Imbalanced expression of functional surface molecules in regulatory and effector T cells in systemic lupus erythematosus

Regulatory T (TREG) cells play an important role in maintaining immune tolerance and avoiding autoimmunity. We analyzed the expression of membrane molecules in TREG and effector T cells in systemic lupus erythematosus (SLE). TREG and effector T cells were analyzed for the expression of CTLA-4, PD1, CD28, CD95, GITR, HLA-DR, OX40, CD40L, and CD45RO in 26 patients with active disease, 31 with inactive disease, and 26 healthy controls. TREG cells were defined as CD25+/highCD127Ø/lowFoxP3+, and effector T cells were defined as CD25+CD127+FoxP3Ø. The ratio of TREG to effector T cells expressing GITR, PD1, HLA-DR, OX40, CD40L, and CD45RO was determined in the three groups. The frequency of TREG cells was similar in patients with SLE and controls. However, SLE patients had a decreased frequency of CTLA-4+TREG and CD28+TREG cells and an increased frequency of CD40L+TREG cells. There was a decrease in the TREG/effector-T ratio for GITR+, HLA-DR+, OX40+, and CD45RO+ cells, and an increased ratio of TREG/effector-T CD40L+ cells in patients with SLE. In addition, CD40L+TREG cell frequency correlated with the SLE disease activity index (P=0.0163). In conclusion, our findings showed several abnormalities in the expression of functionally critical surface molecules in TREG and effector T cells in SLE that may be relevant to the pathogenesis of this disease.

Ocular surface evaluation in patients treated with a fixed combination of prostaglandin analogues with 0.5% timolol maleate topical monotherapy: a randomized clinical trial

OBJECTIVES: To compare ocular surface changes induced via glaucoma treatment in patients using fixed combinations of prostaglandin analogues (travoprost, latanoprost and bimatoprost) with 0.5% timolol maleate METHODS: A prospective, multicenter, randomized, parallel group, single-blind clinical trial was performed in 33 patients with ocular hypertension or open angle glaucoma who had not been previously treated. The ocular surface was evaluated prior to and three months after treatment, with a daily drop instillation of one of the three medications. The main outcome measurements included the tear film break-up time, Schirmer's test, Lissamine green staining, the Ocular Surface Disease Index questionnaire, impression cytology using HE and PAS and immunocytochemistry for interleukin-6 and HLA-DR. Ensaiosclinicos.gov.br: UTN - U1111-1129-2872 RESULTS: All of the drugs induced a significant reduction in intraocular pressure. Decreases in the Schirmer's test results were observed with all of the drugs. Decreases in tear-film break-up time were noted with travoprost/timolol and latanoprost/timolol. An increase in the Lissamine green score was noted with travoprost/timolol and bimatoprost/timolol. The Ocular Surface Disease Index score increased after treatment in the travoprost/timolol group. Impression cytology revealed a significant difference in cell-to-cell contact in the same group, an increase in cellularity in all of the groups and an increase in the number of goblet cells in all of the groups. The fixed combinations induced an increase in IL-6 expression in the travoprost/timolol group, in which there was also an increase in HLA-DR expression. CONCLUSIONS: All of the fixed combinations induced a significant reduction in intraocular pressure, and the travoprost/timolol group showed increased expression of the inflammatory markers HLA-DR and interleukin-6. All three tested medications resulted in some degree of deterioration in ...

Conjunctival inflammation in patients under topical glaucoma treatment with indication to surgery / Inflamação conjuntival em pacientes em tratamento tópico para glaucoma com indicação de cirurgia

PURPOSE: To compare the frequency of conjunctival HLA-DR expression (a surrogate marker for inflammation) in eyes treated with topical prostaglandin analogues versus eyes treated with other topical antiglaucomatous drugs. METHODS: Patients diagnosed with primary open-angle glaucoma presenting indication for trabeculectomy were divided in groups according to the use or not of prostaglandin analogues. All subjects were treated with the maximum tolerated dose of antiglaucomatous drugs until the date of the surgery. At the beginning of the surgical procedure, a 5 x 5 mm biopsy of the bulbar conjunctiva was collected, incubated with monoclonal anti-HLA-DR antibody and processed for histological analysis. RESULTS: Of the 31 eyes included (31 patients), 25 were under topical prostaglandin analogues (Group 1) and six under other topical pharmacological agents (Group 2). Fourteen eyes of Group 1 (56%) and three of Group 2 (50 %) were positive for the inflammatory marker HLA-DR (P=1.0). The percentage of stained cells ranged from 15.49 to 48.09% (median: 27.61) in Group 1, and from 18.35 to 28 (median: 20.71) in Group 2, with no differences statistically significant (p=0.33). CONCLUSION: The use of prostaglandin analogues did not increase conjunctival expression of HLA-DR compared to other topical antiglaucomatous agents.

OBJETIVO: Comparar a frequência da expressão conjuntival de HLA-DR (marcador inflamatório) em olhos tratados com análogos de prostaglandinas de uso tópico com a frequência em olhos tratados com outros medicamentos. MÉTODOS: Pacientes com glaucoma primário de ângulo aberto apresentando indicação de trabeculectomia foram agrupados segundo o uso ou não de análogos de prostaglandinas. Todos os participantes foram tratados com medicação máxima tolerada até o momento da cirurgia. Ao início do procedimento cirúrgico, uma biópsia de 5 x 5 mm da conjuntiva bulbar foi coletada, incubada com anticorpo monoclonal anti-HLA-DR e processada para análise histológica RESULTADOS: Dentre os 31 olhos incluídos (31 pacientes), 25 estavam em uso de análogos de prostaglandinas (Grupo 1) e seis em uso de outros agentes antiglaucomatosos (Grupo 2). Quatorze olhos do Grupo 1 (56%) e três do Grupo 2 (50%) apresentaram positividade para o marcador HLA-DR (p=1,0). A porcentagem de células coradas variou de 15,49 a 48,09% (mediana: 27,61%) no Grupo 1 e de 18,35 a 28% (mediana: 20,71%) no Grupo 2, com diferenças não estatisticamente significativas (p=0,33). CONCLUSÃO: O uso de análogos de prostaglandinas não aumenta a expressão conjuntival de HLA-DR comparado com outros medicamentos tópicos para o tratamento de glaucoma.

Amalgam tattoo: a cause of sinusitis?

Little attention has been paid to the toxicity of silver amalgam fillings, which have been used over the centuries in Dentistry. Amalgam particles may accidentally and/or traumatically be embedded into the submucosal tissue during placement of a restoration and perpetuate in such area. This article presents a case of amalgam tattoo and investigates whether it is related to the patient's repeated episodes of sinusitis. The patient was a 46-year-old woman with a 2 mm diameter radiopaque lesion in the right oral mucosa detected on a panoramic radiograph and presented as a black macula clinically. A complete surgical resection was carried out. The histopathological examination revealed deposits of dark-brownish pigments lining the submucosal tissue with adjacent lymphocytic inflammatory infiltrate and multinucleated giant cells phagocyting pigments. There was a negative staining for both iron and melanin. One year after lesion removal, the patient reported that the sinusitis crises had ceased after repeated episodes for years. It may be speculated that the inflammatory process related to amalgam tattoo seems to lead to a local immune response that causes sinusitis because it enhances the human leukocyte antigen DR (HLA-DR) tissue expression.

Analysis of polymorphism of MHC class II BuLA DRB3 exon 2 gene in north west Iranian populations of the water buffalo [Bubalus bubalis] through PCR-SSCP

The DRB3 gene is a highly polymorphic major histocompatibility complex [MHC] class II gene and plays an important role in variability of immune responsiveness and disease resistance. In the present study, the MHC class II DRB3 gene in water buffalo [Bubalus bubalis] populations from Northwest regions of Iran was investigated through PCR-SSCP. Genomic DNA was extracted from whole blood samples collected from 50 buffaloes. A 284 bp segment of exon 2 of BuLa-DRB3 was amplified by standard PCR, using locus-specific primers. The PCR products were subjected to a non-denaturing gel electrophoresis. A number of 11 different SSCP patterns indicating allelic variation were identified. The three most frequent patterns 1,4 and 10 accounted for 58% of the total patterns. Results indicated that exon 2 of the BuLA-DRB3 gene is highly polymorphic among the examined animals

Análise do lavado broncoalveolar em vítimas de queimaduras faciais graves / Bronchoalveolar lavage analysis in victims of severe facial burns

OBJETIVO: Analisar o lavado broncoalveolar (LBA) de vítimas de queimaduras que inalaram fumaça a fim de identificar alterações que possam estar associadas à morte ou à sobrevida. MÉTODOS: Dezoito vítimas de queimaduras faciais foram submetidas a LBA até 24 h após o evento, sendo realizadas a análise do conteúdo celular e proteico, incluindo TNF-α, HLA-DR, CD14, CD68 e iNOS. RESULTADOS: Dos 18 pacientes submetidos à broncoscopia, 8 (44,4 por cento) morreram durante o seguimento. A média de idade dos pacientes que morreram foi significativamente maior (44,7 vs. 31,5 anos). A superfície corporal queimada foi em média de 60,1 por cento nos pacientes que morreram e de 26,1 por cento nos sobreviventes (p < 0,0001). Entre os 18 pacientes submetidos à broncoscopia, 11 (61,1 por cento) apresentaram sinais endoscópicos de lesão por inalação de fumaça, e 4 (36,4 por cento) destes faleceram. Dos 7 pacientes sem sinais de lesão por inalação de fumaça, 4 (57,1 por cento) faleceram. A média do número de células epiteliais ciliadas no LBA dos pacientes que morreram foi significativamente maior daquela dos sobreviventes (6,6 por cento vs. 1,4 por cento; p = 0,03). Os demais parâmetros analisados não mostraram diferença entre os grupos. CONCLUSÕES: A superfície corporal queimada mostrou ser um fator preditivo de mortalidade. O aumento do número de células epiteliais ciliadas no LBA, denotando descamação epitelial brônquica, esteve associado à maior mortalidade de pacientes com queimaduras faciais.

OBJECTIVE: To analyze bronchoalveolar lavage (BAL) specimens of burn victims who inhaled smoke, in order to identify alterations associated with mortality or survival. METHODS: Eighteen victims of facial burns were submitted to BAL up to 24 h after the event. We investigated cell and protein content, including TNF-α, HLA-DR, CD14, CD68 and iNOS. RESULTS: Of the 18 patients submitted to bronchoscopy, 8 (44.4 percent) died during the follow-up period. The mean age of patients who died was significantly higher (44.7 vs. 31.5 years). On average, the patients who died had burns covering 60.1 percent of the total body surface area, compared with 26.1 percent in the survivors (p < 0.0001). Of the 18 patients submitted to bronchoscopy, 11 (61.1 percent) showed endoscopic signs of smoke inhalation injury, and 4 (36.4 percent) of those 11 died. Of the 7 patients with no signs of smoke inhalation injury, 4 (57.1 percent) died. The mean number of ciliated epithelial cells in the BAL fluid was significantly higher in the patients who died than in the survivors (6.6 percent vs. 1.4 percent; p = 0.03). There were no significant differences between the groups in terms of any of the other parameters evaluated. CONCLUSIONS: The total body surface area burned was a predictive factor for mortality. Increased numbers of ciliated epithelial cells in the BAL fluid, denoting bronchial epithelial desquamation, were associated with higher mortality in patients with facial burns.

Comparison of allele frequency for HLA-DR and HLA-DQ between patients with ECC and caries-free children.

BACKGROUND: Early childhood caries (ECC) is one of the most common diseases of childhood. The etiology of ECC is multifactorial and both genetic and environmental factors play important roles in the pathogenesis of the disease. Genetic variations in the hosts may contribute to changes in the risk for dental caries. Genetic factors such as human leukocyte antigen (HLA) have recently been suggested as a predisposing factor. AIM: The aim of this study was to look for an association between HLA-DRB1 and HLA-DQB1 with ECC for developing new strategies for the diagnosis as well as the prevention of the disease. DESIGN: In this study, we extracted the genomic DNAs from whole blood samples of 44 patients with ECC and 35 caries-free children by the salting-out method. We amplified the genomic DNA by PCR-SSP and then HLA-typing was performed for all alleles. RESULTS: The results revealed a significant increase in the frequency of HLA-DRB1*04 in the patient group (P=0.019). The odds ratio for this allele was detected to be 10. The frequency of HLA-DQB1 alleles was not significantly different between the two groups. CONCLUSION: The above results suggest that HLA-DRB1*04 is associated with the susceptibility to ECC. Thus HLA-DRB1*04 detection as a molecular marker for early diagnosis of ECC may be recommended.

Immunophenotype of hematopoietic stem cells from placental/umbilical cord blood after culture

Identification and enumeration of human hematopoietic stem cells remain problematic, since in vitro and in vivo stem cell assays have different outcomes. We determined if the altered expression of adhesion molecules during stem cell expansion could be a reason for the discrepancy. CD34+CD38- and CD34+CD38+ cells from umbilical cord blood were analyzed before and after culture with thrombopoietin (TPO), FLT-3 ligand (FL) and kit ligand (KL; or stem cell factor) in different combinations: TPO + FL + KL, TPO + FL and TPO, at concentrations of 50 ng/mL each. Cells were immunophenotyped by four-color fluorescence using antibodies against CD11c, CD31, CD49e, CD61, CD62L, CD117, and HLA-DR. Low-density cord blood contained 1.4 ± 0.9 percent CD34+ cells, 2.6 ± 2.1 percent of which were CD38-negative. CD34+ cells were isolated using immuno-magnetic beads and cultured for up to 7 days. The TPO + FL + KL combination presented the best condition for maintenance of stem cells. The total cell number increased 4.3 ± 1.8-fold, but the number of viable CD34+ cells decreased by 46 ± 25 percent. On the other hand, the fraction of CD34+CD38- cells became 52.0 ± 29 percent of all CD34+ cells. The absolute number of CD34+CD38- cells was expanded on average 15 ± 12-fold when CD34+ cells were cultured with TPO + FL + KL for 7 days. The expression of CD62L, HLA-DR and CD117 was modulated after culture, particularly with TPO + FL + KL, explaining differences between the adhesion and engraftment of primary and cultured candidate stem cells. We conclude that culture of CD34+ cells with TPO + FL + KL results in a significant increase in the number of candidate stem cells with the CD34+CD38- phenotype.

A importância dos exames laboratoriais no monitoramento da doença de Graves e artrite reumatóide / The laboratorial tests and its importance in the clinical course of therapeutic monitoring in Graves' disease and rheumatoid arthritis

A auto-imunidade se manifesta quando ocorre uma falha nos mecanismos de auto-tolerância responsáveis pela discriminação entre o próprio e o não próprio, desencadeando uma resposta imune adaptativa específica contra os auto-antígenos. A doença de Graves (DG), uma doença auto-imune associada com atividade excessiva da tireóide, podendo vir associada a outras doenças auto-imunes endócrinas, como Diabetes mellitus tipo 1, e não endócrinas como Lupus Eritematoso Sistêmico (LES) ou Artrite Reumatóide (AR). As doenças auto-imunes são caracterizadas pela etiologia multifatorial, onde fatores genéticos, endócrinos, imunológicos, infecciosos, ambientais e emocionais contribuem para o desencadeamento e agravamento dos processos lesivos. Tem-se pesquisado cada vez mais a influência de fatores genéticos sobre o desenvolvimento de doenças auto-imunes e é neste contexto que se observa a forte associação do HLA-DR, tanto com DG como com AR. O presente trabalho tem como objetivo descrever as alterações laboratoriais observadas em uma paciente do sexo feminino, 22 anos, branca, com o diagnóstico das duas doenças auto-imunes de evolução crônica, a DG e AR, correlacionar os resultados laboratoriais com os sinais e sintomas clínicos apresentados e avaliar os exames laboratoriais realizados para o monitoramento clínico e terapêutico.

Análisis de HLA-DR en pacientes mexicanos con pénfigo / HLA-DR Analysis in Mexican patients with pemphigus

Los pénfigos son un grupo de enfermedades ampollosas de piel y mucosas que histológicamente presentan ampollas intraepidérmicas por acantólisis y anticuerpos fijos y circulantes dirigidos contra la superficie celular de los queratinocitos. En pacientes mexicanos que padecen pénfigo, no se han determinado desequilibrios de presentación de antígenos del HLA, como se ha estudiado en otras poblaciones, por lo que se efectuó un estudio comparativo, prospectivo, transversal y observacional en 25 pacientes; 18 de ellos con pénfigo vulgar y 7 con pénfigo foliáceo a quienes se les tomó muestra de sangre periférica para la extracción de DNA, con el método de expulsión salina ("salting out"). Se realizó determinación de HLA-DR, amplificando la región HLA-DRB1, mediante la técnica de reacción de polimerasa en cadena (PCR) y se determi-naron cada uno de los alelos con oligonucleótidos específicos de alelos (ASO), utilizando el kit Amplicor Hoffman La Roche Basilea, Suiza.Los resultados mostraron que el HLA-DR14 (DR6) es más común en los pacientes con pénfigo, sobre todo pénfigo vulgar, que en la población sana control, esto concuerda con lo descrito en la literatura universal. Por otra parte, el HLA-DR1 representa un riesgo relativo mayor para el desarrollo de pénfigo foliáceo en nuestra población.

Kikuchi histiocytic necrotizing lymphadenitis: clinicopathological and immunohistochemical study

Clinical, morphological and immunohistochemical features of 10 cases having the lymphnodal histological pattern of Kikuchi disease were examined. Two of these were diagnosed as systemic lupus erythematosus [SLE]. Morphologically, Kikuchi disease and SLE were nearly indistinguishable. Plasma cells, neutrophilic infiltration, haematoxyphilic bodies and vasculitis were not useful in differentiating the conditions. Kikuchi lymphadenitis and malignant lymphoma however could be differentiated histologically. Morphological features that exclude malignancy included: polymorphous nature of cellular infiltrate, absence of abnormal mitosis, preservation of sinusoidal pattern on intervening areas and presence of extracellular and intracellular karyorrhectic debris

Preliminary study of HLA-ABCDR antigens in CML, ANLL, thalassemia and severe aplastic anemia in Thais.

The objective of this study was to analyse human leukocyte antigen (HLA) and disease association in common blood diseases [chronic myelogenous leukemia (CML), acute nonlymphocytic leukemia (ANLL), thalassemia and severe aplastic anemia] in Thais. The subjects were patients from the Hematological Clinic, Departments of Medicine and Pediatrics, Ramathibodi Hospital who were referred for HLA typing for bone marrow transplantation (BMT) at the Histocompatibility Laboratory from March 1988 to September 1997. A total of 129 patients had complete HLA-ABC typing. The patients included 45 CML, 40 ANLL, 26 thalassemia (Thal) and 18 severe aplastic anemia (SAA). Of these, 88 patients were typed for HLA class II. The HLA class I (ABC) and II (DR, DQ) typings were performed by microlymphocytotoxicity test. It was found that HLA class I was associated with CML, ANLL and Thal, whereas, HLA class II was associated with SAA. HLA-B8 and HLA-B18 were increased in CML with R.R. values of 12.2 and 3.9, respectively, whereas, HLA-B18 was increased in ANLL with R.R. value of 4.5. In addition, HLA-DR2 and DR3 were increased in SAA with R.R. values of 3.8 and 4.8, respectively. For Thal, HLA-A2 and B46 were increased in Thal in Central Thais with R.R. values of 3.3 and 6.1, respectively, whereas, HLA-B13 was increased in Thal in Northern Thais with R.R. value of 8.5. On the other hand, HLA-B7 was absent in CML. HLA-Cw7 was decreased in CML and SAA, whereas, HLA-DR6 was decreased in ANLL and SAA. Furthermore, HLA-Cw6 was also decreased in CML, whereas, HLA-A33 and Bw4 were decreased in SAA. Although the sample size of each disease was small, the increase of HLA-DR2 was observed in SAA in Thais which was similar to other studies in different ethnic groups. These preliminary data may be useful for further study in HLA and blood disease association.

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.

Autoimmune and non-autoimmune thyroid diseases have different patterns of cellular HLA class II expression

CONTEXT: Surface HLA-DR antigen is usually only expressed by antigen-presenting cells (APC). In autoimmune thyroid disease, follicle cells function as APC, thus expressing HLA-DR. However, non-autoimmune thyroid diseases may also express surface class II antigens. OBJECTIVE: To evaluate the presence and pattern of HLA class II expression in autoimmune and non-autoimmune thyroid disorders. DESIGN: Retrospective: histopathological and immunohistochemical analysis. LOCATION: Referral center, university hospital. SAMPLE: Ten histologically normal thyroids, 11 GravesÆ disease, 7 HashimotoÆs thyroiditis, 10 atoxic multinodular goiter and 3 toxic adenomas were analyzed by immunohistochemistry, using a monoclonal antibody anti-HLA-DR. MAIN MEASUREMENTS: The presence of these antigens in thyroid follicular cells and their relation to inflammatory infiltrate was evaluated. The pattern of HLA-DR expression in thyroid follicular cells was analyzed: membrane, cytoplasmic or both. RESULTS: Although HLA-DR antigens were sparsely present in one of the 8 normal thyroids, in 6 of the 9 atoxic multinodular goiter and in 2 of the 3 toxic adenomas a net positivity could be seen in large areas. In all 5 HashimotoÆs thyroiditis and in 7 of the 10 GravesÆ disease cases. This expression occurred in follicle cells either in contact with inflammatory cells or not. In non-autoimmune thyroid disease, HLA-DR positivity was essentially cytoplasmic, whereas in GravesÆ disease and Hashimoto thyroiditis it was mainly in cell membranes. CONCLUSIONS: It is suggested that the HLA class II expression on the surface of follicle cells could be related to auto-antigen presentation to the immune system by these cells, leading to inflammation

Comparative analysis of serological and molecular results for HLA-DR typing in 120 Thai subjects.

In this study, serological HLA-DR typing results were compared to typing results obtained with sequence-specific primers in the polymerase chain reaction (PCR-SSP). HLA-DR typing was performed on 120 random Thai individuals. Differences in HLA-DR typing results were found in 18 out of 120, which were due to cross reactive antibodies and the lack of potent antisera to define proper HLA-DR splits by serology. Furthermore, PCR-SSP is fast and easy to perform as HLA-DR typing results can be obtained within 2 hours. From the results of this study it can be concluded that PCR-SSP is a reliable and promising technique for HLA-DR typing which can replace the serological technique in routine clinical practice.

Lymphocytotoxic antibodies & immunity in pulmonary tuberculosis.

To understand whether the presence of cold reactive lymphocytotoxic antibodies (LCA) (reactive at 15 degrees C) in the system has any effect on immunity to tuberculosis lymphocytotoxic antibodies to adherent cells (enriched-B cells) and non-adherent cells were studied in active-TB (n = 42) and inactive-TB (cured) patients (n = 49) and healthy controls (n = 32). The plasma samples of inactive-TB patients showed higher percentage of positivity for lymphocytotoxic antibodies (36.7%) than the active-TB patients (21.4%) and control subjects (18.8%). No significant difference on antibody and lymphocyte response against Mycobacterium tuberculosis culture filtrate antigens was observed between LCA positive and LCA negative active-TB patients and normal healthy controls. Further, determination of HLA-DR phenotype of the patients and control subjects showed that individuals positive for lymphocytotoxic antibodies were more among HLA-DR2 positive and DR7 positive active-TB patients and control subjects than non-DR2 and non-DR7 subjects. The present study suggests that the cold reactive lymphocytotoxic antibodies may be against B-lymphocytes and persistent for a longer time. HLA-DR2 and -DR7 may be associated with the occurrence of LCA activity. Further, the presence of LCA has no immunoregulatory role on immunity to tuberculosis.

Estudo imunohistoquímico da expressäo HLA-DR no epitélio superficial e na lâmina própria da mucosa colônica de crianças portadoras de doença de Crohn e retocolite ulcerativa inespecífica / Immunohistochemical study of the HLA-DR expression in the superficial epithelium and in the lamina propria of the colonic mucosa in children with Crohn's disease and nonspecific ulcerative colitis

Alguns estudos têm realçado a importância da expressao HLA-DR no epitélio superficial e na lâmina própria da mucosa dos pacientes portadores de retocolite ulcerativa inespecífica e doença de Crohn. Neste estudo foram quantificadas e analisadas as características histológicas da expressao HLA-DR na mucosa colônica nestas duas doenças. Foram estudados 12 pacientes em doença de Crohn, 19 com retocolite ulcerativa inespecífica e 10 espécimes de mucosa retal representando o grupo controle, de acordo com o seguinte modelo: período I = pé-tratamento; período II = até dois anos de evoluçao e período III = mais de dois anos de evoluçao. A expressao HLA-DR foi identificada pelo monoclonal HLA-DR (DAKO) em espécimes de mucosa colônica pelo método de imunoperoxidase. A quantificaçao da expressao HLA-DR na lâmina própria foi feita através de análise de imagem computadorizada cromática, que expressa em porcentagem a área (mum2) ocupada pelas células HLA-DR positivas e no epitélio superficial quantificada por uma avaliaçao semi-quantitativa. Na lâmina própria da mucosa dos doentes a expressao HLA-DR esteve aumentada em todos osperíodos estudados, quando comparada com o grupo controle, porém sem diferença estatística. A distribuiçao da expressao HLA-DR na lâmina própria foi em localizaçoes correspondentes aos macrófagos e aos linfócitos B. O epitélio colônico superficial dos pacientes com retocolite ulcerativa inespecífica e com retocolite ulcerativa inespecífica e com doença de Crohn foi HLA-DR positivo em 86,95 por cento e 80,96 por cento, respectivamente, nos três períodos estudados. O epitélio superficial de todas as mucosas retais do grupo controle nao mostraram a expressao HLA-DR. A presença da expressao HLA-DR no epitélio colônico superficial dos pacientes com retocolite ulcerativa inespecífica e doença de Crohn e sua ausência no epitélio do grupo controle foi um achado de grande importância neste estudo. É consenso entre os diversos autores que a expressao HLA-DR representa uma intensa açao local das citocinas indutoras desta expressao na mucosa colônica dos pacientes com retocolite ulcerativa inespecífica e donça de Crohn, representando a sua ativaçao em resposta ao antígeno invasor da mucosa intestinal. O estudo da expressao HLA-DR na lâmina própria nao foi conclusivo, pois o resultado nao representou diferença significativa em relaçao ao controle.

Avaliaçäo fenotípica dos infiltrados celulares e da expressäo de HLA-DR em tireóides de pacientes com doença de Graves tratados com propiltiouracil / Phenotype evaluation of the cell infiltrated and of the HLA-DR expression in the thyroid of patients with Graves' disease treated with propylthiouracil

É sabido que as tireóides de pacientes com doença de Graves apresentam uma infiltraçao por mononucleares, principalmente linfócitos. Com a intençao de se avaliar estes infiltrados linfocitários, células T e a associaçao entre estes e o HLA-DR, caso os tireócitos estejam positivos para este antígeno de classe II, oito amostras de tecido tireoideano, obtidas cirurgicamente de oito pacientes com a doença de Graves e tratados com propiltiouracil (PTU), foram estudadas em secçoes criostáticas utilizando-se anticorpos monoclonais específicos (UCHT1: CD3, 16H5: CD4, OKT8: CD8, OKM1: CD14 e MID3: HLA-DR) através de imuno-histoquímica (APAAP). Em sete dos oito pacientes foi identificada a presença de células T, com predominância de células CD4+ sobre as CD8+ em quase todos. A expressao anormal de HLA-DR nos tireócitos foi observada em todos os tecidos estudados, sendo que aqueles com maior densidade de infiltraçao linfocitária foram os que apresentaram a maior positividade para o HLA-DR. Nao houve significância estatística quando testadas as combinaçoes HLA-DR e a infiltraçao linfocitária total, as subpopulaçoes linfocitárias entre si (CD4+ e CD8+) e cada uma destas isoladamente com as células T total (CD3+). Uma provável influência do PTU nos infiltrados celulares intraglandulares, como na expressao anormal de HLA-DR nos tireócitos, nao foi possível de ser avaliada, pois todos os tecidos estudados foram expostos a esta droga antitireoideana.

Ex vivo generation of functional dendritic cells from mobilized CD34+ hematopoietic stem cells

The ability to generate dendritic cells (DCs) in sizeable numbers has enormous implications for the development of clinically-effective antigen presentation procedures for cancer immunotherapy. We evaluated the generation of immunostimulatory DCs from peripheral blood CD34+ cells collected from healthy donors. CD34+ cells purified from leukapheresis product were seeded at 1 x 10(4) cells/mL in complete medium supplemented with GM-CSF, TNF alpha, IL-4, c-kit ligand, and flt3 ligand (FL). By day 14 of culture in the presence of GM-CSF + TNF alpha, the total cell number increased by 23.4 +/- 5.4-fold compared to the starting number of CD34+ cells. When the c-kit and FL were added to GM-CSF and TNF alpha, the cell number increased by 109.8 +/- 11.2-fold without affecting the immunophenotype of recovered cells. Flow cytometric analysis indicated that cells with the markers of mature dendritic cells, i.e., CD1a +CD14 -HLA-DR+, and CD80+CD86+HLA-DR+, constituted 49.0% +/- 7.5%, and 38.9% +/- 6.5%, respectively. This pattern of expression of surface antigen was unchanged whether the c-kit ligand and/or FL was added. The irradiated CD1a+HLA-DR+ cells recovered from in vitro cultures elicit a vigorous proliferation of allogeneic peripheral blood T-cells, irrespective of cytokine combinations. These findings provide advantageous tools for the large-scale generation of DCs that are potentially usable for clinical protocols of immunotherapy or vaccination in patients undergoing cancer treatment.