RESUMO
Chagas disease is a major public health problem in Bolivia. In the city of Cochabamba, 58% of the population lives in peripheral urban districts ("popular zones") where the infection prevalence is extremely high. From 1995 to 1999, we studied the demographics of Chagas infections in children from five to 13 years old (n = 2218) from the South zone (SZ) and North zone (NZ) districts, which differ in social, environmental, and agricultural conditions. Information gathered from these districts demonstrates qualitative and quantitative evidence for the active transmission of Trypanosoma cruzi in urban Cochabamba. Seropositivity was high in both zones (25% in SZ and 19% in NZ). We observed a high risk of infection in children from five to nine years old in SZ, but in NZ, a higher risk occurred in children aged 10-13, with odds ratio for infection three times higher in NZ than in SZ. This difference was not due to triatomine density, since more than 1,000 Triatoma infestans were captured in both zones, but was possibly secondary to the vector infection rate (79% in SZ and 37% in NZ). Electrocardiogram abnormalities were found to be prevalent in children and pre-adolescents (SZ = 40%, NZ = 17%), indicating that under continuous exposure to infection and re-infection, a severe form of the disease may develop early in life. This work demonstrates that T. cruzi infection should also be considered an urban health problem and is not restricted to the rural areas and small villages of Bolivia.
Assuntos
Doença de Chagas/transmissão , Conhecimentos, Atitudes e Prática em Saúde , Insetos Vetores/parasitologia , Triatoma/parasitologia , Trypanosoma cruzi/isolamento & purificação , Adolescente , Animais , Bolívia/epidemiologia , Gatos , Bovinos , Doença de Chagas/diagnóstico , Doença de Chagas/epidemiologia , Criança , Pré-Escolar , Cães , Feminino , Humanos , Masculino , Densidade Demográfica , Prevalência , Coelhos , Fatores de Risco , Estudos Soroepidemiológicos , Ovinos , Fatores Socioeconômicos , População UrbanaRESUMO
Chagas disease is a major public health problem in Bolivia. In the city of Cochabamba, 58 percent of the population lives in peripheral urban districts ("popular zones") where the infection prevalence is extremely high. From 1995 to 1999, we studied the demographics of Chagas infections in children from five to 13 years old (n = 2218) from the South zone (SZ) and North zone (NZ) districts, which differ in social, environmental, and agricultural conditions. Information gathered from these districts demonstrates qualitative and quantitative evidence for the active transmission of Trypanosoma cruzi in urban Cochabamba. Seropositivity was high in both zones (25 percent in SZ and 19 percent in NZ). We observed a high risk of infection in children from five to nine years old in SZ, but in NZ, a higher risk occurred in children aged 10-13, with odds ratio for infection three times higher in NZ than in SZ. This difference was not due to triatomine density, since more than 1,000 Triatoma infestans were captured in both zones, but was possibly secondary to the vector infection rate (79 percent in SZ and 37 percent in NZ). Electrocardiogram abnormalities were found to be prevalent in children and pre-adolescents (SZ = 40 percent, NZ = 17 percent), indicating that under continuous exposure to infection and re-infection, a severe form of the disease may develop early in life. This work demonstrates that T. cruzi infection should also be considered an urban health problem and is not restricted to the rural areas and small villages of Bolivia.
Assuntos
Adolescente , Animais , Gatos , Bovinos , Criança , Pré-Escolar , Cães , Feminino , Humanos , Masculino , Coelhos , Doença de Chagas/transmissão , Conhecimentos, Atitudes e Prática em Saúde , Insetos Vetores/parasitologia , Triatoma/parasitologia , Trypanosoma cruzi/isolamento & purificação , Bolívia/epidemiologia , Doença de Chagas/diagnóstico , Doença de Chagas/epidemiologia , Densidade Demográfica , Prevalência , Fatores de Risco , Estudos Soroepidemiológicos , Ovinos , Fatores Socioeconômicos , População UrbanaRESUMO
We performed a cross-sectional flow cytometric analysis of peripheral blood mononuclear cells to evaluate human immunologic status during early stages of Trypanosoma cruzi infection in children. We identified major immunological features corresponding to three proposed phases of disease: early acute (EA) phase, late acute (LA) phase and recent chronic (RC) phase. EA phase was accompanied by expansion of conventional B cells, up-regulation of CD54 on monocytes and down-regulation of CD54 on T cells and not associated with monocyte-activation phenotypes or changes of natural killer (NK) population. LA phase was characterized by a selective increase in a distinct lineage of NK cells (CD16+CD56-), as well as a persistent expansion of B cells and down-regulation of CD54 on T cells. RC phase showed persistent low levels of CD54 molecule on T cells and an increase of B cells, mainly triggered by expansion of the B1-cell subset, as well as increased expression of human leucocyte antigen (HLA-DR) by monocytes. These findings reinforce the hypothesis that T. cruzi-derived antigens are able to activate NK cells before the development of T-cell-mediated immunity. Moreover, our data support previous findings of increased levels of B1 lymphocytes during human Chagas' disease and show that this event is already present during initial stages of chronic infection.
Assuntos
Doença de Chagas/imunologia , Leucócitos/imunologia , Adolescente , Antígenos CD19/análise , Complexo CD3/análise , Antígeno CD56/análise , Criança , Pré-Escolar , Antígenos HLA-DR/análise , Humanos , Imunofenotipagem , Molécula 1 de Adesão Intercelular/análise , Receptores de IgE/análiseRESUMO
A rapid, sensitive, specific, and reliable enzyme-linked immunosorbent assay (ELISA) is proposed for determination of the levels of anti-Trypanosoma cruzi IgM in acute chagasic sera (ACD). The efficiency of this ELISA as a diagnostic method was compared with that of parasite DNA detection by polymerase chain reaction (PCR) and that of indirect immunofluorescence (iIF) anti-T. cruzi IgM detection. We tested whether this ELISA using fixed epimastigotes (epi) could detect anti-T. cruzi IgM in serum samples from two groups of children with acute Chagas' disease from a hyperendemic area in Bolivia. In a comparison of the ELISA method with other techniques, 95% and 71% of the results correlated with PCR and iIF findings, respectively. At the serum dilution applied (1:250), rheumatoid factor (RF) did not influence the results, and samples from patients carrying leishmaniasis or mixed Leishmania and T. cruzi infection could also be excluded from ACD. Highly specific and reliable results were obtained, a great number of the sera could be tested in only one assay, and a quantitative index of reactivity (IR) could be calculated without serial titration. Using test samples in triplicate, the method provides a useful tool for the detection of early acute-phase T. cruzi infection in humans.
Assuntos
Anticorpos Antiprotozoários/sangue , Doença de Chagas/diagnóstico , Ensaio de Imunoadsorção Enzimática/métodos , Imunoglobulina M/sangue , Doença Aguda , Adolescente , Bolívia , Doença de Chagas/epidemiologia , Criança , Pré-Escolar , Reações Cruzadas , Doenças Endêmicas , Ensaio de Imunoadsorção Enzimática/normas , Fixadores , Técnica Indireta de Fluorescência para Anticorpo/métodos , Formaldeído , Humanos , Testes de Neutralização , Reação em Cadeia da Polimerase/métodos , Fator Reumatoide/imunologia , Sensibilidade e EspecificidadeRESUMO
Here, we analysed the use of Vbeta-TCR regions by CD4+ and CD8+ T cells from acute and chronic chagasic patients using flow cytometry. We determined the Vbeta expression in cells freshly isolated from patients, as well as after in vitro stimulation with antigens derived from epimastigote (EPI) or trypomastigote (TRYPO) forms of Trypanosoma cruzi. Analysis of Vbeta-TCR expression of T cells freshly isolated from patients showed a decrease in Vbeta5 expression in the CD4+ T-cell population from acutely infected individuals, whereas CD4+Vbeta5+ T cells were found to be increased in chronic patients with the cardiac, but not indeterminate, clinical form. After culturing peripheral blood mononuclear cells (PBMC) from chronic patients with EPI or TRYPO, we found that both antigenic preparations led to a preferential expansion of CD4+Vbeta5+ T cells. EPI stimulation also led to the expansion of CD8+Vbeta5+ T cells, whereas TRYPO led to the expansion of this cell population only if PBMC were from cardiac and not indeterminate patients. We observed that TRYPO stimulation led to an increase in the frequency of CD4+Vbeta17+ T cells in cultures of PBMC from indeterminate patients, whereas an increase in the frequency of CD8+Vbeta17+ T cells was found upon TRYPO stimulation of PBMC from cardiac patients. Despite this increase in the frequency of Vbeta17+ T-cell populations upon TRYPO stimulation, the same antigenic preparation led to a much higher expansion of Vbeta5+ T cells. These results show a differential expression of Vbeta5-TCR in cells freshly isolated from chagasic patients in different stages of the disease and that parasite-specific antigens stimulate a portion of the T-cell repertoire with preferential usage of Vbeta5-TCR.
Assuntos
Doença de Chagas/imunologia , Receptores de Antígenos de Linfócitos T alfa-beta/imunologia , Linfócitos T/imunologia , Doença Aguda , Animais , Antígenos de Protozoários/imunologia , Contagem de Linfócito CD4 , Linfócitos T CD4-Positivos/imunologia , Doença Crônica , Cardiopatias/imunologia , Humanos , Trypanosoma cruzi/imunologiaRESUMO
The acute phase of Chagas' disease was classified as early, intermediate, and late based on the levels of anti-Galalpha, 3Gal IgG (Gal) and specific IgM (M) and IgG (G) anti-T. cruzi reactivity. While the early phase was M+G-Gal-, the intermediate phase was M+G-Gal+, M+G+Gal-, or M+G+Gal+, and the late phase was M-G+Gal+. This sequence of stages was consistent with our previous studies on acute-phase proteins. Analysis by the polymerase chain reaction (PCR) of parasite DNA in 65 blood samples of children living in Cochabamba, Bolivia showed a significant correlation (90.8%) between ELISA and PCR positivity. A lower correlation was observed between indirect hemagglutination, PCR (58%), and ELISA. Electrocardiographic analysis of 43 children studied by the PCR did not show any alteration typical of acute chagasic myocarditis. The PCR positivity was observed in eight samples where only Gal was increased, suggesting a very early T. cruzi infection, when specific antibodies were not yet present. By associating anti-Gal IgG with specific serology, early T. cruzi infection can be detected with greater precision. We suggest the use of anti-Gal antibody reactivity as an aid for the detection of recent T. cruzi infections, at least in endemic areas where diseases caused by other trypanosomatids do not overlap.
Assuntos
Anticorpos Antiprotozoários/sangue , Doença de Chagas/classificação , Doença de Chagas/imunologia , Trypanosoma cruzi/imunologia , Adolescente , Animais , Bolívia/epidemiologia , Doença de Chagas/sangue , Doença de Chagas/epidemiologia , Criança , Pré-Escolar , DNA de Protozoário/sangue , Eletrocardiografia , Ensaio de Imunoadsorção Enzimática , Feminino , Galactose/imunologia , Testes de Hemaglutinação , Humanos , Imunoglobulina G/sangue , Masculino , Reação em Cadeia da Polimerase , Testes SorológicosRESUMO
Chagas' disease represents a major public health problem in Latin America. In endemic areas, it is important to detect acute and even asymptomatic infections in children so that specific therapy can be started immediately. We studied 203 sera from children from the region of Cochabamba, Bolivia. A high percentage of seropositive individuals was found in the three villages studies. Levels of alpha-2 macroglobulin (A2M) and C-reactive protein (CRP) increased in a significant number of children with acute Chagas' disease. The combined analysis of serologic and biochemical parameters can define the different stages of acute infection by Trypanosoma cruzi: 1) an early stage, with an increase only in specific immunoglobulin M (IgM) levels; 2) intermediate stages, with high specific IgM and IgG levels and/or high anti-galactose (anti-Gal) levels and increased A2M and/or CRP levels; and 3) a late acute stage, with low IgM levels but high A2M, CRP, anti-Gal, and specific IgG levels. The detection of high IgG levels alone is indicative of the chronic/indeterminate stage of Chagas' disease. We also show serologic differences between seropositive asymptomatic villagers and symptomatic patients undergoing medical care; asymptomatic cases presented higher levels of A2M and lower levels of specific antibodies.