DC-SIGN receptor is expressed by cells from cutaneous leishmaniasis lesions and differentially binds to Leishmania (Viannia) braziliensis and L. (Leishmania) amazonensis promastigotes

BACKGROUND Dendritic cells (DCs) specific intercellular adhesion molecule (ICAM)-3-grabbing non integrin receptor (DC-SIGN) binds to subgenera Leishmania promastigotes mediating its interaction with DC and neutrophils, potentially influencing the infection outcome. OBJECTIVES In this work, we investigated whether DC-SIGN receptor is expressed in cells from cutaneous leishmaniasis (CL) lesions as well as the in vitro binding pattern of Leishmania (Viannia) braziliensis (Lb) and L. (L.) amazonensis (La) promastigotes. METHODS DC-SIGN receptor was labeled by immunohistochemistry in cryopreserved CL tissue fragments. In vitro binding assay with CFSE-labeled Lb or La promastigotes and RAJI-transfecting cells expressing DC-SIGN (DC-SIGNPOS) or mock-transfected (DC-SIGNNEG) were monitored by flow cytometry at 2 h, 24 h and 48 h in co-culture. RESULTS In CL lesion infiltrate, DC-SIGNPOS cells were present in the dermis and near the epidermis. Both Lb and La bind to DC-SIGNPOS cells, while binding to DC-SIGNNEG was low. La showed precocious and higher affinity to DC-SIGNhi population than to DC-SIGNlow, while Lb binding was similar in these populations. CONCLUSION Our results demonstrate that DC-SIGN receptor is present in L. braziliensis CL lesions and interact with Lb promastigotes. Moreover, the differences in the binding pattern to Lb and La suggest DC-SIGN can influence in a difference way the intake of the parasites at the first hours after Leishmania infection. These results raise the hypothesis that DC-SIGN receptor could participate in the immunopathogenesis of American tegumentary leishmaniasis accounting for the differences in the outcome of the Leishmania spp. infection.

Chagas disease in Virgem da Lapa, Minas Gerais, Brazil: left ventricle aneurysm and the risk of death in the 24-year interval

BACKGROUND Left ventricular aneurysm (LVA) is indicator of high morbidity in Chagas' disease. A cross-sectional study performed identified LVA in 18.8% of the chronic chagasic patients (CCP). OBJECTIVE Determine the risk of death of patients with chronic chagasic cardiopathy (CCC) and LVA in 24-year interval. MATERIAL AND METHODS In 1995 a cohort of 298 CCP was evaluated by anamnesis, physical examination, EKG and ECHO and classified in groups: G0 = 86 without cardiopathy; G1 = 156 with cardiopathy without LVA and G2 = 56 with cardiopathy and LVA. 38 patients of G0 and G1 used benznidazole. Information about the deaths was obtained in the notary, death certificates, hospital records and family members. FINDINGS Were registered 113 deaths (37.9%): 107 (35.9%) attributed to cardiopathy and 6 (2.0%) to other causes (p < 0.05). Amongst these 107 deaths, 10 (11.6%) occurred in G0; 49 (31.4%) occurred in G1 and 48 (85.7%) occurred in G2 (p < 0.05). The risk of death was 2.7 and 7.4 times significantly higher in G2, than in G1 and G0, respectively. CONCLUSION Chronic chagasic patients with LVA and ejection fraction < 45% have a higher risk of death than those without.

Severity of tegumentary leishmaniasis is not exclusively associated with Leishmania RNA virus 1 infection in Brazil

Leishmania RNA virus (LRV) has been shown to be a symbiotic component of Leishmania parasites in South America. Nested retro-transcription polymerase chain reaction was employed to investigate LRV1 presence in leishmaniasis lesions from Brazil. In endemic areas of Rio de Janeiro (RJ), no LRV1 infection was observed even with mucosal involvement. LRV1 was only detected in Leishmania (V.) guyanensis cutaneous lesions from the northern region, which were obtained from patients presenting with disease reactivation after clinical cure of their primary lesions. Our results indicated that the severity of leishmaniasis in some areas of RJ, where Leishmania (V.) brazi-liensis is the primary etiological agent, was not associated with Leishmania LRV1 infection.

Primera Red Sudamericana de Biomedicina. Investigación, educación y biotecnología aplicadas a la salud / [First South american network of biomedical research. Education and biotechnology for health].

It is in our interest, in this brief manuscript, to report the creation of the first program of regional integration of a network of research institutes in Biomedicine belonging to members of the MERCOSUR countries. We discuss some of the foundations that gave sustenance to its creation and its objectives in the medium and long term. In addition, we consider the potential of the results of this program in the fields of applied medical research, education and biotechnology.

Polymerase chain reaction of peripheral blood as a tool for the diagnosis of visceral leishmaniasis in children

The diagnosis of visceral leishmaniasis (VL) generally requires the use of invasive tests for the collection of infected tissue (aspirates of bone marrow, spleen, liver or lymph nodes). This difficulty has led to the search for safer and less painful techniques to confirm the occurrence of the disease in children. Polymerase chain reaction (PCR) is a method that is advantageous in that it allows the use of peripheral blood samples for diagnosis. This paper reports the utilisation of PCR on peripheral blood samples to diagnose VL in 45 children in Mato Grosso do Sul, Brazil. This technique is compared with methods carried out using tissue collected by invasive procedures, including direct microscopy, culture and detection of Leishmania DNA by PCR in bone marrow aspirates. The results show that PCR of peripheral blood provides great sensitivity (95.6 percent) that is similar to that from the PCR of bone marrow aspirates (91.1 percent) and higher than that achieved with microscopy (80 percent) or culture (26.7 percent) methods. PCR of peripheral blood proved to be a suitable tool for the diagnosis of VL in children because it is highly sensitive and safe, with tissue collection being less invasive than in traditional tests.

Fritz Koeberle: visão da ciência pelo microscópio / Fritz Koeberle: vision of science microscope

Revisões históricas aos avanços científicos para o controle da doença, o Simpósio Internacional Comemorativo do Centenário da Descoberta da Doença de Chagas (1909-2009).

Sensitivity and specificity of polymerase chain reaction in Giemsa-stained slides for diagnosis of visceral leishmaniasis in children

The aim of this study was to evaluate the sensitivity and specificity of polymerase chain reaction (PCR) in the detection of Leishmania DNA in archived Giemsa-stained bone marrow slides for diagnosis of visceral leishmaniasis (VL), and to compare PCR with conventional diagnostic techniques, like direct microscopy and parasite culture. Specimens of archived Giemsa-stained bone marrow slides from 91 patients with VL and from 79 controls with other diseases or conditions were studied. PCR showed the highest sensitivity (92.3 percent) and had good specificity (97.5 percent). Direct examination detected 79.1 percent and culture 59 percent of positive samples. In addition, PCR was able to detect VL in 16 of 19 patients (84.2 percent) with negative microscopy. PCR in Giemsa-stained bone marrow slides is a suitable tool for confirming diagnosis in patients with VL and may be useful in the diagnosis of difficult cases. Slide smears are easily stored, do not require special storage conditions such as low temperatures, and can be easily mailed to centers where PCR is available, making it an excellent option for diagnosis in the field.

Comparative immunohistological analysis of the Montenegro skin test reaction in asymptomatic infection and in acute and chronic cutaneous leishmaniasis / Análisis inmunohistopatológico comparativo de la reacción a la prueba cutánea de Montenegro en infección asintomática y en leishmaniasis cutánea aguda y crónica

Introducción. La reacción a la prueba cutánea de Montenegro (a leishmanina) y la lesión de leishmaniasis comparten características fundamentales de la reacción de hipersensibilidad de tipo retardado. Objetivos. Determinar si la respuesta cutánea a leishmanina se aproxima y podría modelar la respuesta inflamatoria e inmune temprana a la infección por Leishmania. Materiales y métodos. Este estudio comparó la respuesta inflamatoria de biopsias de lesiones agudas (tiempo de evolución £1 mes) y lesiones crónicas (tiempo de evolución ³6 meses) con su respectiva reacción a la prueba cutánea de Montenegro, y con la reacción a la prueba de Montenegro de individuos infectados asintomáticos. Resultados. La proporción de linfocitos T CD4+ y CD8+, células fagocíticas mononucleares y granulocitos fue similar entre lesiones agudas y sus reacciones a la prueba cutánea a leishmanina. En contraste, los linfocitos CD4+ (32,6 por ciento) representaron una proporción significativamente mas baja, y los linfocitos B (20 por ciento) y macrófagos (27 por ciento) una proporción significativamente más alta del infiltrado celular en lesiones crónicas que en sus correspondientes reacciones a la prueba cutánea de Montenegro (CD4+: 43,7 por ciento, linfocitos B: 0.9 por ciento; macrófagos: 17,5 por ciento). Se encontró una asociación positiva entre linfocitos T CD8+ y macrófagos ( P=0,038) en la reacción a la prueba cutánea de Montenegro de los individuos con infección asintomática, mientras que los linfocitos T CD8+ y CD4+ estuvieron asociados positivamente en las reacciones a la prueba cutánea de Montenegro de pacientes crónicos ( P=0,002). Adicionalmente, la proporción de linfocitos B en lesiones crónicas (20 por ciento) fue más alta que en lesiones agudas (5,3 por ciento) ( P=0,002). Conclusión. La reacción cutánea de Montenegro permitió diferenciar la respuesta celular inmune entre infección asintomática y enfermedad crónica, y simula la respuesta temprana a la infección.

Introduction. The Montenegro skin test reaction and leishmaniasis lesions share fundamental characteristics of a delayed type hypersensitivity reaction. Objectives. To determine whether the Montenegro skin test reaction (response to leishmanin) might approximate and thereby provide insight into the early inflammatory and immune response to Leishmania infection. Materials and methods. We compared the inflammatory response in biopsies of acute (evolution time £one month), and chronic lesions (evolution time ³6 months) with the Montenegro skin test reaction in the corresponding patients, and with the Montenegro skin test of asymptomatically infected volunteers. Results. The proportion of CD4+ and CD8+ T lymphocytes, mononuclear phagocytes and granulocytes were similar in acute lesions and in their corresponding Montenegro skin test reactions. In contrast, CD4+ lymphocytes (32.6%) represented a significantly lower, and B cells (20%) and macrophages (27%) a significantly higher proportion of the cellular infiltrate in chronic lesions as compared to reactions in the corresponding skin test site (CD4+: 43.7%, B cells: 0.9%; macrophages: 17.5%). CD8+ T lymphocytes and macrophages were positively associated ( P=0.038) in the Montenegro skin test of asymptomatically infected individuals whereas CD8+ and CD4+ T cells were positively associated in the Montenegro skin test of chronic patients ( P=0.002). Notably, B cells were markedly more frequent in chronic lesions (20%) than in acute lesions (5.3%) ( P=0.002). Conclusion. The Montenegro skin test distinguished the cellular immune response to Leishmania in asymptomatic infection and chronic disease and may provide a surrogate of the early response to infection.

Paniculite eosinofílica localizada em conseqüência de injeções intramusculares de composto de antimônio para tratamento de leishmaniose tegumentar americana / Localized eosinophilic panniculitis following intramuscular injection of pentavalent antimonium for treatment of american tegumentary leishmaniasis

Paniculite eosinofílica pode ser desencadeada por muitos fatores; aqui os autores descrevem a síndrome em conseqüência de injeções intramusculares de compostos de antimônio para tratamento de leishmaniose tegumentar americana em três pacientes. Todos eles desenvolveram lesões em placa, profundamente infiltradas, no local da injeção antimonial. A histopatologia mostrou acentuado infiltrado inflamatório da hipoderme com numerosos eosinófilos. O estudo imunológico não demonstrou imunoglobulinas ou frações do complemento nas lesões. O diagnóstico final foi de paniculite eosinofílica ocorrendo como efeito colateral da terapia antimonial. O mecanismo patogênico dessa paniculite não pôde ser definido. As hipóteses sugeridas foram de lesão induzida por fenômeno físico - a pressão exercida pelo volume do líquido injetado - ou de uma reação alérgica ao antimônio.

Levantamento de leishmaniose visceral em cäes de Santa Maria e municípios próximos, Estado do Rio Grande do Sul / A survey of visceral leishmaniasis in dogs from Santa Maria and neighbouring municipalities, State of Rio Grande do Sul

A partir de relato de diagnóstico de leishmaniose visceral em cinco cäes dos municípios de Santa Maria, Itaara e Júlio de Castilhos, no centro do Estado do Rio Grande do Sul, foi realizado levantamento sorológico através de teste de imunofluorescência indireta em sangue obtido em papel de filtro de 40 cäes de Santa Maria, 20 de Itaara e 11 de Júlio de Castilhos, em geral relacionados aos cinco cäes considerados como positivos. Além destes, foi também examinado o sangue de 44 cäes vadios de Júlio de Castilhos, 68 de Cachoeira do Sul e 20 de Caçapava do Sul. Soros fracamente reagentes com títulos < 1:40 foram avaliados em ensaios de Ensaio Imuno-enzimático. Amostras de fígado, baço, pulmäo e linfonodos de um sexto cäo de Santa Maria, considerado como positivo à necropsia, foram examinadas através de histopatologia e PCR. Todos os resultados dos 204 cäes foram negativos, indicando que, apesar de näo ser impossível a ocorrência de leishmaniose visceral na regiäo, por vários fatores aqui discutidos, ela deve ser pelo menos extremamente rara, sendo necessários estudos mais detalhados do material dos cinco cäes tidos como positivos

Biodemas de cepas do Trypanosoma cruzi isoladas de humanos de três áreas endêmicas de Minas Gerais / Biodemes of Trypanosoma cruzi strains isolated from humans from three endemic areas in Minas Gerais State

In order to study the biological behavior of T. cruzi strains and to determine a putative association between their biodeme and the clinical forms of Chagas disease, 14 strains isolated from humans were evaluated. The individuals were from the municipalities of Pains, Iguatama and Berilo (Minas Gerais State). The biological behavior was evaluated in albino swiss mice, weighing 10 to 15 grams, which were infected with 1x10(4) blood tripomastigotes. The infectivity, parasitemia, tripomastigote morphology, virulence and the tissue distribution of the protozoan were analyzed. A behavior similar to biodeme II (SÒo Felipe strain) was observed in 9 strains, while 5 stocks were characterized as belonging to biodeme III. It was not possible to establish a relationship between the biodeme strain and the severity of the disease in the patients.

Perfil de citocinas e células imunológicas em pacientes com tuberculose pleural co-infectados ou não pelo HIV / Cytokine and immune cell profile pleural tuberculosis in patients with and without HIV co-infection

Introdução: a tuberculose pleural tem uma evolução benigna, mesmo quando associada à infecção pelo HIV. Com o objetivo de compreender os mecanismos imunológicos envolvidos neste fenômeno, nós comparamos as concentrações de citocinas e subgrupos de células imunológicas no líquido e tecido pleural de pacientes com tuberculose pleural com e sem infecção pelo HIV. Material e métodos: foram incluídos 42 pacientes com o diagnóstico de tuberculose pleural dos quais 12 infectados pelo HIV. A análise imunohistoquímica do tecido pleural foi realizada em 21 pacientes utilizando os seguintes anticorpos monoclonais: anti-CD4, anti-CD8, anti-delta TCR, anti-perforina e anti-fasL. A concentração de citocinas (IL-2,IL-4, IL-10, IL-12 e IFN-) foi medida pelo método ELISA no líquido pleural de 29 pacientes. Resultados: a mediana das proporções de células CD8+ e perforina+ foi superior nos pacientes infectados pelo HIV. A proporção de células CD4+, FasL+ e delta-TCR+ foram semelhantes nos dois grupos. A IL-4 foi indetectável em todos os pacientes. Três de nove pacientes infectados pelo HIV apresentaram uma concentração de IL-2 superior a 40 pg/ml (p=0,02). Conclusões: as concentrações de IFN-, IL-10 e IL-12 foram semelhantes nos dois grupos. A citotoxicidade mediada pela perforina e a IL-2 parecem ter um papel importante na proteção contra Mycobacterium tuberculosis nos estádios iniciais da infecção pelo HIV. As células CD8+ do tecido pleural podem ser uma fonte alternativa de síntese de IFN- em pacientes com tuberculose pleural co-infectados pelo HIV.

Introduction: pleural tuberculosis (TB) has a benign course whether associated or not to HIV infection. To understand the immune mechanisms involved in this phenomenon, we compared cytokine concentrations and subsets of immune cells in the pleural fluid/tissue from patients with TB pleurisy with and without HIV co-infection. Material and methods: forty-two patients diagnosed with pleural TB were included, twelve of whom were HIV-infected. Immunohistochemical analysis of pleural tissue was performed in 21 patients with TB pleurisy with and without HIV co-infection. Material and methods: forty-two patients diagnosed with pleural TB were included, twelve of whom were HIV-infected. Immunohistochemical analysis of pleural tissue was performed in 21 patients using the following monoclonal antibodies: anti-CD4, anti CD-8, anti-delta TCR, anti-perforin and anti FasL. Cytokine (IL-2, OÇ-4, IL-10, IL-12 amd IFN-) concentration was measured by the ELISA method in the pleural fluid of 29 patients. Results: the median proportions of CD8+ and perforin + cells were higher in HIV-infected patients. The proportions of CD4+, FasL+ and delta-TCR+ cells were similar in both groups. IL-4 was undetectable in all patients. Three out of nine HIV-infected patients had IL-2 concentration ouver 40pg/ml (p=0.02). Conclusion: the concentrations of IFN-, IL-10 and IL-12 were similar in both groups. Perforin-mediated cytotoxicity and IL-2 may play an important role in protection against Mycobacterium tuberculosis in the early stages of HIV infection. Pleural CD8+ cells may be an alternative source for IFN- in HIV-infected patients with tuberculosis.

Mucosal leishmaniasis ("espundia") responsive to low dose of N-methyl glucamine (Glucantime) in Rio de Janeiro, Brazil

Response to treatment with antimonial drugs varies considerably depending on the parasite strain involved, immune status of the patient and clinical form of the disease. Therapeutic regimens with this first line drug have been frequently modified both, in dose and duration of therapy. A regimen of 20 mg/kg/day of pentavalent antimony (Sb5+) during four weeks without an upper limit on the daily dose is currently recommended for mucosal disease ("espundia"). Side-effects with this dose are more marked in elderly patients, more commonly affected by this form of leishmaniasis. According to our experience, leishmaniasis in Rio de Janeiro responds well to antimony and, in cutaneous disease, high cure rates are obtained with 5 mg/kg/day of Sb5+ during 30 to 45-days. In this study a high rate of cure (91.4 percent) employing this dose was achieved in 36 patients with mild disease in this same geographic region. Side-effects were reduced and no antimony refractoriness was noted with subsequent use of larger dose in patients that failed to respond to initial schedule

Lesões lupóides na leishmaniose cutânea do Novo Mundo / Lupoid lesions in cutaneous leishmaniasis in the New Word

São apresentados dois casos de lesões lupóides, uma rara forma de apresentação clínica da leishmaniose tegumentar no Novo Mundo. Leishmaniose lupóide (LL) é de relato freqüente no Velho Mundo, mas a maioria dos autores emprega essa designação para casos de leishmaniasis recidiva cutis (LRC), originalmente descrita por Berlin. Neste trabalho, os autores salientam as diferenças existentes entre essas duas entidades: a LL como forma de apresentação clínica inicial da doença, e a LRC como o resultado da persistência de parasitas após a cura de uma lesão incial, geralmente ulcerosa. O valor da reação em cadeia de polimerase (PCR) para o diagnóstico parasitológico de lesões pobres em parasitas, caso das lesões lupóides, é enfatizado

Paleoparasitology: perspectives with new techniques

Paleoparasitologia e o estudo de parasitos encontrados em material arqueologico. O desenvolvimento deste campo da pesquisa teve inicio com a identificacao de ovos de helmintos em tecidos mumificados, analise de coprolitos e, recentemente, atraves da biologia molecular. Neste artigo faz-se uma breve revisao da historia da paleoparasitologia com referencia especial aos estudos de ADN antigo (ancient DNA) em material arqueologico

Autoreactivity in chronic experimental Trypanosoma cruzy infection

The mechanisms by which Trypanosoma cruzi causes cardiomyopathy are discussed in this paper. The bulk of results suggests that autoimmunity is the main cause of the progressive cardiac destruction, and that autoreactivity is restricted to the CD4+ T-cell compartment. The various possibilities involved in the genesis of autoimmunity in chronic T. cruzi infection include molecular mimicry, antigen presentation and imbalance of immune regulation due to suppressor cell activities, idiotypic regulation and/or the influence of cytokines by the two mutually antagonistic type 1 and type 2 T-cell subsets.