Evaluation of HLA-DQ2 and HLA-DQ8 haplotypes in patients with endometriosis, A case-control study

Abstract Objective To evaluate the relationship between genetic haplotypes associated with celiac disease (Human Leucocyte Antigen [HLA] DQ2 and DQ8) with the diagnosis, clinical presentation, and location of endometriosis in Brazilian women. Method A retrospective cross-sectional study, was conducted in a Tertiary hospital. Patients Women aged 18-50 years who underwent HLA-DQ2 and HLA-DQ8 haplotype analysis. Intervention The patients were divided into endometriosis and control groups and evaluated for symptoms; endometriosis location, American Society for Reproductive Medicine (ASRM) stage, and the presence of anti-tissue transglutaminase IgA (anti-TgA), HLA-DQ2, and HLA-DQ8 markers. Results A total of 434 consecutive patients with (n = 315) and without (n = 119) endometriosis were included. Pain and infertility were more frequent in the endometriosis group than in the control group. The presence of HLA-DQ2, HLA-DQ8, and anti-TgA was similar between both groups. The presence of HLA-DQ2 and HLA-DQ8 markers did not differ based on age, pain symptoms, ASRM stage, or endometriosis location. Conclusion Although there are similarities in inflammatory markers and pathophysiology between celiac disease and endometriosis, this study found no significant associations in the presence of HLA-DQ2 or HLA-DQ8 haplotypes and endometriosis.

Role of HLA alleles polymorphism in systemic lupus erythematosus: A prospective study from North India

Background: Systemic lupus erythematosus (SLE) is a chronic autoimmune disorder and has complex etiopathogenesis. The most appropriate hypothesis states that genetic susceptibility in the presence of environmental risk factors predisposes to SLE. HLA class II alleles are critical to immune response and are highly polymorphic. Various alleles in HLA-DR and -DQ regions were analyzed in SLE patients and healthy controls to see their role in susceptibility or protection to SLE. Materials and Methods: This was a prospective observational study, in which a total of 100 SLE patients and 100 controls were analyzed. HLA typing was done by polymerase chain reaction (PCR)-sequence-specific oligonucleotide (SSO) method (SSO probe). Results: DR?1*0301 was significantly increased in SLE patients when compared to controls and had the highest odds ratio. Other risk factor alleles found to be increased were DR?1*0701, DQ?1*0202, and DQ?1*0301, which had a significant positive association with SLE, suggesting their role in susceptibility to SLE. In contrast, DR?1*0401, DR?1*1401, DR?1*1404, DR?1*1501, DQ?1*0501, and DQ?1*0201 showed statistically significant reduction in SLE patients, while these were much more common in controls, suggesting their protective role. Conclusion: This study is only the second study in patients from North India and it determines the role of DR?1*0301, DR?1*0701, DQ?1*0202, and DQ?1*0301 alleles as risk factors in SLE patients.

Seven cases of toxic encephalopathy associated with diquat poisoning / 中华急诊医学杂志

Objective:To improve the understanding of the clinical features of toxic encephalopathy associated with diquat poisoning.Methods:This study collected and analyzed the diagnosis and treatment process of 7 patients with acute diquat poisoning combined with central nervous system complications admitted to the First Affiliated Hospital of Zhengzhou University from April 2021 to April 2022. "Diquat" and "Poisoning" were used as keywords to search in CNKI, Wanfang database and PubMed database, and the literature of previous cases was reviewed for summary analysis.Results:Among the 7 patients in our hospital, there were 2 males and 5 females, with an average age of 31 years (range14-57) and an average dose of 23.14 g [(10-40)g]. During the treatment, 3 patients developed irritability and convulsions, 3 patients occurred coma, and one had generalized tonic-clonic seizures. Four patients died and 3 survived, of which 2 patients returned to normal life and study, and one remained abnormal mental behavior (currently in long-term follow-up). All three survivors developed neurological symptoms later than those who died, and were awake about 30 days after taking the drug.Conclusions:Toxic encephalopathy associated with diquat poisoning has rapid progression, poor prognosis and high mortality. This study found that the survival rate of patients with > 48 h of first onset of neurological symptoms is much higher than that of patients with ≤ 48 h of first onset of neurological symptoms, while sex, age, estimated oral dose, and type of presentation of neurological symptoms for the first time have little effect on the survival rate of hospital discharge. The earlier neurological symptoms appear, the greater the likelihood of a poor prognosis.

HLA-DQA1*04: 01 is related to a higher multiple sclerosis lesion load on T2/Flair MRI sequences / O HLA-DQA1*04: 01 está relacionado com uma alta carga lesional na ressonância magnética em T2/Flair nos pacientes com esclerose múltipla

ABSTRACT Background: The genetic predisposition to multiple sclerosis (MS) is associated with HLA alleles, especially HLA-DRB1*15:01. Objective: To identify associations between findings in magnetic resonance imaging (MRI) and genetic features in a Brazilian cohort of patients with MS. Methods: We retrospectively studied data from 95 consecutive patients with MS. Two independent observers who were blinded to the clinical data identified black holes and enhanced lesions on T1 MRI sequences, and counted and measured contrast-enhanced lesions on T2 and Flair (fluid attenuation inversion recovery) sequences. Cases were classified according to lesion size, number, and volume. The HLA-DRB1, HLA-DQB1, and HLA-DQA1 alleles, and the rs4774, rs3087456, rs6897932, rs731236, and rs1033182 single nucleotide polymorphisms were identified by polymerase chain reaction amplification with sequence-specific primers using the One Lambda Inc. Kit, Canoga Park, CA, USA. Results: Patients with the HLA-DQA1*04:01 allele had lesion load (adjusted for age, sex, and MS duration) above median compared with patients with other HLA-DQA1 alleles (p=0.02). There were no differences among all the other HLA alleles and single nucleotide polymorphisms and lesion load. Conclusions: The correlation of the HLA-DQA1*04:01 allele with a higher lesion load on T2/Flair MRI sequences suggests that the presence of this allele is associated with the risk of greater MS severity.

RESUMO Antecedentes: A predisposição genética para a esclerose múltipla (EM) está associada a alelos HLA, principalmente o HLA-DRB1*15:01. Objetivo: Identificar associações entre lesões na ressonância magnética e características genéticas em uma coorte brasileira de pacientes com EM. Métodos: Estudamos retrospectivamente os dados de 95 pacientes consecutivos com EM. Dois observadores independentes que desconheciam os dados clínicos identificaram "black holes" e lesões realçadas pelo contraste nas sequências de ressonância magnética T1 e contaram e mediram as lesões nas sequências T2 e FLAIR (fluid attenuated inversion recovery). Os casos foram classificados de acordo com tamanho, número e volume da lesão. Os alelos HLA-DRB1, HLA-DQB1 e HLA-DQA1 e os polimorfismos de nucleotídeo único rs4774, rs3087456, rs6897932, rs731236 e rs1033182 foram identificados por amplificação de reação em cadeia da polimerase com iniciadores específicos de sequência usando o kit One Lambda Inc., Canoga Park, CA, EUA. Resultados: Os pacientes com alelo HLA-DQA1*04:01 apresentaram carga de lesão (ajustada para idade, sexo e duração da EM) acima da mediana em comparação com outros pacientes com demais alelos HLA-DQA1 (p=0,02). Não houve diferenças entre todos os outros alelos HLA e polimorfismos de nucleotídeo único e carga lesional. Conclusões: A correlação do alelo HLA-DQA1*04:01 com maior carga de lesão nas sequências de RM em T2 sugere que a presença desse alelo pode estar associada ao risco de maior gravidade da EM.

Clinical effect and mechanism of / 中国针灸

OBJECTIVE@#To observe the effect of @*METHODS@#A total of 60 children with intellectual disability were randomly divided into an observation group (30 cases, 2 cases dropped off) and a control group (30 cases, 2 cases dropped off). In the control group, rehabilitation training and routine acupuncture were adopted, 30 min each time, once a day, 6 times a week for 3 months. On the base of the treatment as the control group, @*RESULTS@#Compared before treatment, the scores of DQ and ADL and the serum levels of DA, NE, 5-HT after treatment were increased (@*CONCLUSION@#On the base of rehabilitation training and routine acupuncture,

Insuficiencias metodológicas lastran el diagnóstico de la enfermedad celiaca / Methodological shortage hampers the diagnostic of celiac disease

Introducción: La enfermedad celiaca es el resultado de una sensibilidad permanente al gluten. Puede conducir principalmente a trastornos intestinales. Cuatro criterios son utilizados para el diagnóstico de esta enfermedad: clínicos, histológicos, serológicos y moleculares. La insuficiente utilización de estos criterios conduce a falsos diagnósticos de dicha enfermedad. Objetivo: Demostrar la existencia de falsos diagnósticos de enfermedad celiaca cuando no se utilizan las herramientas necesarias para ello. Métodos: Se estudiaron 46 niños que fueron remitidos al Servicio de Genética Molecular del Hospital Hermanos Ameijeiras con diagnóstico de enfermedad celiaca basado en criterios clínicos e histopatológicos. Para completar los procederes diagnósticos, a cada paciente se le determinó anticuerpos antitransglutaminasa previa ingesta de gluten, y los alelos HLA DQ2 y HLA DQ8. Se consideraron pacientes con enfermedad celiaca aquellos casos que cumplieron los cuatro criterios. Resultados: De los 46 pacientes, trece (28,3 por ciento) fueron negativos a los alelos HLA DQ2/HLA DQ8, lo que niega estén padeciendo de enfermedad celiaca; ocho (17,39 por ciento) fueron positivos a los alelos HLA y negativos a la presencia de anticuerpos, lo que también niega la enfermedad. Es decir, 21 (45,7 por ciento) eran falsos diagnósticos de enfermedad celiaca. Los 25 (54,3 por ciento) restantes, además de los criterios con que fueron remitidos, cumplieron los serológicos (positividad a anticuerpos antitransglutaminasa) y moleculares (positividad para moléculas HLA DQ2/HLADQ8). Conclusiones: Para un diagnóstico de certeza de enfermedad celiaca es necesario, además de las herramientas clínicas e histopatológicas utilizadas en la red de hospitales pediátricos del país, el uso de procederes serológicos y moleculares(AU)

Introduction: Celiac disease is a caused by a permanent sensitivity to gluten, which results mainly in functional disorders of the small intestine. To successfully diagnose of celiac disease, it is necessary to properly convey four criteria: clinic, histological, serological and molecular. The insufficient utilization of them in the medical practice could conduce to false diagnosis of celiac disease. Objective: To demonstrate the occurrence of mistaken diagnoses of celiac disease when the four criteria are not properly addressed. Methods: Forty-six children were diagnosed with celiac disease based on clinical and histopathological criteria and remitted to the Hermanos Ameijeiras Hospital´s Molecular Genetics service. In order to complete the serological and molecular diagnosis procedure, there were detected antitransglutaminase antibodies after gluten ingestion, and HLA DQ2/HLA DQ8 alleles in every child. Individuals who met the four criteria were considered celiac disease patients. Results: The analysis of 46 patients showed that 13 (28.3 percent) where negative to the presence of both allele HLA DQ2/HLA DQ8, and hence negative for celiac disease diagnosis. Eight patients (17.39 percent) where HLA DQ2/HLA DQ8 positive and antitransglutaminase antibodies negative, so they were considered as negative for diagnosis of celiac disease. According to our results, 21 patients (45.7 percent) were mistakenly diagnosed. The remaining 25 patients (54.3 percent) where positive for all diagnosis criteria. Conclusions: In order to successfully diagnose of celiac disease, in addition to clinical and histopathological tools used in the network of pediatrics hospitals in the country, it is necessary to include the serological and molecular method(AU)

De-novo anti-DQ antibodies as a risk factor for kidney allograft failure / Эрүүл мэндийн лаборатори

Background@#De-novo donor and non-donor specific antibodies could be detrimental to the kidney allograft. Kidney transplantation has being performed in Mongolia since 2006. However there is currently no published data available on post-transplant de-novo antibodies and long-term graft survival. Our aim was to determine immunosuppressive drug through level, its combination, de-novo HLA antibodies and its influence on graft survival in different immunosuppressive protocols. @*Methods@#We analyzed data from 56 adult first kidney transplant recipients at our hospital from August 2006 to May 2013. We determined the level of tacrolimus, cyclosporine A, and the presence of pre and post-transplant anti-HLA antibodies.@*Results@#Post-transplant follow up period was 1-8 years. Mean recipient age on transplantation was 33.9±9.1 years. Male 45 (80.4%). Cadaver donor kidney was 5 (8.9%). Mean donor age on transplantation was 39.98±11.13 years. Rejection occurrence was 12(21.4%). Tacrolimus and cyclosporine A through levels were 3-12.8ng/ml and 65- 324ng/ml respectively. Anti-HLA class I antibodies were detected in 17.9% of pretransplantation (n=10) and in 23.2% of post-transplantation (n=13) cases respectively (p=0.607). On the other hand, anti-HLA class II antibodies were detected in 5.4% of pretransplantation (n=3) and in 33.9% of post-transplantation (n=19) cases (p=0,001). We determined anti-HLA class II antibody specificity. Anti-DQ, DR, DP antibodies were 25% ( n=14), 14.3% ( n=8) and 7.1% ( n=4) respectively on all 56 cases. Two (3.6%) patients’ samples were positive on three loci of HLA class II. Six patient samples (10.7%) were positive on two loci. Nine (64.3%) of anti-DQ positive patients have rejected their grafts and begun hemodialysis treatment. All 9 graft rejected recipients were anti-HLA DQ positive and had taken cyclosporine mono-therapy for the first year after transplantation.@*Conclusion@#The presence of de-novo anti-HLA class II antibodies, especially de-novo anti-DQ were significantly increased on cyclosporine mono-therapy group following transplantation and negatively affected kidney graft survival. The blood through level of cyclosporine was very variable. The graft survival was better in standard triple regimen. Therefore, it is essential to monitor immunosuppressive drug combinations with drug blood level and anti-DSA antibodies as well as to manage antibody removal therapies such as therapeutic plasma exchange, intravenous immunoglobulin and Rituximab therapy on time. HLA –DQ-DP antigen determination is important for the kidney transplantation.

Prevalence of markers of celiac disease in Colombian children with diabetes mellitus type 1 / Prevalencia de marcadores de enfermedad celíaca en niños Colombianos con diabetes mellitus tipo 1

Abstract Introduction: Although the association between diabetes mellitus type 1 (T1DM) and celiac disease (CD) is well established; there are only a few studies that focus on South American children, haplotypes and their possible associations. Objective: To determine the prevalence of CD markers in a group of children with T1DM and to analyze the associated clinical, immunological and genetic manifestations. Methods: A prevalence study focusing on children with T1DM who were assessed based on variables including sociodemographics, anthropometric information, disease characteristics, laboratory results and family medical history. In partitipants a positive tTG2 (Ig A anti-transglutaminase), a duodenal biopsy and genotype were performed. The proportion of children with T1DM and CD was estimated (CI 95%). Determinations of central tendency, univariate and bivariate analysis, were also performed; p <0.05 was considered significant. Results: Thirteen (8.4%) of the 155 children (53.6% girls, 11.0 ±3.6 years, 2-18 years) with T1DM were tTG2 positive, four had CD (2.6%), seven had potential CD (4.5%) and nine were HLA DQ2/DQ8 positive (5.8%). Children with T1DM and CD had their last ketoacidotic episode (21.5 ±30.4 months versus 69.5 ±38.8 months, p= 0.0260) earlier than children with T1DM and potential CD. There were no differences with anthropometry or with the laboratory results regarding glycemic control. Conclusions: The prevalence of CD in these children with T1DM is higher than that reported in other South American countries. The prevalence of CD was found to be associated with the time of presentation of T1DM and its main allele, the DQ2/DQ8. These findings are different from what has been described in other places around the world.

Resumen Introducción: A pesar que la asociación entre diabetes mellitus tipo 1 (DMT1) y enfermedad celíaca (EC) está bien establecida; hay pocos estudios en niños suramericanos sobre haplotipos y sus posibles asociaciones. Objetivo: Determinar la prevalencia de marcadores de EC en un grupo de niños con DMT1, analizando las manifestaciones clínicas, inmunológicas y genéticas. Métodos: Estudio de prevalencia en niños con DMT1 a quienes se les tomaron variables sociodemográficas, antropométricas, de la enfermedad, paraclínicas y familiares metabólicas. A los niños con IgA anti-transglutaminasa (tTG2) positivos, se les realizó biopsia duodenal y genotipo. Se estimó la proporción de niños con DMT1 y EC y su IC 95%; medidas de tendencia central, análisis univariado y bivariado, siendo significativa una p <0.05. Resultados: Trece (8.4%) de los 155 niños (53.6% niñas, de 11.0 ±3.6 años, 2-18 años) con DMT1 fueron tTG2 positivos, cuatro presentaron EC (2.6%), siete EC potencial (4.5%) y nueve HLA DQ2/DQ8 (5.8%). Los niños con DMT1 y EC presentaron más pronto su último episodio cetoacidótico (21.5 ±30.4 meses versus 69.5 ±38.8 meses, p= 0.0260) que los niños con DMT1 y EC potencial. No hubo diferencias con la antropometría ni con los paraclínicos del control glicémico. Conclusiones: La prevalencia de EC en estos niños con DMT1 es superior a la de otros países suramericanos; estando asociada al tiempo de presentación de la DMT1 y su principal alelo el DQ2/DQ8, hallazgos diferentes a lo descrito a nivel mundial.

Prevalência de genótipos predisponentes para doença celíaca, incluindo HLA-DQ2. 2, em crianças brasileiras / Prevalence of celiac disease predisposing genotypes, including hla-dq2. 2 variant, in brazilian children

ABSTRACT BACKGROUND: Celiac disease is an autoimmune enteropathy triggered by the ingestion of gluten in genetically susceptible individuals. Almost all celiac patients carry immune recognition genes coding for HLA-DQ2.5 and DQ8 heterodimers. Over the last few years, great importance has been given to HLA-DQ2.2 as probable predisposing variant, although controversies still exist regarding its relevance. OBJECTIVE: The aim of our study was to determine the possible existence of an association between HLA-DQ2.2 and celiac disease in Brazilian children by analyzing the prevalence of the predisposing variants for celiac disease in a representative group of children of a population in which this determination is still missing. METHODS: HLA-DQ typing was performed in samples from a group of celiac (n=100) and non-celiac children (n=110). All samples were tested for the presence of the following variants: DQA1*05-DQB1*02 (DQ2.5), DQA1*03-DQB1*03:02 (DQ8) and DQA1*02:01-DQB1*02:02 (DQ2.2). Fisher`s exact test was used for statistical analysis. RESULTS: In the group of 100 celiac children, 78 (78%) were positive for DQ2, 13 (13 %) were DQ2/DQ8 and 6 (6%) were DQ8 positives. The HLA-DQ pattern in the 110 non-celiac children was as follows: positive for DQ2 in 33 (29.9%) samples, in 2 (1.8 %) was positive for DQ2/DQ8 and in 15 (13.6%) was positive for DQ8. We found significant differences between the distribution of some but not all of the analyzed alleles when comparing celiac and non-celiac children. CONCLUSION: The genotyping of celiac disease HLA-DQ predisposing alleles showed similarities with HLA-DQ patterns found in both European and non-European populations, which may be a reflection of the miscegenation, which gave origin to the current Brazilian population. No significant association was found between DQ2.2 variant and celiac disease in the studied population.

RESUMO CONTEXTO: A doença celíaca é uma enteropatia autoimune, desencadeada pela ingestão do glúten em indivíduos geneticamente predispostos. Quase todos os pacientes celíacos possuem genes que codificam os heterodímeros HLA-DQ2.5 e DQ8. Nos últimos anos, mesmo com algumas controvérsias a respeito, tem se dado grande importância ao HLA-DQ2.2 como outra provável variante predisponente para doença celíaca. OBJETIVO: O objetivo do nosso trabalho foi determinar a provável associação entre HLA-DQ2.2 e a doença celíaca em crianças brasileiras, mediante a análise da prevalência das variantes predisponentes para doença celíaca em um grupo representativo desta população que ainda carece de dita informação. MÉTODOS: A genotipagem das variantes HLA-DQ foi realizada em populações de crianças celíacas (n=100) e não celíacas (n=110). A presença das seguintes variantes foi testada em todas as amostras: DQA1*05-DQB1*02 (DQ2.5), DQA1*03-DQB1*03:02 (DQ8) e DQA1*02:01-DQB1*02:02 (DQ2.2). A análise estatística foi realizada utilizando o teste exato de Fisher. RESULTADOS: No grupo de 100 crianças celíacas, 78 (78%) foram positivas para DQ2, 13 (13%) para DQ2/DQ8 e 6 (6%) foram DQ8 positivas. O padrão de variantes predisponentes no grupo de 110 crianças não celíacas foi: 33 (29.9%) amostras positivas para DQ2, 2 (1.8%) DQ2/DQ8 positivas e 15 (13.6%) DQ8 positivas. Quando as prevalências de ambos grupos foram compradas, foram achadas diferenças significativas entre algumas, mas não todas as variantes predisponentes. CONCLUSÃO: A genotipagem das variantes HLA-DQ predisponentes para doença celíaca mostrou um padrão similar ao achado em populações europeias e não-europeias, o qual pode ser resultado da miscigenação que deu origem à população brasileira atual. Nosso trabalho não mostrou associação significativa entre a variante DQ2.2 e a doença celíaca na população estudada.

Therapeutic Observation of Electroacupuncture plus Rehabilitation for Cerebral Palsy / 上海针灸杂志

Objective To observe the clinical efficacy of electroacupuncture plus rehabilitation in treating cerebral palsy.Method A total of 104 patients with cerebral palsy were randomized into a treatment group and a control group, 52 cases in each group. The two groups both received routine medications, based on which, the treatment group was given electroacupuncture plus rehabilitation training while the control group was given rehabilitation training alone. Before and after the treatment, the Gross Motor Function Measure (GMFM), muscle tension, comprehensive capability, development quotient (DQ) and cerebral haemodynamics were observed, and the clinical efficacies were compared between the two groups.Result The total effective rate was 94.2% in the treatment group versus 71.2% in the control group, and the between-group difference was statistically significant (P<0.05). After the treatment, the scores of item D and E of GMFM-88 were significantly increased (P<0.01) and the muscle tension scores were significantly decreased in both groups (P<0.01); the scores of item D and E of GMFM-88 and muscle tension in the treatment group were significantly different from those in the control group (P<0.05). The comprehensive capability score and DQ were markedly increased in both groups after the treatment (P<0.01); after the treatment, the comprehensive capability score and DQ in the treatment group were significantly different from those in the control group (P<0.01). The cerebral artery mean flow velocity (Vm) increased significantly (P<0.05), and pulsatility index (PI) and resistance index (RI) decreased significantly after the treatment in both groups (P<0.05); after the treatment, the cerebral artery Vm, PI and RI in the treatment group were significantly different from those in the control group (P<0.05).Conclusion Electroacupuncture plus rehabilitation is an effective approach in treating cerebral palsy, and it can obviously improve the limb dysfunction, DQ, and cerebral haemodynamics of the patients.

Prevalence of genetic susceptibility for celiac disease in blood donors in são paulo, brazil / Prevalência da predisposição genética para doença celíaca nos doadores de sangue em São Paulo, Brasil

ABSTRACT Background Celiac disease is a permanent intolerance induced by gluten, which is expressed by T-cell mediated enteropathy, and has a high prevalence in the general population. There is evidence of a strong genetic predisposition to celiac disease. Objective To determine the prevalence of genetic markers HLA-DQ2 and HLA-DQ8 in blood donors from São Paulo and measure human recombinant tissue transglutaminase antibody IgA class in HLA-DQ2 and HLA-DQ8 positive donors. Methods A total of 404 blood donors from São Paulo city and Jundiaí were included in the study and signed the informed consent form. Information regarding diarrhea, constipation and abdominal pain in the last 3 months was collected. Determination of HLADQ2 and HLADQ8 alleles was performed in all participants and human recombinant tissue transglutaminase antibody class IgA was measured only in blood donors who presentedDQ2 and/or DQ8. Results HLADQ2 and/or HLADQ8 were positive in 49% (198/404) of subjects. Positive samples were associated with alleles DR3, DR4, DR7, DR11 and DR12. The most frequent genotype was DR4-DQ8, which was present in 13.6% of samples, followed by genotypes DR3-DQ2 and DR7-DQ2 with DQB1*02 in heterozygous, which were present in 10.4% and 8.7%, respectively. Eleven out of 198 positive donors (5%) were positive to human tissue transglutaminase test. Conclusion We observed a high prevalence of genetic markers for celiac disease, HLA-DQ2 and HLA-DQ8, in blood donors from São Paulo, similar to prevalence described in Europe. These findings show that the prevalence of celiac disease should not be rare in our country, but underdiagnosed.

RESUMO Contexto A doença celíaca é uma enteropatia imuno mediada causada pela intolerância permanente induzida pelo glúten, que se expressa por enteropatia mediada por linfócitos T, e possui uma alta prevalência na população geral. Há evidências de forte predisposição genética para doença celíaca. Objetivo Determinar a prevalência dos marcadores genéticos HLA-DQ2 e HLA-DQ8 em doadores de sangue da cidade de São Paulo e realizar rastreamento sorológico para doença celíaca com anticorpo antitransglutaminase tissular recombinante humana de classe IgA naqueles doadores de sangue com genotipagem HLA-DQ2 e HLA-DQ8 positivos. Métodos Estudo transversal prospectivo em que participaram 404 doadores de sangue, residentes na cidade de São Paulo e Jundiaí. A determinação dos alelos HLADQ2 e HLADQ8 foi realizada por PCR multiplex e alelo específico em todos os participantes do estudo e o anticorpo antitransglutaminase tissular recombinante humana de classe IgA e dosagem sérica de IgA foi realizada apenas nos doadores de sangue que possuíam DQ2 e/ou DQ8 positivo. Resultados O HLADQ2 e/ou DQ8 foi positivo em 49% (198/404) dos indivíduos, destes, 11 (5%) apresentaram anticorpo antitransglutaminase tissular humana positivo. Conclusão Podemos concluir que a prevalência dos marcadores genéticos para doença celíaca, HLA-DQ2 e DQ8 em São Paulo, mostrou-se elevada e similar à encontrada na Europa, assim como foi elevada a soroprevalênca para doença celíaca nos doadores de sangue com presença HLA-DQ2 e DQ8. Estes achados permitem afirmar que a prevalência da doença celíaca não deve ser rara em São Paulo, mas sim subdiagnosticada.

Prevalence of DQ2, DQ8 and DR4 Alleles in Iraqi Celiac Patients

Celiac disease(CD) is a chronic small intestinal immune-mediated enteropathy precipitated by exposure to dietary gluten in genetically predisposed individuals, many environmental triggering factors are suggested to participate in its pathogenesis, CD is strongly concomitant with specific HLA class II genes known as HLA-DQ2 and HLA-DQ8 which present in 90-95% of celiac patients and the remaining (5-10%) bears DR4-DQ8 haplotype. Also non-HLA genes my influence susceptibility to the disease but their influence has not been confirmed yet. So that this study aims to determine the distribution of DQ2, DQ8, DR4and non DQ2/DQ8 among Iraqi celiac patients. Sample of 80 Iraqi celiac patients (tTg-A,tTg-G and AGA positive)  has been chosen from all suspected patients who attending to Al-Suder- Medical city during the period of  April  2015 to  November 2015, blood samples were obtained from those patients and send for DNA extraction and HLA typing by RT-PCR. The results showed that 70% (56) patients were females and 30% (24) were males, also 37.5% from those patients were lies between 1-10 years old. HLA typing showed that 77.5% of those were had DQ2 genotyping, 7.5% were had D82 genotyping, 10% were had DR4-DQ8 genotyping and 5% were had non DQ2/DQ8 genotyping.

A study on HLA-DR/DQ typing in adult Malay patients with acute amoebic liver abscess

Introduction: Amoebiasis is a parasitic disease caused by Entamoeba histolytica that may lead to death in developing countries. Few important risk factors have been identified in the development of amoebic liver abscess (ALA). There are limited reports that suggest an association between antigens of the major histocompatibility complex (MHC) particularly class II antigens and ALA development. This present work aimed at studying the possible association of HLA antigens with ALA and disease severity. Results of the study may serve as a guide for further immunological studies dealing with E. histolytica. Methods: This preliminary study involved two groups of subjects: 20 ALA patients in the experimental group and 40 healthy individuals in the control group. Cases were selected from adult Malay patients confirmed with ALA based on clinical signs and symptoms, radiological findings, microbiological findings and who were admitted to the medical or surgical ward, Hospital USM, Kelantan. Venous blood was obtained from each patient and HLA typing was then conducted using polymerase chain reaction specific primer sequence. Results: HLA DR12 was most frequently found in the healthy control and ALA groups at 40% and 55% respectively. HLA DQ7 and DQ8 were found to have the highest percentage in the ALA group at 65%. In the control group, HLA DQ8 (57.5%) had the highest percentage. Conclusion: HLA antigens play a role in acquisition of ALA and provide understanding of the disease outcome.

Patrón clínico y sensibilidad de la serología frente a la histología en niños celiacos HLA DQ2/DQ8 / PATTERNS IN CLINICAL PRESENTATIONS, SEROLOGY SENSITIVITY IN FRONT OF HISTOLOGY IN THE CHILDREN CELIACS HLA DQ2/DQ8

Introducción: En áreas tropicales donde el diagnóstico diferencial para enfermedad celíaca está presente, la determinación de HLA DQ2/DQ8 es útil para confirmar la existencia de la enfermedad en pacientes sintomáticos o no con serología negativa y lesión mucosal o anticuerpo positivo y mucosa nor- mal, con la limitante del costo elevado. Objetivo: conocer el patrón clínico en niños celiacos con determinación HLA DQ2/DQ8 e investigar la sensibilidad de la serología frente a la histología en el despistaje diagnóstico de la enfermedad. Pacientes y método: estudio prospectivo y transversal, que incluyo 18 niños celíacos con determinación de DQ2/DQ8. Se registró: edad, sexo, clínica, serología, biopsia y genética. Resultados: edad promedio 3,39 años (8meses-13 años),55,55% hembras. Patrón clínico clásico en 12/18 (66,67%), atípico 3/18 (16,67%), latente 2/18 (11,11%), potencial 1/18 (5,55%). En total 12/18 pacientes (66,67%) con serolo- gía positiva. A la histología: 2/18 mucosa normal (11,11%) y 16/18 alterada (88,89%), de ellos, 4 Marsh I, 5 Marsh II, 7 Marsh III. En todos los niños con serología positiva se observo lesión intestinal, 25% con atrofia de vellosidades. Con serología negativa, 4 con atrofia vellositaria (2/4 con déficit de Ig A) y 2 mucosa normal. Se encontró una sensibilidad de la serología para el diagnóstico en 75%, specificidad 100%. Exactitud diagnóstica en 77,77% de la serología frente a la histología. Conclusiones: la serología resulto con una sensibilidad aceptable para el despistaje diagnóstico de celíaca y la determinación de HLA DQ2/DQ8 fue de utilidad en la caracterización del patrón clínico y la detección de la enfer- medad un grupo de pacientes.

Introduction: In tropical areas where the differential diag- nosis for celiac disease is present, the determination of HLA DQ2/DQ8 is useful to confirm the existence of the disease in symptomatic patients or HIV negative and positive mucosal injury and mucosal antibody or normal with limiting the high cost. Objective: To determine the clinical pattern in celiac children with HLA determination DQ2/DQ8 and investigate the sensitivity of the serology screening histology in the diagnosis of disease. Patients and methods: Prospective, cross-sectional, which included 18 children with celiac DQ2/ DQ8 determination. Was recorded: age, sex, clinical, sero- logy, biopsy and genetics. Results: mean age 3.39 years (8m-13), 55.55% females. Classic clinical pattern in 12/18 (66.67%), atypical 3/18 (16.67%), latent 2/18 (11.11%), potential 1/18 (5.55%). In total 12/18 patients (66.67%) with positive serology. A histology: 2/18 normal mucosa (11.11%) and 16/18 altered (88.89%), of whom 4 Marsh I, 5 Marsh II, 7 Marsh III. In all children with positive serology bowel injury was observed, 25% villus atrophy. With negative serology, 4 with villous atrophy (2/4 with IgA deficiency) and 2 normal mucosa. We found a sensitivity of serology for diagnosis in 75%, specificity 100%. 77.77% diagnostic accuracy of serology against histology. Conclusions: resulted serology with acceptable sensitivity for the screening and diagnosis of celiac HLA determining DQ2/DQ8 was useful in characterizing the clinical pattern and disease detection a group of patients in characterizing the clinical pattern and detection of the disease a group of patients.

Simplifying celaac disease predisposing hla-dq lleles determination by the reaal time pcr method / Simplificação da detecção dos alelos HLA-DQ predisponentes para doença celíaca pelo método de PCR em tempo real

Background Celiac disease is an autoimmune enteropathy triggered by the ingestion of gluten in genetically susceptible individuals. Genetic susceptibility is associated with two sets of alleles, DQA1*05 - DQB1*02 and DQA1*03 - DQB1*03:02, which code for class II MHC DQ2 and DQ8 molecules, respectively. Approximately 90%-95% of celiac patients are HLA-DQ2 positive, and half of the remaining patients are HLA-DQ8 positive. In fact, during a celiac disease diagnostic workup, the absence of these specific DQA and DQB alleles has a near perfect negative predictive value. Objective Improve the detection of celiac disease predisposing alleles by combining the simplicity and sensitivity of real-time PCR (qPCR) and melting curve analysis with the specificity of sequence-specific primers (SSP). Methods Amplifications of sequence-specific primers for DQA1*05 (DQ2), DQB1*02 (DQ2), and DQA1*03 (DQ8) were performed by the real time PCR method to determine the presence of each allele in independent reactions. Primers for Human Growth Hormone were used as an internal control. A parallel PCR-SSP protocol was used as a reference method to validate our results. Results Both techniques yielded equal results. From a total of 329 samples the presence of HLA predisposing alleles was determined in 187 (56.8%). One hundred fourteen samples (61%) were positive for a single allele, 68 (36.3%) for two alleles, and only 5 (2.7%) for three alleles. Conclusion Results obtained by qPCR technique were highly reliable with no discordant results when compared with those obtained using PCR-SSP. .

Contexto Doença celíaca é uma enteropatia autoimmune desencadeada pela ingestão de gluten em indivíduos geneticamente suscetíveis. Essa suscetibilidade genética está associada a dois conjuntos de alelos, DQA1*05 - DQB1*02 e DQA1*03 - DQB1*03:02, que codificam moléculas MHC de classe II DQ2 e DQ8, respectivamente. Aproximadamente 90%-95% dos pacientes celíacos são HLA-DQ2 positivos, e metade dos restantes são HLA-DQ8 positivos. No diagnóstico da doença celíaca, a ausência desses alelos DQA e DQB específicos possui um elevado valor preditivo negativo. Objetivo Nosso objetivo foi melhorar a detecção de alguns alelos predisponentes para doença celíaca, combinando a simplicidade e sensibilidade da técnica de PCR em tempo real (qPCR) e análise da curva de melting com a especificidade dos primers de sequência específica. Métodos Primers de sequência específica para DQA1*05 (DQ2), DQB1*02 (DQ2), e DQA1*03 (DQ8) foram usados para testar a presença de cada alelo em reações independentes. Primers para Hormônio de Crescimento Humano foram usados como controle interno. Em paralelo, foi usado um protocolo de PCR-SSP como um método de referência para validar nossos resultados positivos. Resultados Das 329 amostras testadas, 187 (56.8%) foram positivas para os alelos HLA predisponentes, usando as duas técnicas. Essas 187 amostras positivas foram subdivididas em 114 (61.0%) positivas para apenas um alelo, 68 (36.3%) para dois alelos e apenas 5 (2.7%) para os três alelos. Conclusão Os resultados obtidos pela técnica de qPCR mostraram-se altamente confiáveis, sem resultados discordantes quando comparados àqueles obtidos pelo método PCR-SSP. .

Celiac Disease in a Predisposed Subject (HLA-DQ2.5) with Coexisting Graves' Disease

Celiac disease is an intestinal autoimmune disorder, triggered by ingestion of a gluten-containing diet in genetically susceptible individuals. The genetic predisposition is related to human leukocyte antigen (HLA) class II genes, especially HLA-DQ2-positive patients. The prevalence of celiac disease has been estimated to be ~1% in Europe and the USA, but it is rarer and/or underdiagnosed in Asia. We report a case of celiac disease in a predisposed patient, with a HLA-DQ2 heterodimer, and Graves' disease that was treated successfully with a gluten-free diet. A 47-year-old woman complained of persistent chronic diarrhea and weight loss over a 9 month period. Results of all serological tests and stool exams were negative. However, the patient was found to carry the HLA DQ2 heterodimer. Symptoms improved after a gluten-free diet was initiated. The patient has been followed and has suffered no recurrence of symptoms while on the gluten-free diet. An overall diagnosis of celiac disease was made in a genetically predisposed patient (HLA-DQ2 heterodimer) with Graves' disease.

Increased accumulation of dendritic cells in celiac disease associates with increased expression of autophagy protein LC3.

Background: Celiac disease (CD) an immune-mediated disorder associates with accumulation of dendritic cell (DC) in duodenal mucosa. Autophagy has recently been implicated in autoantigen formation. However, its role in CD is still unknown. Aim: To examine role of autophagic protein LC3 expressed by activated DC in CD. Materials and Methods: Thirty CD patients were analyzed at initial presentation and after 6 months of gluten-free diet (GFD). Duodenal biopsies were studied for histological changes and CD11c, CD86, and MAP1LC3A expressions by double immunohistochemistry (IHC). Masson’s trichrome (MT) staining was used to assess basement membrane (BM) thickness and Oil Red O (ORO) staining for mucosal lipid deposit. Polymerase chain reaction (PCR) was performed for HLA-DQ system. Statistical analysis was done using paired and unpaired t test, chi-square test, Fisher’s exact test, and McNemar-Bowker test. A P-value <0.05 was considered statistically signifi cant. Results: HLA-DQ2 and HLA-DQ8 alleles were present in all studied patients. Increased BM thickness was observed in 63% and 73% had ORO-positive lipid in surface lining epithelium. Pre-treatment biopsies showed increased DCs expressing LC3, which were signifi cantly less in follow-up biopsies. The follow-up biopsies had shown signifi cant reduction in BM thickness and ORO. Conclusion: Histological improvement in duodenal biopsies was associated with reduction in activated DCs expressing autophagic protein, which probably play important role in pathogenesis of an autoimmune disorder like CD.

HLA DR/DQ type in a Malay population in Kelantan, Malaysia

The human leucocyte antigen (HLA) has been documented to be involved in various disease susceptibilities or in resistance against certain diseases. An important element in susceptibility and resistance to disease is ethnic genetic constitution. Cognizant of this, the present study aimed at studying the prevalence of particular HLA class II in a normal healthy Malay population which may serve as a guide for further genetic and immunological studies related to the Malay Malaysian population. The study involved 40 normal healthy Malay persons in Kelantan. HLA typing was conducted on venous blood samples through a polymerase chain reaction-sequence specifi c primer method (low resolution Olerup SSP® HLA Typing Kits). The study found HLA DR12 and HLA DQ8 to be the most frequent HLA class II type. HLA DQ5 was signifi cantly associated with female subjects.

Frecuencia de alelos HLA de clase I y II en una cohorte de pacientes con espondiloartritis provenientes del noroccidente colombiano / Frequency of HLA alleles class I and II in a cohort of northwestern Colombian patients with spondyloarthritis

Introducción. Las espondiloartritis son enfermedades reumatológicas crónicas que afectan el esqueleto axial y las articulaciones periféricas, con varias manifestaciones extraarticulares. La asociación con el HLA-B27 sigue siendo uno de los vínculos más fuertes conocidos entre estas entidades y el complejo mayor de histocompatibilidad; sin embargo, la distribución mundial del HLA-B27 varía considerablemente y se han descrito asociaciones con genes no HLA-B27. Objetivo. Conocer la frecuencia de alelos HLA de clase I y II en pacientes con espondiloartritis provenientes del noroccidente colombiano y su frecuencia en las manifestaciones clínicas y radiológicas específicas. Materiales y métodos. Se condujo un estudio descriptivo, observacional, de corte transversal, retrospectivo y prospectivo entre 2005 y 2008 de 56 pacientes colombianos con espondiloartritis. Se identificaron los alelos correspondientes a los loci HLA de clase I y II (HLA-B, HLADQB1 y HLADRB). Se analizó su frecuencia con las manifestaciones clínicas axiales, periféricas, extraarticulares y radiológicas. Resultados. Se encontró una baja frecuencia de HLA-B27 en la población total (50 %), aunque fue el alelo más frecuente, junto con HLA-DRB4*01 (35,7 %) y HLA-DQB1*0501 (28,6 %), en todos los pacientes en general y en cada una de las manifestaciones clínicas y radiológicas. Se resalta la alta frecuencia de HLA-B27 y HLA-DRB4*01 (64,3 %) en pacientes con dactilitis, hallazgo novedoso sin previa descripción. Conclusión. Los alelos HLA-B27, HLA-DRB4*01 y HLA-DQB1*0501 fueron frecuentes en los diferentes subtipos de espondiloartritis y en las manifestaciones clínicas axiales, periféricas y extraarticulares específicas, además de la sacroiliítis radiológica.

Introduction. Spondyloarthritis is a chronic rheumatic disease that affect the axial skeleton and peripheral joints, along with several extra-articular manifestations. The association with HLA-B27 remains one of the strongest known links between these entities and the major histocompatibility complex. However, the global distribution of HLA-B27 varies considerably and furthermore, associations with non-HLA-B27 genes have been described. Objective. The frequency of HLA class I and II was determined in a population of patients with spondyloarthritis with respect to detection in the clinical setting and by radiology. Materials and methods. A descriptive, observational, cross-sectional, retrospective and prospective study was conducted in 56 patients from northwestern Colombia. Each was diagnosed with spondyloarthritis between 2005 and 2008. In each case, alleles were identified for the loci HLA class I and II (HLA-B; HLADQB1 and HLADRB). The frequency of these alleles in the axial, peripheral, extraarticular and radiological manifestations. Results.The frequency of HLA-B27 was 50% overall, and it was the most frequent allele. The two other alleles were HLA.DRB4*01 at 35.7% and HLA-DQB1*0501 at 28.6%, as detected in each of the clinical and radiological manifestations. A high frequency of HLA-B27 and HLA-DRB4*01 (64.3%) was noted in patients with dactylitis. Conclusion. The alleles HLA-B27, HLA-DRB4*01 and HLA-DQB1*0501 were common in the different subtypes of spondyloarthritis and were frequent in the specific clinical axial, peripheral and extraarticular clinical manifestations, as well as radiological sacroiliitis.