Associação do alelo HLA-DRB1 com suscetibilidade a artrite reumatoide e gravidade da doença na Síria / HLA-DRB1 allele association with rheumatoid arthritis susceptibility and severity in Syria

INTRODUÇÃO: A artrite reumatoide (AR) é uma doença crônica multifatorial complexa. A importância do sistema de antígenos leucocitários humanos (HLA) como fator significativo de risco genético para AR foi estudada no mundo. Embora amplamente distribuídos em diferentes áreas na Síria, faltam estudos sobre o papel dos HLA. OBJETIVO: O objetivo de nosso estudo foi determinar a associação dos alelos HLA-DRB1 com a suscetibilidade a AR e sua gravidade na Síria. PACIENTES E MÉTODOS: Foram genotipados 86 pacientes com AR e 200 controles normais, usando-se reação em cadeia da polimerase com sequência de primer específico (PCR-SSP). Anticorpos anti-CCP foram determinados por ELISA. Fator reumatoide (FR), proteína C-reativa (PCR), velocidade de hemossedimentação (VHS) e o índice de atividade da doença (DAS-28) foram obtidos nos registros médicos e utilizados para avaliar a gravidade clínica dos pacientes. RESULTADOS: Os alelos HLA-DRB1 *01, *04 e *10 mostraram forte associação com suscetibilidade à doença (OR = 2,29, IC 95% = 1,11-4,75, P = 0,022; OR = 3,16, IC 95% = 2,08-4,8, P < 0,0001; e OR = 2,43, IC 95% = 1,07-5,51, P = 0,029, respectivamente), enquanto a frequência dos alelos HLA-DRB1 *11 e *13 foi significativamente mais baixa nos pacientes com AR do que nos controles (OR = 0,49, IC 95% = 0,3-0,8, P = 0,004; OR = 0,32, IC 95% = 0,15-0,69, P = 0,002, respectivamente). Os outros alelos HLA-DRB1 mostraram diferença significativa. A frequência dos anticorpos anti-CCP foi maior em pacientes epítopo compartilhado (EC) positivos do que em pacientes EC-negativos (OR = 5,5, IC 95% = 2-15,1, P = 0,00054). O índice DAS-28 de pacientes com AR não mostrou diferença significativa entre os grupos EC-negativo e EC-positivo. CONCLUSÃO: Nossos resultados indicam que os alelos HLA-DRB1 *01, *04 e *10 estão relacionados com AR, enquanto os alelos HLA-DRB1 *11 e *13 protegem a população síria contra a AR.

INTRODUCTION: Rheumatoid arthritis (RA) is a complex multifactorial chronic disease. The importance of human leukocyte antigen as a major genetic risk factor for RA was studied worldwide. Although it is widely distributed in different Syrian areas, studies of human leukocyte antigen (HLA) alleles' role are absent. OBJECTIVE: The aim of our study was to determine the association of HLA-DRB1 alleles with the susceptibility and severity of RA in Syria. PATIENTS AND METHODS: Eightysix RA patients and 200 healthy controls from Syria were genotyped using polymerase chain reaction with sequencespecific primer (PCR-SSP). Anti-CCP antibodies were measured by ELISA. Rheumatoid factor (RF), C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), and disease activity score 28 (DAS-28) values were obtained from patients' medical records. DAS-28 was used to assess the clinical severity of the patients. RESULTS: The HLA-DRB1*01, *04, and *10 frequencies showed a strong association with the disease susceptibility (OR = 2.29, 95% CI = 1.11-4.75, P = 0.022; OR = 3.16, 95% CI = 2.0 -4.8, P < 0.0001; OR = 2.43, 95% CI = 1.07-5.51, P = 0.029 respectively), while the frequencies of HLA-DRB1*11, and *13 were signifi cantly lower in RA patients than in controls (OR = 0.49, 95% CI = 0.3-0.8, P = 0.004; OR = 0.32, 95% CI = 0.15-0.69, P = 0.002, respectively). The other HLA-DRB1 alleles showed no signifi cant difference. The frequency of anti-CCP antibodies was higher in shared epitope (SE) positive patients compared with SE-negative patients (OR = 5.5, 95% CI = 2-15.1, P = 0.00054). DAS-28 of RA patients didn't show signifi cant difference between the SE negative and the SE positive groups. CONCLUSION: Our results indicate that HLA-DRB1*01, *04, and *10 alleles are related with RA, while HLA-DRB1*11 and *13 protect against RA in the Syrian population.

Role of Citrullinated Fibrinogen Peptides in the Activation of CD4 T Cells from Patients with Rheumatoid Arthritis

This study was conducted to determine whether CD4 T cell responses to citrullinated fibrinogen occur in patients with rheumatoid arthritis (RA), especially in HLA-DR4-positive subjects. Whole peripheral blood mononuclear cells (PBMCs) of RA patients and control subjects were stimulated with citrullinated fibrinogen peptides, and T-cell production of proliferation and proinflammatory cytokines, such as interferon-gamma(IFN-gamma) and interleukin-17A (IL-17A), were measured. In addition, CD4 T cells from RA patients were stimulated with the citrullinated fibrinogen peptide, Fib-alpha R84Cit, identified as a DRB1*0401-restricted T cell epitope in HLA-DR4 transgenic mice, and the degree of T cell activation was examined similarly. No proliferative responses to the citrullinated fibrinogen peptides were observed in whole PBMCs or CD4 T cells from RA patients. Furthermore, no increased production of IFN-gamma or IL-17A was found in whole PBMCs or CD4 T cells stimulated with the citrullinated fibrinogen peptides, although these cells responded to recall antigen, a mixture of tetanus toxoid, purified protein derivative (PPD) from Mycobacterium tuberculosis, and Candida albicans. The results of this study indicate that anti-citrulline immunity in RA patients may be mediated by fibrinogen because there is no evidence of CD4 T cell-mediated immune responses to citrullinated fibrinogen peptides.

Uveíte anterior na ausência de esclerite em paciente com artrite reumatóide: relato de caso / Anterior uveitis in the absence of scleritis in a pacient with rheumathoid arthritis: case report

A artrite reumatóide é a colagenose mais comum, afetando cerca de 0,6 por cento da população brasileira e é uma grande causadora de deformidades articulares em mais variadas formas. A principal manifestação ocular da artrite reumatóide é a ceratoconjuntivite sicca (Sjögren secundária), seguida pela esclerite, úlcera periférica da córnea e uveíte. O objetivo deste trabalho é apresentar um caso de uveíte anterior em paciente com artrite reumatóide, uma apresentação rara em pacientes com esta patologia. Paciente feminina, 55 anos, portadora de artrite reumatóide, apresentando quadro de dor e piora súbita da acuidade visual no olho direito. O exame mostrava reação de câmara anterior com hipópio, úlcera corneana periférica e pressão intraocular de 32 mmHg. Foi realizado o diagnóstico de uveíte anterior hipertensiva e úlcera corneana periférica e realizado tratamento com corticóide oral e tópico, antibiótico tópico, colírio cicloplégico e hipotensores oculares tópicos e sistêmicos. Os casos de uveíte anterior são comuns em doenças reumatológicas, principalmente em artropatias soronegativas relacionadas ao HLA-B27, conferindo grande causa de morbidade a esses pacientes. Neste trabalho relatamos um caso de uveíte anterior em paciente com artrite reumatóide, uma apresentação rara encontrada na literatura médica atual.

Rheumathoid arthritis is the most common collagenosis and affects almost 0.6 percent of brazilian population. It is an important cause of articular deformities. The main ocular manifestation of rheumathoid arthritis is dry eyes (secondary Sjögren's syndrome), followed by scleritis, peripheral ulcerative keratitis and uveitis. The aim of this paper is to present a case of anterior uveitis in the absence of scleritis in a patient with rheumathoid arthritis, a very rare presentation in this type of patient. Female patient, 55 years old, with rheumathoid arthritis, presenting suddenly ocular pain and low vision in the right eye. Her exam showed anterior chamber reaction with hypopion, peripheral corneal keratitis and intraocular pressure of 32 mmHg. She was diagnosed with hypertensive anterior uveitis and peripheral corneal keratitis and treated with systemic and topical corticosteroids, topical antibiotic, topic and systemic ocular hypotensive and mydriatic drops. Anterior uveitis is common in rheumatological diseases, especially in those soronegative arthropathies related to HLA B27. In this paper we present a patient with rheumathoid arthritis and anterior uveitis in the absence of scleritis, a rare presentation in actual medical literature.

Estudo de HLA classes I e II em trinta pacientes equatorianos com artrite reumatoide em comparação com alelos de indivíduos sadios e afetados com outras doenças reumáticas / Study of class I and II HLA alleles in 30 ecuadorian patients with rheumatoid arthritis compared with alleles from healthy and affected subjects with other rheumatic diseases

INTRODUÇÃO: A artrite reumatoide (AR) é uma doença inflamatória crônica sistêmica autoimune que provém de uma desordem incapacitante. Até hoje, a etiologia da AR é desconhecida. No entanto, já se cogitou a existência de indivíduos geneticamente passíveis de tê-la. Muitos estudos já foram realizados em todo o mundo, como, por exemplo, na Polônia, Argentina, Chile, México, Brasil, Colômbia, entre outros países, com relação à influência entre os alelos HLA-DR e a doença, mas não no Equador. OBJETIVO: O principal objetivo deste estudo foi determinar a participação dos alelos de HLA classes I e II em pacientes com AR. PACIENTES E MÉTODOS: Esta pesquisa foi desenvolvida em 30 pacientes adultos com AR, previamente diagnosticados de acordo com os critérios de classificação do Colégio Norte-Americano de Reumatologia (ACR, 1987) e 28 controles. Para a tipificação de HLA classes I e II, adotou-se a técnica PCR-SSP, e as significâncias estatísticas foram avaliadas pelo teste de Qui-Quadrado. RESULTADOS: O HLA-DR4 está presente em 76,7 por cento dos pacientes, com uma frequência alélica de 45 por cento, enquanto apenas 21 por cento dos sujeitos controle o apresentaram. O teste de Qui-Quadrado confirma que as variáveis HLA-DR4 e RA estão altamente vinculadas (X² = 11,38, P = 0,00074). CONCLUSÃO: Há frequência maior de HLA-DR4 e HLA-DR14. Os resultados encontrados são similares aos encontrados em outros estudos. Porém, seria desejável aumentar o tamanho da amostra para encontrar um maior número de perfis genéticos e de alelos envolvidos.

INTRODUCTION: Rheumatoid arthritis (RA) is a chronic systemic inflammatory autoimmune disease that originates from a disabling disorder. To date, the etiology of RA is unknown. However, the existence of genetically susceptible individuals was considered. Many studies have been performed worldwide, for example, in Poland, Argentina, Chile, Mexico, Brazil, and Colombia, among others, regarding the influence between HLA-DR alleles and disease, but not in Ecuador. OBJECTIVE: The aim of this study was to determine the involvement of Class I and II HLA alleles in patients with RA. PATIENTS AND METHODS: This study was conducted in 30 adult patients with RA previously diagnosed, according to the classification criteria of the American College of Rheumatology (ACR, 1987) and 28 controls. For Class I and II HLA typing, we adopted the PCR-SSP, and statistical significances were evaluated by Chi-Square. RESULTS: HLA-DR4 is present in 76.7 percent of patients, with an allele frequency of 45 percent, while only 21 percent of control subjects presented it. The chi-square confirms that HLA-DR4 and RA variables are highly bound (X2 = 11.38, P = 0.00074). CONCLUSION: There is increased frequency of HLA-DR4 and HLA-DR14. The results are similar to those found in other studies. But it would be desirable to increase the sample size in order to find a greater number of genetic profiles and alleles involved.

Relationship between HLA-DR4/1 subtypes and T cell responses to type Ⅱ collagen or anti-CⅡ263-272 antibodies / 北京大学学报(医学版)

Objective: To investigate the relationship between HLA-DR4/1 subtypes and T cell responses to type Ⅱ collagen or anti-CⅡ263-272 antibodies. Methods: Peripheral blood mononuclear cells and sera were obtained from 70 rheumatoid arthritis (RA) patients. T cell proliferative responses to CⅡ263-272 were evaluated by 3H thymidine incorporation assay. Antibodies specific to CⅡ263-272 were determined by ELISA. HLA-DR4/1 subtypes were analyzed by PCR-SSP. Results: In HLA-DR4/1 positive group, T cell proliferative responses to CⅡ263-272 were observed in 55.6% RA patients and anti- CⅡ263-272 antibodies were present in 66.7% RA patients. In HLA-DR4/1 negative group, T cell proliferative responses to CⅡ263-272 were observed in 30.3% RA patients and anti-CⅡ263-272 antibodies were present in 34.8% RA patients. The number of positive antibodies to CⅡ263-272 or T cell proliferative responses to CⅡ263-272 in HLA-DR4/1 positive group were higher than in negative group. Conclusion: There is a relationship between HLA-DR4/1 subtypes and T cell responses to CⅡ263-272 or anti-CⅡ263-272 antibodies.

The relationship between amino-acid motif of HLA-DR?1*04(?chain) and the susceptibility to rheumatoid arthritis / 中华风湿病学杂志

Objective To investigate the susceptibility to rheumatoid arthritis (RA) with HLA- DR?1*04 subtypes.Methods One hundred and thirty-six RA patients and 79 patients with other rheumatic diseases were recruited in this study.HLA typing was performed using high-resolution PCR-SSP DNA techniques.The clinical features and serologic markers between different motifs were analyzed.Results Compared with other rheumatic diseases,the frequency of HLA-DR?1*04 alleles in RA patients was significantly increased(33.8% vs 12.7%)(OR=3.52,95%CI=1.43~5.43,P

Tumor Susceptibility and Digestic System Tumor Susceptible Gene / 基础医学与临床

The carcinogenesis and development is a progress of multi-gene alterations in the human gastric cancer (HGC)．In order to determine the relation between the aberration of these genes and gastric cancer，we chose c-met (7q31)、hMLH1 (3p21)、E-cadherin (16q22.1) and HLA loci DQA1、DR2、DR3、DR4、DR7、DR9 and detected their changes in 32 tumor specimens of intestinal type HGC and 4 cell lines of gastric cancer by performing analysis of SSP/PCR、PCR/SSCP and MSI technigues．Our data show that none point mutation was detected in c-met gene．We examined two microsatellites loci D3S1298 and D3S1561 in hMLH1 gene and detected that 6 cases retain MSI (Microsatellite Instability) and 2 cases of LOH (Loss of Heterozygosity) at D3S1298 and 2 cases of MSI at D3S1561．We also examined E-cadherin gene at two microsatellites loci D16S3083 and D16S3095 close to the gene and detected that 5 cases retain MSI and 1 case of LOH at D16S3083 and no change at D16S3095．The point mutation incidence of HLA-DR4 loci is 9/20 (45%)，higher than the other loci in HLA-Ⅱ．High frequent deletion，expression deregulation and methylation of mts1/p16 gene were detected in cell lines and solid tumors from human gastric cancer patients． 　　Our data showed that the point mutation of c-met gene is not the main pattern of alteration in intestinal type HGC that is consistent with the previous results．E-cadherin and hMLH1 are related to intestinal type HGC but whether they are susceptibility gene still need further study．The point mutation of the HLA-Ⅱ loci DR4 is closely related to intestinal type HGC．Methylation of mts1/p16 gene 5 CpG island might be plays an important role in the carcinogenesis in HGC．

Rheumatoid Arthritis and HLA Class II Genotypes in Korean / 대한정형외과연구학회지

To determine which HLA antigens are associated with rheumatoid arthritis in Korean, we studied the HLA class II genotypes in 40 patients with rheumatoid arthritis and 35 normal healthy persons by the two step polymerase chain reaction. At first we designed 20 pairs of group specific primers followed by the alleles which have the same nucleotide sequences in the exon 2 region. We performed amplification reaction with these 20 pairs of primers for 32 cycles. Reaction products were electrophoresed and only specifically amplified products were reamplified for 15 cycles with total 76 pairs of allele specific primers. We typed the HLA class II genotypes with specific band patterns of each allele. The genotypes in patients were compared with those of normals. HLA-DR4 gave the highest relative risk for rheumatoid arthritis (RR=4), which was statistically significant (P0.05). Among HLA-DR4 subtypes, the frequency of Dwl4(0404) in patient group was relatively higher (19.2%) than that in normal control group(7.7%). The relative risk was high (2.85), although it was not statistically significant (P>0.05). From this study, HLA-DR4 was found to be highly associated while DRI was not associated with rheumatoid arthritis patients in Koreans.

Altered HA308-317 peptides inhibit HLA-DR4 specific T cell response / 中国免疫学杂志

Objective:To evaluate the inhibitory effect of altered HA308-317 peptides on HLA-DR4 restricted CII specific T cell response.Methods:Three altered HA308-317 peptides and CII263-272 were synthesized using solid-phase techniques. The binding of altered HA308-317 peptides for HLA-DR4 molecules was assayed using flow cytometry. The suppressive effect of altered HA308-317 peptides on CII-mediated T cell proliferation was determined using 3H incorporation assay. The level of IL-2 in the supernatants was identified by ELISA. The expression of CD25 and CD69 on T cell surface were studied using flow cytometry.Results:The altered HA308-317 peptides were able to bind to HLA-DR4 molecules and competed with CII263-272. Altered HA308-317 peptides inhibited T cell proliferation and IL-2 production induced by CII263-272( P