Dorlene Maria Cardoso de Aquino

Balance of IL-10 and Interferon-γ plasma levels in human visceral leishmaniasis: Implications in the pathogenesis

BackgroundLeishmaniasis remains a serious public health problem in several parts of the developing world. Effective prophylactic measurements are hampered by imprecise comprehension of different aspects of the disease, including its immunoregulation. A better comprehension of immunoregulation in human VL may be useful both for designing and evaluating immunoprophylaxis.MethodsTo explore immunoregulatory mechanisms, 20 visceral leishmaniasis (VL) patients were evaluated during active disease and at different periods up to one year after treatment determining their plasma cytokine levels, clinical parameters (palpable spleen and liver) and antibody levels.ResultsElevated plasma levels of IFN-γ and of IL-12 p40 were observed during active disease, significantly decreasing after treatment whereas in vitro Leishmania antigen-stimulated IFN-γ production by PBMC exhibited an inverse pattern being low during disease and increasing steadily thereafter. Absence of IFN-γ activity is a hallmark of VL. The main candidate for blunting IFN-γ activity is IL-10, a cytokine highly elevated in plasma with sharp decrease after treatment. Activity of IL-10 is inferred by high levels of anti-Leishmania specific IgG1 and IgG3. TGF-β had elevated total, but not of active, levels lessening the likelihood of being the IFN-γ counterpart. Spleen or liver size presented a steady decrease but return to normal values at only 120 days after treatment. Anti-Leishmania IgG (total and subclasses) levels and DTH or Leishmania-stimulated lymphocyte proliferation conversion to positive also present a slow decrease after treatment. IL-6 plasma levels were elevated in only a few patients.ConclusionTaken together our results suggest that IFN-γ and IL-10 are the molecules most likely involved in determining fate of disease. After treatment, there is a long delay before the immune profile returns to normal what precludes using plasma cytokine levels as criteria of cure as simpler clinical evaluations, as a palpable spleen or liver, can be used.

Using recombinant proteins from Lutzomyia longipalpis saliva to estimate human vector exposure in visceral Leishmaniasis endemic areas.

Background Leishmania is transmitted by female sand flies and deposited together with saliva, which contains a vast repertoire of pharmacologically active molecules that contribute to the establishment of the infection. The exposure to vector saliva induces an immune response against its components that can be used as a marker of exposure to the vector. Performing large-scale serological studies to detect vector exposure has been limited by the difficulty in obtaining sand fly saliva. Here, we validate the use of two sand fly salivary recombinant proteins as markers for vector exposure. Methodology/principal findings ELISA was used to screen human sera, collected in an area endemic for visceral leishmaniasis, against the salivary gland sonicate (SGS) or two recombinant proteins (rLJM11 and rLJM17) from Lutzomyia longipalpis saliva. Antibody levels before and after SGS seroconversion (n = 26) were compared using the Wilcoxon signed rank paired test. Human sera from an area endemic for VL which recognize Lu. longipalpis saliva in ELISA also recognize a combination of rLJM17 and rLJM11. We then extended the analysis to include 40 sera from individuals who were seropositive and 40 seronegative to Lu. longipalpis SGS. Each recombinant protein was able to detect anti-saliva seroconversion, whereas the two proteins combined increased the detection significantly. Additionally, we evaluated the specificity of the anti-Lu. longipalpis response by testing 40 sera positive to Lutzomyia intermedia SGS, and very limited (2/40) cross-reactivity was observed. Receiver-operator characteristics (ROC) curve analysis was used to identify the effectiveness of these proteins for the prediction of anti-SGS positivity. These ROC curves evidenced the superior performance of rLJM17+rLJM11. Predicted threshold levels were confirmed for rLJM17+rLJM11 using a large panel of 1,077 serum samples. Conclusion Our results show the possibility of substituting Lu. longipalpis SGS for two recombinant proteins, LJM17 and LJM11, in order to probe for vector exposure in individuals residing in endemic areas.

Perfil dos pacientes com hanseníase em área hiperendêmica da Amazônia do Maranhão, Brasil

An epidemiological cross-sectional study of 207 patients with leprosy disease, was undertaken between August 1998 to november 2000, aiming at evaluating the socioeconomic, demographic and ambiental profiles of the patients as well as physical incapacity due to the disease. The study was performed in the municipality of Buriticupu-Maranh o state, a hiperendemic leprosy area in the Amazonian Maranh o. The level of incapacity was assessed from parameters established by the Brazilian Health Minister. The clinical evaluation and the results of the physical tests were registered in a standardized form. It was observed a predominance of married people (45,9%), with low level of education (56%), being lend workers (40,1%), with familiar income to the minimum wage (76,3%), aged from 14 to 44 years (63,3%), males (60,9%) and brown (67,6%); 44% living in mud huts, 82,6% deposited their excrements in cesspits and 63,8% do not treat the drinking water, 58% utilized well-water and 51,7% do not use treated water for ingestion. The most affected segments of the body were the feet (62,3%), eyes (51,2%) end hands (7,2%), being the higher percentage of physical incapacitaties found among the patients bearing the borderline form of the disease (93%) mainly hands and feet, and in the virchowian form greatest frequency of eyes incapacities. It is concluded that the hyperendemicity associated with the precarious socioeconomic conditions and with a high level of physical incapacities may be involved with the living quality of the patients.An epidemiological cross-sectional study of 207 patients with leprosy disease, was undertaken between August 1998 to november 2000, aiming at evaluating the socioeconomic, demographic and ambiental profiles of the patients as well as physical incapacity due to the disease. The study was performed in the municipality of Buriticupu-Maranhao state, a hiperendemic leprosy area in the Amazonian Maranhao. The level of incapacity was assessed from parameters established by the Brazilian Health Minister. The clinical evaluation and the results of the physical tests were registered in a standardized form. It was observed a predominance of married people (45,9%), with low level of education (56%), being lend workers (40,1%), with familiar income to the minimum wage (76,3%), aged from 14 to 44 years (63,3%), males (60,9%) and brown (67,6%); 44% living in mud huts, 82,6% deposited their excrements in cesspits and 63,8% do not treat the drinking water, 58% utilized well-water and 51,7% do not use treated water for ingestion. The most affected segments of the body were the feet (62,3%), eyes (51,2%) end hands (7,2%), being the higher percentage of physical incapacitaties found among the patients bearing the borderline form of the disease (93%) mainly hands and feet, and in the virchowian form greatest frequency of eyes incapacities. It is concluded that the hyperendemicity associated with the precarious socioeconomic conditions and with a high level of physical incapacities may be involved with the living quality of the patients.

TGFB1 and IL8 gene polymorphisms and susceptibility to visceral leishmaniasis

Visceral leishmaniasis (VL) or Kala-azar is a serious protozoan infectious disease caused by an obligate intracellular parasite. Cytokines have a major role in determining progression and severity of clinical manifestations in VL. We investigated polymorphisms in the TGFB1and IL8 genes, which are cytokines known to have a role in onset and severity of the disease. Polymorphisms at TGFB1 -509 C/T and +869 T/C, and IL8 -251 A/T were analyzed by a PCR-RFLP technique, in 198 patients with VL, 98 individuals with asymptomatic infection positive for a delayed-type hypersensitivity test (DTH+) and in 101 individuals with no evidence of infection (DTH-). The presence of the T allele in position -509 of the TGFB1 gene conferred a two-fold risk to develop infection both when including those with clinical symptoms (DTH+ and VL, grouped) or when considering DTH+ only, respectively p = 0.007, OR = 1.9 [1.19-3.02] and p = 0.012, OR = 2.01 [1.17-3.79], when compared with DTH- individuals. In addition, occurrence of hemorrhage was associated with TGFB1 -509 T allele. We suggest that the -509 T allele of the TGFB1 gene, a cytokine with a biologically relevant role in the natural history of the disease, may contribute to overall susceptibility to infection by Leishmania and to severity of the clinical disease.

Epidemiological, clinical, and operational aspects of leprosy patients assisted at a referral service in the state of Maranhão, Brazil

INTRODUCTION Leprosy is an infectious disease caused by Mycobacterium leprae. The aim of this study was to describe the epidemiological, clinical, and operational aspects of leprosy carriers. METHODS A cross-sectional study leprosy patients assisted in São Luis, MA, was performed. RESULTS Of the 85 cases analyzed, 51.7% were male participants, and 60% were brown. Concerning the age, 54.8% of women were between 35 and 49 years, and 57.6% of men were between 20 and 34 years. Lepromatous leprosy was found in 42.3% of cases, and the multibacillary form was found in 72.9%. The skin smear was positive in 42.3%. The occurrence of reaction was found in 43.5% of cases, and 83.5% had no Bacillus Calmette-Guérin scar. Leprosy in the family was reported by 44.7% of the patients. Most of the individuals (96.4%) lived in houses made of brick with more than three rooms (72.6%) and two persons per room (65.1%). Concerning the level of education, 41.4% of women and 34.1% of men had more than one to three years of education. The most evaluated age group in the beginning of the treatment was that of 35 to 49 years with a Grade 0 incapability (64.5%), and that in the end was the age group of 20 to 34 (29.9%) with Grade 0, 30.7% Grade 1, and 11.5% Grade 2. CONCLUSIONS The frequency of multibacillary forms found in this study and the cases in family members point out delayed diagnoses. Thus, early diagnosis and appropriate treatment are important in decreasing the outcome of disabilities.

Leishmania infection in humans, dogs and sandflies in a visceral leishmaniasis endemic area in Maranhão, Brazil

Leishmania infection in humans, dogs and sandflies was examined in the endemic visceral leishmaniasis (VL) municipality of Raposa, state of Maranhão, Brazil. In this study, we examined Leishmania chagasi infection in the blood serum of both humans and Canis familiaris and the natural Leishmania sp. infection rate in the sandfly vector, Lutzomyia longipalpis. Enzyme-linked immunosorbent assay, indirect immunofluorescence reaction and polymerase chain reaction were performed to detect Leishmania infections in humans, dogs and sandflies, respectively. Overall, 186 out of 986 studied human beings were infected with L. chagasi parasites, representing an infection prevalence of 18.9%. An even higher infection rate was detected in dogs, where 66 (47.8%) out of 138 were infected. Among all Lu. longipalpis captured (n = 1,881), only 26.7% were females. The Leishmania infection frequency for the vector Lu. longipalpis was 1.56%. Remarkably, all infected sandflies were found in the peridomiciliary area. Furthermore, a high incidence of asymptomatic forms of VL in the human and canine populations was observed. The results of this study suggest autochthonous transmission of L. chagasi in this endemic area for visceral leishmaniasis because infection by Leishmania sp. was identified in all important elements of the transmission chain.

Cytokines and visceral leishmaniasis: a comparison of plasma cytokine profiles between the clinical forms of visceral leishmaniasis

It is not well established whether cytokine production differs in response to different clinical forms of visceral leishmaniasis (VL). In this work, we performed a cross-sectional study to investigate the plasma levels of cytokines [interferon (IFN)-γ, tumour necrosis factor (TNF)-α, interleukin (IL)-2, IL-4, IL-10 and IL-12] involved in the pathogenesis of VL in 80 subjects from VL endemic areas, including subjects with active VL, subjects with asymptomatic infection, subjects with cured VL and uninfected controls. The patients were recruited by sampling from a referral hospital and by random selection from a population-based cohort study. The results showed significant differences in the plasma concentration of all cytokines between the groups (p < 0.05). Patients with the active disease had higher plasma levels of IL-10, IL-4, INF-γ and TNF-α relative to the other groups and they produced more IL-12 than asymptomatic and cured subjects. Only the IL-2 concentration was higher in the asymptomatic and cured subjects relative to the patients with active disease (p < 0.05). Our results suggest that these cytokines can be used as markers in epidemiological studies conducted in endemic areas to distinguish between different clinical forms of VL. However, their usefulness should be confirmed in investigations conducted in other endemic areas.

Epidemiological study of the association between anti-Lutzomyia longipalpis saliva antibodies and development of delayed-type hypersensitivity to Leishmania antigen.

Recent reports from animal models and from cross-sectional studies have suggested that host responses to anti-Lutzomyia longipalpis saliva antibodies may be related to delayed-type hypersensitivity to Leishmania antigen. In a prospective cohort study, we evaluated 1,080 children from two endemic areas for visceral leishmaniasis (VL) by means of Kaplan-Meier analysis. The incidence rate of delayed-type hypersensitivity to Leishmania antigen, measured at the 24th follow-up month, was higher among those reactive to Lu. longipalpis saliva antibodies at the beginning of the study (0.0217 cases per person-month) than among those previously negative (0.0131 cases per person-month) (P value for the log-rank test = 0.0006). It seems that mounting an anti-saliva immune response helps the development of a cell-mediated anti-Leishmania response.

Neural complications and physical disabilities in leprosy in a capital of northeastern Brazil with high endemicity

INTRODUCTION Leprosy is an infectious disease whose etiologic agent is Mycobacterium leprae, manifested by dermatological and neurological signs and symptoms. OBJECTIVE To investigate neural changes and the degree of physical disability in the eyes, hands and feet before and after treatment, as well as sociodemographic and clinical profile of patients affected by leprosy. METHOD A longitudinal epidemiological study comprising 155 patients with leprosy, from a spontaneous demand, diagnosed between March 2010 and February 2011, and treated with multidrug therapy (MDT) between March 2010 and July 2012 in a program for leprosy eradication in São Luis (MA), Brazil. RESULTS Before treatment, 46.5% of patients were considered as borderline, 51.6% had some alteration in the eyes and 52.3% in the feet, and the radial nerve (18.7%) was the most affected. There was a statistically significant difference between the changes in the radial nerve at the beginning of and after treatment. CONCLUSIONS The analysis points to late diagnosis, as some patients have had abnormal neural and physical disabilities before treatment.

Perfil epidemiológico dos indivíduos HIV positivo e coinfecção HIV-Leishmania em um serviço de referência em São Luís, MA, Brasil

A Aids e um problema de saude publica e a Leishmaniose visceral (LV) a forma mais frequente de coinfeccao. O objetivo e descrever aspectos epidemiologicos dos individuos HIV positivo e investigar a ocorrencia de coinfeccao HIV-Leishmania. Trata-se de estudo descritivo realizado com 287 individuos HIV positivo atendidos no Centro de Referencia para DST/Aids em Sao Luis (MA). Foi realizado teste de Intradermorreacao de Montenegro, aspirado de medula ossea para deteccao da infeccao por Leishmania sp e aplicado um questionario na coleta dos dados epidemiologicos. O teste qui-quadrado foi usado para diferencas entre genero, com p < 0,05 de significância. Houve significância estatistica entre os homens relacionados ao uso de fossa septica e renda ate dois salarios. Quanto a exposicao, foi predominante a categoria heterossexual para ambos os sexos, sendo observado um aumento entre as mulheres. Entre os individuos HIV positivo 4,2% apresentavam sintomatologia sugestiva de LV sendo identificadas formas amastigotas para Leishmania sp no aspirado de medula ossea. O perfil dos individuos HIV positivo nao diferiu de outras regioes do Brasil com predominio de jovens em idade produtiva, sendo constatada uma sobreposicao das areas de transmissao para a coinfeccao HIV-LV.Burnoffs (intentional fires for agricultural purposes) and forest fires of large proportions have been observed in various regions of the planet. Exposure to high levels of air pollutants emitted by fires can be responsible for various harmful effects on human health. In this article, the literature on estimating acute effects of air pollution on human health by fires in the regions with the highest number of fires on the planet, using a time series approach is summarized. An attempt was made to identify gaps in knowledge. The study consisted of a narrative review, in which the characteristics of the selected studies were grouped by regions of the planet with a higher incidence of burnoffs: Amazon, America, Australia and Asia. The results revealed a large number of studies in Australia, few studies in the Amazon and great heterogeneity in the results on the significant effects on human health.