HLA among Brazilian patients with spontaneous chronic urticaria and positive autologous serum skin test / HLA em brasileiros com urticária crônica espontânea e teste cutâneo do soro autólogo positivo

BACKGROUND: An association between class I and II alleles of the major histocompatibility complex and idiopathic chronic urticaria has previously been observed in different populations, but there are still no studies on Brazilian populations in this regard. OBJECTIVE: The involvement of the major histocompatibility complex classes I and II (loci A, B and DR) in Brazilian patients with idiopathic chronic urticaria and a positive autologous serum skin test was investigated and compared with a healthy population group. METHODS: DNA was extracted from the blood of 42 patients with idiopathic chronic urticaria and major histocompatibility complex classes I and II alleles were determined using the polymerase chain reaction and a laboratory test for oligonucleotide hybridization using a single-filament probe. The frequencies of these alleles in patients with chronic urticaria were compared with the frequencies in 1000 genetically unrelated voluntary blood donors from the same region of Brazil. The diagnosis of idiopathic chronic urticaria was based on the patients' clinical history and routine laboratory tests. Only the patients with positive autologous serum skin test were selected. The allele distribution resulted from the patient and control groups were analyzed using odds ratios and 95% confidence intervals. RESULTS: No statistically significant differences were found between the positive autologous serum skin test patients with chronic urticaria and the control group. CONCLUSIONS: We found that in this population group, there was no specific association between the HLA alleles studied and chronic urticaria. We believe that further population studies are needed in order to investigate the possible existence of this association.

FUNDAMENTOS: A associação entre os alelos do MHC classe I e II e a urticária crônica idiopática tem sido previamente constatada em diferentes populações, sendo que na população brasileira ainda não existem estudos a este respeito. OBJETIVOS: Foi estudado o envolvimento do MHC classe I e II (locci A, B e DR) em pacientes brasileiros com urticária crônica idiopática e teste cutâneo do soro autólogo positivo, comparando-se com um grupo populacional saudável. MÉTODOS: O DNA foi extraído do sangue de 42 pacientes com urticária crônica idiopática e o MHC classe I e II determinado por reação em cadeia da polimerase e teste laboratorial de hibridização de oligonucleotídeo com sonda de filamento único. A freqüência destes alelos em pacientes com urticária crônica idiopática foi comparada com a de 1000 doadores de sangue voluntários e geneticamente não relacionados, da mesma região do Brasil. O diagnóstico de urticária crônica idiopática foi baseado na história clínica do paciente e exames laboratoriais de rotina; foram selecionados apenas os pacientes com teste cutâneo do soro autólogo positivo. O resultado da distribuição alélica entre o grupo de pacientes e o grupo controle foi analisado através do odds rate com o cálculo do intervalo de confiança de 95% (95% IC). RESULTADOS: Não foram encontradas diferenças com significância estatística entre os pacientes com urticária crônica teste cutâneo do soro autólogo positivos e o grupo controle. CONCLUSÕES: Verificamos que neste grupo populacional estudado não houve associação específica entre os alelos HLA estudados e a urticária crônica; acreditamos na necessidade de outros estudos populacionais, para podermos verificar a possível existência desta associação.

S100, CD68, and MHC class II molecule expression in cervical high- and low-grade HPV-induced lesions / Expressão de S100, CD68 e moléculas de MHC classe II em lesões cervicais de alto e baixo grau induzidas por HPV

INTRODUCTION: Some human papillomavirus (HPV) types are involved in malignant processes in the cervical epithelium, with 99 percent of cases attributed to oncogenic HPV infection. This study aimed to detect S100, CD68, and major histocompatibility complex class II (MHC-II) molecules in cervical uterine epithelial samples in patients with high- and low-grade lesions induced by HPV. METHODS: Fifty-eight samples from patients who were confirmed positive or negative for high-risk oncogenic HPV DNA, had histopathological diagnosis of cervical intraepithelial neoplasia (CIN) of grades I, II, or III, or were negative for intraepithelial lesion or malignancy were subjected to immunohistochemistry reaction to S100 protein, CD68, and MHC-II (HLA-DR alpha chain). RESULTS: The presence of MHC-II predominated in samples exhibiting histopathological alterations (p < 0.05). S100 detection was more numerous in carcinoma samples (CIN III) (75 percent). Presence of this protein correlated significantly (p < 0.05) with histopathological findings and viral load. CONCLUSIONS: A small expression of CD68 was observed, which may be explained by the observation in our study having been made on random microscopic fields and not on specific areas. The findings, such as the presence of S100 protein and MHC-II expression in samples with histological alterations, could suggest that the immune system fails to control HPV replication at the early stages of infection. Further studies with larger prospective data are necessary to confirm this result.

INTRODUÇÃO: Alguns tipos de papilomavirus humano (HPV) estão envolvidos em processos malignos no epitélio cervical, com 99 por cento dos casos atribuídos à infecção por HPV oncogênico. O objetivo deste estudo foi detectar a proteína S100, CD68 e moléculas de MHC-II (complexo principal de histocompatibilidade classe II) em amostras de epitélio cervical uterino, de pacientes com lesões de alto e baixo grau induzidas pelo HPV. MÉTODOS: Cinquenta e oito amostras de pacientes positivos ou negativos, confirmados, para DNA de HPV de alto ou baixo risco oncogênico, e que tiveram diagnóstico histopatológico de neoplasia intraepithelial cervical (NIC) de graus I, II ou III ou foram negativas para lesão intraepithelial e malignidade (NILM), foram submetidas à reação de imunohistoquímica (IHQ) para proteína S100, CD68 e MHC-II (HLA-DR cadeia alfa). RESULTADOS: A presença da molécula MHC-II predominou em amostras exibindo alterações histopatológicas (p < 0,05). A detecção de S100+ foi mais numerosa em amostras com carcinoma (NIC III) (75 por cento). A presença dessa proteína correlacionou-se significantemente (p < 0,05) com achados histopatológicos e a carga viral. CONCLUSÕES: Pequena expressão CD68+ foi observada, uma possível explicação seria que em nosso estudo as observações foram feitas em campo microscópicos aleatórios e não em áreas específicas. Os achados como a presença de S100 e a expressão de MHC-II, em amostras com alterações histológicas, podem sugerir que o sistema imune falha em controlar a replicação do HPV nas fases iniciais da infecção. Maiores estudos, com mais dados prospectivos, são necessários para confirmar esses resultados.

Molecular analysis of human leukocyte antigen class I and class II allele frequencies and haplotype distribution in Pakistani population.

AIM: Distribution of HLA class I and II alleles and haplotype was studied in Pakistani population and compared with the data reported for Caucasoid, Africans, Orientals and Arab populations. MATERIALS AND METHODS: HLA class I and II polymorphisms in 1000 unrelated Pakistani individuals was studied using sequence-specific primers and polymerase chain reaction and assay. RESULTS: The most frequent class I alleles observed were A*02, B*35 and CW*07, with frequencies of 19.2, 13.7 and 20%, respectively. Fifteen distinct HLA-DRB1 alleles and eight HLA-DQB1 alleles were recognized. The most frequently observed DRB1 alleles which represented more than 60% of the subjects were DRB1 *03, *07, *11 and *15. The rare DRB1 alleles detected in this study were HLADRB1 *08 and *09, having frequencies of 0.9 and 1.7%, respectively. In addition, at DRB1-DQB1 loci there were 179 different haplotypes and 285 unique genotypes and the most common haplotype was DRB1*15-DQB1*06 which represented 17% of the total DRB1-DQB1 haplotypes. In our population, haplotype A*33-B*58-Cw*03 comprised 2.8% of the total class I haplotypes observed. This haplotype was seen only in the oriental populations and has not been reported in the African or European Caucasoid. CONCLUSION: Our study showed a close similarity of HLA class I and II alleles with that of European Caucasoid and Orientals. In Pakistani population, two rare loci and three haplotypes were identified, whereas haplotypes characteristic of Caucasians, Africans and Orientals were also found, suggesting an admixture of different races due to migration to and from this region.

Alterations of HLA class I and II antigen expression in preinvasive, invasive and metastatic cervical cancers

HLA expression is altered in a large variety of human cancers. We performed immunohistochemical staining on tissues from normal, preinvasive, invasive and metastatic cervical cancer tissues using anti-HLA class I or class II antibody. In tissues from normal squamous epithelium, carcinoma in situ (CIS) and microinvasive carcinoma (MIC), the expressions of HLA-B, C heavy chains and class II heavy chain were significantly decreased as disease progressed. When the expression patterns were compared between primary and metastatic squamous cell carcinoma (SCC) lesions, statistically significant down-regulation of HLA class I and class II antigen in metastatic lesions was observed. The rates of HLA-B, C heavy chains and class II heavy chain expressions were all significantly down-regulated compared to the down-regulation rate of class I beta2-microglobulin (beta2m) in invasive squamous lesions, and the expressions of class II heavy chain in metastatic lesions was decreased further than that in primary lesions. Unlike SCC, the degree of HLA class I and class II loss was not evident as disease progressed in early stage of adenocarcinoma. In invasive adenocarcinoma lesions, only the expression of HLA-B, C heavy chains was decreased and no differences were seen in HLA-B, C heavy chain expression patterns between primary and metastatic lesions. These results suggest that alterations of HLA class I and II expressions seem to occur at a particular step in cervical cancer development and depend on tissue types: when the tumor becomes invasive and starts to metastasize.

Prevalência de alterações tireoidianas em funcionários da UFRJ e estudo imunogenético anti-tireoidianos positivos / Prevalence of thyroid disorders and imunogenetc study in workers from UFRJ with positive ATPO

Os objetivos da pesquisa foram verificar a prevalência de desordens tireoidianas como bócio, disfunção e anticorpos anti-TPO positivos na população de funcionários da UFRJ e comparar as frequências dos alelos HLA DRB1 e DQB1 entre pacientes com anticorpos positivos e pessoas sadias sem história familiar de doenças auto-imunes. Em todos os 302 indivíduos selecionados por amostragem aleatória simples (210 mulheres e 93 homens, com média de idade de 43,5 e 43 anos, respectivamente) foi feito ratreamento com avaliação clínica, dosagem de TSH e de anticorpos anti-TPO. Quando necessário, outros exames foram realizados como complemento diagnóstico. A tipagem HLA realizada com placas para os alelos DRB1 e DQB1 genéricos utilizou a técnica de SSP baseada na PCR. Alterações à palpação da tireóide foram encontradas em 38 pessoas (12,6 por cento) e em mais 44 a glândula era visível (14,6 por cento), sem diferença significativa entre as frequências nos dois sexos. Vinte e oito (9,3 por cento) tinham anticorpo anti-TPO positivo, com prevalência de 10 por cento, nas mulheres e 7,6 por cento nos homens. Alterações do TSH foram encontradas em 7,3 por cento, sendo TSH baixo em 4 por cento e TSH alto em 3,3 por cento. Em quase todas as comparações, as desordens foram mais prevalentes em pessoas a partir dos 40 anos. Entre os alelos DRB1, o *1131 só foi encontrado em controles (pcorr < 0,05). Para todos os alelos não foram encontrados frequências significativamente diferentes entre os dois grupos. Os resultados da invstigação epidemiológica foram semelhantes aos de outors estudos para algumas alterações, geralmente mais prevalentes em mulheres e a partir dos 40 anos. Entretanto, a prevalência de anticoropos positivos nos homens foi mairo do que o esperado. Os resultados do estudo imunogenético não permitem conclusões definitivas em relação à associação do HLA com doenças auto-imunes da tireóide

HLA class-I and class-II antigen association in rheumatoid arthritis at Varanasi, India.

Clinical presentation of rheumatoid arthritis (RA) and its severity differs in different races. Genetic factors play a significant role in its predeliction. The present study was undertaken to find out association of HLA class I and class II antigens with rheumatoid arthritis prevalent in Asian Indians residing at Varanasi. Ninety rheumatoid arthritis patients strictly fulfilling American Rheumatism Association criteria were screened for prevalent HLA class I and class II antigen by Terasaki Microlympho-cytotoxicity test. Results were compared with 100 healthy controls and 35 Seronegative Spondyloarthritides cases (SSA). Rheumatoid arthritis patients showed increased frequency of HLA-A2 and B40 antigens compared to healthy controls (p < .001). SSA patients showed significantly increased Phenotype frequency (PF) of HLA-B27 (p < .0001) and B40 (p < .001). Significant detection of HLA-A2 exclusively in RA patients suggests a more positive association of A2 in rheumatoid arthritis at Varanasi. HLA-B40 could not be attributed absolute significance of association with SSA or RA as it showed increased frequency in both diseases.

Evaluation in mice of a subunit vaccine produced with the bartha strain of aujeszky s disease virus

Uma vacina de subunidades preparada com a amostra Bartha do vírus da Doença de Aujeszky foi avaliada em camundongos. A vacina de subunidades foi preparada através da solubilizaçäo das glicoproteínas do envelope viral com triton X-100 a 1 por cento, remoçäo das nucleocápsides por ultracentrifugaçäo e adiçäo de Avridine como adjuvante. Em estudos de desafio, esta vacina de subunidades protegeu 83 por cento dos camundongos contra uma infecçäo letal do vírus, após duas doses contendo 500 mg de proteína. A resposta imune humoral induzida por diferentes doses de proteina foi mensurada pelo ELISA. Como vantagem, se aplicada ao suino, a vacina de subunidades descrita poderia permitir a diferenciaçäo sorológica entre animais vacinados e infectados, devido à deleçäo da glicoproteína gI na amostra Bartha

Immunogenetics of the HLA system

The study of the HLA system was primarily initiated to understand the basis for the histocompatibility between recipients and tissue donors. HLA typing methods are being continuously improved and biochemical and molecular typing, in particular, are expected to provide precise typing of the HLA system. Conventional HLA typing methods can define antigen specificities, while biochemical and molecular methods will provide direct allele typing that is based on the actual sequence polymorphism. The precise tissue typing will definitely improve the outcome of transplantation. Structural studies have revealed the highly polymorphic nature of the HLA system and given insight to understanding the molecular basis of the HLA polymorphism. One big immunological puzzle remaining to be answered is how T-cell receptor molecules recognize peptide antigen in conjunction with the HLA molecule. The crystallization of the T-cell receptor molecule, an experiment currently underway, will eventually reveal the structural basis of the trimolecular interaction.

Investigación de anticuerpos anti HLA en suero de puerperas / Research of the anti HLA antibodies in serum of post delivery women

Se estudiaron los sueros de mujeres embarazadas dentro de las 24 horas de puerperio buscando anticuerpos anti HLA; la muestra comprendió un total de 100 pacientes hospitalizadas en la Maternidad Nacional y la Cruz Roja Paraguaya. Las variables estudiadas están con el número de embarazos, abortos, número de transfusiones y de acuerdo a los mismos determinar el grado de sensibilización anti HLA. La metodología laboratorial fue realizada de acuerdo a técnicas standarizadas y preconizadas por P. Terasaki. Una vez enfrentado cada suero con un panel de 50 linfocitos se encontró que 47 de los sueros estudiados tenían anticuerpos anti HLA en diferentes proporciones, siendo 64% mono específicos, así como pudimos observar que las pacientes con 2 y 3 embarazos tenían el porcentaje más elevado de anticuerpos anti HLA. Por lo tanto, contamos en nuestro medio con un material excelente para formar los bancos de sueros tipicadores anti HLA, que nos permitirá en el futuro desarrollar nuevas técnicas y métodos de tratamiento para el avance de la medicina en nuestro país, sobre todo en lo referente a la curación de los enfermos renales que necesitan transpolante renal

Aspecto da resposta imune humoral em camundongos esplenectomizados parcialmente / Aspect of humoral immune response in mice with partial splenectomy

Apresentam-se resultados de pesquisas acerca de resposta imune humoral através de anti-corpos da classe IgG em camundongos submetidos a esplenectomia total e parcial, comparando os resultados com grupos controle, submetidos a operaçäo simulada e intactos, através do teste denominado ELISA. Após duas semanas, o confronto estatístico entre os grupos submetidos as esplenectomias e os grupos controle, revela diferença de níveis de significância que devem ser atribuídos as esplenectomias. Alguns resultados díspares, sugerem a estimulaçäo antigênica de outros órgäos linfóides, bem como a presença de células supressoras no baço, no processo inicial de regeneraçäo do órgäo

In situ characterization of cells in the dermal infiltrates of lepromin reaction using monoclonal antibodies.

A study was made on the in situ characteristics of dermal infiltrates in the early and late lepromin reaction with monoclonal antibodies defining T cell subsets, Langerhan cells and Ia like antigens. The early reaction (24 hrs) was elicited either with standard Dharmendra lepromin or leprosin-A and the late reaction (3-4 weeks) was elicited with standard Dharmendra lepromin. In all, 15 biopsies were studied. Most lymphocytes in the infiltrates of both the lepromin and leprosin reactions were positive for OKT 11, Leu 3a, OKT 8 and Ia like antigens indicating thereby the presence of activated T cells. A high proportion of OKT6 + cells were also noticed in the infiltrates of these reactions. In the late reaction, the lymphocytes in the granulomas were predominantly activated T lymphocytes expressing OKT 11, Leu 3a, OKT 8 and Ia like antigens. Leu 3a + cells were scattered diffusely amidst the epithelioid cells. In contrast, OKT 8 + cells were present mainly in the peripheral region of the granuloma. A small proportion of OKT6 + cells were also seen in these granulomas. Ia like antigens and T6 antigens were not discernible on the epithelioid cells. No difference in the number of OKT6 + epidermal langerhan cells was observed in the various types of reactions.

Fibronectin in leprosy lesions: observations using monoclonal antibodies to human fibronectin.

Croystat sections of dermal lesions from 33 untreated patients with leprosy were studied by indirect immunofluorescence using monoclonal antibodies to human fibronectin. Macrophages in borderline (BL) and lepromatous (LL) leprosy showed intense staining with antifibronectin antibodies. In the tuberculoid lesions cells of the mononuclear phagocyte series in an epithelioid cell granuloma stained for fibronectin. The lymphocytes surrounding these granulomas also showed the presence of fibronectin. These results suggest that the granuloma of leprosy consists of macrophages expressing fibronectin and the lymphocytes in the lesion also appear to express this protein. In six borderline cases, the sub epidermal collagen band showed intense staining with antifibronectin antibodies. In addition, the distribution of Ia-like antigens and fibronectin was studied on the plastic adherent cells obtained from peripheral blood of 11 untreated patients with leprosy. Results indicate that higher percentage of adherent cells from lepromatous patients express fibronectin in comparison to adherent cells from tuberculoid patients or controls. However, no difference was observed in the expression of the Ia-like antigens by the adherent cells, from these patients.

HLA-DR Antigens and HLA-B: DR Haplotypes in Koreans

HLA-DR antigen and gene frequencies were studied in 150 unrelated Koreans in Seoul. HLA-DR4 was the most common DR specificity encountered and HLA-DR1 and -DR3 occurred with the lowest frequencies. The frequency of HLA-DR blank allele was 27.1%. HLA-B:DR haplotypes involving positive delta values differing significantly from zero were DR1:B7, DR2:Bw22, DR3:B17, DR5:Bw35, DRw6:B17, DR7:B12, DR7:B13, and DRw8:Bw16. The supertypic groups (MT1, MT2 and MT3) differ somewhat in frequencies from Other populations. These findings suggested that the Korean population, while having many similarities in HLA-DR antigen frequencies with those of neighboring Orientals, has not only different features in the distribution of HLA-DR antigens but also has unique HLA-B:DR haplotypes.