抗dsDNA抗体两种检测方法的比较

比较抗双链ＤＮＡ（ｄｓＤＮＡ）抗体的两种检测方法。用胶体金快速斑点渗滤技术（ＤＩＧＦＡ）及马疫锥虫间接荧光抗体染以法检测２７名正常儿童和４６例系统性红斑狼疮（ＳＬＥ）病人和非狼疮病人血清抗ｄｓＤＮＡ抗体，２７名正常儿童和１７例非狼疮病人血清抗ｄｓＤＮＡ抗体经两种方法检测均为阴性；２９例ＳＬＥ患者用两种方法检测出抗ｄｓＤＮＡ抗体的阳性符合率为９４．４４％（１７／１８）。说明两种方法具有一致的特异性和     

HLA—II类基因与红斑狼疮自身抗体相关性研究

目的 分析ＨＬＡ－Ⅱ类等位基因与自身抗体的相关性,探讨系统性红斑狼疮的自身抗体反应类型。方法 应用聚合酶链反应／序列特异寡核苷酸探针杂交（ＰＣＲ－ＳＳＯＰＨ）检测技术对１１３例确诊ＳＬＥ病人进行ＨＬＡ－ＤＲ、ＤＱＡ１和ＤＱＢ１基因分型。结果 抗ｄｓＤＮＡ与ＤＱＢ１＊１６０１和ＤＲＢ３＊０３０１负相关;ＡＮＡ与ＤＲＢ３＊０２０２和ＤＱＡ１＊０５０１正相关;抗ＲＮＰ与ＤＱＡ１＊０３０１正相关,与ＤＱＡ     

系统性红斑狼疮活动期血浆降钙素基因相关肽及内皮素-1含量变化

目的：为了探讨肽－降钙素基因相关肽和内上素－１在ＳＬＥ活动期中的作用。方法；采用放射免疫分析法测定了３４例系统性红斑狼疮病人和２０例健康人血浆ＥＴ－１，ＣＧＲＰ水平。结果；有抗ｄｓ－ＤＮＡ抗体升高和补体Ｃ３，Ｃ４下降的ＳＬＥ组，ＥＴ－１水平升高，ＣＧＲＰ水平降低。『     

脱氧核糖核酸抗体的研究进展

在系统性红斑狼疮（ＳＬＥ）患者，由于机体不能适当调节脱氧核糖核酸（ＤＮＡ）抗体的产生，导致特异的致病性双链ＤＮＡ抗体（ｄｓ－ＤＮＡ）过量生成，促进ＳＬＥ的发生发展。本文主要讨论不同类型的ＤＮＡ抗体的致病作用，探讨新的特异性抑制ＤＮＡ抗体产生及其活性的...     

中国汉族系统性红斑狼疮某些易感基因的研究

为了探索我国北方汉族人群人类白细胞抗原－ＤＲ（ＨＬＡ－ＤＲ）及肿瘤坏死因子Ｂ（ＴＮＦＢ）等位基因多态性与系统性红斑狼疮（ＳＬＥ）的易感性关系，对１０６例健康人和４５例ＳＬＥ患者的ＨＬＡ－ＤＲ和对８０例健康人及４５例ＳＬＥ患者的ＴＮＦＢ等位基因，采用多聚酶链反应－限制性酶切片段长度多态性（ＰＣＲ－ＲＦＬＰ）法进行分析。结果显示：ＳＬＥ患者中频率显著升高的等位基因有ＤＲ２［Ｐ＜０．０５，相对危险性（ＲＲ）＝１．５６］及ＤＲ３（Ｐ＜０．０１，ＲＲ＝２．６９）。而ＤＲ５和对照相比呈相反结果（Ｐ＜０．０５，ＲＲ＝０．４３）。ＳＬＥ患者ＴＮＦＢ＊２等位基因频率显著增高（Ｐ＜０．０５，ＲＲ＝１．８４）。提示ＤＲ２、ＤＲ３、ＴＮＦＢ＊２可能是易感等位基因或易感等位基因标记，而ＤＲ５是一拮抗等位基因或拮抗等位基因标记。并研究了ＨＬＡ－ＤＲ、ＴＮＦＢ等位基因与患者血浆中Ｓ蛋白和补体５ｂ－９结合物的复合物（ＳＣ５ｂ－９）的水平和多种自身抗体及狼疮肾炎、狼疮肺炎、狼疮脑病等狼疮并发症之间的相关性，发现ＨＬＡ－ＤＲ２基因与狼疮肾炎的发生存在正相关（Ｐ＜０．０５，ＲＲ＝１．３２）。     

抗dsDNA抗体抗原识别及致病性的分子基础

抗双链ＤＮＡ抗体 (ｄｓＤＮＡ)是系统性红斑狼疮 (ＳＬＥ)的特异性和致病性抗体 ,几乎存在于所有病人血清中 ,并与临床表现密切相关。目前 ,越来越多的证据表明它是特异性抗原驱动的结果。本文就近年来对于抗ｄｓＤＮＡ抗体本身的结构特点、识别的抗原结构以及其致病机制的研究进展作一综述。1　抗ｄｓＤＮＡ抗体的分子结构抗ｄｓＤＮＡ抗体具有许多对外来抗原再次应答的特征 ,如ＩｇＭ向ＩｇＧ的类型转换以及体细胞突变。在抗ｄｓＤＮＡ抗体中 ,Ｖ区基因突变积累的结果常使其产生ｄｓＤＮＡ特异性和 /或增强结合ｓｓＤＮＡ、ｄｓＤＮＡ的…     

活性DNA诱导抗核抗体及系统性红斑狼疮样综合征

目的 寻找系统性红斑狼疮真正的核成分免疫原。方法 从ＣｏｎＡ活化的脾淋巴细胞中提取ＤＮＡ，然后同系免疫ＢＡＬＢ／ｃ小鼠，用ＥＬＩＳＡ方法测定ＩｇＧ类抗ｄｓＤＮＡ，抗组蛋白抗体，用免疫荧光法检测抗核抗体核型和免疫复合物沉积，用免疫印迹法测定抗可溶性抗原抗体，用考马斯亮蓝法检测抗体阳性的动物尿蛋白含量。结果 活性ＤＮＡ能诱导抗ｄｓＤＮＡ、抗组蛋白等多种抗核抗体生成，且能诱发同系小鼠产生ＳＬＥ样综合征。     

HLA-DQB1 基因与中国湖北汉族人 SLE 及其自身抗体的相关性研究

目的：探讨ＨＬＡＤＱＢ１等位基因与系统性红斑狼疮（ＳＬＥ）及其自身抗体的相关性。方法：采用ＰＣＲ／ＳＳＰ技术对５２例中国湖北地区汉族ＳＬＥ患者及１４３例正常对照者进行了ＨＬＡＤＱＢ１基因分型，并采用免疫印迹技术检测患者血清中自身抗体。结果：ＳＬＥ患者ＤＱＢ１０６０８（９６２％，χ２＝１０５１，Ｐ＜０００５）基因频率显著升高，ＤＱＢ１０３０２（５７７％，ＲＲ＝０２６Ｐ＜００５，ＰＦ＝０１４）和ＤＱＢ１０５０１（１９２％，ＲＲ＝０１１，Ｐ＜００１，ＰＦ＝０１３）基因频率显著降低，与正常对照组比较，ＤＱＢ１０６０８在伴抗Ｓｍ（１０３４％，Ｐ＜０００５）、抗ＲＮＰ（１１５４％，Ｐ＜０００５）、抗ｄｓＤＮＡ（２２２２％，Ｐ＜０００５）抗体阳性的ＳＬＥ患者中频率显著升高。结论：ＤＱＢ１０６０８与ＳＬＥ关联，并分别与抗Ｓｍ、抗ＲＮＰ、抗ｄｓＤＮＡ抗体的产生有相关性。而ＤＱＢ１０３０２、ＤＱＢ１０５０１等位基因对ＳＬＥ可能具有保护性。     

配偶同患系统性红斑狼疮二例

例1:先证者,女,34岁。于1992年3月因冻疮样皮疹、光敏、关节痛而就诊。有少尿、口干。体检见口腔溃疡,双手雷诺现象,肝脏肿大,下肢浮肿。实验室检查:尿蛋白(),尿ＦＤＰ(-),ＣＩＣ0105,抗ｄｓＤＮＡ抗体63%,抗Ｓｍ抗体(+),抗ＲＮＰ抗体(+),抗核抗体(+),Ｃ3ａ1485。甲皱微循环中度异常,总积分62分。诊断为系统性红斑狼疮(ＳＬＥ),狼疮肾炎(ＬＮ)。经环磷酰胺及强的松加中药狼疮平治疗,关节痛及皮疹消失,肝脏缩小,尿蛋白降至(±),抗ｄｓＤＮＡ抗体降至14%,恢复轻工作。例2:配偶,男,37岁。于1995年5月因发热、脱发、关节肌肉酸痛而就诊。体检见…     

HLA-Ⅱ类基因与红斑狼疮自身抗体相关性研究

目的分析ＨＬＡⅡ类等位基因与自身抗体的相关性,探讨系统性红斑狼疮的自身抗体反应类型。方法应用聚合酶链反应／序列特异寡核苷酸探针杂交（ＰＣＲＳＳＯＰＨ）检测技术对１１３例确诊ＳＬＥ病人进行ＨＬＡＤＲ、ＤＱＡ１和ＤＱＢ１基因分型。结果抗ｄｓＤＮＡ与ＤＱＢ１１６０１和ＤＲＢ３０３０１负相关;ＡＮＡ与ＤＲＢ３０２０２和ＤＱＡ１０５０１正相关;抗ＲＮＰ与ＤＱＡ１０３０１正相关,与ＤＱＡ１０１０３负相关;抗Ｌａ与ＤＱＡ１０５０１正相关;ＲＦ因子与ＤＱＢ１０５０１负相关。结论不同的ＨＬＡ等位基因介导不同的自身抗体反应类型,同一基因位点的不同等位基因对抗体反应的影响作用有明显差异,甚至作用相反。     

DNA antibodies and complement in SLE patients

Sixty-seven patients with systemic lupus erythematosus (SLE) were followed up for 3-19 months (mean 12) in a prospective study. The activity of SLE was estimated on clinical grounds and correlated with DNA antibody and complement levels. The disease reactivations consisted mostly of articular and cutaneous symptoms. There were 17 relapses and 22 complicating infections during the follow-up period. The levels of antibodies to native, double-stranded (ds) DNA (P less than 0.001) and antibodies to denatured, single-stranded (ss) DNA of IgG class (P less than 0.001) and C3 (P less than 0.001) correlated best with disease activity, which was estimated on the clinical symptoms and signs. These assays were not reliable, however, in predicting minor exacerbations. The levels of IgM class ss-DNA antibodies were significantly higher in SLE patients without nephritis than in SLE nephritis patients. In most cases, the combination of IgG class ss-DNA antibody and complement (C3 and CH50) determinations differentiated SLE relapse from infection.     

The role of the immunoglobulin heavy chain in human anti-dna antibody binding specificity

Objective. To investigate the structural basis for DNA binding of the natural human IgMλ monoclonal antibody KIM4.6. Methods. An IgMλ, non–DNA-reactive variant hybridoma was derived during in vitro subcloning of the anti-DNA antibody KIM4.6. The variable (V)-region heavy (H) and light (L) chain genes expressed by the variant hybridoma were amplified by polymerase chain reaction, cloned, sequenced, and compared with those of the KIM4.6 parent and other DNA-binding and non–DNA-binding antibodies. Results. The VL chain of the variant was identical to that of KIM4.6. In contrast, the VH chain was completely different from the VH chain of the parent but was similar or identical, except in the diversity (D) and joining regions, to the VH chain of the systemic lupus erythematosus (SLE) IgG anti-DNA antibody T14 and SLE IgM nephritogenic anti-DNA antibodies NE-1 and NE-13. Conclusion. The expression of the KIM4.6 VL chain is not sufficient for DNA specificity. The VH chain and its D region play a key role in conferring DNA binding of the KIM4.6 anti-DNA antibody.     

Antigenic triggers and molecular targets for anti-double-stranded DNA antibodies

While anti-double-stranded (ds)DNA antibodies are a characteristic serologic hallmark for SLE, the triggering antigen is unknown. Using phage display libraries, we identified DWEYSVWLSN as a peptide mimic of DNA for a pathogenic anti-dsDNA antibody. Peptide immunization of non-autoimmune mice induced anti-dsDNA as well as other lupus-associated antibodies. Molecular analysis of the induced anti-dsDNA antibodies revealed several similarities with anti-dsDNA antibodies that appear spontaneously in lupus mice. Furthermore, lupus-prone mice immunized with this peptide DNA mimic had higher autoantibody titers as well as more severe nephritis. Anti-DNA antibodies may contribute to lupus nephritis via cross-reactivity with renal antigen. Using western blotting of lysates of mesangial cells from a lupus mouse, we found that a pathogenic anti-DNA antibody binds to alpha-actinin. High titers of anti-alpha-actinin antibodies were present in the sera and kidney eluates of lupus mice with active disease. Binding to alpha-actinin was diminished in mesangial cells derived from BALB/c mice, suggesting that target antigen expression may play a role in determining autoantibody binding to the kidney. We conclude that a pathogenic, lupus-like autoantibody response can be induced by a peptide antigen, and that alpha-actinin is a cross-reactive renal target for the pathogenic anti-dsDNA autoantibody response in lupus mice.     

血清C1q抗体水平与系统性红斑狼疮活动性和狼疮肾炎的关系

目的分析探讨血清C1q抗体水平与系统性红斑狼疮（SLE）活动性以及狼疮肾炎之间的关系。方法采用ELISA方法检测92例SLE患者C1q抗体水平。并与其他SLE活动性指标进行相关分析。结果SLE患者C1q抗体阳性率为67.4%。活动性狼疮组的C1q抗体阳性率及C1q抗体水平显著高于非活动性狼疮组（P〈0.001）。活动性狼疮肾炎组C1q抗体阳性率（P〈0.05）和C1q抗体水平（P〈0.01）显著高于非活动性狼疮肾炎组。联合抗dsDNA抗体检测,没有1例活动性狼疮肾炎患者的C1q抗体和抗dsDNA抗体同时阴性。结论血清C1q抗体与狼疮活动以及活动性狼疮肾炎关系密切,C1q抗体的检测有助于活动性狼疮的诊断,联合抗dsDNA抗体的检测是活动性狼疮肾炎的特异性检测指标。     

Lymphocytotoxic antibody activity in cryoprecipitates from serum of patients with SLE

Complement-dependent cytotoxic activity against normal human peripheral blood lymphocytes was detected in 11 of 17 cryoprecipitates from the serum of patients with systemic lupus erythematosus (SLE). The lymphocytotoxicity was eliminated by treatment with 2-mercaptoethanol and iodoacetamide, and it was inhibited by antibody to human IgM but not anti-IgG. The titers of lymphocytotoxic activity in the cryoprecipitates were roughly proportional to the corresponding serum titers, but when they were normalized for IgM concentration it was apparent that selective concentration of lymphocytotoxic antibody occurred in the cryoglobulins. The relationship between cryoprecipitable lymphocytotoxicity and a number of laboratory and clinical parameters of SLE was studied. The amount of protein in the cryoprecipitates, which was greatest in patients with significant renal disease, correlated with a reduction of serum complement and the amount of antibody to DNA. However the lymphocytotoxic activity of the cryoglobulins did not correlate with the severity of SLE. The titer of lymphocytotoxic antibody was independent of a) the presence or absence of active lupus nephritis, b) the total protein or immunoglobulin content of the cryoprecipitates, c) serum complement levels, and d) the amount of circulating antibody to DNA. These findings cast doubt upon the pathogenetic significance of cryoprecipitable lymphocytotoxic antibody.     

Lymphocytotoxic antibody activity in cryoprecipitates from serum of patients with SLE.

Complement-dependent cytotoxic activity against normal human peripheral blood lymphocytes was detected in 11 of 17 cryoprecipitates from the serum of patients with systemic lupus erythematosus (SLE). The lymphocytotoxicity was eliminated by treatment with 2-mercaptoethanol and iodoacetamide, and it was inhibited by antibody to human IgM but not anti-IgG. The titers of lymphocytotoxic activity in the cryoprecipitates were roughly proportional to the corresponding serum titers, but when they were normalized for IgM concentration it was apparent that selective concentration of lymphocytotoxic antibody occurred in the cryoglobulins. The relationship between cryoprecipitable lymphocytotoxicity and a number of laboratory and clinical parameters of SLE was studied. The amount of protein in the cryoprecipitates, which was greatest in patients with significant renal disease, correlated with a reduction of serum complement and the amount of antibody to DNA. However the lymphocytotoxic activity of the cryoglobulins did not correlate with the severity of SLE. The titer of lymphocytotoxic antibody was independent of a) the presence or absence of active lupus nephritis, b) the total protein or immunoglobulin content of the cryoprecipitates, c) serum complement levels, and d) the amount of circulating antibody to DNA. These findings cast doubt upon the pathogenetic significance of cryoprecipitable lymphocytotoxic antibody.     

Idiotypic vaccination with a murine anti-dsDNA antibody: phase I study in patients with nonactive systemic lupus erythematosus with nephritis

OBJECTIVE: To determine the safety and immunogenicity of an idiotypic anti-dsDNA vaccine in patients with nonactive systemic lupus erythematosus (SLE) and stable lupus nephritis. METHODS: Patients with SLE with a history of nephritis were randomized for vaccination with the murine anti-dsDNA monoclonal antibody (Mab) 3E10 in a dose ranging, double blind, placebo controlled study (phase I). RESULTS: Of the 9 patients injected with Mab 3E10, 5 showed a human anti-mouse antibody (HAMA) response, in large part antiidiotypic, which developed within the first 3 months in 3 strong HAMA responders, and more than one year after immunization in an initially weak HAMA responder. All but one nonresponder were receiving low dose prednisone. No adverse events, in particular no evidence of lupus flares, and no untoward laboratory findings were reported over a followup of 2 years. CONCLUSION: In patients with stable lupus nephritis, immunization with Mab 3E10 appears safe and can generate a significant antiidiotypic response. Idiotypic vaccination may be an approach to specific immunotherapy of autoimmune lupus nephritis.     

肿瘤坏死因子α基因多态性及其与系统性红斑狼疮的相关性

目的 探讨人类肿瘤坏死因子α基因多态性与中国汉族人系统红斑狼疮易感性及临床表现的关系。方法 采用聚合酶链反应单链构象多态性方法,检测８９例ＳＬＥ病人及７０例正常人ＴＮＦα基因的ＴＮＦ１及ＴＮＦ２基因亚型,同时用序列特异性引物方法,检测人类白细胞抗原ＨＬＡ－ＤＲ２基因频率,并分析它们之间的相互关系。     

系统性红斑狼疮与CD19基因多态性的相关性研究

目的研究CD19基因第4外显子705位点(以下简称CD19基因705位点)多态性在中国南方地区汉族人群中的分布及其与系统性红斑狼疮(SLE)和狼疮肾炎(LN)的相关性。方法103例患者诊断均符合1982年美国风湿病学会修订的SLE分类标准,男13例,女90例,其中62例伴有LN。正常对照组110例,男21例,女89例。全部研究对象均为无血缘关系的中国南方地区汉族人群。应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)方法,对所有SLE患者和正常对照者进行CD19基因705位点多态性检测。结果CD19基因705位点多态性在中国南方地区汉族人群中普遍存在。SLE患者CD19基因705位点基因型和等位基因频率分布与正常对照组比较差异无统计学意义(P>0.05);CD19基因705位点基因型和等位基因频率分布,按性别分层后,男性和女性SLE患者分别与正常对照组比较及LN患者和正常对照组比较以及伴有LN患者与未伴有LN的SLE患者间比较,差异均无统计学意义(P>0.05)。结论CD19基因705位点多态性与SLE及LN无相关性。     

Association of increased interferon-inducible gene expression with disease activity and lupus nephritis in patients with systemic lupus erythematosus

OBJECTIVE: To study 5 type I interferon (IFN)-inducible genes (LY6E, OAS1, OASL, MX1, and ISG15) in patients with systemic lupus erythematosus (SLE) and to correlate expression levels with disease activity and/or clinical manifestations. METHODS: Peripheral blood cells were obtained from 48 SLE patients, 48 normal controls, and 22 rheumatic disease controls, and total RNA was extracted and reverse transcribed into complementary DNA. Gene expression levels were measured by real-time polymerase chain reaction, standardized to a housekeeping gene, and summed to an IFN score. Disease activity was determined by the Safety of Estrogens in Lupus Erythematosus: National Assessment-Systemic Lupus Erythematosus Disease Activity Index (SELENA-SLEDAI) composite. RESULTS: Each gene was highly expressed in SLE patients compared with normal controls (P < or = 0.0003) or disease controls (P < or = 0.0008 except for MX1). IFN scores were positively associated with the SELENA-SLEDAI instrument score (P = 0.001), the SELENA-SLEDAI flare score (P = 0.03), and the physician's global assessment score (P = 0.005). Compared with patients without nephritis, lupus nephritis patients had higher IFN scores (overall P < 0.0001), especially during active renal disease. IFN scores were weakly associated with neurologic manifestations. Elevated IFN scores were positively associated with the current presence of anti-double-stranded DNA (anti-dsDNA) antibodies (P = 0.007) or hypocomplementemia (P = 0.007). LY6E expression levels distinguished active from inactive lupus nephritis (P = 0.02) and were positively associated with proteinuria (P = 0.009). CONCLUSION: The 5 IFN-inducible genes were highly expressed in SLE patients, and increased levels were correlated with disease activity defined by several methods. IFN scores, or LY6E levels, were elevated in lupus nephritis patients, especially during active renal disease, and in patients with anti-dsDNA antibody positivity and hypocomplementemia. IFN scores, or LY6E levels, may be useful as a biomarker for lupus nephritis therapy.