Increased platelet distribution width and reduced IL-2 and IL-12 are associated with thrombocytopenia in Plasmodium vivax malaria

BACKGROUND Thrombocytopenia in malaria involves platelet destruction and consumption; however, the cellular response underlying this phenomenon has still not been elucidated. OBJECTIVE To find associations between platelet indices and unbalanced Th1/Th2/Th17 cytokines as a response to thrombocytopenia in Plasmodium vivax infected (Pv-MAL) patients. METHODS Platelet counts and quantification of Th1/Th2/Th17 cytokine levels were compared in 77 patients with uncomplicated P. vivax malaria and 37 healthy donors from the same area (endemic control group - ENCG). FINDINGS Thrombocytopenia was the main manifestation in 55 patients, but was not associated with parasitaemia. The Pv-MAL patients showed increases in the mean platelet volume (MPV), which may be consistent with larger or megaplatelets. Contrary to the findings regarding the endemic control group, MPV and platelet distribution width (PDW) did not show an inverse correlation, due the increase in the heterogeneity of platelet width. In addition, the Pv-MAL patients presented increased IL-1β and reduced IL-12p70 and IL-2 serum concentrations. Furthermore, the reduction of these cytokines was associated with PDW values. MAIN CONCLUSIONS Our data demonstrate that an increase in MPV and the association between reductions of IL-2 and IL-12 and PDW values may be an immune response to thrombocytopenia in uncomplicated P. vivax malaria.

New perspectives on traumatic bone infections / 中华创伤杂志(英文版)

In this paper, we review the results of previous studies and summarize the effects of various factors on the regulation of bone metabolism in traumatic bone infections. Infection-related bone destruction incorporates pathogens and iatrogenic factors in the process of bone resorption dominated by the skeletal and immune systems. The development of bone immunology has established a bridge of communication between the skeletal system and the immune system. Exploring the effects of pathogens, skeletal systems, immune systems, and antibacterials on bone repair in infectious conditions can help improve the treatment of these diseases.

Establishing th reference range for t lymphocyes subpopulations in adults andd children from Brazil / Estabelecimento de valores de referencias para subpopulacoes de linfocitos T em adultos e criancas no Brasil

SUMMARY In Brazil, the existing reference values for T-lymphocytes subsets are based on data originated in other countries. There is no local information on normal variation for these parameters in Brazilian adults and children. We evaluated the normal variation found in blood donors from five large Brazilian cities, in different regions, and in children living in Salvador, and Rio de Janeiro. All samples were processed by flow cytometry. The results were analyzed according to region, gender, and lifestyle of blood donors. A total of 641 adults (63% males), and 280 children (58% males) were involved in the study. The absolute CD3+, and CD4+ cells count were significantly higher for females (adults and children). Higher CD4+ cell count in adults was associated with smoking, while higher CD8+ count was found among female children. Higher counts, for all T-cells subsets, were detected in blood donors from southeast / south regions while those living in the northern region had the lowest values. Individuals from midwestern and northeastern regions had an intermediate count for all these cells subsets. However, these differences did not reach statistical significance. In Brazil, gender and smoking, were the main determinants of differences in T-lymphocytes reference values. .

RESUMO Os valores de referências de linfócitos T existentes no Brasil são baseados em dados originados de outros países. Não existem dados locais da variação normal para estes parâmetros em adultos e crianças brasileiras. Avaliamos a variação normal encontrada em doadores de sangue de cinco grandes cidades brasileiras em diferentes regiões e em crianças residentes em Salvador e Rio de Janeiro. Todas as amostras foram processadas por citometria de fluxo. Os resultados foram analisados de acordo com região, gênero e estilo de vida dos doadores. Um total de 641 adultos (63% homens) e 280 crianças (58% meninos) participaram do estudo. Valores absolutos de CD3+ e CD4+ foram significantemente maiores no gênero feminino (adultos e crianças). Maiores valores de CD4+ em adultos foram associados com tabagismo, enquanto que maiores valores de CD8+ foram encontrados entre crianças do sexo feminino. Adultos das regiões sul e sudeste apresentaram maiores valores absolutos para todas as células T enquanto que adultos da região norte, apresentaram menores valores. Indivíduos residentes no nordeste e centro-oeste obtiveram contagens intermediárias para todas as populações de células T. Entretanto, estas diferenças entre as regiões, não demonstraram diferença estatística. No Brasil, gênero e tabagismo foram os principais determinantes para diferenças em valores de referências de linfócitos T. .

Efeito da suplementação dietética com L-arginina em subpopulações linfocitárias de ratos submetidos a esplenectomia total isolada ou combinada com autoimplante esplênico / Effect of dietary supplmentation with L-arginine on lymphocyte subpopulations of rats submitted to total splenectomy isolated or combined with splenic autoimplantation

A L-aginina é reconhecida como um nutriente de fundamental importância na resposta imune, apesar de seus efeitos serem, por vezes, considerados inconstantes. O autoimplante esplênico tem sido proposto como alternativa à esplenectomia total isolada, mas existem preocupações quanto à eficácia do restabelecimento da resposta imune, haja vista que o paciente pode permanecer com risco aumentado de desenvolvimento de infecção fulminante pós esplenectomia, mesmo após a regeneração morfológica do órgão. O objetivo deste estudo foi avaliar a participação da suplementação dietética com L-arginina em subpopulações linfocitárias no sangue, no baço e nos autoimplantes esplênicos de ratos submetidos a esplenectomia isolada ou combinada com autoimplante esplênico. Foram utilizados 42 ratos Sprague-Dawlay machos, randomicamente distribuídos em seis grupos: 1 - Controle - operação simulada; 2 - esplenectomia total; 3 - esplenectomia total combinada com autoimplante esplênico; 4 - Controle - operação simulada, com suplementação de L-arginina; 5 - esplenectomia total, com suplementação de L-arginina; e 6 - esplenectomia total combinada com autoimplante esplênico, com suplementação de L-arginina. Os animais dos grupos 4, 5 e 6 receberam suplementação de L-arginina, uma vez ao dia, durante 15 dias anteriores a coleta sangüínea realizada imediatamente antes dos procedimentos operatórios (semanas 0 e 12). A dose utilizada foi de 1,0 g/kg/dia, administrada por via intragástrica em bolus. As avaliações foram realizadas por meio de hemograma e citometria de fluxo. A análise estatística utilizou testes paramétricos e não-paramétricos, sendo p<0,05 considerado para a rejeição da hipótese nula. A suplementação com L-arginina acarretou elevação da contagem relativa e absoluta de neutrófilos periféricos, 12 semanas após a realização de esplenectomia total combinada com autoimplante esplênico. A esplenectomia total ocasionou diminuição da contagem relativa de linfócitos T totais, T CD4+...

L-arginine is recognized as a nutrient of fundamental importance in immune implants has been proposed as an alternative to total splenectomy isolated, but there are concerns about the effectiveness of the restoration of the immune response, considering that the patient can remain at increased risk of developing overwhelming postsplenectomy infection, even after morphological regeneration of the organ. The aim of this study was to determine the role of dietary supplementation with L-arginine in lymphocyte subsets in blood, spleen, and splenic auto-transplantation in rats subjected to total splenectomy alone or in combination with splenic auto-transplantation. Forty two male Sprague-Dawley rats, were randomly divided into six groups: 1 - Control - sham operation, 2 - total splenectomy, 3 - total splenectomy combined with splenic auto-implants, 4 - Control - sham operation, with L-arginine supplementation, 5 - total splenectomy, supplemented with L-arginine, and 6 - total splenectomy combined with splenic auto-implants, supplemented with L-arginine. Animals in groups 4, 5 and 6 were supplemented with L-arginine, once daily for 15 days before blood sample was collected immediately before the operative procedures (weeks 0 and 12). The dose was 1.0 g/kg/day administered by intragastric bolus. The laboratory evaluations were made by blood count and flow cytometry. Statistical analysis used parametric tests and nonparametric, p<0.05 was considered to reject the null hypothesis. Supplementation with L-arginine led to increase in relative and absolute count of peripheral neutrophils, 12 weeks after completion of total splenectomy combined with splenic auto-implants. Total splenectomy caused a decrease in relative count of T lymphocytes, CD4+ and CD8B in blood, but dietary supplementation with L-arginine prevented the decrease in the percentage of total T cells and CD8B in the blood of animals subjected to splenic auto-transplantation...

Profile of immune cells in lymph nodes draining human malignant tumors / Perfil de las células inmunes en los ganglios linfáticos que drenan tumores malignos humanos

The purpose of this study was to characterize and quantify cells involved in immune response in metastasis-free regional lymph nodes (RLNs) draining different human epithelial tumors and compare them (by immunohistochemistry) with control lymph nodes from patients with non malignant diseases. We showed that T cells number was decreased in RLNs as compared to the controls with reduction in both CD4+ T cells and CD8+ T cells subsets and an inverted ratio (CD4+: CD8+). B lymphocytes and follicular dendritic cells were decreased with respect to the controls. S100+ dendritic cells (DCs) and mature DCs were detected in T dependent areas. Their mean number was significantly lower as compared to control. Immature DCs were significantly diminished compared to RLN and control nodes. CD57+ cells, follicular T helper cells and/or NK cells, were localized in the clear zone of germinal centres and their mean number was significantly increased. There were no CD57+ cells in hypoplastic follicles. In this study we show that RLNs draining human cancer present reduction in almost all immune cells, except CD57+ cells. These findings may be related to the deficient anti-tumor immune response in patients with cancer and subsequent tumor progression.

El objetivo del trabajo fue caracterizar y cuantificar utilizando inmuno-histoquímica, las células involucradas en la respuesta inmune en ganglios linfáticos regionales (GLRs) que drenan distintos tumores epiteliales malignos humanos y compararlas con ganglios controles (GLCs) provenientes de pacientes sin enfermedad neoplásica maligna. Determinamos que los GLRs presentaban una marcada depleción de linfocitos B y T, células dendríticas (CD) foliculares y CD interdigitantes maduras respecto a los controles. En los linfocitos T, además de estar disminuidos, se observó una inversión de la relación T CD4+: T CD8+, a favor de los T CD8+. La depleción de CD inmaduras fue mayor respecto a las maduras. Las células CD57+, células foliculares T helper y/o células NK, localizadas en las zonas claras de los centros germinativos, presentaron un marcado incremento en los GLRs comparados con los GLCs, excepto en los casos de ganglios linfáticos con folículos hipoplásicos. En este estudio, demostramos que los GLRs que drenan carcinomas humanos presentan una significativa reducción en casi todas las células de la respuesta inmune, excepto las células NK. Estos hallazgos podrían estar relacionados con la deficiente respuesta antitumoral de los pacientes con cáncer y la subsiguiente progresión tumoral.

Characterization of atypical lymphocytes and immunophenotypes of lymphocytes in patients with dengue virus infection.

To characterize the immunophenotypes of lymphocytes in patients with dengue infection, we performed flow cytometric analysis of peripheral blood mononuclear cells collected from 49 dengue hemorrhagic fever (DHF), 25 dengue fever (DF), and 26 dengue-like syndrome (DLS) cases. The mean total atypical lymphocytes in DHF (916.1 +/- 685.6 cells/microl) and DF (876.2 +/- 801.9 cells/microl) were higher than those of DLS (310.5 +/- 181.4 cells/microl). An atypical lymphocyte count of 10% or higher was a good indicator of dengue infection (sensitivity 50% and specificity 86%). Flow cytometric studies showed that the percentages of atypical lymphocytes correlated with those of CD19+ B lymphocytes and inversely correlated with the percentages of CD69+ lymphocytes. The mean absolute counts of atypical lymphocytes and CD19+ cells on the discharge day were significantly higher than those on the admission day. Low percentages of TdT+ cells were found in all groups of patients. We concluded that atypical lymphocyte and CD19+ cell counts may be a useful diagnostic tool for dengue infection and the recovery from the disease could be judged when numbers of both cell types are significantly elevated.

Comparative Analysis of Bronchoalveolar Lavages in Interstitial Lung Diseases / 대한진단검사의학회지

BACKGROUND: This study was purposed to find out the differences in the lymphocyte subsets and differential cell counts of the bronchoalveolar lavage (BAL) fluid in patients with interstitial lung disease (ILD) and to analyze the differences according to their ages, gender and smoking habits. METHODS: BAL fluid samples of 141 ILD patients were examined for lymphocyte subsets and differential cell counts, and the differences among the patients were analyzed according to their diseases. Then, within the three most common disease groups, the differences were further analyzed by the age, gender and smoking habit of the patients. RESULTS: There were no statistically significant differences in total cell counts (per millimeters of BAL fluid) among the patient groups with each ILD. However, significant differences were observed in the percentages of neutrophils, lymphocytes, eosinophils, and macrophages of BAL fluid. Also, in lymphocyte subset analyses, the percentages of total T cells, B cells, CD4+ T cells, CD8+ T cells, CD4/CD8 T cell ratios, and NK cells were significantly different among the patients with each ILD. However, within the same disease group, there were no differences in differential cell counts and lymphocyte subset analyses according to the age, smoking habit, and gender of the patients. CONCLUSIONS: Although the age, smoking habit and gender did not have an effect on the BAL fluid analyses among the patients with the same ILD, there were significant differences among the patients with each ILD; therefore, the differential cell counts and lymphocyte subset analyses of BAL fluid can be useful in differential diagnosis for determining the types of ILD.

Tumor-infiltrating CD4+ T lymphocytes in early breast cancer reflect lymph node involvement

INTRODUÇÃO: O papel do sistema imunológico na patogênese e progressão do câncer de mama ainda é controverso, e isto nos estimulou a verificar a associação do imunofenótipo dos linfócitos tumor infiltrantes do câncer de mama inicial com a disseminação de células tumorais para os linfonodos axilares. MÉTODOS: Amostras tumorais de 23 pacientes com câncer de mama inicial do Departamento de Ginecologia e Obstetrícia da Faculdade de Medicina de Ribeirão Preto (USP) foram obtidas no momento da biópsia e depois submetidas ao método de digestão enzimática para a extração dos linfócitos tumor infiltrantes. Os linfócitos extraídos foram analisados por citometria de fluxo com anticorpos monoclonais nas seguintes combinações: CD3 FITC/CD19 PE, CD3 FITC/CD4 PE, CD3 FITC/CD8 PE, e CD16/56 PerCP, específicos para imunofenotipagem de linfócitos T e B, linfócitos T helper, linfócitos citotóxicos, e células Natural Killer. Os valores médios destas subpopulações leucocitárias foram comparados entre grupos de pacientes com ou sem metástases linfonodais. RESULTADOS: O valor médio do infiltrado por linfócitos T foi 24,72±17,37%, para o infiltrado por linfócitos B foi 4,22±6,27%, e para o infiltrado por células Natural Killer foi 4,41±5,22%, e para o infiltrado por linfócitos T CD4+ e CD8+ foram, respectivamente, 12,43±10,12% e 11,30±15,09%. Os valores médios do infiltrado por células T e T CD4+ foram maiores no grupo de pacientes com metástase axilar, enquanto nas outras subpopulações nada foi encontrado.CONCLUSÃO: A associação dos linfócitos T CD4+ tumor infiltrantes com metástases linfonodais sugere um papel destas células na disseminação das células neoplásicas aos linfonodos dos pacientes com câncer de mama inicial.

Lymphocyte subset analyses in healthy adults vaccinated with yellow fever 17DD virus

In this study the kinetics of humoral and cellular immune responses in first-time vaccinees and re-vaccinees with the yellow fever 17DD vaccine virus was analyzed. Flow cytometric analyses were used to determine percentual values of T and B cells in parallel to the yellow fever neutralizing antibody production. All lymphocyte subsets analyzed were augmented around the 30th post vaccination day, both for first-time vaccinees and re-vaccinees. CD3+ T cells increased from 30.8 percent (SE ± 4 percent) to 61.15 percent (SE ± 4.2 percent), CD4+ T cells from 22.4 percent (SE ± 3.6 percent) to 39.17 percent (SE ± 2 percent) with 43 percent of these cells corresponding to CD4+CD45RO+ T cells, CD8+ T cells from 15.2 percent (SE ± 2.9 percent) to 27 percent (SE ± 3 percent) with 70 percent corresponding to CD8+CD45RO+ T cells in first-time vaccinees. In re-vaccinees, the CD3+ T cells increased from 50.7 percent (SE ± 3 percent) to 80 percent (SE ± 2.3 percent), CD4+ T cells from 24.9 percent (SE ± 1.4 percent) to 40 percent (SE ± 3 percent) presenting a percentage of 95 percent CD4+CD45RO+ T cells, CD8+ T cells from 19.7 percent (SE ± 1.8 percent) to 25 percent (SE ± 2 percent). Among CD8+CD38+ T cells there could be observed an increase from 15 to 41.6 percent in first-time vaccinees and 20.7 to 62.6 percent in re-vaccinees. Regarding neutralizing antibodies, the re-vaccinees presented high titers even before re-vaccination. The levels of neutralizing antibodies of first-time vaccinees were similar to those presented by re-vaccinees at day 30 after vaccination, indicating the success of primary vaccination. Our data provide a basis for further studies on immunological behavior of the YF 17DD vaccine.

Lymphocyte Subsets and Mood States in Panic Disorder Patients

This study was conducted to examine lymphocyte subset counts and mood states in panic disorder patients. Twenty patients with panic disorder and 20 age- and gendermatched normal healthy subjects were recruited for the study. We used the Spielberger State (STAIS) & Trait (STAIT) Anxiety Inventory, Hamilton Depression Rating scale (HAMD) and Hamilton Anxiety Rating scale (HAMA) to measure mood states in all subjects. Lymphocyte subsets counts were made by flow cytometry. Panic patients showed significantly higher scores for anxiety and depression than normal subjects. Panic patients showed no differences in terms of the numbers of immune cells, as compared with normal healthy subjects, other than a lower proportion of T suppressor cells and a higher T helper cell/T suppressor cell ratio. HAMA and STAIS scores were common factors that could predict T cell numbers and proportions, T helper cell numbers, and natural killer cell proportions in panic disorder patients. We suggest that anxiety levels are related to the T-cell population in panic disorder patients and that quantitative immune differences may reflect altered immunity in this disorder.

Anticuerpos anti-CCP en artritis reumatoidea: relación con características clínicas, citocinas Th1/Th2 y HLA-DRB1 / Anti-cyclic citrullinated peptide antibodies in rheumatoid arthritis: relation with clinical features, cytokines and HLA-DRB1

En el presente estudio se examinó la especificidad y la sensibilidad de los anticuerpos antipéptidos citrulinados cíclicos (CCP) en pacientes latinoamericanas con artritis reumatoidea (AR), así como su relación con la actividad de la enfermedad, manifestaciones extraarticulares (MEA), síntesis de citocinas (IL-4, IL-10, IL-12, TNF-µ e IFN-gamma ) y factor reumatoideo (FR) IgM e IgA, y con el polimorfismo del HLA-DRB1. Se examinaron 79 pacientes con AR (69 con AR establecida y 10 con AR temprana sin previo tratamiento), 56 pacientes con espondilitis anquilosante (EA), 25 con lupus eritematoso sistémico (LES), 50 con síndrome de Sjögren primario (SSp) y diez individuos sanos. De las 69 pacientes con AR establecida, 36 fueron reevaluadas 2 años después. La actividad de la AR se examinó según los criterios del Colegio Americano de Reumatología. Los anticuerpos anti-CCP2, el FR y los niveles de citocinas se determinaron mediante inmunoensayo, y la genotipificación del HLA se llevó a cabo por reacción en cadena de la polimerasa utilizando mezclas de iniciadores específicos. Los anticuerpos anti-CCP se observaron en 96% de los pacientes con AR en la primera evaluación y en 86% en la segunda ( p=0,12), sin modificación significativa en los valores (131±58,7 vs. 130,6±67,1 UI). Su sensibilidad y especificidad global fue de 94% y 92%, respectivamente, pero cuando sólo se consideraron los niveles altos (>60 UI) fueron de 84% y 95%, respectivamente. La razón de probabilidades (RP) positiva fue de 12 y la RP negativa de 0,06. El valor predictivo (VP) positivo fue de 87% y el VP negativo de 96%. Los anticuerpos anti-CCP se observaron en 12% de los pacientes con LES y con SSp, en 2% de los de EA y en 10% de los controles sanos. En los pacientes con AR no se asociaron con la actividad de la enfermedad, MEA y alelos del HLA-DRB1. Tampoco se observaron correlaciones significativas entre sus valores y los niveles de citocinas. En conclusión, los anticuerpos anti-CCP tienen un interés diagnóstico para la AR en nuestra población, pero su utilidad en el seguimiento clínico es limitada y su síntesis es independiente del HLA-DRB1 y no se correlacionan con niveles de citocinas Th1/Th2.

The specificity and sensitivity of anti-cyclic citrullinated peptide antibodies (anti-CCP) was examined in Latin-American patients with rheumatoid arthritis (RA). The variables considered included: 1) relation with the activity of disease, 2) extra-articular manifestations (EAM), 3) synthesis of cytokines (IL-4, IL-10, IL-12, TNF-alpha , and IFN-gamma ) and IgM and IgA rheumatoid factor (RF), and 4) the association with HLA-DRB1 polymorphism. Seventy-nine RA patients were assessed (69 with established RA, and 10 with recent-onset RA not receiving any treatment), 56 with ankylosing spondylitis (AS), 25 with systemic lupus erythematosus (SLE), 50 with primary Sjögren’s syndrome (pSS), and 10 healthy individuals. Of the 69 patients with established RA, 36 were reexamined 2 years later. The activity of the RA was measured by criteria adopted by the American College of Rheumatology. Anti-CCP2, RF and cytokines levels were determined by ELISA. HLA genotypes were established by first, PCR sequence amplification using sequence-specific primers and then, complete sequencing of the product. Anti-CCP antibodies were observed in 96% of patients with RA during the first evaluation and in 86% at the second evaluation ( p=0.12). No significant change in antibody titre was observed between the two evaluations (131±58.7 and 130.6±67.1 IU, respectively). The overall sensitivity and specificity was 94% and 92%, respectively; however, at titres >60 IU, the values were 84% and 95%, respectively. The anti-CCP likelihood ratio positive test was 12 and the likelihood ratio negative test was 0.06. The positive predictive value was 87%, and the negative predictive value was 96%. Anti-CCP antibodies were observed in 12% of SLE and pSS patients, in 2% of AS patients, and in 10% of healthy controls. In RA patients, these antibodies were not associated with the activity of disease, EAM or HLA-DRB1 alleles; no significant correlation was observed between antibody titre and cytokines level. Although anti-CCP antibodies have potential as a diagnostic tool for RA, they are not useful for monitoring clinical activity or predicting the clinical course of disease. Antibody synthesis is HLA-DRB1 independent and not correlated with Th1/Th2 cytokines.

Characterization of lymphocyte subpopulations and major histocompatibility complex haplotypes of mastitis-resistant and susceptible cows

Bovine mastitis is an infectious disease with a major economic influence on the dairy industry worldwide. Many factors such as environment, pathogen, and host affect susceptibility or resistance of an individual cow to bovine mastitis. Recently, there has been considerable interest in defining genetic and immunological markers that could be used to select for improved disease resistance. In this study we have analyzed the lymphocyte subpopulations of mastitis-resistant and susceptible cows using monoclonal antibodies specific for bovine leukocyte differentiation antigens and flow cytometry. We have also used a microarray typing technique to define the bovine leukocyte antigen (BoLA) class I and class II haplotypes associated with resistance or susceptibility to bovine mastitis. A striking finding of the present study is that susceptibility to mastitis was associated with major histocompatibility complex (MHC) haplotypes that have only a single set of DQ genes. The study also revealed that susceptible cows had CD4:CD8 ratios of less than one in both their mammary gland secretions and peripheral blood. These results raise the possibility that the number of DQ genes that a cow has and/or a cow's CD4:CD8 ratio could be used as indicators of susceptibility to bovine mastitis.

Longitudinal follow-up of lymphocyte subsets during the first year of life.

A longitudinal study of lymphocyte subsets during infancy was evaluated by using the flow cytometric immunophenotyping method. Two hundred and thirteen blood samples were obtained from 92 healthy, full-term infants of the following ages: 1-7 days old (n = 43), 3 months old (n = 55), 6 months old (n = 57) and 11 months old (n = 58). The absolute numbers of CD3+ and CD3+/CD4+ T lymphocytes increased from birth to 3 months of age, and remained stable thereafter. The absolute number of CD3+/CD8+ T lymphocytes increased from birth to 11 months of age. The absolute number of CD19+ B lymphocytes and NK cells increased rapidly (3 months) after birth and continued to increase throughout the study period. However, the changes in the relative counts of lymphocyte subsets did not always correspond with the changes in their absolute numbers. These results demonstrate the age-related changes in lymphocyte subpopulations and provide reference ranges for lymphocyte subsets during infancy.

Células natural killer y el sistema inmune innato en la patología infecciosa / Natural killer cells and the innate immune system in infectious diseases

Natural killer (NK) cells form a unique third group of lymphocytes that differs from T and B cells in surface phenotype, target cell recognition and function. NK cells have two relevant functions, related to the innate immune response against pathogens microorganisms. One is cytotoxicity, mediated by the recognition and lysis of target cells such as virus and bacteria infected-cells. The second NK cell function is to produce cytokines, mainly IFN-g, that can modulate innate and specific immune responses. Cytotoxicity and cytokine secretion contribute to host resistance against microorganisms and both functions are significantly altered in infectious diseases

Avaliaçäo de testes imunológicos em pacientes com aborto recorrente / Evaluation of immunologic tests in recurrent abortion

INTRODUÇÃO: O aborto recorrente é um grande enigma, pois apesar de várias causas que lhe säo atribuidas poucas tem um real valor. O papel do fator imunológico tem sido pesquisado amplamente. OBJETIVO: Avaliar alguns testes imunológicos em pacientes com aborto recorrente. TIPO DE ESTUDO: prospectivo LOCAL: Setor de Reproduçäo Humana do Departamento de Ginecologia e Obstetrícia da Faculdade de Medicina de Ribeiräo Preto. PARTICIPANTES: Num estudo piloto foram selecionados nove casais com duas perdas fetais ou mais sem causa aparente e nove casais voluntários com pelo menos dois filhos, sem história de aborto e com idade inferior a 40 anos VARIAVEIS ESTUDADAS: A freqüência de compartilhamento de antígenos HLA e de anticorpos linfocitotóxicos contra antígenos paternos foram avaliados por métodos sorológicos, a variaçäo de fenótipos celulares (CD4, CD8, CD19, CD16, CD56 e HLA-DR) por citometria de fluxo, a atividade "natural killer" (NK) pela liberaçäo de 51Cr e a dosagem de progesterona por radioimunoensaio. RESULTADOS: Näo houve diferença de compartilhamento entre casais com aborto recorrente tanto em relaçäo à classe I quanto II e näo se detectou o aparecimento de anticorpos citotóxicos no grupo investigado. Houve um número absoluto maior de células CD8+(587 vs 448 linfócitos/mmü, p=0,01) e das CD19+(215 vs 182 linfócitos/mmü, p=0,05) nas pacientes. A atividade NK näo foi estatisticamente diferente entre os dois grupos estudados, mas houve uma tendência à reduçäo da atividade NK entre pacientes com aborto recorrente. Näo houve correlaçäo da atividade NK com um número de células CD16+ e CD56+ nem com dosagem de progesterona nos dois grupos estudados. CONCLUSÃO: Estes dados sugerem que o compartilhamento de antígenos HLA, o aparecimento de anticorpos linfocitotóxicos e elevaçäo da atividade NK podem näo ser importantes para a ocorrência de abortos repetidos. o aumento de células CD8+ e CD19+ circulantes pode ocorrer independente de qualquer terapêutica e a citotoxicidade contra antígenos fetais pode ser mediada por células T e näo por células NK.

Relation between lymphocyte subpopulations of peripheral blood and immune responses of modified live hog cholera virus vaccine in pigs treated with an ionized alkali mineral complex

Thirty-nine healthy pigs (28-32 days old) were purchased from a commercial swine farm and housed at swine pens of the College. The animals were vaccinated intramuscularly (1 ml) with an attenuated live hog cholera virus (HCV, LOM strain) and then boostered at 5 weeks after the first vaccination. The animals were divided into 4 experimental groups: 0.05% (w/w) PowerFeel-supplemented diet (T-1, n = 10); 3% (w/w) SuperFeed-supplemented diet (T-2, n = 10); diluted PowerFeel solution (1 : 500, v/v) as drinking water (T-3, n=9); control (n=10). PowerFeel is an original form of ionized alkali mineral complex (IAMC) and SuperFeed is a commercial product of IAMC. The subpopulation of lymphocyte in blood was assayed by a flow cytometry and HCV-specific antibody was determined by an indirect immunofluorescence assay. In IMAC-treated groups, the proportions of subpopulation expressing MHC-class II, CD2+, CD4+, CD8+, and surface IgM+ B lymphocytes were significantly decreased at 5-weeks after the first vaccination. Significant decreases were also observed in the proportions of MHC-class II, CD2+ and CD8+ lymphocyte at 3-weeks after the booster injection. The humoral immune responses in T-1 and T-2 groups were greater than those in T-3 or control group. These results suggest that IAMC-supplemented diets may have an HCV-specific immunostimulatory effect in pigs.

Decreased CD4+/CD8+ ratio in major type of recurrent aphthous ulcers: comparing major to minor types of ulcers.

The etiology of recurrent aphthous ulcers (RAU) has not been clearly defined. However, the results of several studies indicated the evidence of the role of immunological factors. The association between the regulator and effector component of the immune system in RAU needs clarifying by comparing major and minor type of RAU patients. The proportion of peripheral blood lymphocyte subsets were enumerated during active ulcer phase and analyzed in relation to ulcer types. Nineteen patients with RAU (12 minor type and 7 major type) and 8 healthy volunteers, of both sexes, aged 24-54 years old were tested. CD3+ (T cell), CD4+ (helper T cell), CD8+ (suppressor/cytotoxic T cell), CD19+ (B cell), and CD16+/CD56+ (NK cell) were determined by using appropriate monoclonal antibodies in double colored flow cytometry. The results showed that CD4+ was lower in RAU than control (P < 0.01). Comparing both types of RAU, it appeared that CD8+ was higher in the major type than the minor type (p < 0.01); CD4+/CD8+ ratio in the major type was lower than the minor type (P < 0.01). There was no difference in CD19+ and CD16+/CD56+ between any groups compared. The finding indicated that RAU was associated with abnormal proportions of CD4+ and CD8+ cells which was dependent on the severity of the lesion.

Patología por Trypanosoma cruzy. Cepa dependiente? / Is Trypanosoma cruzy pathology strain dependent?

En la actualidad existe consenso en el papel que las características de la población de Trypanosoma cruzi tienen en la patogésesis de las distintas formas clínicas de la enfermedad de Chagas. En nuestro laboratorio, estudiamos en el modelo murino las consecuencias de la infección con dos poblaciones de características biológicas polares: RA y CA-I. Demostramos que el daño neuromuscular es, en parte, mediato por diferentes subpoblaciones de linfocitos T. Además, observamos que el fenotipo celular responsable de la patologia y los distintos blancos tisulares dependen de la población parasitaria. A pesar de no haber observado diferencias en la reactividad jugaría un papel adicional en el desarrollo de la patología neuromuscular: sueros de ratones infectados con RA, pero no con CA-I, desencadenaron alteraciones en el potencial de acción nerviosa. También hemos detectado, en el modelo murino, una disminución en la fertilidad de las hembras infectadas con CA-I/K-98, mientras las hembras infectadas con RA se comportaron igual a los controles. Sin embargo, sólo pudo comprobarse pasaje transplacentario de T. cruzi en la cepa RA. Las diferencias encontradas en la fertilidad, en la sobrevida de las crías nacidas de madres infectadas, así como en el número de resorciones fetales detectadas con la cepa miotrópica, podrían atribuirse a la respuesta inflamatoria hallada en el útero de estas hembras, ya que no se observaron alteraciones en el estro de ambos grupos infectados con respecto a los controles.

Comparison of CD34+ subsets and clonogenicity in human bone marrow, granulocyte colony-stimulating factor-mobilized peripheral blood, and cord blood

To compare the clonogenicity and distribution of CD34+ subsets in bone marrow (BM), granulocyte colony-stimulating factor (G-CSF) mobilized peripheral blood (PB) and cord blood (CB), we analyzed in vitro colony formation and CD34+ cells co-expressing differentiation molecules (CD38, HLA-DR), myeloid associated molecules (CD13, CD33), a T-cell associated molecule (CD3), and a B-cell associated molecule (CD19) from mononuclear cells (MNCs) in the three compartments. The proportions of CD34+CD38- cells (BM: 4.4+/-2.8%, PB: 5.3+/-2.1%, CB: 5.9+/-3.9%) and CD34+HLA-DR cells (BM: 4.7+/-3.4%, PB: 5.5+/-2.3%, CB: 6.1+/-3.7%) did not differ significantly among the compartments. In contrast, a significantly higher proportion of CD34 cells of PB and CB co-expressed CD13 (75.0+/-11.4%, 77.7+/-17.3%) and CD33 (67.1 +/-5.7%, 56.8+/-10.3%) compared with those of BM (43.0+/-6.3%, 27.6+/-5.1%) and a significantly higher number of granulocyte-macrophage colony-forming units (CFU-GM) and erythroid burst-forming units (BFU-E) were detected in MNCs derived from PB and CB compared with those from BM (p<0.01). The proportion of CD34+CD19+ cells was higher in BM (34.9+/-11.9%) than those in PB (5.6+/-3.0%) and CB (4.7=2.1%) (p<0.05). The proportion of CD34+CD3+ was comparable in all three compartments. In conclusion, our findings show that MNCs of mobilized PB and CB display similar phenotypic profiles of CD34+ subsets and clonogenicity, different from those of BM.

Abnormal immunological response to Mycobacterium tuberculosis antigens in a patient with chronic myelocytic leukemia and active tuberculosis

The pathogenic mechanisms of immunosuppression leading to susceptibility of Mycobacterium tuberculosis (MT) infection in chronic myelocytic leukemia (CML) are not clear. To address this issue, we measured the proliferative response, variation of T cell subpopulations (CD4+, CD8+, TCR-V delta 2 and TCR-V beta 8 T cells) and the cytokine profile (IL-1 beta, IL-2, IL-4, IL-6, IL-10, TNF-alpha, IFN-gamma) after MT stimulation of peripheral blood mononuclear cells (PBMC) in a patient with concomitant CML and active pulmonary tuberculosis. The results were compared to four patients with active pulmonary tuberculosis and no other coexistent diseases. The immunologic response to phytohemagglutinin (PHA) was also evaluated. In contrast to controls, the CML PBMC failed to proliferate in response to MT antigens. Mycobacterium-reactive CD4+, V delta 2 and V beta 8 T cells did not expand after MT stimulation of the CML PBMC. In MT antigens-stimulated cultures from the CML patient, IL-2 was not produced and mild reduction of IL-1 beta and INF-gamma were observed. In contrast, IL-10 was markedly elevated in these cultures. Similarly, PHA-stimulated PBMC from the CML patient showed no expansion of CD4+ and CD8+. T cells. In these cell cultures, INF-gamma concentration in supernatants was decreased and IL-10 was significantly elevated. This study suggests that patients with CML may present a profound immunosuppression of essential cellular and molecular immune effectors, a scenario which might contribute to the development of active tuberculosis. These findings further support the need of establishing immunotherapeutic modalities with potential value for myeloproliferative disorders.