Myalgic Encephalomyelitis/Chronic Fatigue Syndrome – Evidence for an autoimmune disease

Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS) is a frequent and severe chronic disease drastically impairing life quality. The underlying pathomechanism is incompletely understood yet but there is convincing evidence that in at least a subset of patients ME/CFS has an autoimmune etiology. In this review, we will discuss current autoimmune aspects for ME/CFS. Immune dysregulation in ME/CFS has been frequently described including changes in cytokine profiles and immunoglobulin levels, T- and B-cell phenotype and a decrease of natural killer cell cytotoxicity. Moreover, autoantibodies against various antigens including neurotransmitter receptors have been recently identified in ME/CFS individuals by several groups. Consistently, clinical trials from Norway have shown that B-cell depletion with rituximab results in clinical benefits in about half of ME/CFS patients. Furthermore, recent studies have provided evidence for severe metabolic disturbances presumably mediated by serum autoantibodies in ME/CFS. Therefore, further efforts are required to delineate the role of autoantibodies in the onset and pathomechanisms of ME/CFS in order to better understand and properly treat this disease.     

人精子特异性乳酸脱氢酶在大肠杆菌中的表达及其初步应用

目的 构建人精子特异性乳酸脱氢酶(hLDH-CA)的原核表达载体,在大肠杆菌中进行表达,将重组hLDH-CA应用于抗精子抗体(ASA)的检测.方法 以人睾丸TripIEx cDNA文库为模板,PCR扩增hLDH-CA编码序列.PCR产物经Hind Ⅲ-Xho I酶切后,克隆至表达载体pET-28a(+)中,在E.coli BL21(DE3)中诱导His-Tag融合的重组蛋白表达.用免疫印迹、酶活性测定等方法鉴定表达产物.以纯化的重组hLDH-CA为基质,建立检测ASA的ELISA方法.结果 构建了hLDH-C4原核表达载体pET-28a(+).hLDHC.在IPTG的诱导下,重组菌可高效表达相对分子质量35 000的产物,与预期大小相符.免疫印迹显示,重组蛋白可被抗His-Tag单克隆抗体和兔抗人LDH-C4抗体识别.重组菌在IPTG诱导后,其裂菌液的乳酸脱氢酶活性是诱导前的11.2倍.用基于hLDH-C4抗原的间接ELISA法,在一组不育症患者中检测血清抗hLDH-CA抗体,阳性率达30.51%.结论 成功地克隆了hLDH-C4编码序列,并在E.coli BL21(DE3)中获得高效的表达,重组hLDH-C4在ASA检测中得到初步应用.     

Pathogenetic aspects of organ-specific autoimmunity

Summary In the last few years a great deal of information on the etiopathogenetic aspects of organ-specific autoimmune diseases (OSADS) has been obtained. It has been shown that genetic factors play an etiologic fundamental role. They are responsible for the dysregulation of the immune system and for the target organ susceptibility which favour the onset of the diseases. Putative environmental factors, such as viral infections, can act as initiating or precipitating events only in genetically predisposed individuals. Immunological mechanisms capable of triggering autoimmune responses have been demonstrated. Data obtained from experimental models and from humans suggest that the ongoing expansion of autoreactive T cells with specificity for autoantigens (AAgs) can be considered as the main immunological event capable of inducing and maintaining the target organ damage. These cells can activate different effector systems, i.e., autoantibody (AAb)-producing B cells, cells with cytotoxic activity, etc., by releasing different combinations of lymphokines. In overt diseases AAbs are directly involved in the pathogenesis of lesions due to autoimmune responses against functional molecules and cellular receptors. The pathogenesis of the common inflammatory destructive lesions of the target organs is more complex and not yet clarified. A large proportion of T cells present in the inflammatory infiltrates are apparently not directed to the AAgs. Most cells display cytolytic activity and may contribute to tissue damage by releasing lymphokines which activate other cells and cascade the process. Vicious cycles, i.e., upregulation of class II and I molecules, alterations of the cytokine network, etc., are supposed to be involved in the maintenance of target organ lesions.     

哮喘患者循环型β2受体自身抗体检测及其意义

为探讨β２受体自身抗体在哮喘发病中的作用，在观察高度选择性β２受体激动剂克伦特罗（ｃｌｅｎｂｕｔｅｒｏｌ）对体外培养的乳鼠心肌细胞的正性变时反应基础上，利用哮喘患者血清中β２受体自身抗体对克伦特罗正性变时反应的抑制作用，以及羊抗人免疫球蛋白对抑制作用的影响，来检测哮喘患者血清β２受体自身抗体。结果受检的１６例哮喘患者显示血清中均存在β２受体自身抗体，２０例健康对照者血清中均未检测出β２受体自身抗体，这种抑制性的自身抗体可能系ＩｇＧ型。结果表明，β２受体自身抗体可能是哮喘发病机制的一个重要环节。     

The effect of HLA class II, insulin and CTLA4 gene regions on the development of humoral beta cell autoimmunity

Aims/hypothesis The aim of this study was to explore the contribution of genetic factors to the emergence of beta-cell-specific humoral autoimmunity.Subjects and methods We analysed the effect of HLA class II, insulin (INS; –23 HphI variant) and cytotoxic T-lymphocyte-associated protein 4 (CTLA4 [+49 and CT60]) genes on the appearance of beta-cell-specific autoantibodies in a large population-based birth cohort recruited in Finland. Infants carrying increased risk HLA DQB1 genotypes were monitored for the appearance of autoantibodies (islet cell autoantibodies [ICA], insulin autoantibodies [IAA], glutamic acid decarboxylase autoantibodies [GADA] and islet antigen 2 antibodies [IA–2A]). Those who developed beta-cell-specific autoantibodies were studied (n=574, mean follow-up time: 4.9 years; range 0.5–9.3).Results IAA emerged at a higher rate in children with the –23 HphI AA INS genotype than in those carrying AT or TT variants (hazard ratio 2.1, 95% CI 1.4–2.9, p<0.001). This effect of the INS locus was present in both HLA DQB1 risk groups. The appearance of IAA showed a strong association also with the HLA DRB1*0401 allele (hazard ratio 13.1, 95% CI 1.8–93.4, p<0.001). The development of IA–2A was also somewhat accelerated by the DRB1*0401 variant (p=0.03). Isolated ICA positivity was independent of the HLA and INS genotypes. None of the humoral immune markers showed association with the CTLA4 gene.Conclusions/interpretation The INS and the DRB1 loci appear to contribute to the pathogenesis of type 1 diabetes by initiating/modifying insulin-specific autoimmunity. The emergence of IAA represents a crucial step in the development of beta cell autoimmunity in young children, in whom the appearance of GADA and IA–2A is linked to IAA.     

白细胞介素-33的测定及其与早期类风湿关节炎相关性的研究

目的 通过检测早期类风湿关节炎(RA)患者血清中的白细胞介素(IL)-33水平,分析其与早期RA之间的相关性.方法 收集病程<1年的早期RA患者100例,骨关节炎(OA)患者40例以及健康对照者70名.采用双抗体夹心酶联免疫吸附试验(ELISA)测定血清中的IL-33水平,并分析血清IL-33水平与RA各临床和实验室指标的相关性.计量资料的比较采用Kruskal-Wallis检验和(或)Mann-Whitney U检验,计数资料比较采用X2检验,相关性分析采用Spearman相关分析.结果 RA患者血清IL-33水平为(282±871)pg/ml,显著高于健康对照组[(7±38)pg/ml,P(0.01)和OA患者[(8±35)pg/ml,P<0.01].血清IL-33水平与类风湿因子(RF)、隐性类风湿因子IgG(HRF-IgG)、抗环瓜氨酸肽(CCP)抗体、抗突变型瓜氨酸化波形蛋白(MCV)抗体呈正相关(r分别为:0.312,0.277,0.213,0.302,P<0.01或P<0.05). IL-33阳性组患者的RF阳性率、HRF-IgG阳性率、抗CCP抗体阳性率、抗MCV抗体阳性率(86%、31%、86%、94%)较IL-33阴性组患者(54%、11%、42%、72%)显著升高(P均<0.05). 结论 IL-33在RA患者血清中显著升高,并与多种自身抗体(包括RF、抗CCP抗体、抗MCV抗体和HRF-IgG)显著相关,可能是RA预后不良的因素之一.     

血清抗神经节苷脂抗体水平与系统性红斑狼疮患者临床特征的相关性研究

目的 初探血清抗神经节苷脂(GM1)抗体与系统性红斑狼疮(SLE)临床特征的相关性.方法 采用改良酶联免疫双抗夹心(ELISA)法,对48例SLE患者血清中的抗GM1-IgM/IgG抗体和抗GQ1b-IgM/IgG抗体水平进行检测,同时检测10例类风湿关节炎(RA)、5例干燥综合征(SS)、1例混和性结缔组织病(MCTD)、1例多发性肌炎(PM)及97名健康对照(HC)的血清样品.对其中5例SLE患者随访3个月,比较前后血清抗体水平.48例SLE患者包括7例神经精神性SLE(NPSLE)与41例非神经精神性SLE(non-NPSLE)患者.以健康对照组99%参考值上限作为阳性界值,进行Fisher确切概率法x2检验、单因素方差分析、Dunnett t检验.结果 首先确立了中国人血清抗GMI-IgM/IgG抗体与抗GQ1b-IgM/IgG抗体的阳性界值,取99%参考值上限,分别为0.411、0.408、0.481和0.441.SLE患者血清抗GM1-IgG抗体水平显著高于对照组(0.33±0.09与0.27±0.05,P<0.05),而血清抗GM1-IgM抗体、抗GQ1b-IgM抗体、抗GQ1b-IgG抗体水平在各组中差异无统计学意义(P>0.05).根据我们所建立的阳性界值,SLE患者血清抗GM1抗体阳性率为19%(抗GM1-IgM抗体为4%,抗GM1-IgG抗体为15%).血清抗GM1-IgM/IgG抗体、抗GQ1b-IgM/IgG抗体水平与SLE患者是否伴发神经精神性表现、抗dsDNA-IgG抗体阳性与否及治疗3个月前后SLE疾病活动指数均无明显关联(P>0.05).结论 SLE患者外周循环中存在对神经节苷脂GM1的免疫应答,并可能参与了SLE的发病过程.血清抗GM1抗体和抗GQ1b抗体水平和SLE临床特征无明显关联.     

原发性胆汁性肝硬化36例临床和免疫学特征

目的:分析原发性胆汁性肝硬化(PBC)患者的临床特征及免疫学特征。方法:回顾性分析我院2007年8月~2010年7月收治的36例PBC患者的临床资料。结果:36例PBC患者中黄疸33例(91.7%)、皮肤瘙痒27例(75.0%)、腹水22例(61.1%)。血生化指标:总胆红素(149.2±65.6)μmol/L、碱性磷酸酶(298.6±128.1)U/L,γ-谷氨酰转肽酶(172.6±84.9)U/L;免疫学指标:IgM(4.98±3.1)g/L,抗线粒体抗体AMA-M2阳性30例(83.3%),GP210阳性5例(13.9%),SP100阳性2例(5.6%)。肝肾微粒体LKM-1阳性2例(5.6%)肝细胞溶质抗原Ⅰ型(LC-1)抗体(2.8%),抗核抗体(ANA)5例(13.9%)。36例患者经熊去氧胆酸等治疗,31例病情好转,有效率达86.1%。结论:PBC以中年女性多见,黄疸、皮肤瘙痒为患者最主要表现,AMA-M2、GP210、SP100等自身抗体检测是诊断P B C的重要依据之一,熊去氧胆酸治疗效果确切。     

抗血管紧张素Ⅱ1受体和α1肾上腺素受体抗体与高血压患者心脏重构的观察

目的观察原发性高血压患者血清抗血管紧张素Ⅱ1受体（抗AT1受体）和α1肾上腺素受体自身抗体（抗α1受体抗体）对高血压患者心脏重构的影响。方法采用酶联免疫吸附法（ELISA）对553例原发性高血压患者进行血清抗α1受体和抗AT1受体抗体检测并记录所有患者的超声心动图检查结果。比较不同滴度组血清抗体对心脏重构的影响。结果553例高血压病患者血清抗α1-受体和抗AT1受体抗体阳性率分别为32．3％（179／553）,42．3％（234／553）。抗劬受体抗体1：40（滴度）阳性组左心房扩大比率明显高于阴性组（P=0．021）;≥1：80阳性组左心室扩大比率明显高于阴性组（P=0．006）。抗AT1受体抗体1：40阳性组右心房扩大比率明显高于阴性组（P=0．011）;≥1：80阳性组左心室扩大比率明显高于阴性组（P=0．004）。双抗阳性组左心室和右心房扩大比率明显高于双抗阴性组（P分别为0．006和0．044）。结论抗α1-受体和AT1受体抗体与高血压痛患者心脏扩大有关,可能是导致高血压病患者心脏重构的重要因素之一,且两种抗体可能有相互促进作用。     

Peroxynitrite-induced modification of H2A histone presents epitopes which are strongly bound by human anti-DNA autoantibodies: role of peroxynitrite-modified-H2A in SLE induction and progression

Peroxynitrite is a potent oxidant and nitrating agent and has in vivo existence. It is a powerful proinflammatory substance and may increase vascular permeability in inflamed tissues. Systemic lupus erythematosus (SLE) is an autoimmune inflammatory disease of unknown etiology. Since its discovery, numerous self- and non-self, nuclear, and cytoplasmic antigens have been suggested as stimuli for SLE initiation, but the exact trigger is yet to be identified. In this study, an attempt has been made to investigate the binding characteristics of SLE anti-DNA autoantibodies to native DNA and native and peroxynitrite-modified H2A histone to explore the possible role of modified protein antigen(s) in SLE initiation and progression. The nuclear protein (H2A histone) was modified by peroxynitrite synthesized in our laboratory. The peroxynitrite-modified H2A revealed generation of nitrotyrosine, dityrosine, and carbonyls when subjected to investigation by physicochemical methods. Binding characteristics and specificity of SLE anti-DNA antibodies were analyzed by direct binding and inhibition enzyme-linked immunosorbent assay. The data show preferential binding of SLE autoantibodies to peroxynitrite-modified H2A histone in comparison with native H2A histone or native DNA. A band shift assay further substantiated the enhanced recognition of peroxynitirite-modified H2A histone by anti-DNA autoantibodies. The results suggest that peroxynitrite modification of self-antigen(s) can generate neoepitopes capable of inducing SLE characteristic autoantibodies. The preferential binding of peroxynitrite-modified H2A histone by SLE anti-DNA antibodies points out the likely role of oxidatively modified and nitrated H2A histone in the initiation/progression of SLE. Moreover, oxidatively modified and nitrated nuclear protein antigen, rather than nucleic acid antigens, appear to be more suitable as a trigger for SLE.