Expression of the chemokine receptor CXCR4 on lymphocytes of leprosy patients

Leprosy is caused by Mycobacterium leprae, which induces chronic granulomatous infection of the skin and peripheral nerves. The disease ranges from the tuberculoid to the lepromatous forms, depending on the cellular immune response of the host. Chemokines are thought to be involved in the immunopathogenesis of leprosy, but few studies have investigated the expression of chemokine receptors on leukocytes of leprosy patients. In the present study, we evaluated 21 leprosy patients (M/F: 16/5) with a new diagnosis from the Dermatology Outpatient Clinic of the University Hospital, Federal University of Minas Gerais. The control group was composed of 20 healthy members (M/F: 15/5) of the community recruited by means of announcements. The expression of CCR2, CCR3, CCR5, and CXCR4 was investigated by flow cytometry on the surface of peripheral blood lymphocytes. There was a decrease in percentage of CD3+CXCR4+ and CD4+CXCR4+ lymphocytes in the peripheral blood of leprosy patients (median [range], 17.6 [2.7-41.9] and 65.3 [3.9-91.9], respectively) compared to the control group (median [range], 43.0 [3.7-61.3] and 77.2 [43.6-93.5], respectively). The percentage of CD4+CXCR4+ was significantly lower in patients with the tuberculoid form (median [range], 45.7 [0.0-83.1]) of the disease, but not in lepromatous patients (median [range], 81.5 [44.9-91.9]). The CXCR4 chemokine receptor may play a role in leprosy immunopathogenesis, probably directing cell migration to tissue lesions in tuberculoid leprosy patients.

Talidomida usada por pacientes com eritema nodoso hansênico / Thalidomide used by patients with erythema nodosum leprosum

INTRODUÇÃO: A talidomida é um fármaco utilizado atualmente no tratamento do eritema nodoso hansênico no Brasil. MÉTODOS: Estudo prospectivo para acompanhar a evolução clínica, registrar os eventos adversos e determinar as concentrações plasmáticas de talidomida em dose diária de 100mg/dia, em 20 pacientes com manifestações clínicas de eritema nodoso hansênico, divididos em dois grupos: após ou em curso da poliquimioterapia para hanseníase. RESULTADOS: Não foram observadas diferenças significativas nos grupos no decorrer do estudo, tanto na evolução clínica favorável dos pacientes, de 70 por cento e 90 por cento, quanto nos eventos adversos registrados que foram tontura e sonolência. Os teores plasmáticos de talidomida em D7 e D14 foram de 0,82±0,4μg/mL e 0,79±0,3μg/mL no grupo 1 e de 0,82±0,4 e 1,55±1,0 no grupo 2, respectivamente. CONCLUSÕES: Na amostra estudada, a poliquimioterapia não interferiu na evolução clínica, na incidência dos efeitos adversos e nos níveis plasmáticos de talidomida.

INTRODUCTION: Thalidomide is a drug currently used in Brazil for treating erythema nodosum leprosum. METHODS: This was a prospective study to follow up clinical evolution, record adverse events and determine plasma thalidomide levels from a dose of 100 mg/day, among 20 patients with clinical manifestations of erythema nodosum leprosum, divided into two groups: during or after leprosy multidrug therapy. RESULTS: No significant differences between the groups were seen during the study, either in relation to favorable clinical evolution among the patients (70 percent and 90 percent), or in relation to the adverse events recorded, which were dizziness and somnolence. The plasma thalidomide levels on D7 and D14 were 0.82 ± 0.4μg/ml and 0.79 ± 0.3 μg/ml in group 1 and 0.82 ± 0.4 and 1.55 ± 1.0 in group 2, respectively. CONCLUSIONS: In this sample, the multidrug therapy had no effect on the clinical evolution, incidence of adverse events and plasma thalidomide levels.

Estudo da microcirculação na hanseníase virchowiana com o uso da microscopia de luz ortogonal polarizada e com laser-doppler fluxometria / Study of microcirculation in lepromatous leprosy with the use of orthogonal polarized light microscopy and laser doppler flowmetry

A hanseníase é uma doneça infecciosa com características únicas, dentre elas o fato de atingir intensamente a inervação da pele e seus anexos. Entremeando estes anexos, está a microcirculação cutânea, que a principio também tem sua inervação comprometida. Vários artigos apontam para alterações de disautonomia microcirculatória, citando como exemplo as alterações no reflexo vasomotor. O presente estudo se propõe a avaliar a microcirculação cutânea na hanseníase virchowiana, tanto em sua morfologia quanto em sua reatividade vascular. Para isto, utilizamos a tecnologia de luz ortogonal polarizada através do equipamento Cytoscan, a análise de Fourier do sinal do laser Doppler para estudo da vasomotricidade e o laser Dopplerfluxometria associado à iontoforese de substâncias vasoativas (acetilcolina, nitroprussiato de sódio e noradrenalina) para avaliação da reatividade vascular. Dez pacientes portadores de hanseníase virchowiana sem outras comorbidades que pudessem alterar os parâmetros microcirculatórios, foram avaliados pelos métodos descritos e seus resultados foram comparados aos de dez controles sem hanseníase ou qualquer outra comorbidade. Em relação à vasomotricidade não foram observadas alterações estatisticamente significativas entre os grupos, o que fala a favor da teoria de origem miogênica para a vasomotricidade. Em relação à iontoforese de substâncias vasoativas constatou-se uma diminuição da resposta vasodilatadora à acetilcolina e ao nitroprussiato nos pacientes com hanseníase. Os exames com o Cytoscan mostraram aumento no tamanho dos capilares, bem como alterações em sua morfologia. Os resultados apresentados sugerem que, provavelmente devido ao longo período de alteração inervatória decorrente da hanseníase virchowiana, estes pacientes apresentam uma alteração significativa tanto morfológica quanto na reatividade vascular da microcirculação cutânea.

Leprosy is an infectious disease with unique characterístics. One of them is the fact that it compromises not only the cutaneous and adnexial innervation, but also the innervation of the cutaneous microcirculation. Several articles indicate the impact of disautonomy on the microcirculatory level, citing the example of changes in vasomotor level. The present study proposes to evaluate morphology and microvascular reactivity of the cutaneous microcirculation of the virchowian leprosy. Methods employed in the study were: the Cytoscan, which uses the orthogonal polarized light, the Fourier analysis of the laser Doppler signal to study vasomotion, and the laser Doppler flowmetry associated with iontophoresis of vasoactive substances (acetylcholine, sodium nitroprusside and norepinephrin). Ten patients with virchowian leprosy, without any other comorbidity that could modify the microvascular parameters were evaluated and their results were compared to ten controls without leprosy or any other comorbidity. Regarding the vasomotion, no statistical significant differences were noticed between the groups. Our data are in agreement with the vasomotion's miogenic origin theory. According to iontophoresis of vasoactive substances, it was found that there is a reduced endothelial-dependent and endothelial-independent vasodilation in patients with leprosy while tests by direct visualization we observed an increase in the size of capillaries, as well as changes in their morphology. The results suggest that the significant changes in morphology and vascular reactivity of skin microcirculation are probably due to the long period of innervatory changes arising from leprosy.

Serum adenosine deaminase levels in reactional and non-reactional leprosy.

BACKGROUND AND AIMS: Altered serum adenosine deaminase (ADA) levels have been recorded in various diseases involving lymphocytes and/or lymphoreticular system including leprosy. The study was planned to evaluate alterations in serum ADA levels, if any, in reactional and non-reactional leprosy. METHODS: Eighty patients of leprosy, comprising 60 patients of non-reactional leprosy and 20 patients of reactional leprosy were studied along with 20 normal healthy controls. Five milliliters of venous blood was collected and ADA levels were estimated by the method of Giusti (1974). RESULTS: There were 54 males and 26 females. The age of the patients ranged from 5 years to 62 years. The duration of leprosy ranged from 15 days to 3 years. The mean serum ADA level in normal controls was 10.31 +/- 0.58 u/L. The serum ADA levels were raised in leprosy patients, significantly so in multibacillary patients. The serum ADA levels were higher in patients of leprosy with reaction. CONCLUSIONS: The study showed significantly high serum ADA levels in multibacillary leprosy and this was further increased in patients of leprosy with reaction. This may be because of increased lymphoreticular activity during the reactional phases.

Quantificacao do estresse oxidativo no sangue de hansenianos sob o efeito ou nao de tratamento especifico / Quantification of the oxidative stress in the blood of people with Hansen's disease undertaking or not a specific treatment

Neste estudo avaliamos a quantificaçäo do estresse oxidativo no sangue de hasenianos sob efeito ou näo de tratamento específico. Foram coletadas amostras de sangue de 62 portadores de hanseníase e 13 sadios. Destes 62 pacientes, 35 eram de forma multibacilar e faziam tratamento com clofazimina, dapsona e rifampicina; 16 de forma paucibacilar tratados com dapsona e rifampicina; 11 dos pacientes estavam sem tratamento, sendo 5 da forma multibacilar e 6 da forma paucibacilar. As amostras foram levadas para análises laboratoriais onde foram feitas as dosagens de metahemoglobina e contagem de corpos de Heinz. Os resultados encontrados foram submetidos à análise estatística com cálculo da Odds Ratio e Intervalo de Confiança 95 por cento. Concluimos que a doença haseníase näo foi a causadora do estresse oxidativo, e sim a terapêutica administrada, e o grupo que fazia uso da clofazimina apresentou piores resultados, ou seja, maior estresse oxidativo.

Avaliacao da lesao e disfuncao endotelial atraves da analise da trombomodulina e fator de von Willebrand, no grupo multibacilar da hanseniase / ?

A hanseniase e uma doenca infecciosa cronica, cujo curso evolutivo, lento e insidioso, pode ser interrompido por episodios agudos ou subagudos, caracterizando os periodos reacionais. O comprometimento vascular e muito frequente, sendo a vasculite cronica um achado comum no doente multibacilar. Durante os surtos de reacao tipo II foram relatados episodios de tromboses venosa e/ ou arterial, muitas vezes constituindo a causa de obito desses doentes. Analisamos as dosagens plasmaticas de trombomodulina e do antigeno do fator de von Willebrand como marcadores de lesao e disfuncao endotelial, respectivamente, em doentes multibacilares, durante episodios de reacao tipo II e fora dos periodos reacionais. Apos consentimento previo informado, foram coletadas 163 amostras de um total de 34 doentes com hanseniase, formas multibacilares de sofrerem reacao tipo II

Perfil hematológico de pacientes de hanseníase com episódios reacionais de eritema nodoso / Hematologic profile of leprosy patients in reactional episode of erythema nodosum leprosum

Neste trabalho foram avaliados os parâmetros hematológicos de dezoito pacientes com eritema nodoso hansênico.(ENH). Oito desses pacientes tiveram ENH intenso e moderado e dez pacientes tiveram ENH leve. Os resultados mostraram que os pacientes com ENH intenso e moderado, tiveram alteraçoes significativas nos resultados hematológicos: deficiência de ferro, um tempo aumentado de lise de euglobulina e hiperfibrinogenemia. Contudo, nenhuma alteraçao foi observada nos outros parâmetros estudados (tempo de protrombina, corpos de Heinz e produtos de degradaçao do fibrinogênio) em todos os casos de ENH.

Correlation of serum lipids and lipoproteins in leprosy.

Serum lipids and lipoproteins were assessed in sixty leprosy and forty age and sex matched healthy controls. The study subjects included cases of LL with reactions, LL without reactions, BL with reactions, BL without reactions, BT and TT types of leprosy. The levels of serum phospholipids, triglycerides, total cholesterol, LDL and VLDL fractions were significantly decreased in leprosy patients compared to control subjects. The levels of serum HDL cholesterol and HDL fraction were significantly elevated in leprosy patients. Maximum elevation in serum HDL cholesterol level and HDL fraction and maximum reduction in the levels of serum phospholipids, triglycerides, total cholesterol and LDL and VLDL fractions were observed in lepromatous leprosy (LL) patients with reactions.

Fibrinolytic phenomenon in leprosy.

Fibrinolytic activity in eighty-one patients with different types of leprosy and thirty-two normal healthy controls was studied by Euglobulin Lysis Time Method, Fibrinolytic activity was markedly decreased in patients with lepromatous leprosy and those with ENL reaction. Decline in fibrinolytic activity during ENL was independent of frequency of attacks. Fibrinolytic activity was partly restored after subsidence of ENL reaction, though it failed to attain normal levels. Cutaneous vasculitis seems to be most probable cause of fall in fibrinolytic activity in lepromatous leprosy and ENL reaction.

Glucose tolerance test in leprosy.

Glucose tolerance test was carried out in 43 cases of leprosy. They included cases of tuberculoid, borderline, lepromatous leprosy and those with lepra reaction. It was observed that normal curve was common in tuberculoid leprosy. Flat glucose tolerance curve was observed in borderline and lepromatous leprosy. However, the diabetic curve was common in Lepra reaction. Fasting blood sugar was low in lepromatous leprosy and it tended to be marginally high in lepra reaction. Normal GTT response was observed in those with duration of disease between 0-6 months, flat curves in those with duration of disease between 7-12 months while diabetic curve was more common in those with disease duration of more than 2 years.

Interleukin-1 beta production by monocytes from leprosy patients

The cause responsible for the lack of an efficient cell-mediated immunity or a delayed type hypersensitivity to M. leprae in lepromatous patients is poorly understood. But the resistance to M. leprae infection in humans is likely mediated by the activated macrophages to present M. leprae antigen to T cells for cell-mediated immunity. Phenolic glycolipid-I (PGL-I) is a M. leprae-specific antigen and is supposed to play a significant role in the long lasting unresponsiveness in lepromatous leprosy. In this study, IL-1 activities were tested among leprosy patients to evaluate monocyte function and the role of IL-1 in the immunosuppression in leprosy. We found that peripheral blood mononuclear cells (PBMCs) from tuberculoid patients were strongly reactive to M. leprae (mean cpm; 28,853 +/- 28,916), but the proliferative responses of PBMCs from lepromatous patients (mean cpm; 6,051 +/- 803) were significantly lower. IL-1 concentration in culture supernatant of monocytes from lepromatous patients was similar to that from tuberculoid patients with stimulation of M. leprae (lepromatous: 1,014 +/- 637 pg/ml, tuberculoid: 1,012 +/- 167 pg/ml) or lipopolysaccharides (IPS) (lepromatous: 3,479 +/- 2,188 pg/ml, tuberculoid: 4,246 +/- 2,432 pg/ml). The IL-1 concentration is sera from lepromatous patients (42 +/- 30 pg/ml) tended to be higher than those from tuberculoid patients (28 +/- 69 pg/ml). And there was no significant difference in IL-1 production between peritoneal macrophages from mice sensitized with PGL-1 and those from nonsensitized mice. In conclusion, this study suggests that the immunosuppression in lepromatous patients may not be due to the decreased production of IL-1. And the increased IL-1 activity in sera may affect the inflammatory response of lepromatous patients.

Hormone profile in leprosy.

Hormone profile were carried out in 35 male cases of leprosy. They were divided into tuberculoid leprosy, borderline leprosy, lepromatous leprosy and lepra reaction. Serum testosterone, follicle-stimulating hormone, Luteinizing hormone, and Oestradiol level were measured in these cases of leprosy. It was observed that serum testosterone were significantly low in lepromatous leprosy (P less than 0.001) and lepra reaction (P less than 0.01). The serum levels of follicle-stimulating hormone and luteinizing hormone were significantly high in lepromatous leprosy (P less than 0.02) and lepra reaction (P less than 0.05). Serum Oestradiol was raised in approx. 60% cases in borderline leprosy, lepromatous leprosy and lepra reaction.

Conformational changes in the IgG molecule of lepromatous sera using laser Raman spectroscopy.

In the present work we report our studies on IgG separated from the serum of lepromatous patients and non-lepromatous leprosy cases using Laser Raman Spectroscopy. Striking spectral changes in LL cases have been observed in the following special regions: (a) the amide I and III, (b) the S-S and C-S stretching (c) the skeletal bending and (d) skeletal stretching regions. These changes indicate a decrease in the amount of beta-structure and a transition towards alpha-helical conformation.

Undernutrition and lepromatous leprosy. Serum vitamin A and E levels in leprosy spectrum.

Serum vitamins A and E were estimated by spectrophotocolorimetric methods in 67 leprosy patients comprising 9 BT, 10 BB, 15 BL, 27 LL, including 12 Histoid cases. These findings were evaluated in comparison to 55 normal subjects serving as controls. A significant reductions in the mean serum levels of Vitamins 'A' and 'E' were observed in the leprosy groups as compared to normal controls. These findings are of considerable importance and need to be taken note of in the light of delineating these alterations to the cause or effect of the disease. As far as we know, this is the first report describing serum levels of Vitamins 'A' and 'E' in the leprosy spectrum.