系统性红斑狼疮血清抗核小体抗体水平及意义的探讨

目的 研究抗核小体抗体(AnuA)在系统性红斑狼疮(systemic lupus erythematosus，SLE)患者血清中的水平及其相关影响因素，探讨AnuA在SLE诊治中的作用和意义。方法 采用酶联免疫吸附法(EUSA)测定120例初诊SLE患者、55例其他风湿性疾病和30名健康对照血清中AnuA水平。同时记录各种临床表现，检测并分析其治疗前的其他自身抗体和实验室指标。结果 自身抗体在SLE及其他风湿病对照组的阳性率分别为AnuA56％和7％，抗dsDNA抗体35％和1％，抗Sm抗体24％和0。AnuA与SLE患者性别、年龄、病程无相关性，对sLE的诊断敏感性和特异性分别为55．8％、9513％，对狼疮肾炎(1upus nephritis，LN)的诊断敏感性和特异性分别为77．％、64．5%。AnuA与肝脏损害和疾病活动呈线性相关(t=3．152，2．171，P＜O．05)。AnuA分别与抗dsDNA、抗Sm联合检测对SLE诊断敏感性提高27％、40％（X^2值分别为38．930、18．161，P＜O．01)。结论AnuA在SLE血清中水平显著增高，AnuA测定是SLE诊断和治疗监测中有价值的新的实验室检测指标之一，与抗dsDNA抗体联合检测可提高诊断SLE、LN的敏感性和特异性。     

Anti-nucleosome antibodies in the diagnosis of systemic lupus erythematosus

OBJECTIVE: To study the prevalence and diagnostic significance of antibodies against nucleosomes in patients with systemic lupus erythematosus (SLE) as compared to five anti-nuclear antibody (ANA) assays. METHODS: The study included 305 patients with SLE, 125 patients with other autoimmune rheumatic diseases, and 415 healthy controls. Anti-nucleosome antibodies were measured by an enzyme-linked immunosorbent assay (ELISA) and ANA by immunofluorescence (IF) using Hep-2 cells. Anti-double-stranded DNA (anti-dsDNA) antibodies were measured by three commercial ELISAs and by IF using Crithidia luciliae as antigen. RESULTS: Compared to three ELISAs for anti-dsDNA, the anti-nucleosome assay was less sensitive (30% vs. 29-69%) but equally specific (90% vs. 77-95%) for SLE. The most sensitive test was ANA (76%), and the least sensitive was Crithidia (13%). The correlations between the different assays were good (p < 0.001 for all comparisons). CONCLUSION: The anti-nucleosome antibody assay does not offer additional information compared to conventionally used anti-dsDNA tests in the differential diagnosis of SLE.     

Anti-nucleosome antibodies may predict lupus nephritis and severity of disease in systemic lupus erythematosus

Background/ObjectiveDetection of anti-nucleosome antibodies in patients with systemic lupus erythematosus (SLE) has been well established and it is claimed that their presence is associated with disease activity. The objective of this study is to determine the diagnostic value of anti-nucleosome antibodies in the assessment of lupus nephritis and clinically active SLE.MethodsThe anti-nucleosome antibodies were evaluated in the serum of 200 Tunisian SLE patients at disease onset by a sensitive immunodot assay. Serum samples from each patient were also tested for ANA and anti-ds DNA antibody by IIF on Hep 2 cells and Crithidia luciliae respectively. During the follow-up, the patients were regularly monitored for clinical parameters. Global SLE activity was measured by the European Consensus Lupus Activity Measurement (ECLAM).ResultsThe prevalence of anti-nucleosome and anti-dsDNA antibodies was 69% and 63.5% respectively. Anti-nucleosome antibodies were found to be 30.1% positive in SLE patients lacking anti-dsDNA antibody. 79.5% patients had active SLE at the first clinical examination. Anti-nucleosome antibodies were more sensitive than anti-dsDNA antibodies to detect active SLE (78% vs. 71.7%, P =0.19). 52.5% of SLE patients had renal involvement. Among these patients, the rate of anti-nucleosome positivity and anti-dsDNA were 77.1% and 67.6% respectively. The positivity of anti-nucleosome antibodies was significantly higher in patients with renal disease than the subjects without renal disease (P = 0.009). Anti-nucleosome and anti-ds DNA antibodies were significantly correlated with disease activity (P < 0.001 and P < 0.001 respectively).ConclusionAnti-nucleosome antibody reactivity may be a useful marker in the diagnosis and assessment of active SLE.     

Anti-nucleosome antibodies in systemic lupus erythematosus patients: Relation to anti-double stranded deoxyribonucleic acid and disease activity

Aim of the work

To measure the level of anti-nucleosome (anti-NCS) antibodies in systemic lupus erythematosus (SLE) patients and to evaluate their relation with anti-double stranded deoxyribonucleic acid (anti-dsDNA) antibodies and SLE disease activity.

Platelet antibodies in systemic lupus erythematosus

In systemic lupus erythematosus (SLE), the precise cause of the thrombocytopenia is unknown. Since platelet associated IgG is increased in many patients, it has been suggested that the destruction of platelets might be dependent on specific antibodies. In nine patients with SLE, platelet associated immunoglobulins were found together with free serum antibody which bound to platelets from all normal subjects. Using an immunoblotting technique with membrane proteins from normal platelets incubated with patient sera, target antigens were localized on a band of mol wt 108,000 in two cases (B. and N.) and on a band of mol wt 66,000 in a third (M.). When the same technique was applied to autologous platelets of patient N., autoantibody binding to the protein of mol wt 108,000 was demonstrated. The antigenic determinants were not removed from the platelets by enzyme treatment or by disulphide bond reduction, and were localized in the cytoplasmic fraction of the platelets.     

Neurofilament antibodies in systemic lupus erythematosus

Autoantibodies against neuronal antigens occur in sera of patients with systemic lupus erythematosus (SLE). These antibodies may have significance in the pathogenesis of neurological complications of SLE. However, the neuronal structures containing the corresponding autoantigens are poorly known. In our study we assayed circulating antibodies against defined neuronal components--neurofilaments--by an enzyme-linked immunosorbent assay (ELISA) using purified neurofilament polypeptides as targets. Circulating neurofilament antibodies (anti-NF) of IgG class were detected in 21% of 28 patients with SLE and in 6% of 17 patients with rheumatoid arthritis and in none of the 14 patients with primary sicca syndrome and 40 blood donors. The presence of anti-NF could also be confirmed by the indirect immunofluorescence technique using frozen sections of rat spinal cord. In one serum, anti-NF cross reacted with vimentin type of intermediate filaments. The antibodies bound both to the 70 kilodalton and the 200 kilodalton polypeptides of neurofilaments as judged by the immunoblotting technique. Two of 6 anti-NF positive patients had neurological complications.     

Anti-insulin antibodies and the natural autoimmune response in systemic lupus erythematosus

Systemic lupus erythematosus (SLE) is characterized by the finding of ample serum autoantibodies. The role and the origin of many of these antibodies are still obscure. The aim of this work was to study the occurrence of anti-insulin antibodies (AIA) in SLE, and to postulate, based on AIA determination, on the mechanisms involved in the production of some autoantibodies in SLE. IgG and lgM AIA, anti-DNA antibodies (ADA) and anti-tetanus toxoid antibodies (ATA) were determined using ELISA in sera and B-lymphocytes culture media of 24 SLE patients, 10 healthy controls and 19 insulin-dependent diabetes mellitus (IDDM) patients. B- and T-lymphocytes were isolated using Ficoll gradient, depleted of T-cells using cyclosporin A, EBV infected and grown in medium. The frequencies of IgM-AIA and IgG-ADA were higher in SLE patients than in healthy controls (P < 0.02 and P < 0.05, respectively). The rate of IgM-AIA in SLE and IDDM was comparable, while IgG-AIA was significantly less common in SLE than in IDDM (P < 0.05). The prevalence of ATA in SLE patients and healthy controls was similar. These findings increase the spectrum of the humoral autoimmune response in SLE and suggest that part of it (natural autoantibodies) is independent of antigen driven response.     

Antilymphocyte antibodies in systemic lupus erythematosus

Antilymphocyte antibodies in SLE bind to normal lymphocytes and are not always associated with lymphocytotoxic activity. Detection of these antibodies on the surface of normal lymphocytes by immuno-fluorescent techniques is enhanced by short incubation periods, reduced temperature, and metabolic inhibition. The same effects occur in SLE blood and May-June influence the enumeration of B lymphocytes as determined by surface immunoglobulin. The percent of T lymphocytes forming sheep erythrocyte rosettes is not affected by these antibodies.     

Anti-DNA antibodies in systemic lupus erythematosus.

Anti-DNA antibodies are the serologic hallmark of systemic lupus erythematosus and important markers for diagnosis and prognosis. Although a number of mechanisms for anti-DNA production have been proposed, recent evidence from human as well as murine lupus indicate a prominent role of DNA antigen drive. To identify further the basis of this response, current research is focusing on the structural features of DNA associated with immunogenicity in both the normal and autoimmune settings. Elucidating these mechanisms should thus provide insights into the pathogenesis of systemic lupus erythematosus as well as the immunologic activity of nucleic acids.     

Antineutrophil cytoplasmic antibodies in systemic lupus erythematosus

Objective. To examine the prevalence, subspecificities, and clinical associations of antineutrophil cytoplasmic antibodies (ANCA) in patients with systemic lupus erythematosus (SLE). Methods. One hundred fifty-seven sera from 120 patients with SLE were examined for classic (c) and perinuclear (p) pattern ANCA by indirect immunofluorescence. Antibody subspecificities were determined by enzyme-linked immunosorbent assay (ELISA). Serologic results were correlated with clinical manifestations as categorized by the BILAG (British Isles Lupus Assessment Group) index. Results. ANCA were found in 40 of the 157 sera (25%). Only a pANCA, not a cANCA, pattern of fluorescence was seen. By ELISA testing, 16 sera reacted to lactoferrin, 8 to elastase, and 4 to lysozyme. There was no reactivity to proteinase 3 (PR3) or myeloperoxidase (MPO). No correlation of pANCA, or any of the ANCA subspecificities, with organ system involvement, as categorized by the BILAG index, was found. Notably, there was no correlation of ANCA results with lupus vasculitis. Conclusion. The absence of cANCA, anti-PR3, and anti-MPO shows that with appropriate assay conditions, ANCA testing assists in the differentiation between SLE and the ANCA-associated vasculitides. The lack of a correlation between pANCA or any ANCA subspecificity and clinical manifestations suggests that ANCA do not identify particular clinical subsets among SLE patients, including those with lupus vasculitis.     

Antiendothelial cell antibodies in systemic lupus erythematosus

Objective: Anti‐endothelial cell antibodies (AECAs) are a heterogeneous group of antibodies against a variety of antigenic determinants on endothelial cells (EC). AECAs are known to play an immunopathogenic role in triggering EC activation, leading to vascular damage. The purpose of this study was to assess: (i) the incidence of AECAs in systemic lupus erythematosus (SLE) patients with nephritis (LN) and to compare this with SLE patients without clinical evidence of nephritis; and (ii) to understand the association of AECAs with disease severity based on renal histopathological findings. Method: Fifty‐three clinically and histopathologically proven cases of LN were studied along with 20 patients without evidence of nephritis. AECAs were detected by immunofluorescence using cultured human umbilical vein endothelial cells (HUVECs). The titres and immunoglobulin subclass of AECAs were also identified. Other autoantibodies were also detected. Results: In the LN group, 21 (39.6%) were AECA positive and 19 (35.8%) were antineutrophil cytoplasmic antibody (ANCA) positive. Autoantibodies to double‐stranded DNA (anti‐dsDNA) were present in 49 (92.4%) cases. In patients without nephritis, seven (35%) tested positive for AECA, five for ANCA and all were antinuclear antibody (ANA) positive. Anti‐dsDNA was detected in 16 patients (80%), higher incidence of AECAs was noted in diffuse proliferative glomerulonephritis (41.2%) as compared to focal proliferative glomerulonephritis (37.5%) and membranoproliferative glomerulonephritis (33.3%). IgG‐AECA subclass was noted in 85.7% patients, IgM‐AECA and IgG + M AECA subclasses of AECA were detected in 7.1% cases. AECAs were also found to be associated with other autoantibodies such as ANA, anti‐dsDNA and ANCA. Conclusion: No significant differences in AECA positivity was found between SLE with and without nephritis.     

Antineuronal antibodies in neuropsychiatric systemic lupus erythematosus

The diagnosis of neuropsychiatric systemic lupus erythematosus (NP-SLE) is clinical and one of exclusion. Brain cross-reactive lymphocytotoxins or neuronal antibodies have been proposed as a mechanism underlying NP-SLE. We assessed the clinical relevance of neuronal cell binding antibodies using a standardized clinical definition of NP-SLE. Serum from 54 SLE patients and 77 controls were tested for binding to 3 neuroblastoma and 3 glioblastoma cell lines. Thirty-three SLE patients (61%) fulfilled clinical criteria for the diagnosis of NP-SLE; of these, 55% had serum binding activity to both neuroblastoma and glioblastoma cell lines, compared with 33% of the other SLE patients. When reactivity to neuroblastoma cell lines only was assessed, 43% of NP-SLE patient sera demonstrated binding activity, versus 14% of sera from the remaining SLE patients. Control subjects' reactivity to neuroblastoma cell lines was positive in 12% of sera. Analysis of serum reactivity using non-neuronal cell lines revealed that neuroblastoma, but not glioblastoma, cell binding was specific. NP-SLE patients with evidence of diffuse symptomatology had a higher mean titer of neuroblastoma cell line binding than those with focal symptomatology. Using a panel of substrates, one can identify a significant proportion of patients who are independently defined as having NP-SLE, who demonstrate specific serum neuronal antibodies.     

Antiribosomal antibodies in systemic lupus erythematosus.

ARA occur in approximately 10% of randomly selected SLE patients but in up to 40% of patients with active disease. Anti-P antibodies appear to be a highly specific diagnostic marker for SLE because they are rarely detected in other multisystem autoimmune disorders. ARA are most frequently directed against the P proteins, and the shared conserved C-terminus of the P proteins is immunodominant in almost all sera tested. Anti-P antibodies increase in titer in patients with active disease and have been reported to be detected more frequently in patients with severe behavioral disturbances. This may be particularly true of patients with affective disorders. The clinical utility of serologic tests for anti-P in central nervous system lupus must await large, prospective studies. Other ARA antibodies have been detected in patients with SLE. These antibodies include anti-28S rRNA, anti-S10, and anti-L12. In all cases, the frequency with which these antibodies are detected is increased in sera containing anti-P. The P proteins and the 28S rRNA epitope play essential, but as yet undefined, roles in GTPase activity on the ribosome. The L12 protein is the mammalian homologue of the E. coli and yeast proteins known to bind to the 28S rRNA epitope. These findings indicate that some SLE patients produce autoantibodies against multiple components of a functionally related domain of the ribosome. This, in turn, supports the notion that the ribosome initiates and/or maintains autoantibody production. Despite these findings, attempts to induce anti-P antibodies by immunization with autologous ribosomes in the autoimmune strain of mouse, MRL, have been unsuccessful. It therefore seems likely that the ribosomal components must be altered to break tolerance or that other abnormalities of the immune system are necessary for autoantibody production.     

Antineuronal antibodies and neuropsychiatric systemic lupus erythematosus

Determination of antineuronal antibodies was carried out by a solid phase radioimmunoassay, in which the neuronal cells SK-N-SH were cultured as the target antigens, the positive rate in sera and CSF of neuropsychiatric SLE patients being 85% and 77.8% respectively, all with quite high level of antibodies. Although 66.7% sera of SLE patients without CNS involvement were also positive for the antibodies, yet the antibody levels of them were distinctly lower, and only 3 of them showed weak positive reactions in their CSF. It was shown that 90% of neuropsychiatric SLE patients with diffuse CNS manifestations had increased antineuronal antibodies, compared with only 20% in cases with local CNS involvements. The antibody levels both in sera and CSF decreased remarkably following the patients' recovery from the neuropsychiatric attack. It is concluded that the antineuronal antibodies might play a role in the pathogenesis of neuropsychiatric lupus and the determination of them in CSF might be useful in the diagnosis and differential diagnosis of neuropsychiatric lupus.     

Analysis of T-cell clones in systemic lupus erythematosus

BACKGROUND AND OBJECTIVE: It is fairly well established that T-helper (TH)((1)) cells play a role in the pathogenesis of organ-specific autoimmune diseases, while their role and their relationship with TH((2)) cells is far from being defined in systemic lupus erythematosus (SLE). To address this issue, six female patients who fulfilled the American Rheumatism Association criteria for the diagnosis of SLE were studied. DESIGN AND METHODS: We analyzed the intracellular production of cytokines by T-cells from the peripheral blood (PB). Then, we established T-cell clones (TCC) from the peripheral blood (PB) of all cases as well as from the synovial fluid of one patient with an articular flare-up. RESULTS: The percentages of IL-4 positive and IFN-g positive PB T-cells were not different between SLE patients and normal controls. When 93 TCC (67 CD4(+), 23 CD8(+)) from the PB of 5 different SLE patients were compared to 118 TCC (94 CD4(+), 23 CD8(+)) from 5 healthy controls no statistical difference was observed between SLE and controls in terms of TH((1)), TH((2)) or TH((0)) phenotype. However, SLE clones showed a reduced ability to secrete IL-10 (p = 0. 002). In contrast, the analysis of the 30 clones obtained from synovial fluid revealed that 11/23 CD4(+) clones were TH((1)), 12/23 were TH((0)), 2/7 CD8(+ )clones were TH((1)) and 5/7 were TH((0)). No TH((2)) clones were obtained from the synovial fluid. INTERPRETATION AND CONCLUSIONS: The data suggest that the T-cell subsets operating in actively inflamed organs of SLE may belong to the TH((1)) and TH((0)) subsets.     

Significance of anti-lymphocyte antibodies in systemic lupus erythematosus

Sera from patients with SLE frequently contain IgM and IgG antibodies with multiple specificities for lymphocyte surface determinants, including autologous antigens. The IgM antibodies are of relatively low binding avidity and exhibit broad reactivity with B and T lymphocytes from most individuals. IgG antibodies are reactive selectively with PBL from different individuals and appear to be more specific for B cell and a minor proportion of T cells. The molecular nature of the surface determinants involved and their relationship with known antigens and receptors remain largely undefined. Interest in anti-lymphocyte antibodies in SLE relates in part to data suggesting a causal role in the abnormal immune system function in this disorder. In this regard, possible mechanisms that are supported by indirect data include: a) antibody-mediated lymphocyte depletion in vivo, perhaps involving functional subsets specifically; b) antibody blockade of surface receptors operant in cell-cell and in cell-soluble antigen interactions. Certain data have raised the possibility that anti-lymphocyte antibodies represent serum markers for infection with virus as etiologic in SLE, but this question is controversial. Nevertheless, further investigation may yet reveal viral or genetically determined “SLE-specific” lymphocyte surface antigens. Clinically, anti-lymphocyte antibodies may have potential for mediating tissue injury in SLE, either directly or indirectly as circulating complexes in association with “shed” lymphocyte surface antigen. Direct evidence in support of such a role in the natural history of this disorder has not been forthcoming.