The association of PTPN22 (rs2476601) and IL2RA (rs11594656) polymorphisms with T1D in Egyptian children

BackgroundType 1 diabetes mellitus (T1D) is a T cell-mediated autoimmune disease characterized by the destruction of pancreatic β cells. PTPN22 and IL2RA polymorphisms have been found to be associated with several autoimmune diseases including T1D.AimsWe aimed to elucidate the role of PTPN22 and IL2RA polymorphisms in predisposition of T1D in Egyptian children.MethodsWe studied 150 children and adolescents with T1D and 165 healthy controls. The PTPN22 (rs2476601) and IL2RA (rs11594656) polymorphisms were genotyped using polymerase chain reaction.ResultsWe found that carriers of the T allele of PTPN22 were significantly more likely to develop T1D (OR = 2.2, 95% CI = 1.2–4, P = 0.01). Also, the carrier of TT genotype and T allele of IL2RA more likely to develop T1D (OR = 2.8, 1.4, respectively, P = 0.03). There was a statistically significant association between T allele of PTPN22 gene and females ⩽10 years old at the onset of diabetes (OR = 4, 95% CI = 1.2–13.4, P = 0.019).ConclusionThis study suggests a possible association between the T allele of PTPN22 gene and TT genotype of IL2RA with T1D in studied Egyptian children, especially, females with early onset diabetes who carried the 1858T allele.     

红细胞分布宽度和中性粒细胞淋巴细胞比值对类风湿性关节炎诊断价值的评价

目的 探讨红细胞分布宽度(RDW-SD)和中性粒细胞淋巴细胞比值(NLR)在类风湿关节炎(RA)诊断中的价值.方法 选取104例RA患者、104例其他风湿性疾病患者(骨关节炎25例,系统性结缔组织病69例,强直性脊柱炎10例)和99例健康体检者,比较各组血沉、超敏C反应蛋白、抗环瓜氨酸肽抗体、免疫球蛋白M、免疫球蛋白G、免疫球蛋白A、补体C3、补体C4,红细胞分布宽度,计数中性粒细胞、淋巴细胞数量、中性粒细胞淋巴细胞比值的差异,Spearman相关分析及多元逐步回归分析RDW-SD、NLR与其他指标的关系,ROC曲线比较各指标的诊断性能.结果 RA患者和其他风湿性疾病患者RDW-SD、NLR均高于健康对照组(P＜0.05);RA患者RDW-SD与CCP、C4相关,标准化回归系数分别为0.267(P＜0.05)、-0.189(P ＜0.05),NLR与CRP独立相关,标准化回归系数分别为0.616(P ＜0.05);ROC曲线分析显示,抗环瓜氨酸肽抗体是诊断RA最佳指标,AUC为0.832(P ＜0.05).结论 RDW-SD和NLR可以作为一个炎症反应指标,在RA诊断及疾病观察中有一定的价值.     

鸦胆子苦醇通过调节细胞骨架力学性质抑制类风湿关节炎成纤维样滑膜细胞的侵袭行为

目的研究鸦胆子苦醇对类风湿关节炎(rheumatoid arthritis,RA)成纤维样滑膜细胞(fibroblast-like synoviocytes,FLS)细胞骨架力学性质的调控以及对RA FLS侵袭行为的影响。方法细胞骨架染色法检测鸦胆子苦醇对RA FLS细胞骨架力学性质的调控;Transwell小室法检测鸦胆子苦醇对RA FLS细胞骨架的调控和对RA FLS侵袭行为的影响;酶谱和Western blotting法检测鸦胆子苦醇对RA FLS中MMP-2、MMP-3表达的影响。结果通过细胞骨架染色并在显微镜下观察发现,鸦胆子苦醇可显著减少RA FLS伪足数量的产生,通过调节细胞骨架网络的力学性质抑制细胞的运动能力,鸦胆子苦醇可抑制RA FLS的侵袭并能下调MMP-2、MMP-3的表达水平。结论鸦胆子苦醇能调节RA FLS的细胞骨架的力学性质并抑制细胞的侵袭行为,同时鸦胆子苦醇可通过降低MMP-2、MMP-3的表达抑制RA FLS的侵袭。研究结果为进一步开发治疗RA的新药物提供了相应实验依据。     

Preferential recognition of epitopes on AGE–IgG by the autoantibodies in rheumatoid arthritis patients

Incubation of proteins with glucose lead to their non-enzymatic glycation ultimately resulting in the formation of advanced glycation end products (AGEs) in vivo. AGEs alter unique three dimensional structures of various plasma proteins such as IgG. The role of oxidative stress in the pathogenesis of rheumatoid arthritis (RA), a chronic inflammatory autoimmune disease, is well established. In view of this, commercially available human IgG was glycated in vitro with physiological concentration of glucose (5 mM) and the possible involvement of glycated IgG (AGE–IgG) in RA was evaluated. The RA patients were divided into two groups on the basis of disease onset with respect to age: group I (early onset: 20–32 years) and group II (late onset: 36–54 years). AGE–IgG and oxidative stress levels were detected in RA patients and normal healthy individuals by nitroblue tetrazolium (NBT) assay and carbonyl content estimation respectively. Binding characteristics and specificity of RA antibodies were analyzed by enzyme-linked immunosorbent assay (ELISA). We observed preferential binding of RA antibodies to AGE–IgG in comparison to native IgG. Band shift assay further substantiated the enhanced recognition of AGE–IgG by RA antibodies. The results suggest that glycation of IgG results in the generation of neo-epitopes, making it a potential immunogen. Our findings project AGE–IgG as one of the factors for induction of circulating RA autoantibodies.     

Altered Th17 cells and Th17/regulatory T-cell ratios indicate the subsequent conversion from undifferentiated connective tissue disease to definitive systemic autoimmune disorders

A shift in the balance between Th17-cells and regulatory T-cells (Treg) is an important feature of systemic autoimmune diseases (SAID), and may also contribute to their development. Hereby, we assessed the distribution of peripheral Th17 and Treg-cells in patients with undifferentiated connective tissue disease (UCTD), the forerunner of SAIDs and followed these parameters during the development towards definitive SAIDs. Fifty-one UCTD patients were investigated and followed-up for 3 years. Flow cytometry was used to identify and follow three cell-populations: Th17-cells (CD4+IL-17+ T-cells), natural regulatory T-cells (CD4⁺CD25^brightFoxP3⁺; nTregs) and IL-10 producing Type-1 regulatory T-cells (CD4+IL-10+ T-cells; Tr1). Altogether 37.3% of these patients progressed into SAIDs. Th17-cells were increased in UCTD vs. controls, which further increased in those, whom developed SAIDs eventually. The Th17/nTreg ratio gradually increased from controls through UCTD patients, reaching the highest values in SAID-progressed patients. Regarding the Th17/Tr1 ratios, a similar tendency was observed moreover Th17/Tr1 could distinguish between UCTD patients with, or without subsequent SAID progression in a very early UCTD stage. Various immunoserological markers showed association with Th17 and Th17/nTreg at baseline, indicating the consecutive development of a distinct SAID. The derailed Th17/Treg balance may contribute to disease progression therefore could function as a prognostic marker.     

High frequencies of activated B cells and T follicular helper cells are correlated with disease activity in patients with new‐onset rheumatoid arthritis

This study aimed to examine the frequency of different subsets of circulating B and T follicular helper (Tfh) cells in patients with new-onset rheumatoid arthritis (RA) and following standard therapies. Twenty-five RA patients and 15 healthy controls (HC) were recruited for characterizing the frequency of CD27⁺, immunoglobulin (Ig)D⁺, CD86⁺, CD95⁺, Toll-like receptor (TLR)-9⁺ B cells and inducible T cell co-stimulator (ICOS) and programmed death 1 (PD-1)-positive Tfh cells and the level of serum interleukin (IL)-21. The potential correlation between the frequency of different subsets of B and Tfh cells and the values of clinical measures in RA patients was analysed. In comparison with HC, significantly higher percentages of circulating IgD⁺CD27⁻CD19⁺ naive B, CD86⁺CD19⁺ and CD95⁺CD19⁺ activated B, CD3⁺CD4⁺CXCR5⁺, CD3⁺CD4⁺CXCR5⁺ICOS⁺, CD3⁺CD4⁺CXCR5⁺PD-1⁺ and CD3⁺CD4⁺CXCR5⁺ICOS⁺PD-1⁺ Tfh cells but lower IgD⁺CD27⁺CD19⁺ preswitch memory B cells were detected, accompanied by significantly higher levels of serum IL-21 in the RA patients. Furthermore, the percentages of CD95⁺ B cells were correlated positively with the frequency of PD-1⁺ Tfh cells, but negatively with ICOS⁺ Tfh cells. The percentages of CD86⁺ B cells and ICOS⁺ Tfh cells were correlated positively with the values of disease activity score 28 (DAS28). Following the drug therapies for 1 month, the percentages of CD86⁺ B and PD-1⁺ Tfh cells were reduced significantly in the drug-responding patients. Our data suggest that activated B and Tfh cells may contribute to the pathogenesis of RA and the frequency of activated B and Tfh cells may be used as biomarkers for evaluating the therapeutic responses of individual patients with RA.     

Novel insights into Tim-4 function in autoimmune diseases

Abstract

T-cell immunoglobulin domain and mucin domain-4 (Tim-4) was first recognized as a costimulatory molecule regulating T-cell activation. Dysregulation of Tim-4 has been found in some autoimmune conditions, particularly in the immune cells. Recently, Tim-4 was found to be critical for regulating T cells, with the ability of inhibiting naïve CD4⁺ T cells and Th17 cells, increasing Th2 cell development. Tim-4 can also enhance T cell expansion via linker for activation of T cells, extracellular signal-regulated kinase (ERK) and Protein kinase B (PKB, also known as Akt) signaling pathways. Moreover, the Tim-4 signaling pathway may affect multiple molecular processes in autoimmune diseases. A number of previous studies have demonstrated that Tim-4 influences chronic autoimmune diseases, such as rheumatoid arthritis (RA) and systemic lupus erythematosus. In addition, an association between Tim-4 polymorphisms and susceptibility to several autoimmune diseases have been identified, such as RA. Taken together, recent works have indicated that Tim-4 may represent a novel target for the treatment of autoimmune diseases. In this article, we will discuss the Tim-4 function and the therapeutic potential of modulating the Tim-4 in autoimmune diseases.     

The Distinct Subgroup of Patients with Rheumatoid Arthritis Shown by IgG3-Reactive Rheumatoid Factor

The reactivity of rheumatoid factor (RF) with immunoglobulins of the IgG3 subclass was examined in 49 patients with rheumatoid arthritis (RA) using two types of IgG3 myeloma (routine and IgG3m-15 allotype). Among 49 patients, serum from eight cases showed positive reactivity with both types of IgG3 myeloma by radio-immunoassay (RIA). The isotype of IgG3-reactive RF was not specific; it belonged to the IgM class as well as the IgG subclasses IgG1, IgG2 and IgG4. The patients with IgG3-reactive RF belonged to the clinically-severe classification of RA, having a high erythrocyte sedimentation rate (ESR), high titre in the RA hemagglutination (RAHA) test, and above all they had low levels of complement. Generally, it is concluded that patients with IgG3-reactive RF have serious arthritis and that IgG3-reactive RF might play an important role in the inflammatory process. Furthermore, it was also shown that the RF-reactive site was not associated with the protein-A binding site of IgG3, since RF reacting with IgG3m-15 reacted similarly with routine IgG3, regardless of the difference of the protein-A binding activity. This was confirmed by adding protein-A to the reaction of RF and IgG3m-15 which binds with protein-A. ‘This suggests that the actual reactive site of RF is different to the site that binds protein-A.     

A Comparative Study of Interleukin-1β Production and P2x7 Expression After Atp Stimulation by Peripheral Blood Mononuclear Cells Isolated From Rheumatoid Arthritis Patients and Normal Healthy Controls

Interleukin 1 beta (IL-1β) is a proinflammatory cytokine that is considered to play an important role in the progression of rheumatoid arthritis (RA). A stimulus such as ATP is necessary to cause the release of mature IL-1β, via activation of the P2X₇ receptor on monocytes. In this study, the production of IL-1β in whole blood after ATP stimulation and expression of P2X₇ receptors in RA and healthy subjects were examined. Blood samples from RA patients or healthy controls were stimulated with ATP in the presence of lipopolysaccharide (LPS). Supernatants were harvested and IL-1β levels were measured by enzyme-linked immunosorbent assay (ELISA). Expression of P2X₇ receptors was measured using flow cytometry. ATP induced significantly higher levels of IL-1β in LPS-activated RA blood samples compared to controls. A significant up-regulation of P2X₇ receptor expression on mononuclear cells was observed after overnight incubation with ATP without any significant differences between RA patients and normals. These data suggest that RA patient mononuclear cells are more sensitive to ATP stimulation than healthy individuals perhaps due to genetic polymorphism in the P2X₇ gene.     

重症肌无力CD45RA、CD45RO胸腺细胞亚群及相关细胞因子表达分析

目的:探讨胸腺细胞异常分化与重症肌无力发生的关系.方法:采用基因芯片对多种白细胞介素、干扰素及其受体mRNA表达进行分析, 采用流式细胞术测定胸腺细胞CD45RA、 CD45RO的表达率, 采用免疫组化对胸腺组织切片CD45RA、 CD45RO表达及分布进行检测.结果:MG患者IL-1R、 IL-4R、 IFNγR1、 IFNγR2、 IL-6、 IL-8表达水平显著低于对照组; IL-10RB表达水平显著高于对照组; IL-1、 IL-2、 IL-4、 IL-10、 IL-7、 IFN-α、 IFN-β、 IFN-γ表达水平在MG和对照组均很低, 无显著差异; MG患者CD56 胸腺细胞百分率(0.56±0.33)显著低于对照组(1.78±0.69), MG患者CD45RO 、 CD1a 细胞显著高于对照组.免疫组化和RT-PCR也有相同结果.结论:MG患者胸腺细胞发育过程中CD45RO 细胞向CD45RA T细胞转变存在异常.