Enhanced Migratory Capacity of T Lymphocytes in Severe Chagasic Patients Is Correlated With VLA-4 and TNF-α Expression.

Trypanosoma cruzi infection in humans leads to progression to chronic chagasic myocarditis (CCM) in 30% of infected individuals, paralleling T cell inflammatory infiltrates in the heart tissue. T-cell trafficking into the hearts of CCM patients may be modulated by in situ expression of chemotactic or haptotactic molecules, as the chemokine CXCL12, the cytokine tumor necrosis factor-alpha (TNF-α), and extracellular matrix proteins (ECM), such as fibronectin. Herein we evaluated the expression of fibronectin, CXCL12, and TNF-α in the myocardial tissue of T. cruzi seropositive (asymptomatic or with CCM), as well as seronegative individuals as healthy controls. Hearts from CCM patients exhibited enhanced expression of these three molecules. CXCL12 and TNF-α serum levels were also increased in CCM individuals. We then evaluated T lymphocytes from chronic chagasic patients by cytofluorometry, in terms of membrane expression levels of molecules involved in cell activation and cell migration, respectively, HLA-DR and the VLA-4 (very late antigen-4, being one integrin-type fibronectin receptor). Indeed, the expression of HLA-DR and VLA-4 was enhanced on T lymphocytes from chagasic patients, especially in the CCM group. To further approach the dynamics of T cell migratory events, we performed fibronectin-, TNF-α-, and CXCL12-driven migration. Peripheral blood mononuclear cells (PBMCs) and T cells from CCM patients presented an ex vivo enhanced migratory capacity driven by fibronectin alone when this ECM protein was placed in the membrane of transwell migration chambers. When TNF-α was previously placed upon fibronectin, we observed a further and significant increase in the migratory response of both PBMCs and T lymphocytes. Overall, these data suggest the existence in patients with chronic Chagas disease of a cardiac inflammatory infiltrate vector that promotes the recruitment and accumulation of activated T cells, driven in part by enhanced tissue expression of fibronectin and TNF-α, as well as the respective corresponding VLA-4 and TNF receptors.

Estudio preliminar de la miocarditis chagásica aguda experimental y su relación con la administración de esteroides sexuales / Preliminary study of experimental acute Chagasic cardiomyopathy and its relation to sex steroid administration / Estudo preliminar de miocardite chagásica aguda experimental e sua relação com a administração de esteróides sexuais

Introducción. En nuestro modelo experimental, la infección aguda por Trypanosoma cruzi (T. cruzi) en ratas adultas cursa con una parasitemia poco evidente, mientras que en los animales prepúberes (PP) se registran parasitemias más elevadas. Estas discrepancias podrían asociarse a la inmadurez inmunológica que exhiben los animales más jóvenes, pero asimismo, a la diferente madurez sexual del huésped al momento de la infección. Siendo la testosterona (T) una hormona capaz de influir sobre las células del sistema inmune y en consecuencia modificar el curso de las infecciones parasitarias, nos propusimos evaluar el efecto de dosis fisiológicas de T sobre la miocarditis aguda en fase temprana en ratas PP. Material y métodos. Se inocularon 1 millón de T. cruzi al destete y dos dosis de T de 1 mg/kg de peso, previo al T. cruzi. Se realizaron los controles respectivos, incluido un grupo experimental que recibió bicalutamida, antagonista de la T (5 mg/kg/día) +T+T. cruzi. Se evaluó parasitemia a los 7, 10 y 14 días post infección (pi) y se realizó el estudio anátomo-patológico de corazón, timo y bazo a los 4, 7 y 14 días pi. Resultados. A los 14 días pi, los animales que recibieron dosis fisiológicas de T presentaron un incremento significativo en la parasitemia y desarrollaron una mayor esplenomegalia que el resto de los grupos infectados. El estudio histológico de esos grupos reveló una miocarditis de intensidad similar -moderada a intensa- y nidos de amastigotes, mientras que en los animales sacrificados al día 4 y 7, se observó nidos de amastigotes sin reacción inflamatoria. Los controles no presentaron alteraciones histológicas. Conclusiones. La administración de T en los animales PP, previo a la infección con T. cruzi, propició la replicación temprana del parásito, evidenciada por el aumento en la parasitemia; sin embargo, no fue capaz de modificar la lesión cardíaca aguda en fase temprana ni tardía.

Background. According to our experimental model, along the acute phase of Chagas illness, adult rats infected with Trypanosome cruzi (T. cruzi) presents very low and almost undetectable parasitemias, whereas in prepubertal animals (PP) parasitemias reported were higher than in the adult ones. These differences could be associated with the immunological immaturity exhibited by younger animals, but also owing to the different sexual maturity of the host at the time of infection. As testosterone (T) is a hormone that can influence immune system cells and thus modify the course of parasitic infections, we have evaluated the effect of physiological doses of T on early acute stage of myocarditis in rats PP. Methods and material. Two doses of T (1 mg/kg) were inoculated to weaning rats (prior infection) followed by the inoculation of 1 million of T. cruzi trypomastigotes. The proper controls were performed, including an experimental group which received a concomitant therapy of bicalutamide, an antagonist of T (5 mg/kg/day), plus T and T. cruzi inoculation. Parasitemia was assessed at 7, 10 and 14 days post infection (pi) and the anatomopathological studies of heart, thymus and spleen were also performed at 4, 7 and 14 days pi. Results. At 14 day pi, those animals receiving physiological doses of T showed a significant increase in parasitemia and developed a higher splenomegaly compare to the rest of the infected groups. The histological examination of these groups presented a myocarditis of similar intensity -moderate to intense-, and amastigotes nests, while at days 4 and 7, amastigotes nests were observed without inflammatory reaction. Controls did not present histological alterations. Conclusions. Administration of T in the PP animals prior to T. cruzi infection led to an early parasite replication, as evidenced by the increase in parasitemia, however, it was not able to modify the acute cardiac injury during the early or late stage.

Introdução. Em nosso modelo experimental, a infecção aguda pelo Trypanosoma cruzi (T. cruzi) em ratos adultos cursos com parasitemia evidente pouco, enquanto que em animais pré-púberes (PP) são registrados parasitemias mais elevadas. Estas discrepâncias podem estar associadas à imaturidade imunológica exibidos por animais mais jovens, mas também para a diferente maturidade sexual do parasitado no momento da infecção. Como o hormônio testosterona (T) que pode influenciar as células do sistema imunológico e, assim, modificar o curso das infecções parasitárias, nós avaliamos o efeito de doses fisiológicas de T em estágio inicial miocardite aguda em ratos PP. Material e métodos. Um milhão de T. cruzi foram inoculados a desmame e duas doses de T de 1 mg / kg, antes a T. cruzi. Respectivos controles foram realizados, incluindo um grupo experimental que receberam bicalutamida, um antagonista da T (5 mg/kg/dia) + T + T. cruzi. Parasitemia foi avaliada aos 7, 10 e 14 dias após a infecção (ai) e realizado o estudo patológico do coração, timo e baço a 4, 7 e 14 dias ai. Resultados. Aos 14 dias ai, os animais que receberam doses fisiológicas de T mostraram um aumento significativo na parasitemia e desenvolveram uma maior esplenomegalia que outros grupos infectados. Exame histológico desses grupos revelou uma intensidade similar de miocardite -moderada a intensa-, e ninhos de amastigotas, enquanto em animais sacrificados nos dias 4 e 7, ninhos de amastigotas foram observados sem reação inflamatória. Os controles não apresentaram alterações histológicas. Conclusões. A administração de T nos animais PP antes da infecção com T. cruzi, levou à replicação inicial do parasita, como evidenciado pelo aumento da parasitemia, entretanto, não foi capaz de modificar a lesão cardíaca aguda na fase precoce ou tardia.

Enfermedad de chagas sistémico en fase aguda por transmisión oral: diagnóstico integral de un caso autopsiado / Acute phase of systemic chagas disease due to oral transmission: integral diagnosis of an autopsied case

Se presenta el primer caso autopsiado en Venezuela con enfermedad de Chagas por transmisión oral. Se trata de una joven de 24 años de edad con 8 semanas de embarazo quien contrajo la enfermedad conjuntamente con 71 niños y 14 adultos, la mayoría integrantes de la comunidad escolar “Rómulo Monasterios” en la localidad de Chichiriviche de la costa Vargas, donde ocurrió el segundo brote agudo de enfermedad de Chagas por transmisión oral registrado en Venezuela. El diagnóstico epidemiológico, clínico, serológico y parasitológico en sangre y líquido pleural extraídos en vida, fueron confirmados con los hallazgos histopatológicos y moleculares (PCR) de los tejidos. Se plantearon las características peculiares del caso así como algunos aspectos de la transmisión oral, ampliamente estudiados en nuestro país en animales de experimentación.

This first autopsy case with Chagas disease by oral transmissions in Venezuela is presented. This is a 24 year old girl with 8 weeks of pregnancy who contracted the disease together with 71 and 14 adults, the majority members of the school community “Romulo Monasterios” in the town of Vargas State, where occurred the second acute Chagas disease outbreak by oral transmission in Venezuela.The epidemiological, clinical, serological and parasitological diagnosis in the blood and pleural fluid extracted in life, were confirmed with the histopathology and molecular (PCR) finding in the tissues, Special features of the case are shown, as well as, some aspects of oral transmission widely studied in our country in experimental animals.

Miocardite chagásica em caninos no Estado do Rio Grande do Sul / Chagasic myocarditis in dogs in Rio Grande do Sul

Neste trabalho, são relatados dois casos de morte súbita por doença de Chagas aguda em caninos da zona rural de Porto Alegre, Rio Grande do Sul, Brasil. Os cães, um macho Pit Bull com nove meses (canino 1) e uma fêmea Labrador Retriever com dois anos (canino 2), morreram em janeiro de 2005 e maio de 2008, respectivamente. As necropsias revelaram aumento cardíaco em ambos os casos. O coração do canino 2 apresentou formato globoso com múltiplas áreas pálidas na musculatura cardíaca, mais evidentes no ventrículo direito e câmaras cardíacas dilatadas, principalmente as da direita. Ao exame histológico, ambos os casos apresentaram alterações semelhantes caracterizadas por infiltrado inflamatório difuso não-purulento acentuado, predominantemente linfocitário intersticial. Nas fibras miocárdicas, havia grande número de pseudocistos, repletos de formas amastigotas do Trypanosoma cruzi. Ao teste sorológico TESA-blot, amostra do canino 2 foi positiva para anticorpos IgM e IgG anti-T.cruzi, achado característico da fase aguda da miocardite chagásica. Os resultados indicam que a doença de Chagas deve ser investigada em casos de morte súbita em cães na região Sul do Brasil e que a espécie pode servir como reservatório e sentinela da doença em humanos.

Acute Chagas disease caused sudden death in two dogs from Porto Alegre rural zone of, Rio Grande do Sul, southern Brazil. A 9-month-old Pit Bull male (dog 1) and a 2-year-old Labrador Retriever female (dog 2) died in January 2005 and May 2008, respectively. At necropsy, the hearts were enlarged. In dog 2, heart was remarkably globoid with multiple pale areas scattered in the myocardium, especially in the right ventricle. Heart chambers, especially in the right side, were dilated. Histological findings were similar in both cases and consisted of diffuse non suppurative myocarditis predominantly with lymphocytic interstitial infiltrates. Within myocardial fibers were observed pseudocysts filled with amastigotes forms of Trypanosoma cruzi. Serologic test TESA-blot resulted positive in samples from dog 2 and showed IgM e IgG anti-T.cruzi antibodies characteristic of acute Chagas disease. The results indicate that Trypanosoma cruzi infection must be considered in the differential diagnosis of sudden death in dogs in southern Brazil and that the specie may act as a reservoir and sentinel for the disease in human beings.

Variabilidade espontânea da extra-sistolia ventricular na cardiopatia chagásica crônica / Spontaneous variabitity of ventricular extrasystole in chronic Chagas cardiopathy

Purpose - To study the spontaneous variability of single (VPCs) and coupled (CVPCs) in patients with chronic Chagas' disease (CCD). Patients and Methods - Twenty patients with CCD, 14 male, in class I and III NYHA, with frequent VPCs and VCPCs, free of drug therapy were studied.21 hour Holter monitoring was done for 4 subsequent days. The data analysis assessed the variation in the frequency of VPCs and CVPCs between patients, seven hour periods one hour periods in a hierarchical model by a Poisson process. Results - a) the frequency of VPCs follows a circadian rhythm, closely related to the hourly variations of the mean heart rate; b) disregarding the heart rate influence on the variability of the ventricular arrhythmia, its behavior was at random and unpredictable; c) the minimal percentual reduction in VPCs/h that discriminated between antiarrhythmic effect and spontaneous between antiarrhythmic effect and spontaneous variability was 121.86% for sevenhour, 58.42% for 21-hour and 38.45% for 42-hour electrocardiographic monitoring periods; d) the same approch for the VCPCs revealed values of 133,8%, 63.21% and 41.3% respectively

A lesão apical do ventrículo esquerdo na evoluçäo clínica da cardiopatia chagásica crônica / Apical lesion of the left ventricle in clinical evolution of chronic Chagas

Purpose - To study the spontaneous variability of single (VPCs) and coupled (CVPCs) in patients with chronic Chagas' disease (CCD). Patients and Methods - Twenty patients with CCD, 14 male, in class I and III NYHA, with frequent VPCs and VCPCs, free of drug therapy were studied.21 hour Holter monitoring was done for 4 subsequent days. The data analysis assessed the variation in the frequency of VPCs and CVPCs between patients, seven hour periods one hour periods in a hierarchical model by a Poisson process. Results - a) the frequency of VPCs follows a circadian rhythm, closely related to the hourly variations of the mean heart rate; b) disregarding the heart rate influence on the variability of the ventricular arrhythmia, its behavior was at random and unpredictable; c) the minimal percentual reduction in VPCs/h that discriminated between antiarrhythmic effect and spontaneous between antiarrhythmic effect and spontaneous variability was 121.86% for sevenhour, 58.42% for 21-hour and 38.45% for 42-hour electrocardiographic monitoring periods; d) the same approch for the VCPCs revealed values of 133,8%, 63.21% and 41.3% respectively...

Terapêutica empírica com amiodarona em portadores de miocardiopatia chagásica crônica e taquicardia ventricular sustentada / Long term results of empiric amiodarone therapy in patients with sustained ventricular tachycardia and chronic chagasic myocarditis

Objetivo: Avaliar eficácia e tolerância do uso empírico de amiodarona, a longo prazo, em portadores d e Miocardiopatia Chagásica Crônica (MCC) e Taquicardia Ventricular Sustentada (TVS). Casuística e Métodos: Trinta e cinco portadores de MCC e TVS, 18 (51% ) dos quais, refratários a outras drogas antiarritmicas. A dose de impregnação variou de 600 a 1200 (média de 883 ± 239) mg/dia, por período de uma a quatro semanas. A dose de manutenção foi reduzida progressivamente à média de 356 ± 125 mg/dia ao final de seis a 80 (média de 27 ± 20) meses. Foram construídas curvas de estimativa de recorrência clínica, morte súbida e morte cardíaca, segundo o método de Kaplan e Meier e comparadas variáveis clínicas na avaliação de risco de recorrências e morte súbita, através dos testes do Qui-quadrado de Pearson e t de Student.

Purpose: To evaluate the efficacy and safety of long-term empiric amiodarone therapy in patients with recurrent Sustained Ventricular Tachycardia (SVT) and Chronic Chagasic Myocarditis (CCM). Patiens and Methods: Thirty-five patients with CCM and SVT, eighteen (51%) of them were refratary to other antiarrhythmic drugs. The Amiodarone loading dose was between 600 and 1200 mg/day, mean of 883 ± 239 mg/day, from a period of one to four weeks. The maintenance dose was decreasing in the follow-up period, it fell down to 356 ± 125 mg/day at the end of six to 80 (mean = 27 ± 20) months. Cumulative, event-free interval curves were generated by the Kaplan-Meier method. Clinicals variables were compared with the use of the Student t-test or by means of chi-square tests.