Killer Cell Immunoglobulin-like Receptors and Their HLA Ligands are Related with the Immunopathology of Chagas Disease.

The aim of this study was to investigate the influence of killer cell immunoglobulin-like receptor (KIR) genes and their human leucocyte antigen (HLA) ligands in the susceptibility of chronic Chagas disease. This case-control study enrolled 131 serologically-diagnosed Chagas disease patients (59 men and 72 women, mean age of 60.4 ± 9.8 years) treated at the University Hospital of Londrina and the Chagas Disease Laboratory of the State University of Maringa. A control group was formed of 165 healthy individuals - spouses of patients or blood donors from the Regional Blood Bank in Maringa (84 men and 81 women, with a mean age of 59.0 ± 11.4 years). Genotyping of HLA and KIR was performed by PCR-SSOP. KIR2DS2-C1 in the absence of KIR2DL2 (KIR2DS2+/2DL2-/C1+) was more frequent in Chagas patients (P = 0.020; Pc = 0.040; OR = 2.14) and, in particular, those who manifested chronic chagasic cardiopathy-CCC (P = 0.0002; Pc = 0.0004; OR = 6.64; 95% CI = 2.30-18.60) when compared to the control group, and when CCC group was compared to the patients without heart involvement (P = 0.010; Pc = 0.020; OR = 3.97). The combination pair KIR2DS2+/2DL2-/KIR2DL3+/C1+ was also positively associated with chronic chagasic cardiopathy. KIR2DL2 and KIR2DS2 were related to immunopathogenesis in Chagas disease. The combination of KIR2DS2 activating receptor with C1 ligand, in the absence of KIR2DL2, may be related to a risk factor in the chronic Chagas disease and chronic chagasic cardiopathy.

Genetic susceptibility to Chagas disease: an overview about the infection and about the association between disease and the immune response genes.

Chagas disease, which is caused by the flagellate parasite Trypanosoma cruzi, affects 8-10 million people in Latin America. The disease is endemic and is characterised by acute and chronic phases that develop in the indeterminate, cardiac, and/or gastrointestinal forms. The immune response during human T. cruzi infection is not completely understood, despite its role in driving the development of distinct clinical manifestations of chronic infection. Polymorphisms in genes involved in the innate and specific immune response are being widely studied in order to clarify their possible role in the occurrence or severity of disease. Here we review the role of classic and nonclassic MHC, KIR, and cytokine host genetic factors on the infection by T. cruzi and the clinical course of Chagas disease.

Inhibitory effect of anethole in nonimmune acute inflammation.

Anethole [1-methoxy-4-(1-propenyl)benzene] occurs naturally as a major component of the essential oil of star anise (Illicium verum Hook.f., family Illiciaceae), comprising more than 90 % of its volatile components. Studies showed that this substance has antioxidant, antibacterial, antifungal, and anesthetic properties. In this study, the anti-inflammatory properties of anethole in animal models of nonimmune acute inflammation such as croton oil-induced ear edema and carrageenan-induced pleurisy were investigated. The investigated parameters were edema formation, leukocyte migration, and inflammatory mediators involved. Oral administration of anethole at a dose of 250 and 500 mg/kg reduced both the volume of pleural exudates and the number of migrated leukocytes. Levels of nitric oxide (NO) and prostaglandins (PGE2) in the inflammatory exudate were reduced by treatment with anethole, but levels of tumor necrosis factor-α and interleukin-1ß were not significantly altered. In ear edema, the oral treatment with anethole inhibited the formation of exudate and the activity of myeloperoxidase, but not after topical administration. These results suggest that the anethole may be effective in controlling some nonimmune acute inflammation-related disease, probably by an inhibitory action on production and/or release of PGE2 and NO.

Anti-arthritic effect of eugenol on collagen-induced arthritis experimental model.

This study was designed to test the efficacy of eugenol, a compound obtained from the essential oil of cloves (Syzygium aromaticum) in collagen-induced arthritis (CIA), a well characterized murine model of rheumatoid arthritis. Macroscopic clinical evidence of CIA manifests first as periarticular erythema and edema in the hind paws. Treatment with eugenol starting at the onset of arthritis (day 25) ameliorated these clinical signs of CIA. Furthermore, eugenol inhibited mononuclear cell infiltration into the knee joints of arthritic mice and also lowered the levels of cytokines (tumor necrosis factor (TNF)-α, interferon (IFN)-γ and tumor growth factor (TGF)-ß) within the ankle joints. Eugenol treatment did not affect the in vitro cell viability as assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Therefore, eugenol ameliorates experimental arthritis and could be useful as a beneficial supplement in treating human arthritis.

Production of TNF-α, nitric oxide and hydrogen peroxide by macrophages from mice with paracoccidioidomycosis that were fed a linseed oil-enriched diet

Omega-3 polyunsaturated fatty acids (n-3 PUFA) can modulate the immune system and their primary effect is on macrophage function. Paracoccidioidomycosis (PCM) is an endemic systemic mycosis in Latin America that is caused by the dimorphic fungus Paracoccidioides brasiliensis (Pb). Macrophages are the main defence against this pathogen and have microbicidal activity that is dependent on interferon-Γ and tumour necrosis factor (TNF)-α. These cytokines stimulate the synthesis of nitric oxide (NO) and hydrogen peroxide (H2O2), leading to the death of the fungus. To study the effect of n-3 PUFA on the host immune response during experimental PCM, macrophages that were obtained from animals infected with Pb18 and fed a diet enriched by linseed (LIN) oil were cultured and challenged with the fungus in vitro. The macrophage function was analysed based on the concentrations of TNF-α, NO and H2O2. LIN oil seems to influence the production of TNF-α during the development of disease. A diet enriched with LIN oil influences the microbicidal activity of the macrophages by inducing the production of cytokines and metabolites such as NO and H2O2, predominantly in the chronic phase of infection.

Production of TNF-α, nitric oxide and hydrogen peroxide by macrophages from mice with paracoccidioidomycosis that were fed a linseed oil-enriched diet.

Omega-3 polyunsaturated fatty acids (n-3 PUFA) can modulate the immune system and their primary effect is on macrophage function. Paracoccidioidomycosis (PCM) is an endemic systemic mycosis in Latin America that is caused by the dimorphic fungus Paracoccidioides brasiliensis (Pb). Macrophages are the main defence against this pathogen and have microbicidal activity that is dependent on interferon-Γ and tumour necrosis factor (TNF)-α. These cytokines stimulate the synthesis of nitric oxide (NO) and hydrogen peroxide (H2O2), leading to the death of the fungus. To study the effect of n-3 PUFA on the host immune response during experimental PCM, macrophages that were obtained from animals infected with Pb18 and fed a diet enriched by linseed (LIN) oil were cultured and challenged with the fungus in vitro. The macrophage function was analysed based on the concentrations of TNF-α, NO and H2O2. LIN oil seems to influence the production of TNF-α during the development of disease. A diet enriched with LIN oil influences the microbicidal activity of the macrophages by inducing the production of cytokines and metabolites such as NO and H2O2, predominantly in the chronic phase of infection.

Induction of phagocytic activity and nitric-oxide production in natural populations of Trypanosoma cruzi I and II from the state of Paraná, Brazil.

Twelve strains of Trypanosoma cruzi isolated from wild reservoirs, triatomines, and chronic chagasic patients in the state of Paraná, southern Brazil, and classified as T. cruzi I and II, were used to test the correlation between genetic and biological diversity. The Phagocytic Index (PI) and nitric-oxide (NO) production in vitro were used as biological parameters. The PI of the T. cruzi I and II strains did not differ significantly, nor did the PI of the T. cruzi strains isolated from humans, triatomines, or wild reservoirs. There was a statistical difference in the inhibition of NO production between T. cruzi I and II and between parasites isolated from humans and the strains isolated from triatomines and wild reservoirs, but there was no correlation between genetics and biology when the strains were analyzed independently of the lineages or hosts from which the strains were isolated. There were significant correlations for Randomly Amplified Polymorphic Deoxyribonucleic acid (RAPD) and biological parameters for T. cruzi I and II, and for humans or wild reservoirs when the lineages or hosts were considered individually.

Induction of phagocytic activity and nitric-oxide production in natural populations of Trypanosoma Cruzi I and II from the state of Paraná, Brazil / Indução da atividade fagocitária e produção de óxido nítrico numa população natural de Trypanosoma cruzi I e II do Estado do Paraná, Brasil

Twelve strains of Trypanosoma cruzi isolated from wild reservoirs, triatomines, and chronic chagasic patients in the state of Paraná, southern Brazil, and classified as T. cruzi I and II, were used to test the correlation between genetic and biological diversity. The Phagocytic Index (PI) and nitric-oxide (NO) production in vitro were used as biological parameters. The PI of the T. cruzi I and II strains did not differ significantly, nor did the PI of the T. cruzi strains isolated from humans, triatomines, or wild reservoirs. There was a statistical difference in the inhibition of NO production between T. cruzi I and II and between parasites isolated from humans and the strains isolated from triatomines and wild reservoirs, but there was no correlation between genetics and biology when the strains were analyzed independently of the lineages or hosts from which the strains were isolated. There were significant correlations for Randomly Amplified Polymorphic Deoxyribonucleic acid (RAPD) and biological parameters for T. cruzi I and II, and for humans or wild reservoirs when the lineages or hosts were considered individually.

Doze cepas de Trypanosoma cruzi isoladas de reservatórios silvestres, triatomíneos e de pacientes chagásicos crônicos do Estado do Paraná, Brasil, classificadas como Tc I e II foram usadas para avaliar a correlação entre genética e diversidade biológica. Índice fagocítico (IF) e produção de óxido nítrico (ON) in vitro foram os parâmetros biológicos utilizados. O IF de cepas T. cruzi I e II não diferiram significativamente assim como o IF de cepas isoladas de humanos, triatomíneos ou de reservatórios silvestres. Há diferença estatística na inibição da produção de ON entre T. cruzi I e II e entre parasitos isolados de humanos e de cepas isoladas de triatomíneos e reservatórios silvestres, mas não foi observada correlação entre genética e biologia quando as cepas foram analisadas independentemente da linhagem ou hospedeiros das quais elas foram isoladas. Observou-se correlação significativa para amplificação aleatória do DNA polimórfico e parâmetros biológicos de Tc I ou II e para os seres humanos ou reservatório silvestre quando linhagens ou hospedeiros são consideradas separadamente.

Bioterápico 17DH de Trypanosoma cruzi aumenta a apoptose em camundongos experimentalmente infectados / Biotherapic of Trypanosoma cruzi 17d increases apoptosis in experimentally infected mice

Introduction: the mechanism of action of ultradiluted medicines has not yet been established[1,3]. Many basic research studies have focused on isopathic models using in vitro or in vivo designs [4,5]. Recent studies indicate that an ultradiluted (isopathic) antigen can transfer signals to the immune system and modulate its response when an organism is challenged against this same antigen [6]. Some studies on experimental infection of mice by T. cruzi identified apoptotic cells and showed that the increase of their number is associated with an increase also in the number of parasites in the blood of the infected animals, while blockage of apoptosis can be the target of therapeutic intervention [7,8].Conclusion: these results show that apoptosis is increased in animals treated with biotherapic of T. cruzi 17d.n(AU)

Introdução: O mecanismo de ação de medicamentos ultradiluídos ainda não está elucidado [1-3]. Muitos estudos em pesquisas básicas concentraram-se nos modelos de isopatia, utilizando protocolos in vivo ou in vitro [4,5]. Estudos recentes relatam que um antígeno ultradiluído (isopático) pode transferir sinais para o sistema imunológico e modular a sua resposta quando o organismo é desafiado contra este antígeno [6]. Outros trabalhos mostraram que na infecção experimental de camundongos pelo T. cruzi foram detectadas células apoptóticas e que um aumento das células apoptóticas está relacionado ao aumento da parasitemia em animais infectados e que o bloqueio da apoptose pode ser alvo de intervenção terapêutica [7,8].Conclusão: A apoptose está aumentada em animais tratados com bioterápico 17DH de T. cruzi e infectados pelo protozoário.(AU)

Ácidos graxos poli-insaturados n-3 e n-6: metabolismo em mamíferos e resposta imune / Omega-3 and omega-6 polyunsaturated fatty acids: metabolism in mammals and immune response

A experimentação animal apresenta uma grande importância para o desenvolvimento da ciência. O uso de camundongos em experimentos ocorre devido à semelhança destes animais com os seres humanos, fácil criação e manutenção e resposta experimental bastante rápida. Esses animais possuem as mesmas enzimas dessaturases e elongases que os humanos, por isso são usados em pesquisas envolvendo incorporação e síntese de ácidos graxos em tecidos. Os ácidos graxos da família ômega-3 e ômega-6 são de suma importância na dieta humana, pois estes não são sintetizados pela síntese de novo e são precursores dos ácidos graxos poli-insaturados de cadeia muito longa, como os ácidos eicosapentaenóico, docosahexaenóico e araquidônico. Estes desempenham funções importantes no organismo, como a síntese de eicosanóides que estão envolvidos diretamente no sistema imune e nas respostas inflamatórias. A razão entre o consumo de ácidos graxos n-6 e n-3 na dieta é um importante fator para determinar a ingestão adequada de ácidos graxos bem como prevenir o aparecimento de doenças. Este artigo tem como objetivo avaliar a incorporação de ácidos graxos em tecidos de animais e discutir a importância dos ácidos da família n-3 e seus metabólitos no sistema imunológico.

Experiments with animals are very important for the improvement of science. The use of mice in experiments is due to their similarity with humans, the easy of raising and maintaining them and their very fast response. These animals have the same desaturase and elongase enzymes as humans and so they are used in research involving the incorporation and synthesis of fatty acids in tissues. The fatty acids omega-3 and omega-6 are extremely important in the human diet because they are not synthesized de novo and are precursors of very long-chain polyunsaturated fatty acids, such as the eicosapentaenoic, docosahexaenoic and arachidonic acids. These acids play important roles in animals, such as precursors of eicosanoids, which are directly involved in the immune system and inflammatory response. The dietary n-3:n-6 intake ratio is important for assessing proper fatty acid intake and for preventing the development of diseases. Thus, this article assessed the incorporation of fatty acids in animal tissues and discussed the importance of n-3 fatty acids and its metabolites for the immune system.

Produção de fator de necrose tumoral-alfa e peróxido de hidrogênio na infecção pelo Trypanosoma cruzi em camundongos submetidos ao exercício / Production of tumoral necrosis factor-alpha and hydrogen peroxide during Trypanosoma cruzi infection in mice submitted to exercise

Este trabalho teve como objetivo determinar os níveis de fator de necrose tumoral alfa (TNF-alfa) sérico e produção de peróxido de hidrogênio (H2O2) por macrófagos peritoneais em camundongos experimentalmente infectados pelo Trypanosoma cruzi e submetidos ao treinamento físico realizado antes da infecção ou ao exercício físico durante o período de infecção. Foram utilizados camundongos BALB/c fêmeas com 30 dias de idade, inoculados com 1.400 tripomastígotas sanguíneos da cepa Y. O exercício físico consistia em atividade de intensidade moderada em esteira rolante. As dosagens foram realizadas com material coletado no 13º dia de infecção. Para a dosagem de TNF-alfa, foi utilizada a técnica de ELISA de captura. A produção de H2O2 foi expressa por coloração produzida após a incubação de macrófagos peritoneais com peroxidase e a leitura realizada em aparelho de ELISA. Não foram encontradas diferenças significativas nas concentrações de TNF-alfa e produção de H2O2 entre os grupos infectados. O treinamento físico realizado antes da infecção e o exercício físico realizado após a infecção não foram capazes de alterar significativamente os níveis de TNF-alfa e a produção de H2O2 na infecção pelo T. cruzi.

This study aimed to determine the levels of seric tumor necrosis factor-alpha (TNF-alpha) and production of hydrogen peroxide (H2O2) by peritoneal macrophages in mice experimentally infected with T. cruzi and submitted to pre-infection exercise training and to post-infection acute exercise. Female 30-day-old BALB/c mice were inoculated with 1,400 blood trypomastigotes of Y-strain T. cruzi. Exercise programs consisted in moderate-intensity activity and were carried out in a treadmill. The measurements were performed with material collected at the 13th day after infection Serum TNF-alpha was evaluated using capture ELISA. H2O2 production was expressed by coloration produced after incubation of peritoneal macrophages and the measurement was performed using an ELISA reader. There were no statistically significant differences in TNF-alpha levels and H2O2 production between the trained and non-trained infected groups. Thus, the physical training performed before infection and physical exercise performed after the infection were not able to change the levels of TNF-alpha and production of H2O2 in the infection by T. cruzi.

Action of Canova® medicine onto peritoneal resident macrophages infected with Trypanosoma cruzi / Ação do medicamento Canova® em macrófagos peritoniais residentes infectados por Trypanosoma cruzi

Approximately 20 million of people are chronically infected with Trypanosoma cruzi in Latin America. The present work investigated the action of the homeopathic medicine Canova® in in vitro experimental infections with T.cruzi, type Y, using Swiss mice peritoneal resident macrophages. Our results demonstrated that Canova® induced a decrease in the production of H2O2 and TNF-alpha at 20% and 40% concentrations when compared to the control RPMI. However, when compared with this medicine excipient, there was a significant decrease of these nediators at 40% concentration only. The production of NO and the phagocytic activity were not affected. TNF-alpha inhibits the T.cruzi replication in peritoneal macrophages in vitro, becoming an important agent of infection control by this parasite. Within this context, Canova®, unlike what has been reported to other infections, would function as a stimulator of the infection, since it inhibited the production of TNF-alpha by peritoneal resident macrophages in vitro. Further studies should be carried out with elicited macrophages, in order to confirm the Canova® inhibitory activity on the production of TNF-alpha and other mediators in macrophages infected by T.cruzi.

Aproximadamente 20 milhões de pessoas são cronicamente infectadas pelo Trypanosoma cruzi na América Latina. O presente trabalho investigou a ação do medicamento homeopático Canova® em infecções experimentais in vitro com Trypanosoma cruzi, cepa Y, usando macrófagos residentes peritoniais de ratos Swiss. Os resultados mostraram que Canova® induz uma diminuição significativa da produção de H2O2 e TNF-alfa em concentrações de 20% e 40%, quando comparado com o controle RPMI. Quando comparado com o excipiente do medicamento, observou-se uma diminuição na concentração destes mediadores apenas na concentração de 40%. A produção de NO e a atividade fagocítica não foram afetadas. TNF-alfa inibe a replicação do protozoário em macrófagos peritoniais in vitro, mostrando-se um importante agente para o controle da infecção pelo parasita. Portanto, o medicamento Canova® poderia estimular o processo de infecção, pois promoveu inibição da produção de TNF-alfa por macrófagos peritoniais residentes in vitro. Estudos adicionais devem ser realizados com macrófagos elicitados, a fim de confirmar a atividade inibitória da Canova® sobre a produção de TNF-alfa e outros mediadores em macrófagos infectados por T. cruzi.

Análise de associação entre cardiopatia chagásica crônica e antígenos HLA de classe I (HLA-A, -B e -C) e de classe II (HLA-DR e -DQ) / Analysis of association between chronic Chagas Cardiomyopathy and antigens HLA of class I (HLA It, - B and - C) and of class II (HLA-DR and - DQ)

Considerando que o sistema HLA contem genes que controlam a resposta imune, bem como genes de susceptibilidade genética a diversas doenças, temos como objetivo a realização de um estudo de associação entre os antígenos HLA e a doença de CHAGAS, forma cardíaca. Foram analisadas as freqüências dos antigenos HLA-A, -B, -C, -DR E -DQ em 47 pacientes com cardiopatia chagásica crônica e 95 indivíduos controles. Essas amostras iniciais são constituídas por caucasóides e negroides. As analises estatísticas mostram um aumento estatisticamente significante da freqüência de HLA-DR2 nos pacientes, quando comparados com os controles. a significativo ns freqüências de dr-2, cujas freqüências nos pacientes e controles são de 48,3por cento e 12,3por cento, respectivamente (pc=0,0058). Os resultados sugerem uma associação positiva do antígeno DR-2 com cardiopatia chagásica crônica. Embora os resultados indiquem uma possível associação com DR-2 também em negroides, os nossos dados não são conclusivos para esse grupo racial, devido ao pequeno tamanho da amostra analisada

Utilizaçäo do marcador genético HLA na investigaçäo de paternidade / The use of HLA genetic marker in paternity investigation

O sistema HLA é excelente marcador genético por ser altamente polimórfico, expressar-se por codominância e apresentar baixa freqüência de seus antígenos na populaçäo. Em 35 casos de exames de investigaçäo de paternidade, foram realizadas 110 tipagens HLA. Cada qual foi testada com uma bateria de 240 anti-soros HLA para os locos HLA-A, B, C, DR e DQ. A maioria dos casos (85,71 por cento ) apresentou probabilidade de inclusäo de paternidade e 14,29 por cento apresentaram exclusäo total de paternidade. A média de p foi de 93,00ñ4,38 por cento . Nenhum caso analisado ficou com probabilidade inferior a 85,00 por cento . O estudo do sistema HLA é reconhecido como exame de exclusäo de paternidade pela área jurídica