High-avidity anti-DNA antibody removal from the serum of systemic lupus erythematosus patients by adsorption using dextran sulfate cellulose columns

The purpose of this study is to determine whether immunoadsorption treatment using a dextran sulfate (DS) column can remove high-avidity anti-double-stranded DNA (anti-dsDNA) antibody from the blood of patients with systemic lupus erythematosus (SLE). Before and after each immunoadsorption therapy routine, titers of the high-avidity anti-dsDNA antibody of 11 SLE patients were measured by using a newly developed assay kit to exclusively detect high avidity anti-DNA antibody. Patients with active SLE showed significantly higher titers of high-avidity antibody than did those with inactive SLE, and their titers were significantly reduced by immunoadsorption procedures. Removal of high-avidity antibodies in vitro was also confirmed by mixing patients' sera and DS gel. Immunoadsorption therapy using DS columns is effective in the removal of high-avidity anti-dsDNA antibodies that are closely associated with pathogenicity in SLE. © 1996 Wiley-Liss, Inc.     

Anti-DNA antibody kinetics following selective removal by adsorption using dextran sulphate cellulose columns in patients with systemic lupus erythematosus

The aim of this study is to determine by mathematical analysis which of two models, the one- or the two-compartment model, more closely approximates the kinetics of anti-dsDNA following immunoadsorption procedures in patients with systemic lupus erythematosus. Titers of anti-dsDNA were measured at specified intervals after apheresis to each model by nonlinear least-squares methods, and Akaike's Information Criterion (AIC) was calculated to determine which model most approximately described the kinetics. The A1C of the two-compartment model was larger than that of the one-compartment model in all 14 SLE patients (P < .001). Therefore, the one-compartment model is thought to be suitable. The generation rate and catabolic rate of anti-dsDNA were obtainable using this model. The anti-dsDNA replenishment curve after an immunoadsorption session was defined by only two parameters: the generation and catabolic rates of anti-dsDNA. © 1996 Wiley-Liss, Inc.     

Therapeutic selective adsorption of anti-DNA antibody using dextran sulfate cellulose column (Selesorb) for the treatment of systemic lupus erythematosus.

The Selesorb therapeutic dextran sulfate cellulose column (Kaneka Corporation, Osaka, Japan) selectively adsorbs anti-DNA antibody, anti-cardiolipin antibody, and immune complex from the plasma of patients with systemic lupus erythematosus (SLE). The Selesorb system is composed of twin columns attached to an automated regeneration apheresis unit. Anti-DNA antibody in plasma is continuously removed through 1 of the 2 columns alternately. Clinical application of the Selesorb system to SLE patients with high titers of anti-DNA antibody showed improvement of proteinurea, arthralgia, rash, lymphocytopenia, etc., with the concurrent use of steroid and/or immnosuppressant. Angiotensin converting enzyme inhibitor should not be administered to patients treated with the Selesorb system to avoid anaphylactoid reactions based on rapid increases of bradykinin concentrations in the blood.     

Adsorption of anti-annexin V using dextran sulfate bound cellulose beads

Anti-annexin V (Anx V) antibodies are detected in SLE patients and patients with habitual fetal loss or preeclampsia. Several case reports have indicated that recurrent abortion based on antiphospholipid syndrome (APS) could be successfully treated with immunoadsorption by using dextran sulfate (DS) columns. The purpose of this study is to clarify whether or not anti-Anx V is also adsorbed by DS-bound cellulose beads. Sera from anti-Anx V-positive patients were mixed with DS-bound cellulose beads in vitro, and the titers of anti-Anx V were measured both before and after incubation. The anti-Anx V titers significantly decreased after incubation. The Anx V also bound to bovine serum albumin-conjugated DS immobilized on microtiter plates. The results of the present study lend support to the basic rationale for immunoadsorption therapy using DS columns in the treatment of habitual abortion closely associated with anti-Anx V antibodies.     

The role of immunoadsorption using dextran-sulfate cellulose columns in the treatment of systemic lupus erythematosus.

Immunoadsorption using dextran sulfate (DS)-cellulose columns is reviewed. An extracorporeal selective adsorption system using such columns has been developed and clinically used to remove anti-DNA from the circulating blood of systemic lupus erythematosus (SLE) patients. These columns can adsorb pathogenic anti-DNA subgroups of high avidity and/or cationic antibodies, anticardiolipin, anti-CLbeta2GPI, and anaphylatoxins. An open clinical study on 19 SLE cases (the mean number of apheresis sessions totaled 3.7 times; the mean dose of prednisolone, 38 mg/day) revealed that the mean SLE disease activity index (SLEDAI) score significantly decreased from the pretreatment level of 10.2 to 4.5 after treatment. Several case reports have indicated that this modality might also be useful for treating patients with antiphospholipid syndrome. Compartment model analyses showed the one-compartment model to be the most suitable for the kinetics of anti-DNA during and following the apheresis procedure. The indications for immunoadsorption in the treatment of SLE remain controversial. A steroid-sparing effect might be one of them, but further controlled studies are necessary to verify this hypothesis.     

系统性红斑狼疮患者抗双链DNA抗体检测及其与免疫学指标的相关性

目的探讨系统性红斑狼疮患者抗双链DNA（dsDNA）抗体与免疫学指标的相关性。方法收集77例系统性红斑狼疮患者血清,采用短膜虫间接免疫荧光法（CLIFT）及酶联免疫吸附测定（ELISA）检测血清抗dsDNA抗体、总补体、补体C3、补体C4水平。结果 77例系统性红斑狼疮患者50例抗dsDNA抗体阳性,27例抗dsDNA抗体阴性。抗dsDNA抗体阳性组和阴性组患者血清IgG、IgM、IgA水平的差异没有统计学意义（P〉0.05）,抗dsDNA抗体阳性组患者血清总补体、补体C3、补体C4水平明显低于阴性患者（P〈0.05）。抗dsDNA抗体阳性患者血清抗dsDNA抗体水平与总补体、补体C3显著相关（P〈0.05）。结论抗dsDNA抗体可能通过补体旁路途径而非经典途径抑制补体生成。     

抗ds-DNA定量检测在系统性红斑狼疮诊断和治疗中的应用观察

目的探讨定量检测抗双链脱氧核糖核酸(ds-DNA)在系统性红斑狼疮(SLE)诊断和治疗中的应用价值。方法 77例SLE患者(活动期37例、缓解期40例)、74例其他结缔组织病患者和68例体检健康者血清用间接酶联免疫吸附试验定量检测抗ds-DNA,以观察比较各组差异;另对其中36例接受免疫治疗的活动期SLE住院治疗患者于治疗前和治疗后第3、7、14、20天各抽血检测抗ds-DNA、补体C3、C4、红细胞沉降率(ESR)等,以比较各指标的变化敏感性。结果活动期和缓解期SLE患者抗ds-DNA浓度与健康对照组比较差异有统计学意义(P<0.01),其他结缔组织病患者与健康对照组比较差异无统计学意义(P>0.05);36例活动期SLE患者接受免疫治疗后的第3天抗ds-DNA含量较治疗前明显下降,差异有统计学意义(P<0.05),C3和ESR治疗后第14天较治疗前有明显差别,差异有统计学意义(P<0.05),C4在治疗前后差异无统计学意义(P>0.05)。结论抗ds-DNA对SLE的诊断具有较高的敏感性和特异性,有效治疗时其下降比C3、C4和ESR等非特异性炎性指标恢复快而明显,定量检测抗ds-DNA更能显示其临床应用价值。     

蛋白A免疫吸附治疗系统性红斑狼疮1例

报告1例应用蛋白A免疫吸附治疗系统性红斑狼疮病人的个案,总结治疗的护理要点,包括治疗前病人评估与用物准备,治疗中预冲、吸附、回浆、洗脱、平衡、二次预冲循环过程的操作、病情观察及吸附柱的储存,治疗结束后的治疗效果评价。通过该病例的护理总结,为临床应用蛋白A免疫吸附治疗系统性红斑狼疮的护理提供参考。     

血清抗核小体抗体表达与系统性红斑狼疮疾病活动相关性评价

目的研究抗核小体抗体与系统性红斑狼疮(SLE)疾病活动的相关性、灵敏度、特异度等。方法收集SLE病例58例,按SLE疾病活动指数分为非活动组、轻度、中度、重度活动组,同时设对照组25例。采用间接免疫荧光法、免疫印迹法、速率散射比浊法、全自动血沉分析仪等分别检测抗核心抗体(ANA)、抗核小体抗体、抗-ds-DNA、抗-Sm、C3、红细胞沉降率(ESR)。结果 (1)抗核小体抗体阳性检出率与SLE疾病活动程度呈正相关(r=0.638,χ2=40.235,P=0.00)。(2)抗核小体抗体对SLE疾病诊断的灵敏度(51.72%)高于抗-ds-DNA(20.69%)和抗-Sm(41.38%),其特异度(92%)也与抗-ds-DNA(96%)和抗-Sm(92%)接近。(3)SLE患者血清抗核小体抗体表达水平与C3呈负相关(r=-0.595,P=0.00),与ESR呈正相关(r=-0.783,P=0.00),而与ANA核型、滴度水平无明显相关性(P>0.05)。结论抗核小体抗体检测作为一项新的与SLE疾病活动密切相关、灵敏度高、特异性强且操作简便的实验室评价指标,将在SLE疾病诊断、合理治疗及疗效评估方面发挥重要价值。     

Development of an in vitro miniature model to simulate immunoadsorption in patients with systemic lupus erythematosus

In SLE, immunoadsorption is used as an adjuvant therapy; however, adsorption profiles and binding mechanisms have not yet been completely investigated. Using a minicolumn filled with the sorbent IMPH with or without the ligand phenylalanine, we developed a model simulating clinical conditions in a reduced scale with a constant ratio of plasma to column volume and a constant plasma flow at room temperature. By desorbing the column, the adsorption efficacy for different antibodies could be measured directly. We demonstrate that the adsorption rate can be increased by a low plasma flow and by covering the column surface. Double perfusion of the same column did not increase the amount of adsorbed antibodies. We further demonstrate that the carrier material without a ligand is unable to bind antibodies or protein. In the IMPH sorbent anti-dsDNA antibodies were significantly better adsorbed than total IgG or total protein. After a single perfusion of 21 samples, we estimated a mean anti-dsDNA antibody adsorption rate of 22.5% (+/-13.6). A group of ten responders with a medium adsorption rate of 35.4% (+/-6.5) clearly differed from a second group of eleven nonresponders (10.9% +/- 4.2). Anti-cardiolipin antibodies (ACA) were adsorbed in a wide range (IgG type, 2.5-52.7%, IgM type, 1.1-37.8%) while anti-Ro (SSA) antibody adsorption was negligible. This in vitro minimodel provides a precise simulation of therapeutic immunoadsorption and helps to analyze the binding characteristics of the sorbent IMPH and shows its effectiveness in several antibody subsets of different patients.     

Abatacept for systemic lupus erythematosus: the outlook

Abatacept is a selective T-cell co-stimulation modulator that inhibits full T-cell activation and subsequent antibody production by B cells. Despite the efficacy of abatacept in murine lupus, randomized controlled trials in human SLE do not reveal benefit of abatacept in non-renal and renal lupus. While problems in the study design and the primary efficacy end points may contribute to the negative results of these trials, post hoc analyses using alternative definitions for clinical response suggest the possibility that abatacept may have beneficial effects in active lupus arthritis and proliferative nephritis. Future clinical trials of abatacept should target on defined subsets of SLE patients, utilize multiple pre-defined outcomes based on experience from previous studies and determine the best timing of adding abatacept on a background of minimal immunosuppressive therapies.     

系统性红斑狼疮患者血浆部分抗凝:物质与血栓形成关系的研究

目的　研究系统性红斑狼疮 (SLE)患者血浆蛋白C(PC)、蛋白S(PS)活性水平、活化蛋白C抵抗 (APCR)和狼疮样抗凝物质 (LA)阳性率 ,探讨SLE患者发生血栓的机制。方法　用凝固法在SysmexCA 15 0 0、CA 6 0 0 0全自动血凝仪上检测 4 4例SLE患者和 4 0例正常对照的血浆PC、PS活性、活化蛋白C反应性和LA。结果　SLE组血浆PC、PS活性分别为 (10 6 5± 14 3) %、(98 8± 19 6 ) % ,与对照组相比差异均无显著性意义 (均P >0 0 5 )。SLE组APCR阳性率为 36 4 % ,明显高于对照组 2 5 % (P <0 0 1) ,SLE组LA阳性率为 18 2 % ,明显高于对照组 0 % (P <0 0 5 )。相关性分析表明LA、APCR阳性均与SLE患者血栓形成有关 (均P <0 0 5 ) ,LA和APCR之间有相关性 (P <0 0 5 )。结论　SLE患者血栓形成与血浆PC、PS活性无关 ,与APCR、LA有关 ,且APCR与LA密切相关 ,APCR、LA是SLE患者血栓形成危险性增高的指标。     

Antiglycolipid autoantibody detected in the sera from systemic lupus erythematosus patients.

A high incidence of autoantibody against the neutral glycolipid "asialo GM1" was observed in sera from patients with systemic lupus erythematosus (SLE) with neurological disorders, using an immunoflocculation test. The sera from 14 out of 17 cases of SLE with neurological disorders showed antibody activity against asialo GM1 but not against the following glycolipids: asialo GM2 GM1, and galactocerebroside. In another 87 cases of SLE without any history of seizures, as well as 61 cases of other autoimmune diseases (rheumatoid arthritis, progressive systemic sclerosis, mixed connective tissue disease, etc.) and 20 cases of various neurological diseases (epilepsy, multiple sclerosis, etc.), no antibody could be detected. In general, the antibody titer was high several months, even years, before and/or after the seizure, though the titer was low at the time that patients showed definite neurological symptoms. Immunochemical characterization with Sephadex G-200 chromatogrphy and protein A-Sepharose CL-4B affinity column indicated that the antiasialo GM1 was probably an autoantibody belonging to the immunoglobulin G class. The above results suggest that this newly found autoantibody plays a role in the pathogenesis of neurological disorders accompanying SLE.     

Effectiveness of testing for anti-DNA and the complement components iC3b, Bb, and C4 in the assessment of activity of systemic lupus erythematosus

Seventy-one patients with systemic lupus erythematosus (SLE), seen in an outpatient setting for follow-up evaluation during a 3-mo period, were tested (in addition to routine lupus monitoring studies) with enzyme immunoassays (EIAs) for anti-DNA, iC3b, and factor Bb to determine the relationship of these test results to disease activity. SLE activity was scored by four previously reported scoring systems, and six patients were identified as active by all four systems. We found that the EIA for anti-DNA was the best indicator of disease activity and that iC3b and Bb were not informative for this purpose in this group of patients. Mean iC3b levels were higher in a subset of seven patients with past biopsy evidence of severe (WHO class IV) glomerulonephritis than in the rest of the study population, even though none of these patients had active disease at the time of this study.     

Complement breakdown products in plasma from patients with systemic lupus erythematosus and patients with membranoproliferative or other glomerulonephritis.

A dynamic estimation of the involvement of the complement system in various diseases was obtained by the direct quantitation of breakdown products of C3 and of properdin factor B. The methods used were based, first on the separation of native and fragmented molecules according to their molecular size through a precipitation with polyethylene glycol and, secondly, on an immunochemical quantitation, using specific antisera for the major antigens of C3 and factor B. The sensitivity and the specificity of these methods were demonstrated by activation of complement in vitro with generation of C3 and factor B fragments. A clinical investigation was carried out in 41 patients with systemic lupus erythematosus (SLE), 31 with membranoproliferative glomerulonephritis (MPGN), 26 with other types of glomerulonephritis, and 6 with severe alcoholic cirrhosis of the liver. The following observations were made: (a) an elevated plasma level of C3d fragment of C3 was found in 68% of SLE patients, in 87% of MPGN patients, in 62% of patients with other hypocomplementemic nephritis, and in 15% of those with normocomplementemic nephritis, but in only 33% of patients with liver cirrhosis and very low levels of C3; (b) a significant difference was observed between the levels of C3 obtained with either anti-"native" C3 or anti-C3c sera for immunochemical quantitation, in patients with SLE or MPGN, indicating the presence of "altered" or fragmented C3 in plasma; (c) an elevated plasma level of Ba fragment of properdin factor B was found in 46% of SLE patients, in 67% of MPGN patients, in 50% of patients with other hypocomplementemic nephritis, and in 9% of patients with normocomplementemic nephritis, while the level of properdin factor B was only slightly decreased in these diseases; (d) in SLE and MPGN there was an inverse correlation between the levels of C3d and Ba and the level of C3 in plasma. The level of these fragments was directly correlated with the clinical manifestations of SLE; (e) some patients with a normal C3 level exhibited an elevated plasma concentration of C3 and factor B fragments, suggesting the coexistence of an increased synthesis with a hypercatabolism of complement components. Therefore, the quantitation of complement breakdown products by simple immunochemical methods provides additional information concerning the involvement of complement in disease and new features for the evaluation of the intensity of immune reactions during immune complex diseases.     

系统性红斑狼疮抗活化的蛋白C研究

目的了解抗活化的蛋白C(APCR)在系统性红斑(SLE)患者中的发生情况,并进一步探讨SLE患者发生血栓的机制。方法采用APG-KPTT法,ELISA法和PTT-LA法分别对36例SLE患者及20例正常对照(NC)进行APCR、抗心磷脂抗体(ACA)和狼疮抗凝物(LA)检测。结果SLE患者APCR阳性率(58%)明显高于NC组(5%)(P<0.005),APCR阳性患者中血栓发生率(27.6%)明显高于APCR阴性患者(4.8%)(P<0.01),患者LA阳性率(22%)明显高于对照组(0/10)(P<0.05),患者ACA-IgG及IgM明显高于对照组(P<0.05),而IgA与对照组差异不显著(P>0.05),LA阳性组中的APCR阳性率(90.9%)明显高于LA阴性组(53.8%)(P<0.05),ACA阳性组中的APCR阳性率(64.7%)与ACA阴性组(60.6%)未发现明显相关性(P>0.05)。结论APCR在国人SLE患者中有较高的发生率且与LA有明显相关性。APCR可能是SLE患者诱发血栓的主要原因之一。     

Spectrotype distributions of circulating IgG from patients with systemic lupus erythematosus.

Pathogenic autoantibodies from patients with systemic lupus erythematosus (SLE) may represent a relatively cationic fraction of IgG. We compared the spectrotype distributions of affinity-purified IgG from the sera of 10 SLE patients and 10 age- and sex-matched control subjects. Purified IgG was subjected to isoelectric focusing between pI 3 and 9. No significant difference was observed for pI 6.0-6.5 and 7.5-8.0. However, control subjects had a higher percent of total IgG at 6.5-7.0 (15.4 +/- 4.0 vs 11.1 +/- 2.0, P = 0.008) and at 7.0-7.5 (22.4 +/- 4.8 vs 18.2 +/- 3.8, P = 0.04) whereas SLE patients had a higher percent of total IgG at 8.0-8.5 (24.3 +/- 3.0 vs 20.5 +/- 4.0, P = 0.03) and at 8.5-9.0 (21.9 +/- 5.9 vs 15.1 +/- 3.7, P = 0.006). Spectrotype distributions of circulating IgG from SLE patients are skewed toward higher pI, providing further evidence of proliferation of B-cell clones that express more cationic IgG in patients with SLE. Longitudinal studies of serum IgG from several patients for > 1 year reveal distinct changes in both cationic and anionic clonotypes, suggesting clonal expansion of antiidiotypes to autoantibodies.     

系统性红斑狼疮患者th1/th2失衡的研究

目的探讨系统性红斑狼疮(SLE)病人th1/th2的失衡状况。方法利用三荧光标记法流式细胞术检测抗细胞表面抗原,抗细胞因子。检测出CD4+干扰素(IFN)-γ+白细胞介素(IL)4-及CD4+IFN-γ-IL-4+细胞作为Thl及Th2细胞,从单细胞水平研究初发SLE患者Thl/Th2平衡。结果未用药治疗的SLE病人其Thl细胞明显低于正常对照组(P=0.02)。31例SLE患者按Thl/Th2可分成两组,Thl/Th2降低组与Thl/Th2升高组。Thl/Th2降低组其Thl细胞的降低十分明显。Thl/Th2升高组中Thl细胞仅轻度下降。结论Thl型反应与Th2型反应均参与了SLE的发病。不同患者可能免疫系统的异常并不同。