Márcia Machado de Oliveira Dalalio

Association of human leukocyte antigen DQ1 and dengue fever in a white Southern Brazilian population

Dengue is an infectious disease of viral etiology transmitted by the mosquitoes Aedes aegypti, A. albopictus, and A. scutellaris. It can develop either as a benign form or as a severe hemorrhagic form. Previous work showed an association of the hemorrhagic form with human leukocyte antigens (HLA), suggesting a role of genetic factors in disease susceptibility. Nevertheless, data on HLA association with the classical form of the disease is scarce in literature. Sixty-four patients and 667 normal individuals, living in the state of Parana, Southern Brazil, were used as test and control group, respectively. The patients developed the disease during a virus 1 dengue outbreak either in Maringa city in 1995 (47) or in Paranavai city in 1999 (17). The diagnostic was confirmed through serology and/or viral culture. HLA class I and II typing was performed by the classical microlynfocitotoxicity test using monoclonal antisera and fluorobeads. Qui-square statistical analysis confirmed a positive association with HLA-DQ1 (76.6% vs 57.7%; p = 0.005243; pc = 0.026215). HLA-DR1 also presented an increased frequency in the test group, not statistically significant after p correction though (32.8% vs 15.9%; p = 0.005729; pc = 0.080206). In conclusion, genetic factors may play a role on the susceptibility to the classical dengue, virus 1, in the Brazilian population. Further independent studies should be performed in the Brazilian population to confirm these preliminary data.

Ácidos graxos poli-insaturados n-3 e n-6: metabolismo em mamíferos e resposta imune

A experimentacao animal apresenta uma grande importância para o desenvolvimento da ciencia. O uso de camundongos em experimentos ocorre devido a semelhanca destes animais com os seres humanos, facil criacao e manutencao e resposta experimental bastante rapida. Esses animais possuem as mesmas enzimas dessaturases e elongases que os humanos, por isso sao usados em pesquisas envolvendo incorporacao e sintese de acidos graxos em tecidos. Os acidos graxos da familia omega-3 e omega-6 sao de suma importância na dieta humana, pois estes nao sao sintetizados pela sintese de novo e sao precursores dos acidos graxos poli-insaturados de cadeia muito longa, como os acidos eicosapentaenoico, docosahexaenoico e araquidonico. Estes desempenham funcoes importantes no organismo, como a sintese de eicosanoides que estao envolvidos diretamente no sistema imune e nas respostas inflamatorias. A razao entre o consumo de acidos graxos n-6 e n-3 na dieta e um importante fator para determinar a ingestao adequada de acidos graxos bem como prevenir o aparecimento de doencas. Este artigo tem como objetivo avaliar a incorporacao de acidos graxos em tecidos de animais e discutir a importância dos acidos da familia n-3 e seus metabolitos no sistema imunologico.

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.

Influence of KIR genes and their HLA ligands in susceptibility to dengue in a population from southern Brazil.

Killer cell immunoglobulin-like receptors (KIR) form a group of regulatory molecules that specifically recognise human leukocyte antigen (HLA) class I molecules, modulating the cytolytic activity of natural killer cells. The purpose of this study was to investigate the influence of KIR genes and their class I HLA ligands in susceptibility to dengue fever in a population from southern Brazil through a case-control study. One hundred four subjects with confirmed diagnoses of dengue participated in this study, along with a control group of 172 individuals from the same geographic area. HLA and KIR genotyping was performed by polymerase chain reaction with sequence-specific oligonucleotide probes (PCR-SSOP) and with sequence-specific primer (PCR-SSP) techniques, respectively. Data analysis showed significant differences for the KIR2DS1 (54.8% vs 40.7%, P = 0.03), KIR2DS5 (50.0% vs 36.0%, P = 0.03) and KIR2DL5 (76.0% vs 56.4%, P = 0.001) genes. With regard to KIR-ligand pairs, positive associations with dengue were observed in KIR3DS1-Bw4 (45.2% vs 29.7%, P = 0.01), KIR3DL1-Bw4 (80.7% vs 65.1%, P < 0.001), KIR2DL1-C2 (75.0% vs 62.2%, P = 0.03) and KIR2DS1-C2 (40.4% vs 25.6%, P = 0.01) interactions, and a negative association in KIR2DL3-C1/C1 (18.2% vs 33.1%, P = 0.01). Furthermore, the analysis of KIR haplogroups showed a possible protective factor against dengue fever in individuals with the AA genotype. Taken together, these results suggest the existence of genetic predisposition to dengue fever in the population from southern Brazil.

Preinfection aerobic treadmill training improves resistance against Trypanosoma cruzi infection in mice

Exercise performed before infections has been linked to improvement of the immune response against infections. The purpose of this study was to evaluate the influence of preinfection moderate-intensity treadmill training on acute Trypanosoma cruzi infection in mice. Ninety-nine female BALB/c mice were divided into 4 groups, as follows: training + infection (T+I) (n = 41); no training + infection (NT+I) (n = 38); training + no infection (T+NI) (n = 10); and no training + no infection (NT+NI) (n = 10). The exercise program for trained groups was carried out on a motorized treadmill for 8 weeks. Infected groups were inoculated with the Y strain of T. cruzi. Infectivity, prepatent period, patent period, parasitemia peak, mortality, survival time, weight, food intake, tumor necrosis factor-alpha serum levels, and peritoneal macrophage hydrogen peroxide production were evaluated. We found that preinfection training induced statistically significant reductions in parasitemia peak (p < 0.03) and weight loss (p < 0.04). However, no statistically significant differences were found for the other parameters evaluated when trained and nontrained infected groups were compared. We conclude that preinfection aerobic training induces some improvement in the immune response to T. cruzi infection in female BALB/c mice.

Physical exercise protects myenteric neurons and reduces parasitemia in Trypanosoma cruzi infection.

To evaluate the parasitemia, nitrergic neurons, and cytokines in Trypanosoma cruzi-infected mice subjected to moderate physical exercise, forty male Swiss mice, 30days of age, were divided: Trained Control (TC), Trained Infected (TI), Sedentary Control (SC), and Sedentary Infected (SI). The moderate physical exercise program on a treadmill lasted 8weeks. Three days after completing the moderate physical exercise program, the TI and SI groups were inoculated with 1300 blood trypomastigotes of the Y strain of T. cruzi, and parasitemia was evaluated from day 4 to day 22 after inoculation. After 75days of infection, cytokines were measured and colonic neurons were quantified using immunofluorescence to identify neuronal nitric oxide synthase (nNOS). The results were analyzed using analysis of variance - Tukey and Kruskal-Wallis tests, to 5% significance. Moderate physical exercise reduced the parasite peak on day 8 of infection and total parasitemia (p<0.05), contributed to survival of number of nNOS-immunoreactive neurons (p<0.01) and promoted neuronal hypertrophy of the neurons (p<0.05), increased the synthesis of tumor necrosis factor-α (p<0.01) and transforming growth factor-β (p>0.05), providing beneficial effects to the host by acting on the immune system to preserve nitrergic neurons.

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.

Role of Omega-3 Polyunsaturated Fatty Acids in the Production of Prostaglandin E2 and Nitric Oxide during Experimental Murine Paracoccidioidomycosis

There has recently been increased interest in the potential health effects of omega-3 polyunsaturated fatty acids on the immune system. Paracoccidioidomycosis is the most important endemic mycosis in Latin America. Macrophages have a fundamental role and act as first line of organism defense. The purpose of this study was to analyze the effect of n-3 fatty acids on the production of PGE2 and NO by mice infected with Pb18 and fed a diet enriched with LNA for 8 weeks. To study the effect of omega-3 fatty acids on macrophage activity during experimental paracoccidioidomycosis, mice were infected with Pb18 and fed a diet supplemented with LNA. PGE2 in the serum of animals was analyzed and NO in the supernatants of macrophages cultured and challenged in vitro with Pb18 was measured. Omega-3 fatty acids seemed to decrease the production of PGE2 in vivo in the infected group fed an LNA-supplemented diet during the 4th and 8th weeks of the experiment. At the same time, we observed an increase in synthesis of NO by peritoneal macrophages in this group. Omega-3 fatty acids thus appear to have an immunomodulatory effect in paracoccidioidomycosis.

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.

A novel HLA allele, HLA-DRB1*13:204, detected in a Brazilian unrelated hematopoietic stem cell donor.

The HLA-DRB1*13:204 allele differs from HLA*13:64 by two nucleotide substitutions at positions 181 and 189 in the exon 2.