Congenital transmission of Mexican strains of Trypanosoma cruzi TcIa: interaction between parasite and human placental explants.

Congenital transmission of Chagas disease plays an important role in endemic countries because it is not a diagnosis that is encountered frequently in prenatal care. Due to limited information regarding congenital transmission of Trypanosoma cruzi in Mexico, the present study aimed to investigate protozoan infectivity and modulation of immune responses in human placental explants infected with T. cruzi Ia Mexican strains. The Inc-5 strain showed increased infectivity and modulated IL-1ß, IL-10 and TLR-4, decreasing their expression after 24 h of infection. Both strains (Inc-5 and Ninoa) stimulated the production of TNF-α and decreased IL-6 levels 96 h after infection. An important detachment of the syncytiotrophoblast caused by infection with T. cruzi was observed after 24 h of infection. In this study, ex vivo infection of human placental villi was performed to better understand interactions involving parasitic T. cruzi and human placental tissue. It was concluded that the strains of TcIa present parasitism in placental tissue, modulation of the innate immune system of the placenta, and cause intense detachment of the syncytiotrophoblast, a fact that may be more associated with abortion and premature birth events than the congenital transmission itself, justifying the low rate of this transmission mechanism by this genotype.

Analysis of regulatory T cells and CTLA-4 expression in pregnant women according to seropositivity to Toxoplasma gondii.

Pregnancy is considered a period in which immunomodulation occurs, although it is important for the maintenance of the foetus, could contribute to infections as Toxoplasma gondii. Immune response cells such as regulatory T cells participate in this immunomodulation, and surface molecules such as CTLA-4 develop an immunosuppressive role, could contribute to the establishment of the parasite. This study aimed to evaluate the presence of regulatory T cells and the expression of CTLA-4 in parturient and non-pregnant seropositive and seronegative for anti-T. gondii antibodies. Sixty-two participants were evaluated, 14 parturient with negative serology, 23 parturient with positive serology, 16 non-pregnant women seronegative and 9 non-pregnant women seropositive. Immunophenotyping was performed for characterize TCD4+Foxp3+ cells, T CD4+CD25-Foxp3+, TCD4+CD25highFoxp3+, TCD4+CTLA-4+, TCD4+CD25-CTLA-4+ and TCD4+CD25highCTLA-4+. We observed a lower level of CD4+CD25highFoxp3+ cells from seropositive parturient compared with seropositive non-pregnant cells. Significative levels of CD4+CD25-Foxp3+ cells from seronegative pregnant were observed compared with seropositive pregnant cells. Furthermore, the higher level of CD4+CD25-CTLA-4+ cells populations was detected in seropositive pregnant cells compared with seropositive non-pregnant. Although a significant increase in CTLA-4 cells was observed in pregnant women positive for anti-T. gondii antibodies, this increase did not cause a risk of reactivation of the infection.

Identification of T3 and T4 Genotypes of Acanthamoeba sp. in Dust Samples Isolated from Air Conditioning Equipment of Public Hospital of Ituiutaba-MG.

The free-living amoebae are currently considered an emerging parasitic infection. The infection by this protozoan can generate serious infection and even cause death. Due to their amphizoic and opportunistic characters of these microorganisms, one should give more attention, not only in swimming pools but also where there are immunologically susceptible patients such as those found in intensive care units and surgical centers. Due to their difficult diagnosis often postmortem, because they are considered to be an emerging parasitic infection and their diagnosis is difficult, often performed post mortem. This study aimed to evaluate the safety of these protozoa in air conditioners by taking samples of dust from both the surgical center and the intensive therapy unit. We analyzed 48 dust samples that were collected from six air conditioners equipment located in the Intensive Care Unit (ICU) and Surgical Center (SC) of a public hospital. We found 10.4% of the samples collected in the SC, and 75% of the samples collected in the ICU presented free-living amoeba cysts by light microscopy analysis. In total, 35.4% (17/48) of the air conditioning samples of the hospital were positive and, by PCR, were identified to belong to the genus Acanthamoeba spp and Balamuthia mandrillaris species. By DNA sequencing analysis, it was possible to classify the Acanthamoeba samples as belonging to the T3 and T4 genotypes. These genotypes are the main cause of keratitis in humans, and Balamuthia may cause amoebic encephalitis, and together are emerging parasitic infections. Our results show the presence of the two most important amoebas Acanthamoeba (T3 and T4 genotypes) and Balamuthia in the SC and the ICU, and these necessary precautions these sites could be propagating cysts of these amoebas and patients during their stay or discharge could present ocular and NSC alterations without perhaps arriving to the diagnosis of free-living amoeba infection.

Evaluation of the multispecies coalescent method to explore intra-Trypanosoma cruzi I relationships and genetic diversity.

Chagas Disease is a zoonosis caused by the parasite Trypanosoma cruzi. Several high-resolution markers have subdivided T. cruzi taxon into at least seven lineages or Discrete Typing Units (DTUs) (TcI-TcVI and TcBat). Trypanosoma cruzi I is the most diverse and geographically widespread DTU. Recently a TcI genotype related to domestic cycles was proposed and named as TcIDOM. Herein, we combined traditional markers and housekeeping genes and applied a Multispecies Coalescent method to explore intra-TcI relationships, lineage boundaries and genetic diversity in a random set of isolates and DNA sequences retrieved from Genbank from different countries in the Americas. We found further evidence supporting TcIDOM as an independent and emerging genotype of TcI at least in Colombia and Venezuela. We also found evidence of high phylogenetic incongruence between parasite's gene trees (including introgression) and embedded species trees, and a lack of genetic structure among geography and hosts, illustrating the complex dynamics and epidemiology of TcI across the Americas. These findings provide novel insights into T. cruzi systematics and epidemiology and support the need to assess parasite diversity and lineage boundaries through hypothesis testing using different approaches to those traditionally employed, including the Bayesian Multispecies coalescent method.

Oral infection of mice and host cell invasion by Trypanosoma cruzi strains from Mexico.

Oral infection by Trypanosoma cruzi has been responsible for frequent outbreaks of acute Chagas disease in the north of South America and in the Amazon region, where T. cruzi genetic group TcI predominates. TcI strains from different geographical regions have been used in oral infection in mice, but there is no information about strains from Mexico where TcI is prevalent. Here, we analyzed four Mexican strains as concerns the course of oral infection, the ability to invade host cells in vitro, and the profile of metacyclic trypomastigote surface molecules gp82 and gp90 that are implicated in parasite internalization. Oral infection of mice with metacyclic forms of all strains resulted in reduced blood and tissue parasitism, and mild to moderate inflammatory process in the heart/skeletal muscle. They expressed pepsin-resistant gp82 and gp90 molecules at high levels and invaded host cells poorly in full nutrient medium and efficiently under nutrient-deprived condition. The properties exhibited by Mexican strains were similar to those displayed by TcI strains from other geographical regions, reinforcing the notion that these features are common to the genetic group TcI as a whole.

Leishmania infection in bats from a non-endemic region of Leishmaniasis in Brazil.

Leishmaniasis is a complex of zoonotic diseases caused by parasites of the genus Leishmania, which can develop in domestic as well as wild animals and humans throughout the world. Currently, this disease is spreading in rural and urban areas of non-endemic regions in Brazil. Recently, bats have gained epidemiological significance in leishmaniasis due to its close relationship with human settlements. In this study, we investigated the presence of Leishmania spp. DNA in blood samples from 448 bats belonging to four families representing 20 species that were captured in the Triangulo Mineiro and Alto Paranaiba areas of Minas Gerais State (non-endemic areas for leishmaniasis), Brazil. Leishmania spp. DNA was detected in 8·0% of the blood samples, 41·6% of which were Leishmania infantum, 38·9% Leishmania amazonensis and 19·4% Leishmania braziliensis. No positive correlation was found between Leishmania spp. and bat food source. The species with more infection rates were the insectivorous bats Eumops perotis; 22·2% (4/18) of which tested positive for Leishmania DNA. The presence of Leishmania in the bat blood samples, as observed in this study, represents epidemiological importance due to the absence of Leishmaniasis cases in the region.

IL-17-expressing CD4⁺ and CD8⁺ T lymphocytes in human toxoplasmosis.

This study aimed to measure the synthesis of Th1 and Th2 cytokines by mononuclear cells after culture with live T. gondii and identified Th17 (CD4(+)) and Tc17 (CD8(+)) cells in toxoplasma-seronegative and toxoplasma-seropositive parturient and nonpregnant women. Cytometric bead arrays were used to measure cytokine levels (IL-2, TNF-α, IFN-γ, IL-4, IL-5, and IL-10); immunophenotyping was used to characterize Th17 and Tc17 cells, and the cells were stained with antibodies against CD4(+) and CD8(+) T cells expressing IL-17. The addition of tachyzoites to cell cultures induced the synthesis of IL-5, IL-10, and TNF-α by cells from seronegative parturient women and of IL-5 and IL-10 by cells from seropositive, nonpregnant women. We observed a lower level of IL-17-expressing CD4(+) and CD8(+) T lymphocytes in cultures of cells from seronegative and seropositive parturient and nonpregnant women that were stimulated with tachyzoites, whereas analysis of the CD4(+) and CD8(+) T cell populations showed a higher level of CD4(+) T cells compared with CD8(+) T cells. These results suggest that the cytokine pattern and IL-17-expressing CD4(+) and CD8(+) T lymphocytes may have important roles in the inflammatory response to T. gondii, thus contributing to the maintenance of pregnancy and control of parasite invasion and replication.

Effects of Meglumine Antimoniate Treatment on Cytokine Production in a Patient with Mucosal Leishmaniasis and Chagas Diseases Co-Infection.

The influence of antimoniate treatment on specific anti-protozoan T-cell responses was evaluated in a 48-year-old male patient diagnosed with mucosal leishmaniasis and Chagas disease infection. Before and after treatment, PBMC (peripheral blood mononuclear cells) were cultured in the absence or presence of Leishmania braziliensis or Trypanosoma cruzi live parasites, their soluble antigens, or PHA (phytohaemagglutinin). Cytokines were measured and Treg (T regulatory) cell percentages were quantified. Before treatment, PBMC were able to produce higher amounts of TNF-α, IL-6 (Interleukin-6), and IL-10 (Interleukin-10) but lower amounts of IL-12 (Interleukin-12) in response to culture stimulation. However, after treatment, there was a down-modulation of TNF-α, IL-6, and IL-10 cytokines but an up-modulation in IL-12 production. PBMC had the ability to produce TNF-α only against live parasites or PHA. There was an overall decrease of circulating Treg cells after treatment. In mixed Leishmaniasis and Chagas disease infection, treatment with antimoniate could modulate immune responses toward a more protective profile to both diseases.

[Prevalence of triatomines (Hemíptera: Reduviidae: Triatominae) infected by Trypanosoma cruzi: seasonality and distribution in the Ciénega region of the State of Jalisco, Mexico]. / Prevalência de triatomíneos (Hemíptera: Reduviidae: Triatominae) infectados por Trypanosoma cruzi: sazonalidade e distribuição na região Ciénega do Estado de Jalisco, México.

The physical and geographical characteristics of the Ciénega region, Jalisco, Mexico make it suitable for transmission of Trypanosoma cruzi, the causative agent for Chagas disease. This study characterizes the prevalence of triatomines infected by this parasite, their seasonality and their distribution in this region. A total of 328 triatomines were evaluated between January 2005 and June 2007, from 13 municipalities in the region. April, May and June were the months with the highest capture levels. Among the triatomines examined, 57.3% were positive for Trypanosoma cruzi, corresponding to 15.4% in urban areas and 84.6% in rural areas. The species with greatest prevalence was Triatoma longipennis and the species with the highest parasitism rate was Triatoma barberi, with an infection rate of 83.3%, whereas the rate for Triatoma longipennis was 67.5% (p<0.05). This natural infection in the captured vectors may indicate that individuals in this region have high exposure to Trypanosoma cruzi. The recent findings of positive Triatoma dimidiata in this region suggest that new species are becoming adapted to the ecological conditions of these populations.

Toxoplasma gondii Infection Promotes Epithelial Barrier Dysfunction of Caco-2 Cells.

After oral infection, Toxoplasma gondii invades intestinal cells, induces breakdown of intestinal physiology and barrier functions, and causes intestinal pathology in some animal species. Although parasites' invasion into host cells is a known phenomenon, the effects of T. gondii infection in the intestinal barrier are still not well established. To evaluate morphological and physiological modifications on the colorectal adenocarcinoma-derived Caco-2 cell line during T. gondii infection, microvilli, tight junction integrity, and transepithelial electrical resistance (TEER) were investigated under infection. It was observed that the dextran uptake (endocytosis) and distribution were smaller in infected than in noninfected Caco-2 cells. The infection leads to the partial loss of microvilli at the cell surface. Claudin-1, zonula occludens-1 (ZO-1), and occludin expressions were colocalized by immunofluorescence and presented discontinuous net patterns in infected cells. Immunoblotting analysis at 24 hr postinfection revealed decreasing expression of occludin and ZO-1 proteins, whereas claudin-1 presented similar expression level compared with noninfected cells. T. gondii decreased TEER in Caco-2 cells 24 hr after infection. Our results suggest that T. gondii infection may lead to the loss of integrity of intestinal mucosa, resulting in impaired barrier function.

PREVALENCE OF CHAGAS DISEASE AMONG BLOOD DONOR CANDIDATES IN TRIANGULO MINEIRO, MINAS GERAIS STATE, BRAZIL.

Despite public health campaigns and epidemiological surveillance activities, Chagas disease remains a major health problem in Latin America. According to data from the World Health Organization, there are approximately 7-8 million people infected with Trypanosoma cruzi worldwide, a large percentage of which in Latin America. This study aims to examine the serological profile of blood donors in blood banks of Hemominas hematology center, in the town of Ituiutaba, Minas Gerais State, Brazil. The study sample consisted of 53,941 blood donors, which were grouped according to gender and age. Sample collections were performed from January 1991 to December 2011, and 277 donors (0.5%) were considered serologically ineligible due to Chagas disease. Analysis of data showed no significant difference between genders. As for age, the highest proportion of ineligible donors was from 40 to 49 years (30%), and there was a positive correlation between increasing age and the percentage of patients seropositive for Chagas disease. Therefore, adopting strategies that allow the safe identification of donors with positive serology for Chagas disease is essential to reduce or eliminate indeterminate serological results.

Parasite detection in sand from bays on the north coast of São Paulo state, Brazil

Soil contamination by protozoan parasites and soil-transmitted helminths (STH) is common in beach sand due to a number of factors such as pets, pluvial water, garbage, etc. These pathogens may cause many diseases in humans and animals and become a public health problem. Thus, systematic evaluation and inspection are necessary to develop control strategies regarding public contamination. For this purpose, our aims were to evaluate the parasitic profile of sandy soils on an urban beach and an untouched beach and correlate this with environmental and seasonal characteristics in Ubatuba, Brazil, in two seasons (winter and summer). 132 soil samples were collected for parasite analysis utilizing Rugai's method and the sedimentation adapted method. Our results showed positivity in 62% of the samples for at least one parasite in the urban beach and no parasitic structures on the untouched beach. The positivity was higher in summer (85%) than in winter (51.7%). All samples were positive for both, helminths and protozoa. Seasonal influence was noted regarding the presence of STH while for protozoa this influence was not observed. The parasitic structures most found were larvae of hookworms (35%) and Toxocara eggs (31.7%). We also noted the presence of Strongyloides sp, Ascaris lumbricoides, coccidia, Dipylidium caninum, Entamoeba hystolitica/dispar, Endolimax nana, Entamoeba coli, Giardia sp, Toxascaris leonina, Trichuris sp and Dibothricephalus latus. There was a positive correlation between temperature and the presence of STH in the sand samples. Most of the collection sites on the urban beach presented dogs or canine traces and garbage in both seasons. There was an association between the presence of dogs or their traces and parasitic structures. In conclusion, seasonality, urbanization and the presence of pets on beaches can potentially favor environmental contamination by parasites increasing the risk of transmission of zoonotic and parasitic diseases.

Production of cytokine and chemokines by human mononuclear cells and whole blood cells after infection with Trypanosoma cruzi.

INTRODUCTION: The innate immune response is the first mechanism of protection against Trypanosoma cruzi, and the interaction of inflammatory cells with parasite molecules may activate this response and modulate the adaptive immune system. This study aimed to analyze the levels of cytokines and chemokines synthesized by the whole blood cells (WBC) and peripheral blood mononuclear cells (PBMC) of individuals seronegative for Chagas disease after interaction with live T. cruzi trypomastigotes. METHODS: IL-12, IL-10, TNF-α, TGF-ß, CCL-5, CCL-2, CCL-3, and CXCL-9 were measured by ELISA. Nitrite was determined by the Griess method. RESULTS: IL-10 was produced at high levels by WBC compared with PBMC, even after incubation with live trypomastigotes. Production of TNF-α by both PBMC and WBC was significantly higher after stimulation with trypomastigotes. Only PBMC produced significantly higher levels of IL-12 after parasite stimulation. Stimulation of cultures with trypomastigotes induced an increase of CXCL-9 levels produced by WBC. Nitrite levels produced by PBMC increased after the addition of parasites to the culture. CONCLUSIONS: Surface molecules of T. cruzi may induce the production of cytokines and chemokines by cells of the innate immune system through the activation of specific receptors not evaluated in this experiment. The ability to induce IL-12 and TNF-α contributes to shift the adaptive response towards a Th1 profile.

Molecular characterization of Trypanosoma cruzi Mexican strains and their behavior in the mouse experimental model.

INTRODUCTION: For a long time, the importance of Chagas disease in Mexico, where many regarded it as an exotic malady, was questioned. Considering the great genetic diversity among isolates of Trypanosoma cruzi, the importance of this biological characterization, and the paucity of information on the clinical and biological aspects of Chagas disease in Mexico, this study aimed to identify the molecular and biological characterization of Trypanosoma cruzi isolates from different endemic areas of this country, especially of the State of Jalisco. METHODS: Eight Mexican Trypanosoma cruzi strains were biologically and genetically characterized (PCR specific for Trypanosoma cruzi, multiplex-PCR, amplification of space no transcript of the genes of the mini-exon, amplification of polymorphic regions of the mini-exon, classification by amplification of intergenic regions of the spliced leader genes, RAPD (random amplified polymorphic DNA). RESULTS: Two profiles of parasitaemia were observed, patent (peak parasitaemia of 4.6×10(6) to 10(7) parasites/mL) and subpatent. In addition, all isolates were able to infect 100% of the animals. The isolates mainly displayed tropism for striated (cardiac and skeletal) muscle. PCR amplification of the mini-exon gene classified the eight strains as TcI. The RAPD technique revealed intraspecies variation among isolates, distinguishing strains isolated from humans and triatomines and according to geographic origin. CONCLUSIONS: The Mexican T. cruzi strains are myotrophic and belong to group TcI.

PREVALENCE OF CHAGAS DISEASE AMONG BLOOD DONOR CANDIDATES IN TRIANGULO MINEIRO, MINAS GERAIS STATE, BRAZIL / Prevalência de casos de doença de Chagas em candidatos a doação de sangue na região do Pontal do Triângulo Mineiro, Brasil

Despite public health campaigns and epidemiological surveillance activities, Chagas disease remains a major health problem in Latin America. According to data from the World Health Organization, there are approximately 7-8 million people infected with Trypanosoma cruzi worldwide, a large percentage of which in Latin America. This study aims to examine the serological profile of blood donors in blood banks of Hemominas hematology center, in the town of Ituiutaba, Minas Gerais State, Brazil. The study sample consisted of 53,941 blood donors, which were grouped according to gender and age. Sample collections were performed from January 1991 to December 2011, and 277 donors (0.5%) were considered serologically ineligible due to Chagas disease. Analysis of data showed no significant difference between genders. As for age, the highest proportion of ineligible donors was from 40 to 49 years (30%), and there was a positive correlation between increasing age and the percentage of patients seropositive for Chagas disease. Therefore, adopting strategies that allow the safe identification of donors with positive serology for Chagas disease is essential to reduce or eliminate indeterminate serological results.

A doença de Chagas, apesar das campanhas de saúde pública e das ações de vigilância epidemiológica, ainda constitui-se um sério problema de saúde na América Latina. De acordo com dados divulgados pela Organização Mundial de Saúde, existem cerca de 7 a 8 milhões de pessoas infectadas com Trypanosoma cruzi em todo o mundo, principalmente na América Latina. Este estudo tem por objetivo analisar o perfil sorológico de doadores de sangue dos bancos de sangue do Hemominas de Ituiutaba, Minas Gerais. Os doadores também foram separados de acordo com o sexo e a idade. A amostra do estudo foi composta por 53.941 doadores de sangue durante o período de janeiro de 2001 a dezembro de 2011. Duzentos e setenta e sete doações (0,5%) foram considerados sorologicamente inaptas para a doença de Chagas. Quanto à idade, a maior proporção de doadores impróprios foi de 40 a 49 anos (30%). Os dados não revelaram diferença significativa entre os sexos (p < 0,05). Houve correlação positiva entre o aumento da idade e o percentual de pacientes soropositivos para doença de Chagas. É imprescindível a adoção de estratégias que permitam a identificação segura de um doador com sorologia positiva para doença de Chagas, tentando assim minimizar ou eliminar resultados sorológicos indeterminados.

Frequência de enteroparasitos em amostras de fezes de cães em um município do pontal do Triângulo Mineiro, Minas Gerais, Brasil / Frequency of intestinal parasites in dog stool samples in a town of the Pontal do Triângulo region, Minas Gerais state, Brazil

Os cães são considerados importantes reservatórios de parasitos intestinais com potencial zoonótico associado à íntima convivência com o ser humano. O objetivo deste trabalho foi determinar a frequência de cistos de Giardia sp. e oocistos de coccídeos em amostras de fezes de cães comercializados em pet shops e daqueles recolhidos pelo Centro de Controle de Zoonoses (CCZ) do município de Ituiutaba, Minas Gerais. Foram coletadas 80 amostras de fezes de cães de ambos os sexos (40 porcento filhotes e 60 porcento adultos) em canis individuais e coletivos. Os cistos foram isolados das amostras utilizando-se solução de Sheather e os oocistos, segundo o protocolo de Ortega-Mora modificado (1999). Verificou-se que 34 (42,5 porcento) amostras apresentaram positividade para cistos de Giardia sp., sendo 26 (51,0 porcento) de cães do CCZ e 8 (27,6 porcento) dos pet shops. Quanto à presença de oocistos nas amostras de fezes, foi observada a frequência de 13,7 porcento (7) nos cães oriundos do CCZ e de 34,5 porcento (10) naqueles de pet shops. A elevada frequência de cistos de Giardia sp. e de oocistos de coccídeos nas amostras de fezes de animais de ambos os locais pode estar relacionada à exposição dos cães às formas evolutivas dos parasitos e à ausência de tratamento antiparasitário. A aglomeração de animais em canis/pet shops favorece o risco de transmissão de parasitos intestinais com elevado potencial zoonótico para os animais e pessoas que com eles convivem...

Dogs are considered important reservoirs of intestinal parasites due to their coprophagic behavior and close interaction with humans. The objective of this study was to determine the prevalence of Giardia spp. cysts and of coccidia oocysts in dog stool samples collected at the Zoonosis Control Center (CCZ) and at pet shops in Ituiutaba town, in the state of Minas Gerais, Brazil. Eighty stool samples (40 percent puppies and 60 percent adult dogs) were collected from dogs of both genders in individual and collective kennels. Cysts were then isolated from the samples using Sheather’s solution, and the oocysts were isolated in accordance with a modified version of the protocol by Ortega-Mora (1999). Thirty-four (42.5 percent ) of dog samples were found to be positive for Giardia spp., 26 (51.0 percent ) of which came from the CCZ, and 8 of which (27.6 percent ) came from the pet shops. There was a prevalence of 13.72 percent (7) of oocysts in stool samples of CCZ dogs, and of 34.48 percent (10) oocysts in pet shop animals. The high prevalence of Giardia spp. cysts and oocysts in the samples may be associated with exposure to parasites from stray dogs. Clustering of animals in collective kennels or pet shops promotes the risk of intestinal parasite transmission with high zoonotic potential for the animals and for the people living with them...

Production of cytokine and chemokines by human mononuclear cells and whole blood cells after infection with Trypanosoma cruzi / Produção de citocinas e quimiocinas por células mononucleares e células do sangue total humano após infecção com Trypanosoma cruzi

INTRODUCTION: The innate immune response is the first mechanism of protection against Trypanosoma cruzi, and the interaction of inflammatory cells with parasite molecules may activate this response and modulate the adaptive immune system. This study aimed to analyze the levels of cytokines and chemokines synthesized by the whole blood cells (WBC) and peripheral blood mononuclear cells (PBMC) of individuals seronegative for Chagas disease after interaction with live T. cruzi trypomastigotes. METHODS: IL-12, IL-10, TNF-α, TGF-β, CCL-5, CCL-2, CCL-3, and CXCL-9 were measured by ELISA. Nitrite was determined by the Griess method. RESULTS: IL-10 was produced at high levels by WBC compared with PBMC, even after incubation with live trypomastigotes. Production of TNF-α by both PBMC and WBC was significantly higher after stimulation with trypomastigotes. Only PBMC produced significantly higher levels of IL-12 after parasite stimulation. Stimulation of cultures with trypomastigotes induced an increase of CXCL-9 levels produced by WBC. Nitrite levels produced by PBMC increased after the addition of parasites to the culture. CONCLUSIONS: Surface molecules of T. cruzi may induce the production of cytokines and chemokines by cells of the innate immune system through the activation of specific receptors not evaluated in this experiment. The ability to induce IL-12 and TNF-α contributes to shift the adaptive response towards a Th1 profile.

INTRODUÇÃO: A resposta imune inata é o primeiro mecanismo de proteção contra o Trypanosoma cruzi e a interação de células inflamatórias com moléculas do parasita pode ativar esta resposta e modular a resposta adaptativa. O objetivo deste trabalho foi analisar os níveis de citocinas e quimiocinas sintetizados por células do sangue total (WBC) e células mononucleares do sangue periférico (PBMC) de voluntários soronegativos para doença de Chagas depois da interação com Trypanosoma cruzi. MÉTODOS: IL-12, IL-10, TNF-α, TGF-β, CCL5, CCL2, CCL3, CXC-9 foram avaliados por ELISA. Níveis de nitrito foram determinados pelo método de Griess. RESULTADOS: Foram produzidos altos níveis de IL-10 por WBC quando comparado aos sintetizados por PBMC, inclusive após incubação com tripomastigotas. A produção de TNF-α foi significativamente maior nas culturas de PBMC e WBC após estímulo com o parasita. O aumento significativo dos níveis de IL-12 foi observado apenas em PBMC depois do estímulo com tripomastigotas. A adição de tripomastigotas nas culturas induziu aumento dos níveis de CXCL9 produzidos por WBC. Os níveis de nitrito produzidos pelos PBMCs de todos os voluntários após a adição de parasito nas culturas aumentaram. CONCLUSÕES: Moléculas de superfície do parasito podem induzir a produção de citocinas e quimiocinas pelas células da resposta imune inata através da ativação dos receptores específicos não avaliados neste experimento. A habilidade de induzir IL-12 e TNF-α contribui para direcionar uma resposta imune adaptativa de perfil Th1.

Molecular characterization of Trypanosoma cruzi Mexican strains and their behavior in the mouse experimental model / Caracterização molecular de cepas mexicanas de Trypanosoma cruzi e seu comportamento no modelo experimental do camundongo

INTRODUCTION: For a long time, the importance of Chagas disease in Mexico, where many regarded it as an exotic malady, was questioned. Considering the great genetic diversity among isolates of Trypanosoma cruzi, the importance of this biological characterization, and the paucity of information on the clinical and biological aspects of Chagas disease in Mexico, this study aimed to identify the molecular and biological characterization of Trypanosoma cruzi isolates from different endemic areas of this country, especially of the State of Jalisco. METHODS: Eight Mexican Trypanosoma cruzi strains were biologically and genetically characterized (PCR specific for Trypanosoma cruzi, multiplex-PCR, amplification of space no transcript of the genes of the mini-exon, amplification of polymorphic regions of the mini-exon, classification by amplification of intergenic regions of the spliced leader genes, RAPD - (random amplified polymorphic DNA). RESULTS: Two profiles of parasitaemia were observed, patent (peak parasitaemia of 4.6×10(6) to 10(7) parasites/mL) and subpatent. In addition, all isolates were able to infect 100 percent of the animals. The isolates mainly displayed tropism for striated (cardiac and skeletal) muscle. PCR amplification of the mini-exon gene classified the eight strains as TcI. The RAPD technique revealed intraspecies variation among isolates, distinguishing strains isolated from humans and triatomines and according to geographic origin. CONCLUSIONS: The Mexican T. cruzi strains are myotrophic and belong to group TcI.

INTRODUÇÃO: Durante muito tempo, foi questionada a importância da doença de Chagas no México onde muitos a consideravam um padecimento exótico. Considerando a grande diversidade genética existente, entre os isolados de Trypanosoma cruzi, a importância da caracterização biológica desses e o escasso número de informações sobre os aspectos clínicos e biológicos da doença de Chagas no México, o objetivo deste trabalho foi realizar a caracterização biológica e molecular de isolados de Trypanosoma cruzi originários de diferentes áreas endêmicas deste país, principalmente do Estado de Jalisco. MÉTODOS: Oito cepas mexicanas de Trypanosoma cruzi foram caracterizadas biologicamente e geneticamente (PCR específica para Trypanosoma cruzi, PCR-multiplex, amplificação do espaço não transcrito dos genes do mini-exon, amplificação das regiões polimórficas do gene do mini-exon, classificação pela amplificação de regiões intergênicas dos genes do spliced leader, RAPD - random amplified polymorphic DNA). RESULTADOS: Foram observados dois tipos de parasitemia: patente com picos máximos de parasitemia entre 4,6x10(6) e 10(7) parasitas/mL e subpatente. Além disso, todos os isolados foram capazes de infectar 100 por cento dos animais. Observou-se tropismo predominante pelo músculo estriado (cardíaco e esquelético). As técnicas de PCR do gene do mini-éxon classificaram as oito cepas como TcI e a técnica de RAPD mostrou variação intra-especifica das mesmas, separando as cepas isoladas de humanos daquelas de triatomíneos e por origem geográfica. CONCLUSÕES: As cepas mexicanas de Trypanosoma cruzi são miotrópicas e correspondem ao TcI.

Prevalência de triatomíneos (Hemíptera: Reduviidae: Triatominae) infectados por Trypanosoma cruzi: sazonalidade e distribuição na região Ciénega do Estado de Jalisco, México / Prevalence of triatomines (Hemíptera: Reduviidae: Triatominae) infected by Trypanosoma cruzi: seasonality and distribution in the Ciénega region of the State of Jalisco, Mexico

As características físico-geográficas da região Ciénega, Jalisco, México a tornam propícia para transmissão do Trypanosoma cruzi, causador da doença de Chagas. Este trabalho caracteriza a prevalência de triatomíneos infectados pelo parasita, sua sazonalidade e distribuição nesta região. Foram analisados 328 triatomíneos no período de janeiro de 2005 a junho de 2007 procedentes de 13 municípios da região, sendo abril, maio e junho os meses de maior captura. Dos triatomíneos analisados, 57,3 por cento foram positivos para Trypanosoma cruzi, correspondendo 15,4 por cento para a área urbana e 84,6 por cento para a área rural. A espécie mais freqüente foi Triatoma longipennis e a mais parasitada foi Triatoma barberi com índice de infecção de 83,3 por cento quando comparada a Triatoma longipennis (67,5 por cento) (p<0,05). A infecção natural dos vetores capturados pode indicar alta exposição das pessoas ao Trypanosoma cruzi. O achado recente do Triatoma dimidiata positivo nesta região, sugere a adaptação de novas espécies às condições ecológicas destas populações.

The physical and geographical characteristics of the Ciénega region, Jalisco, Mexico make it suitable for transmission of Trypanosoma cruzi, the causative agent for Chagas disease. This study characterizes the prevalence of triatomines infected by this parasite, their seasonality and their distribution in this region. A total of 328 triatomines were evaluated between January 2005 and June 2007, from 13 municipalities in the region. April, May and June were the months with the highest capture levels. Among the triatomines examined, 57.3 percent were positive for Trypanosoma cruzi, corresponding to 15.4 percent in urban areas and 84.6 percent in rural areas. The species with greatest prevalence was Triatoma longipennis and the species with the highest parasitism rate was Triatoma barberi, with an infection rate of 83.3 percent, whereas the rate for Triatoma longipennis was 67.5 percent (p<0.05). This natural infection in the captured vectors may indicate that individuals in this region have high exposure to Trypanosoma cruzi. The recent findings of positive Triatoma dimidiata in this region suggest that new species are becoming adapted to the ecological conditions of these populations.