Efficacy of novel benznidazole solutions during the experimental infection with Trypanosoma cruzi.

Chagas' disease is caused by the protozoan parasite Trypanosoma cruzi. About 8 million people throughout Latin America are infected causing approximately 10,000 deaths annually. Benznidazole, available as unique 100 mg tablets in many of the endemic countries, is currently the drug of choice for the specific treatment of this condition. Despite of the large number of pediatric patients infected, there are no commercial liquid dosage forms available to treat this trypanosomiasis. This work showed that novel benznidazole-water-polyethylene glycol 400 solutions are active against T. cruzi in a murine model of Chagas' disease. Present results constitute the first demonstration on the usefulness of benznidazole solutions in infected mice.

TNF-α is involved in the abnormal thymocyte migration during experimental Trypanosoma cruzi infection and favors the export of immature cells.

Previous studies revealed a significant production of inflammatory cytokines together with severe thymic atrophy and thymocyte migratory disturbances during experimental Chagas disease. Migratory activity of thymocytes and mature T cells seem to be finely tuned by cytokines, chemokines and extracellular matrix (ECM) components. Systemic TNF-α is enhanced during infection and appears to be crucial in the response against the parasite. However, it also seems to be involved in disease pathology, since it is implicated in the arrival of T cells to effector sites, including the myocardium. Herein, we analyzed the role of TNF-α in the migratory activity of thymocytes in Trypanosoma cruzi (T. cruzi) acutely-infected mice. We found increased expression and deposition of TNF-α in the thymus of infected animals compared to controls, accompanied by increased co-localization of fibronectin, a cell migration-related ECM molecule, whose contents in the thymus of infected mice is also augmented. In-vivo studies showed an enhanced export of thymocytes in T. cruzi-infected mice, as ascertained by intrathymic injection of FITC alone or in combination with TNF-α. The increase of immature CD4(+)CD8(+) T cells in secondary lymphoid organs was even more clear-cut when TNF-α was co-injected with FITC. Ex-vivo transmigration assays also revealed higher number of migrating cells when TNF-α was added onto fibronectin lattices, with higher input of all thymocyte subsets, including immature CD4(+)CD8(+). Infected animals also exhibit enhanced levels of expression of both mRNA TNF-α receptors in the CD4(+)CD8(+) subpopulation. Our findings suggest that in T. cruzi acute infection, when TNF-α is complexed with fibronectin, it favours the altered migration of thymocytes, promoting the release of mature and immature T cells to different compartments of the immune system. Conceptually, this work reinforces the notion that thymocyte migration is a multivectorial biological event in health and disease, and that TNF-α is a further player in the process.

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.

A high corticosterone/DHEA-s ratio in young rats infected with Trypanosoma cruzi is associated with increased susceptibility.

We have previously established that young male rats are more susceptible to the effects of Trypanosoma cruzi infection than adult rats. To explore underlying age-associated differences in disease outcome, we simultaneously assessed hormone levels and cytokine release throughout the acute infection period in young and adult rats infected with T. cruzi. Young rats were inoculated with 1 x 10(6) and adult rats with 7 x 10(6) blood trypomastigotes, according to their relative body weight. At zero, seven, 14, 21 and 28 days after infection, blood was collected for the determination of gonadal and adrenal hormones, tumor necrosis factor α (TNF-α), interleukin (IL)-10 and specific IgM and IgG subtypes. Young animals displayed significantly higher parasitaemia values and an endocrine pattern that was characterised by elevated values in corticosterone (CT) and the CT/dehydroepiandrosterone-sulfate ratio, which favours immunosuppression and susceptibility. In contrast, adult male rats were able to restrict the parasite burden, which likely resulted from increased IgG antibody synthesis and oestradiol levels. Adult rats also showed a reduced TNF-α/IL-10 ratio and less tissue damage. We conclude that young animals exhibited increased vulnerability to T. cruzi infection compared with adults and this is associated with an unsuitable immunoendocrine milieu.

A high corticosterone/DHEA-s ratio in young rats infected with Trypanosoma cruzi is associated with increased susceptibility

We have previously established that young male rats are more susceptible to the effects of Trypanosoma cruzi infection than adult rats. To explore underlying age-associated differences in disease outcome, we simultaneously assessed hormone levels and cytokine release throughout the acute infection period in young and adult rats infected with T. cruzi. Young rats were inoculated with 1 x 10(6) and adult rats with 7 x 10(6) blood trypomastigotes, according to their relative body weight. At zero, seven, 14, 21 and 28 days after infection, blood was collected for the determination of gonadal and adrenal hormones, tumor necrosis factor α (TNF-α), interleukin (IL)-10 and specific IgM and IgG subtypes. Young animals displayed significantly higher parasitaemia values and an endocrine pattern that was characterised by elevated values in corticosterone (CT) and the CT/dehydroepiandrosterone-sulfate ratio, which favours immunosuppression and susceptibility. In contrast, adult male rats were able to restrict the parasite burden, which likely resulted from increased IgG antibody synthesis and oestradiol levels. Adult rats also showed a reduced TNF-α/IL-10 ratio and less tissue damage. We conclude that young animals exhibited increased vulnerability to T. cruzi infection compared with adults and this is associated with an unsuitable immunoendocrine milieu.

Benznidazole treatment attenuates liver NF-κB activity and MAPK in a cecal ligation and puncture model of sepsis.

Recent studies have shown that Benznidazole (BZL), known for its antiparasitic action on Trypanosoma cruzi, modulates pro-inflammatory cytokines and nitric oxide (NO) release in activated macrophages by blocking NF-κB through inhibition of IKK in vitro. As so far, little is known about the mechanism by which BZL provokes the inhibition of inflammatory response in sepsis in vivo, we aimed to delineate the possible role of BZL as a modulator in liver inflammation in mice with sepsis induced by cecal ligation and puncture (CLP). Specifically, we analyzed leukocytes, liver production of TNF-α and NO and the intracellular pathways modulated by these mediators, including NF-κB and MAPKs, in the liver of mice 24 h post-CLP. Our results show that BZL reduces leukocytes in peripheral blood accompanied by an increase in peritoneal macrophages 24h after CLP. In the liver of these septic mice, BZL decreased expression of mRNA and protein for TNF-α and NOS-2 by inhibition of NF-κB and MAPK (p-38 and ERK). The body of evidence suggests that the immunomodulatory effects of BZL could act selectively, as it is able to decrease the systemic inflammatory reaction and the hepatic response but it can increase the number of cells in the site of infection.

Benznidazole blocks NF-kappaB activation but not AP-1 through inhibition of IKK.

Previously, we demonstrated that benznidazole (BZL), known for its antiparasitic action on Trypanosoma cruzi, modulates pro-inflammatory cytokines and NO release in macrophages by inhibiting NF-kappaB. We now proceeded to elucidate the molecular mechanisms by which BZL exerts its inhibitory action on NF-kappaB. We demonstrated that the inhibitory effect of BZL is not extended to other macrophage responses, since it did not inhibit other typical hallmarks of macrophage activation such as phagocytosis, MHC-II molecules expression or production of reactive oxygen species (ROS) by NADPH oxidase. BZL was able to interfere specifically with the activation of NF-kappaB pathway without affecting AP-1 activation in RAW 264.7 macrophages, not only in LPS-mediated activation, but also for other stimuli, such as pro-inflammatory cytokines (IL-1beta, TNF-alpha), PMA or H(2)O(2). Also, BZL delayed the activation of p38 MAPK, but not that of ERK1/2 and JNK. Finally, treatment with BZL inhibited IkappaBalpha phosporylation and hence its degradation, whereas it did not block IkappaB kinase (IKK) alpha/beta phosphorylation. Collectively, BZL behaves as a broad range specific inhibitor of NF-kappaB activation, independently of the stimuli tested.

Chagas' disease: an update on immune mechanisms and therapeutic strategies.

* Introduction * Chagas' disease * Chemotherapy * Immune response in experimental T. cruzi infection * Immune response in human beings infected with T. cruzi * Immune response in the treatment of chagasic infection * The need for new therapeutic alternatives for Chagas' disease * Conclusions The final decade of the 20th century was marked by an alarming resurgence in infectious diseases caused by tropical parasites belonging to the kinetoplastid protozoan order. Among the pathogenic trypanosomatids, some species are of particular interest due to their medical importance. These species include the agent responsible for Chagas' disease, Trypanosoma cruzi. Approximately 8 to 10 million people are infected in the Americas, and approximately 40 million are at risk. In the present review, we discuss in detail the immune mechanisms elicited during infection by T. cruzi and the effects of chemotherapy in controlling parasite proliferation and on the host immune system.

Tumor necrosis factor alpha induced by Trypanosoma cruzi infection mediates inflammation and cell death in the liver of infected mice.

Trypanosoma cruzi (T. cruzi) infected C57BL/6 mice developed a progressive fatal disease due to an imbalance in the profile of circulating related compounds accompanying infection like tumor necrosis factor alpha (TNFalpha). TNFalpha has been proposed as an important effector molecule in apoptosis. In this work, we evaluate inflammation and the proteins involved in apoptotic process in liver of infected mice and the role of TNFalpha. C57BL6/mice were infected subcutaneously with 100 viable trypomastigotes of Tulahuén strain of T cruzi. One set of these animals were treated with 375 microg of antihuman TNFalpha blocking antibody. Animals were sacrificed at 14 days post-infection (p.i).The analyses of Bcl-2 family proteins revealed an increase of the pro-apoptotic proteins Bax and tBid in T. cruzi-infected mice. Compared with control animals, cytochrome c release was increased. Apoptosis was also induced in infected mice. Anti-TNFalpha treatment decreases hepatic apoptosis. Our results suggest that T. cruzi infection induces programmed cell death in the host liver by increase of TNFalpha production, associated with TNF-R1 over-expression, that set in motion the Bid cleavage and mitochondrial translocation, Bax mitochondrial translocation, cytochrome c release, and ultimately apoptosis induction.

Novel cytostatic activity of the trypanocidal drug Benznidazole.

We have shown that Benznidazole (BZL), a compound with well documented trypanocidal activity, possesses anti-inflammatory properties and inhibits the nuclear factor kappaB (NF-kappaB). Given the relationship between this transcription factor and cell growth, in this study we address the role of NF-kappaB blockade by BZL in the proliferation of different cell lines. Our studies demonstrate that this compound significantly reduced proliferation of RAW 264.7 macrophage cell line, as assessed by trypan blue exclusion, MTT reduction and [(3)H]-thymidine incorporation, at a concentration shown to inhibit NF-kappaB. Treatment with BZL also led to growth arrest in CHO, MDCK and HeLa cells. Interestingly, growth inhibition was found to be a reversible process, not accompanied by significant cell death, indicating that the drug behaves mainly as a cytostatic compound. As this effect might be related to NF-kappaB inhibition, we next evaluated whether other NF-kappaB inhibitors could induce growth arrest in RAW 264.7 and HeLa cells. We found that IKK inhibition led to growth arrest in both cell lines, indicating that NF-kappaB inhibition may be the potential mechanism by which BZL inhibits cell proliferation. To the best of our knowledge, this is the first report of an anti-proliferative activity of the trypanocidal drug against different cell lines and provides a mechanistic insight that may help understand some of the adverse effects associated with prolonged treatment.

Beneficial effects of benznidazole during an infectious-based situation of systemic inflammatory response: cecal ligation and puncture.

We have shown that benznidazole (BZL), a drug used to treat Chagas disease, markedly reduced the production of pro-inflammatory cytokines and NO-derived metabolites in experimentally Trypanosoma cruzi-infected rats. Treatment with BZL exerted beneficial effects in a model of inflammation-based pathology like murine experimental endotoxemia. Based on these findings, we wished to ascertain the effect of BZL in a closer situation to sepsis: the cecal ligation and puncture (CLP) model in C57BL/6 mice. We analyzed clinical course, survival, circulating levels of inflammation-related compounds (NO, tumor necrosis factor [TNF]-alpha), and bacteriemia. Recipients of BZL, 25 mg/kg, had an increased survival rate at 24 hours after CLP, showing a better clinical situation and a significant reduction of TNF-alpha levels and bacteriemia, with respect to the other groups. BZL failed to inhibit in vitro bacterial growth, suggesting that these effects may be partly caused by the immunomodulatory effects of BZL.

Immunization with an engineered mutant trans-sialidase highly protects mice from experimental Trypanosoma cruzi infection: a vaccine candidate.

Chagas' disease is a major tropical disease for which a cure for chronic phase does not exist yet. Trypanosoma cruzi trans-sialidase (TS) seems to be involved in relevant processes such as infectivity, host survival and, very importantly, disease pathogenesis. In this study, we show that mice vaccinated with an engineered enzymatically deficient mutant TS containing the catalytic domain without the immunodominant SAPA (Shed Acute Phase Antigen) repeats, were highly protected against T. cruzi infection. Adult male BALB/c mice were immunized with mutant protein, purified from Pichia pastoris yeast, using three inoculations in Freund's adjuvant. All immunized mice were protected against challenge with a lethal dose of T. cruzi trypomastigotes. The protected immunized mice developed no clinical or tissue evidence of infection throughout the study. In contrast, 60-90% mortality and 100% occurrence of myocardial lesions were observed in the non-immunized counterparts. Titers of circulating antibody against TS did not correlate with protection, while anti-SAPA antibodies were coincident with disease severity. Further studies indicated that a single inoculation of mutant recombinant protein in Freund's complete adjuvant was not associated with blood or organic alterations, per se. Mutant TS vaccination seems to be a promising tool for immune intervention strategies in Chagas' disease, aimed at preventing T. cruzi-related heart tissue damage.

Benznidazole, a drug used in Chagas' disease, ameliorates LPS-induced inflammatory response in mice.

Benznidazole (BZL) is a drug currently used for treating Chagas' disease. Given our earlier demonstration in which BZL downregulated cytokine and nitric oxide (NO) synthesis by LPS and/or IFN-gamma-stimulated murine macrophages, we have now analysed whether this compound could exert beneficial effects in a model of LPS-induced inflammation in C57BL/6 mice. The lethal model consisted of two LPS intraperitoneal injections, 200 microg each separated by 2 h, with BZL given orally at a dose of 200 mg/kg, 18 and 2 h before the first challenge and 20 and 44 hr following the second one. In this model, BZL treatment led to a significantly decreased mortality in comparison with untreated counterparts. Remaining experiments were carried out in mice given a unique LPS dose, pretreated with BZL or not, since those subjected to the lethal protocol were unsuitable for laboratory handling. Analysis of IL-1beta, IL-6, TNF-alpha, IL-12 and iNOS mRNA expression in liver samples taken at 90 min post-LPS showed a marked reduction of the two latter mRNAs in BZL-treated mice. These animals also displayed significantly decreased peaks levels of serum TNF-alpha and IL-6, accompanied by a diminished number of IL-6-producing peritoneal macrophages. Present effects may broaden the potential usefulness of BZL in situations accompanied by an excessive inflammatory response.

Thymocyte depletion during acute Trypanosoma cruzi infection in C57BL/6 mice is partly reverted by lipopolysaccharide pretreatment.

Infection with Trypanosoma cruzi in C57BL/6 mice leads to a progressive fatal disease accompanied by thymocyte depletion, which is not related with a higher parasite burden but with increased serum levels of tumour necrosis factor alpha (TNF- alpha). Because this situation may result from an excessive inflammatory syndrome, mice were now given anti-TNF-alpha mAbs throughout their acute infection, or subjected to a LPS desensitization protocol before parasite challenge. Treatment with anti-TNF-alpha mAbs failed to ameliorate thymocyte depletion but shortened survival time and increased parasite load. Pretreatment with LPS (desensitization followed by a sublethal LPS dose) prolonged survival time with a trend to reduce parasitemias and TNF-alpha serum concentrations. Given that pentoxifylline (PTx) interferes with in vitro LPS tolerance, experiments by administering PTx in combination with the tolerance-inducing LPS doses were also performed. Such schedule significantly reduced mortality, TNF-alpha and IL-6 serum concentrations, and CD4+ CD8+ thymocyte loss. LPS pretreatment allowed a better infection control and protected from the accompanying tissue damage.

Evaluation of an attenuated Trypanosoma cruzi strain in rats: analysis of survival, parasitemia and tissue damage

El objetivo de este trabajo fue detectar infección y daño tisular en una línea endocriada de ratas, inducidos por formas vivas de una cepa de T. cruzi cultivada in vitro (TCC). Se inocularon: a) ratas cultivadas in vitro (TCC). Se inocularon: a) ratas lactantes (S), de 3-5 días de edad por vía i.p. con 10**6 TCC (S1), 10**7 TCC (S2) y 10**8 TCC (S3); b) ratas al destete, de 21-25 días de edad por vía s.c. con 10**6 TCC (W1), 10**7 TCC (W2) y 10**8 TCC (W3). Los cultivos tenían como máximo un 2// de trypomastigotes; c) testigos por vía i.p. o s.c. con 10**6 trypomastigotes sanguiíneos de la cepa Tulahuén (SC y WC); se incluyeron testigos normales. La sobrevida fue del 100// en S1, S2 y S3 y 0// en SC en el día 13 post-infección (p.i.). Estos animales murieron con signos de enfermedad de Chagas aguda. La sobrevida fue del 100// en el grupo W. En los primeros 30 días p.i. los parásitos fueron detectados en los grupos S1, S2, S3 y W1, W2 y W3 por búsqueda exhaustiva. Los parásitos se encontraron rápidamente en los testigos SC y WC hasta el día 13. Los xenodiagnósticos fueron positivos (5/5) a los 2 meses p.i. y negativos a los 6 meses p.i. (W1, W2, W3, 0/23; WC, 0/5) Tabla 1). Se observó un leve aumento de la frecuencia de miocarditis crónica focal en algunos animales infectados, independientemente a la dosis (S1, S2, S3, 26//; W1, W2, W3, 46//) que no fue significativo en comparación con la de los testigos y no alcanza el porcentaje de lesión obtenida habitualmente en WC (61,3//). La menor virulencia y patogenicidad observadas en las ratas, especie no estudiada hasta ahora, sugieren que la cepa TCC tiene una importante atenuación después de largo tiempo de cultivo in vitro

Evaluation of an attenuated Trypanosoma cruzi strain in rats: analysis of survival, parasitemia and tissue damage

El objetivo de este trabajo fue detectar infección y daño tisular en una línea endocriada de ratas, inducidos por formas vivas de una cepa de T. cruzi cultivada in vitro (TCC). Se inocularon: a) ratas cultivadas in vitro (TCC). Se inocularon: a) ratas lactantes (S), de 3-5 días de edad por vía i.p. con 10**6 TCC (S1), 10**7 TCC (S2) y 10**8 TCC (S3); b) ratas al destete, de 21-25 días de edad por vía s.c. con 10**6 TCC (W1), 10**7 TCC (W2) y 10**8 TCC (W3). Los cultivos tenían como máximo un 2// de trypomastigotes; c) testigos por vía i.p. o s.c. con 10**6 trypomastigotes sanguiíneos de la cepa Tulahuén (SC y WC); se incluyeron testigos normales. La sobrevida fue del 100// en S1, S2 y S3 y 0// en SC en el día 13 post-infección (p.i.). Estos animales murieron con signos de enfermedad de Chagas aguda. La sobrevida fue del 100// en el grupo W. En los primeros 30 días p.i. los parásitos fueron detectados en los grupos S1, S2, S3 y W1, W2 y W3 por búsqueda exhaustiva. Los parásitos se encontraron rápidamente en los testigos SC y WC hasta el día 13. Los xenodiagnósticos fueron positivos (5/5) a los 2 meses p.i. y negativos a los 6 meses p.i. (W1, W2, W3, 0/23; WC, 0/5) Tabla 1). Se observó un leve aumento de la frecuencia de miocarditis crónica focal en algunos animales infectados, independientemente a la dosis (S1, S2, S3, 26//; W1, W2, W3, 46//) que no fue significativo en comparación con la de los testigos y no alcanza el porcentaje de lesión obtenida habitualmente en WC (61,3//). La menor virulencia y patogenicidad observadas en las ratas, especie no estudiada hasta ahora, sugieren que la cepa TCC tiene una importante atenuación después de largo tiempo de cultivo in vitro (AU)

Influencia de la edad de la rata en la evolución de la infección com trypanosoma cruzi / Influence of the age of the rat on the course of infection with trypanosoma cruzi

En la primera parte de este trabajo se estudió en la fase aguda, la parasitemia y respuesta inmune humoral específica, en ratas de línea "1", inoculadas al destete con 1x10**6 T. cruzi vivos de la cepa Tulahuén (GD), y adultos jóvenes con 1x10**6 T. cruzi (GA) y con 7x10**6 T. cruzi, cantidad equivalente a la inoculada en los destetes según peso (GA-1). La parasitemia fue significativamente mayor en el GD que en los GA y GA-1. Por otro lado, el GD presentó bajos títulos de anticuerpos, en relación a los adultos con ambas concentraciones de T. cruzi, que tuvieron un comportamiento similar. En el período crónico se evaluaron, a los 180 días post-infección, los ECG y la histolatología de corazón en animales al destete y adultos, con sólo 1x10**6 T. cruzi. La frecuencia cardíaca en los animales infectados fue mayor que la de los testigos. La proporción de lesión cardíaca (miocarditis, pericarditis y/o fibrosis) fue superior en el GD y en el GA, con respecto al GT, y el GA presentó un 35,6% de fibrosis, significativamente diferente de los otros grupos. La lesión fue sobre todo leve, ventricular y no se visualizaron parásitos in situ. En síntesis, en la rata, en la fase aguda de la infección, se encontraron diferencias marcadas en serología y parasitemia según la edad del huésped; sin embargo, la patología miocárdica crónica fue similar, con la excepción de que 1/3 de las ratas adultas inoculadas presentaron fibrosis

Influencia de la edad de la rata en la evolución de la infección com trypanosoma cruzi / Influence of the age of the rat on the course of infection with trypanosoma cruzi

En la primera parte de este trabajo se estudió en la fase aguda, la parasitemia y respuesta inmune humoral específica, en ratas de línea "1", inoculadas al destete con 1x10**6 T. cruzi vivos de la cepa Tulahuén (GD), y adultos jóvenes con 1x10**6 T. cruzi (GA) y con 7x10**6 T. cruzi, cantidad equivalente a la inoculada en los destetes según peso (GA-1). La parasitemia fue significativamente mayor en el GD que en los GA y GA-1. Por otro lado, el GD presentó bajos títulos de anticuerpos, en relación a los adultos con ambas concentraciones de T. cruzi, que tuvieron un comportamiento similar. En el período crónico se evaluaron, a los 180 días post-infección, los ECG y la histolatología de corazón en animales al destete y adultos, con sólo 1x10**6 T. cruzi. La frecuencia cardíaca en los animales infectados fue mayor que la de los testigos. La proporción de lesión cardíaca (miocarditis, pericarditis y/o fibrosis) fue superior en el GD y en el GA, con respecto al GT, y el GA presentó un 35,6% de fibrosis, significativamente diferente de los otros grupos. La lesión fue sobre todo leve, ventricular y no se visualizaron parásitos in situ. En síntesis, en la rata, en la fase aguda de la infección, se encontraron diferencias marcadas en serología y parasitemia según la edad del huésped; sin embargo, la patología miocárdica crónica fue similar, con la excepción de que 1/3 de las ratas adultas inoculadas presentaron fibrosis (AU)

La enfermedad del adyuvante en ratas infectadas experimentalmente con Trypanosoma cruzi / Adjuvant disease in rats experimentally infected with Trypanosoma cruzi

Se estudió la evolución de la artritis por adyuvante en ratas que habían sido infectadas previamente con Trypanosoma cruzi, con el objeto de evaluar su competencia inmunológica a través de la respuesta artrítica. La artritis por adyuvante se indujo en ratas adultas, endocriadas de ambos sexos, con 0.1 ml de adyuvante completo de Freund en la almohadilla plantar, en 2 lotes: a) inyectadas 90 días antes con 1 x 10***6 T. cruzi y b) testigos normales simultáneos. Se midieron, la lesión artrítica macroscópicamente con una escala semicuantitativa, y con microscopía óptica la histopatología de la lesión local y la del corazón, a los 180 días post-infección. La magnitud de las lesiones artríticas en las ratas con T. cruzi fue significativamente menor (p < 0.001) que la de los testigos, en todo el período. El infiltrado inflamatorio local, formado por linfocitos, plasmocitos y macrófagos fue significativamente menor (p < 0.001) en las ratas chagásicas, con respecto al de los testigos. Se postula que en las ratas que recibieron T. cruzi la respuesta artrítica menor podría deberse a una competición antigénica con los determinantes del parásito o a mecanismos inmunosupresores que interfieren en la producción de la entidad experimental

Efecto de extractos ricos en ARN inmune sobre algunos parámetros inmunopatológicos de ratas infectadas con Trypanosoma cruzi / Effect of extracts rich in immune RNA on some immunopathological parameters of rats infected with Trypanosoma cruzi

Se estudió el efecto de una inyección única de un extracto rico en ARN inmune (Ext-ARNi) (4 mg/100 g peso), en ratas inoculadas con T. cruzi viables, realizada 5 días antes que el desafío con los parásitos. Se midieron parasitemia en la fase aguda, nivel y cinética de anticuerpos en forma crónica; ECG y estudio histopatológico de corazón por microscopía óptica &P, a los 6 meses de evolución. No se visualizaron cambios en la parasitemia, el nivel de anticuerpos específicos mostró diferenciass puntuales y el ECG no se modificó con la presencia del Ext-ARNi. Sólo el porcentaje de lesión cardíaca (miocarditis, pericarditis y/o fibrosis) se redujo significativamente en las ratas que recibieron Ext-ARNi y T. cruzi (17,6%), en comparación con el grupo desafiado con T. cruzi (66,7%), y sin diferencias con los controles de Ext-ARNi (34,4%), y testigos (15,6%). Se postula que el extracto rico en ARN inmune podría modificar la "calidad" de los anticuerpos que se forman, los que protegerían al parásito haciéndolo menos inmunogénico provocando, consecuentemente, menos lesión cardíaca. No se descarta la participación de componentes celulares u otros efectos en la fisiopatología de este mecanismo