their spastic gait after prednisone treatment. Among patients of the second generation, all had dacrysialoadenitis, three had leukemia like lymphocytes in the blood smear, two had mycosis fungoides and one had hepatic cirrhosis.
Assuntos
Paraparesia Espástica Tropical/genética , Adolescente , Adulto , Feminino , Seguimentos , Anticorpos Anti-HTLV-I/imunologia , Vírus Linfotrópico T Tipo 1 Humano , Humanos , Masculino , Pessoa de Meia-Idade , Paraparesia Espástica Tropical/imunologia , Paraparesia Espástica Tropical/virologiaRESUMO
El virus linfotrópico humano 1 (HTLV-1) ha sido asociado con la leucemia de células T del adulto (ATL) y la paraparesia espástica tropical (PET) o mielopatía asociada con HTLV-1 (HAM). Aunque la prevalencia de anticuerpos contra el HTLV-1 ha sido descrita en diversos países, especialmente en Japón, poco se conoce acerca de este marcador de contacto viral en nuestra población. En este artículo describimos la ausencia de niveles de anticuerpos anti-HTLV-1 en el suero, medidos por la prueba de elisa, en una población negra de la costa atlántica de Colombia. Nuestros hallazgos sugieren que: a) este grupo descendiente de Africa no se ha expuesto al virus; b) puede ser que algunos de los individuos incluídos en el estudio estén en periódo de incubación y, que debido a su juventud, no fue posible demostrar una respuesta humoral contra el virus; y c) como ha sido descrito en otros estudios, la prevalencia de anticuerpos anti-HTLV- 1 varía entre las diferentes regiones de Colombia
Assuntos
Humanos , População Negra/genética , Anticorpos Anti-HTLV-I/imunologia , Colômbia , Infecções por HTLV-I/epidemiologia , PrevalênciaRESUMO
During 1985 and 1986, the authors measured antibodies to human T-lymphotropic virus type I (HTLV-I) in a cohort of 13,260 Jamaicans from all parts of the island who applied for food-handling licenses. HTLV-I seroprevalence was strongly age and sex dependent, rising from 1.7% (10-19 years) to 9.1% (greater than or equal to 70 years) in men and from 1.9% (10-19 years) to 17.4% (greater than or equal to 70 years) in women. In a logistic regression analysis, women were more likely to be seropositive than were men, and farmers, laborers, and the unemployed were more likely to be HTLV-I seropositive than were those reporting student or professional occupations. In men, African ethnicity was associated with HTLV-I seropositivity in the univariate analysis but was not a risk factor after adjustment for age and sex. There was a trend toward higher age-stratified HTLV-I seroprevalence among younger women who reported more pregnancies, but older multigravidas had lower rates of HTLV-I seropositivity. Persons born outside Jamaica had significantly lower seroprevalence than did those born in Jamaica, but they were of slightly different ethnic and occupational compositions than those born in Jamaica.
Assuntos
Anticorpos Anti-HTLV-I/análise , Vírus Linfotrópico T Tipo 1 Humano/isolamento & purificação , Adolescente , Adulto , Fatores Etários , Idoso , Criança , Análise por Conglomerados , Estudos de Coortes , Ensaio de Imunoadsorção Enzimática , Feminino , Anticorpos Anti-HTLV-I/imunologia , Humanos , Jamaica , Masculino , Pessoa de Meia-Idade , Fatores Sexuais , Inquéritos e QuestionáriosRESUMO
During 1985 and 1986, the authors measured antibodies to human T-lymphotropic virus type I (HTLV-I) in a cohort of 13,260 Jamaicans from all parts of the island who applied for food-handling licenses. HTLV-I seroprevalence was strongly age and sex dependent, rising from 1.7 percent (10-19 years) to 9.1 percent (o70 years) in men and from 1.9 percent (10-19 years) to 17.4 percent (o70 years) in women. In a logistic regression analysis, women were more likely to be seropositive than were men, and farmers, laborers, and the unemployed were more likely to be HTLV-I seropositive than were those reporting student or professional occupations. In men, African ethnicity was associated with HTLV-I seropositivity in the univariate analyssis but was not a risk factor after adjustment for age and sex. There was a trend toward higher age-stratified HTLV-I seroprevalence among younger women who reported more pregnancies, but older multigravidas had lower rates of HTLV-I seropositivity. Persons born outside Jamaica had significantly lower seroprevalence than did those born in Jamaica, but they were of slightly different ethnic and occupational compositions than those born in Jamaica. (AU)