Serum cytokine responses over the entire clinical-immunological spectrum of human leishmania (l.) infantum chagasi infection

The clinical-immunological spectrum of human Leishmania (L.) infantum chagasi infection in Amazonian Brazil was recently reviewed based on clinical, DTH, and IFAT (IgG) evaluations that identified five profiles: three asymptomatic (asymptomatic infection, AIsubclinical resistant infection, SRIand indeterminate initial infection, III) and two symptomatic (symptomatic infection, SIAmerican visceral leishmaniasis, AVLand subclinical oligosymptomatic infection, SOI). TNF-alpha, IL-4, IL-6, and IL-10 serum cytokines were analyzed using multiplexed Cytometric Bead Array in 161 samples from endemic areas in the Brazilian Amazon: SI [AVL] (21 cases), III (49), SRI (19), SOI (12), AI (36), and a control group [CG] (24). The highest IL-6 serumlevels were observed in the SI profile (AVL)higher IL-10 serum levels were observed in SI than in SOI or CG and in AI and III than in SOIhigher TNF-alpha serum levels were seen in SI than in CG. Positive correlations were found between IL-6 and IL-10 serum levels in the SI and III profiles and between IL-6 and TNF-alpha and between IL-4 and TNF-alpha in the III profile. These results provide strong evidence for associating IL-6 and IL-10 with the immunopathogenesis of AVL and help clarify the role of these cytokines in the infection spectrum.Instituto Evandro Chagas (Secretaria de Vigilancia em Saude, Ministerio da Saude, Brazil)Nucleo de Medicina Tropical (Universidade Federal do Para, Brazil)Laboratorio de Investigacao Medica (LIM)-50 (Hospital de Clinicas (HC)-Faculdade de Medicina (FM)-Universidade de Sao Paulo (USP), Brazil)Sao Paulo Research Foundation (FAPESP) [2006/56319-1]Parasitology Department, Evandro Chagas Institute, Surveillance Secretary of Health, Ministry of Health, Ananindeua, PA, BrazilAlbert Einstein Israelite Hospital, São Paulo, SP, BrazilDivision of Immunology, Federal University of São Paulo, São Paulo, SP, BrazilPathology Department, Medical School of São Paulo University, São Paulo, SP, BrazilTropical Medicine Nucleus, Federal University of Pará, Belém, PA, BrazilDivision of Immunology, Federal University of São Paulo, São Paulo, SP, BrazilFAPESP: 2006/56319-1Web of Scienc

Serum Cytokine Responses over the Entire Clinical-Immunological Spectrum of Human Leishmania (L.) infantum chagasi

The clinical-immunological spectrum of human Leishmania (L.) infantum chagasi infection in Amazonian Brazil was recently reviewed based on clinical, DTH, and IFAT (IgG) evaluations that identified five profiles: three asymptomatic (asymptomatic infection, AI; subclinical resistant infection, SRI; and indeterminate initial infection, III) and two symptomatic (symptomatic infection, SI; American visceral leishmaniasis, AVL; and subclinical oligosymptomatic infection, SOI). TNF-α, IL-4, IL-6, and IL-10 serum cytokines were analyzed using multiplexed Cytometric Bead Array in 161 samples from endemic areas in the Brazilian Amazon: SI [AVL] (21 cases), III (49), SRI (19), SOI (12), AI (36), and a control group [CG] (24). The highest IL-6 serum levels were observed in the SI profile (AVL); higher IL-10 serum levels were observed in SI than in SOI or CG and in AI and III than in SOI; higher TNF-α serum levels were seen in SI than in CG. Positive correlations were found between IL-6 and IL-10 serum levels in the SI and III profiles and between IL-6 and TNF-α and between IL-4 and TNF-α in the III profile. These results provide strong evidence for associating IL-6 and IL-10 with the immunopathogenesis of AVL and help clarify the role of these cytokines in the infection spectrum

Estudo in vitro da infectividade de espécies de Leishmania do subgênero Viannia em macrófagos peritoneais de camundongo BALB/c

Parasites ofthe genus Leishmania show both intra- and inter-specific variations of infectivity. There is little available information, however, regarding the infective behavior of New World species particularly those of the Amazon Region of Brazil, where there occur at six species of the subgenus Viannia causing human cutaneous leishmaniasis (ACL). The aim of the present study was to investigate the infectivity of 5 of these species for the peritoneal macrophages of BALB/c mice, and their role in the host-cell's production of nitric oxide (NO). Thirty strains of Leishmania were divided into 6 groups of the fol1owing species: I-L. (V.) braziliensis (from cases of localized skin lesions-LCL); II- L. (V.) braziliensis (from cases of mucocutaneous lesions- LeM); III- L. (V.) guyanensis; IV- L. (V.) shawi; V- L. (V.) naiffi and VI- L. (V.) lainsoni. They were cultivated in RPMI 1640 medium supplemented with 10% fetal bovine serum and 1% penicillin/gentamicin, until reaching the stationary phase of development. These promastigotes were used to infect the macrophage cell cultures, in the proportion of 4 flagellates/macrophage. The cultures were then incubated at 35°C with 5% CO2 and, after 24 hours following inoculation, the corvelips were stained by Giemsa's method to determine the infection index. The nitric oxide (nitrite) concentration was measured in the culture supernatant by Griess' s method to determine the infection index. The nitric oxide (nitrite) concentration was measured in the culture supernatant by Griess' s method. It was found that LCM strains of L. (V.) braziliensis showed the highest infection index (385), significance p<0.05,compared with L. (V.) braziliensis LCL strains (264). L. (V). naiffi and L. (V.) lansoni had the lowest infection indices of (215) and (272), respectively, but, were not significally different from index of L. (V.) guyanensis (300). Rrgarding the NO levels, the highest was that for L. (V.) naiffi (4,1µM), and the lowest for L. (V.) braziliensis-LCM strains (2, 15µM). The other readings were (3,14µM) for L. (V.) lainsoni, 2,96µM for L. (V.) shawi, 2,76µM for L. (V.) guyanensis and 3,1µM for L. (V.) braziliensis-LCL strains. It is conc1uded that L. (V.) natffi. In this way it may be noted the NO levels for infected macrophage were inversely proportional to the degree of infectivity of the species studied.Os parasitos do gênero Leishmania apresentam variações de infectividade intra e inter específicas. Entretanto, são escassas as informações a respeito da infectividade das espécies de Leishmania do Novo Mundo, principalmente, daquelas encontradas na região Amazônica brasileira, onde, até o presente momento são conhecidas seis espécies pertencentes ao subgênero Viannia causadoras de LTA. Diante disso, o objetivo do presente trabalho foi investigar, in vitro, o comportamento da infectividade de 5 espécies de Leishmania do subgênero Viannia em macrófagos peritoneais de camundongos BALB/c e sua correlação com a produção de óxido nítrico pelos macrófagos infectados. Trinta cepas de Leishmania foram distribuídas em seis grupos iguais, de acordo com as espécies seguintes: I- L. (V) braziliensis/LCL, II- L. (V) braziliensis/LCM, III- L. (V) guyanensis, IV- L. (V) shawi, V -L. (V) naiffi e VI- L. (V) lainsoni. As cepas foram cultivadas em meio RPMI suplementado com 10% de soro bovino fetal e 1% de penicilina-gentamicina, até atingir a fase estacionária de cultivo, quando foram usadas para infectar macrófagos na proporção de 4 parasitos/macrófago. As culturas foram incubadas a 35°C e 5% de CO2 e após 24h, as lamínulas foram coradas com Giemsa para contagem do número de parasitos e determinação do índice de infecção, enquanto a concentração de NO (nitrito) foi calculada pelo método de Griess. Observou-se que as cepas de L. (V) braziliensis/LCM apresentaram o maior índice de infecção (385), sendo este significativamente maior (P<0,005) que as cepas de L. (V) braziliensis/LCL (264), L. (V) naiffi (215) e L. (V) lainsoni (272), porém, não significativamente maior que os índices das espécies L. (V) shawi (292) e L. (V) guyanensis (300). Quanto aos níveis de NO (nitrito), detectou-se maior concentração para a espécie L. (V) naiffi (4,1µM e menor concentração para as cepas de L. (V) braziliensis/LCM (2,15µM). As demais espécies tiveram concentrações de: L. (v:) lainsom (3,14µM), L. (V) shawi (2,96µM), L. (V) guyanensis (2,76µM) e cepas de L. (V) braziliensis/LCL (3,1µM). Diante do exposto, concluímos que cepas de L. (V) braziliensis/LCM são mais infectivas do que as demais espécies estudadas e, também, mais infectivas que cepas homólogas isoladas de casos clínicos de LCL. Além disso, observou-se menor infectividade da espécie L. (V) naiffi. Desse modo, notou-se que os níveis de NO produzidos pelos macrófagos infectados foram inversamente proporcionais ao grau do parasitismo

Reviewing the trajectory of American visceral leishmaniasis in Brazilian Amazon: from Evandro Chagas to the current days

Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Leishmanioses Prof. Dr. Ralph Lainson. Ananindeua, PA, Brasil / Universidade Federal do Pará. Núcleo de Medicina Tropical. Belém, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Leishmanioses Prof. Dr. Ralph Lainson. Ananindeua, PA, BrasiMinistério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Leishmanioses Prof. Dr. Ralph Lainson. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Leishmanioses Prof. Dr. Ralph Lainson. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Leishmanioses Prof. Dr. Ralph Lainson. Ananindeua, PA, Brasil.O presente estudo reviu a trajetória da leishmaniose visceral americana (LVA) na Amazônia, Brasil, desde os tempos do dr. Evandro Chagas, fundador do Instituto de Pathologia Experimental do Norte, em 1936, o qual, após a morte trágica do seu patrono, em 1940, passou a chamar-se Instituto Evandro Chagas até os dias atuais, objetivando melhor visibilidade a cerca do legado médico-científico deixado pelo ilustre personagem, além de descrever os caminhos que fizeram essa endemia sair do anonimato epidemiológico cinco décadas atrás, para surgir como um dos maiores agravos parasitários no início do século atual. Nesse contexto, Chagas e seus colaboradores deixaram três contribuições marcantes: i) descreveram a espécie parasitária responsável pela LVA, a Leishmania chagasi (= Leishmania (Leishmania) infantum chagasi); ii) incriminaram a espécie flebotomínica Phlebotomus longipalpis como o provável vetor da LVA; e iii) postularam que a origem da doença humana deveria estar em algum animal silvestre. A situação da LVA na Amazônia brasileira não mudou muito nas décadas seguintes, porém, a partir dos anos 1980, assumiu um perfil novo, reaparecendo com maior frequência nos focos rurais e em zonas suburbanas e urbanas de cidades de médio porte, como Santarém, no Estado do Pará. Nas duas últimas décadas, o processo de expansão intensificou-se face aos fatores ambiental (desflorestamento), socioeconômico e a ocupação desordenada na periferia das cidades, onde a presença do vetor (Lutzomyia longipalpis) no peridomicílio humano, e do cão doméstico altamente suscetível à infecção, facilitaram sua disseminação. Hoje, a LVA já alcança a Região Metropolitana de Belém (ilha de Cotijuba), capital do Pará.This study reviewed the trajectory of American visceral leishmaniasis (AVL) in Brazilian Amazon, since that time of Dr. Evandro Chagas, who founded the Instituto de Pathologia Experimental do Norte, in 1936, which following the tragic death of its patron, in 1940, was renamed Instituto Evandro Chagas till the actual days, aiming the best visibility on the medical-scientific legacy left by that distinguished person, as well as trying to describe the ways that made AVL leaves the epidemiologic anonymity five decades ago for arising as one of greatest parasitic disease at the beginning of this century. In this context, Chagas et al have left three marked contributions: i) described a new parasitic species responsible for AVL, Leishmania chagasi; ii) incriminated the phlebotomine species Phlebotomus longipalpis as the likely vector of AVL; and iii) postulated that the human disease origin should be in any forest animal. The AVL situation in Brazilian Amazon has not changed in the following decades, however, in the early 1980s the disease resurfaced with a greater frequency in rural foci and in the suburban and urban areas of medium-size cities as Santarém, Pará State. In the last two decades, the expansive process increased due to the deforestation, socio-economic factor and unorganized occupation in the outskirts of cities, where the presence of Lutzomyia longipalpis in the peridomiciliar human area and the domestic dog highly susceptible to infection have facilitated its dissemination. Actually, AVL has already arrived in the Metropolitan Region of Belém (Cotijuba island), capital of Pará

In vitro infectivity of species of Leishmania (Viannia) responsible for American cutaneous leishmaniasis

There is little available information regarding the infectivity of New World Leishmania species, particularly those from the Amazonian Brazil, where there are six species of the subgenus Viannia causing American cutaneous leishmaniasis (ACL). The aim of this study was to compare, in vitro, the potential infectivity of the following Leishmania (Viannia) spp.: L. (V.) braziliensis from localized cutaneous leishmaniasis (LCL) and mucocutaneous leishmaniasis (MCL) patients, L. (V.) guyanensis, L. (V.) shawi, L. (V.) lainsoni and L. (V.) naiffi from LCL patients only, in cultured BALB/c mice peritoneal macrophage, as well as the production of NO by the infected cells. The infectivity of parasites was expressed by the infection index and, the nitric oxide (NO) production in the macrophage culture supernatant was measured by the Griess method. It was found that L. (V.) braziliensis from MCL, the more severe form of disease, showed the highest (p <= 0.05) infection index (397), as well as the lowest NO production (2.15 mu M) compared with those of other species. In contrast, L. (V.) naiffi which is less pathogenic for the human showed the lowest infection index (301) and the highest NO production (4.11 mu M). These results demonstrated a negative correlation between the infectivity and the ability of these parasites to escape from the microbicidal activity of the host cell

First report on feline leishmaniasis caused by Leishmania (Leishmania) amazonensis in Amazonian Brazil

Ministério da Saúde; Iowa Energy CenteMinistério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro Nacional de Primatas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro Nacional de Primatas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro Nacional de Primatas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro Nacional de Primatas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro Nacional de Primatas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro Nacional de Primatas. Ananindeua, PA, Brasil / Universidade Federal do Pará. Núcleo de Medicina Tropical. Belém, PA, Brazil.In the present study, we reported the natural infection by Leishmania sp. in a domestic cat, in which the amastigote forms of the parasite were observed within a lesion on its ear-tip. Fragment of the lesion was obtained and cultured in NNN medium, and PCR-RFLP analysis of the isolated sample was performed, which revealed that the profile was compatible with Leishmania (L.) amazonensis. This is the first proven case of a cat infected by L. (L.) amazonensis reported in Belém city, Pará state, northern Brazil

Correlation between the components of the insulin-like growth factor I system, nutritional status and visceral leishmaniasis

University of São Paulo Medical School. Department of Pathology. São Paulo, SP, Brazil.University of São Paulo Medical School. Division of Endocrinology. São Paulo, SP, Brazil.Federal Universty of Maranhão. Department of Pediatrics. São Luis, MA, Brazil.University of São Paulo. Department of Epidemiology. São Paulo, SP, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Belém, PA, Brasil.University of São Paulo Medical School. Department of Pathology. São Paulo, SP, Brazil.The role of the insulin-like growth factor I (IGF-I) system and nutritional status was studied in 241 children from a Brazilian area endemic for visceral leishmaniasis (VL). Thirtynine children had the active form, 20 were oligosymptomatic, 38 were asymptomatic and 144 were not infected. Serum concentrations of growth hormone (GH), total and free IGF-I and IGF binding-protein 3 (IGFBP3) were measured by radioimmunoassay. Nutritional status was evaluated by anthropometric indicators and biochemical measurements. Total and free IGF-I and IGFBP3 were significantly reduced in the active form. Z scores for total and free IGF-I and for IGFBP3 were found to be significantly lower for active VL and oligosymptomatic individuals than for asymptomatic individuals, but never reached values ≤2 SD. Median values of weight-for-age Z and height-for-age Z (HAZ) scores and albumin concentration were significantly different in the active VL group compared with the other groups. Multiple discriminant analysis selected albumin and HAZ score as predictors of active and oligosymptomatic VL. The lack of correlation between auxological data and serum concentrations of the GH/IGF axis components suggested that the primary cause of retarded growth in children with active VL is not dependent on IGF-I or IGFBP3, but rather on VL intrinsic factors that might secondarily involve the GH/IGF axis