Clinical significance of serum properdin levels and properdin deposition in the dermal-epidermal junction in systemic lupus erythematosus.

61 biopsies of normal skin from the deltoid area and lesional skin from various sites from 48 patients with systemic lupus erythematosus (SLE) were studied for the presence of properdin, C3, C4, and immunoglobulins (IgG, IgM, and IgA) in the dermal-epidermal junction (DEJ) using direct and indirect immunofluorescence. Properdin was present in 50% of normal and 40% of lesional skins. Properdin was present without C4 in only 2 of 38 nonlesional skin biopsies and in only 2 of 20 lesions. There was no significant difference in incidence of deposition of any of the six proteins studied between nonlesional and lesional skin. The frequency of deposition of each of the proteins correlated with clinical disease activity. The presence of proteins in the DEJ did not correlate with the presence of active renal disease at the time of biopsy nor with previously documented active nephritis. In addition, no other single clinical manifestation correlated with the presence of DEJ deposition of any protein studied. IgA was not demonstrated in the DEJ of nonlesional skin of 16 patients in remission and was present in 7 of 23 patients with active disease (P less than 0.05). Deposition of properdin in lesional skin correlated with the presence of extracutaneous disease activity (P less than 0.05). Analysis of serologic studies on serum obtained at the time of biopsy revealed a statistically significant correlation between C4 and C3 (r = 0.67). This correlation was stronger than that between properdin and C4 (r = 0.37). Titer of antinuclear antibody and percent of DNA binding correlated better with C4 levels than with properdin levels. Serum properdin levels were significantly lower in patients with active disease than in those in remission (P less than 0.05). Serum properdin levels were significantly lower in patients with properdin deposits in lesional skin than in those without properdin deposits. The data suggest that both alternative and classical pathways are activated in patients with clinically active SLE.     

Light chain ratios of serum immunoglobulins in disease

We have determined the individual kappa (kappa)/lambda (lambda) ratios of serum IgG, IgA, and IgM in normal subjects and patients with rheumatoid arthritis, systemic lupus erythematosus, hepatic cirrhosis and IgA nephropathy--40 in each group. Serum samples were first screened by agarose electrophoresis to exclude paraproteinaemia. Concentrations of IgG, IgA, and IgM were determined by enzyme-linked immunosorbent assay (ELISA). The kappa and lambda chain concentrations of each immunoglobulin class were assayed by an ELISA method first developed by us for the determination of kappa/lambda ratios. Our results showed that kappa/lambda ratios of serum IgA and IgM were significantly different from that of IgG in normal subjects and the 4 groups of patients studied (p less than 0.01). The kappa/lambda ratios of individual immunoglobulins in patients with rheumatoid arthritis, systemic lupus erythematosus and liver cirrhosis were similar to those of normal subjects. However, patients with IgA nephropathy displayed a distinctly lower IgA kappa/lambda ratio, suggesting a unique antibody response in the immunopathogenesis of this disease.     

Hypercatabolism of normal IgG; an unexplained immunoglobulin abnormality in the connective tissue diseases

The metabolism of radioiodinated IgG was studied in a series of 42 patients with connective tissue diseases (16 systemic lupus erythematosus, nine rheumatoid arthritis, five polymyositis, five vasculitis, and seven miscellaneous diagnoses). Fractional catabolic rates were increased and survival half-lives were shortened in all diagnostic categories indicating hypercatabolism of IgG. This hypercatabolism was masked by increased IgG synthesis, resulting in elevated serum concentrations of IgG in patients with systemic lupus erythematosus and rheumatoid arthritis and in generally normal concentrations in the others.The metabolism of iodinated IgM was also studied in eight patients with systemic lupus erythematosus, in seven with rheumatoid arthritis, and in 12 controls. The fractional catabolic rates were normal in both groups of patients. Serum concentrations of both IgM and IgA were moderately elevated in all diagnostic categories. Serum albumin metabolism was entirely normal in the nine subjects studied who were not receiving corticosteroids; in three who were receiving them, moderate hypercatabolism was observed.The hypercatabolism of IgG could not be accounted for by factors previously known to alter IgG metabolism. It was not observed in 15 patients with other chronic, inflammatory diseases and was not explained by concomitant administration of adrenal corticosteroids to some patients. Identical results were obtained whether the IgG was obtained from a patient himself or from a normal donor, demonstrating that the hypercatabolism is a host defect and not an abnormality of the protein. Thus, patients with connective tissue disease of several different diagnostic categories have been shown to have an unexplained immunoglobulin abnormality: they catabolize normal IgG at an accelerated rate.     

Low molecular mass IgM in urine of patients with primary and secondary renal diseases

Urine specimens from 50 patients with various renal diseases and from 10 normal subjects were examined for the presence of low molecular mass (7S) IgM by double diffusion, using a specific anti-7S IgM serum. 7S IgM in urine was also quantitated by radial immunodiffusion in 5% polyacrylamide gel. It was found with high frequency in the urine of patients with secondary renal diseases associated with multi-system immunological disorders, especially in systemic lupus erythematosus. Two of the patients with lupus nephritis and positive results for 7S IgM in urine were initially diagnosed as nephrotic syndrome due to a primary renal disease; the symptoms and signs of systemic lupus erythematosus developed later. However, 7S IgM was not detected in the urine of patients with primary renal diseases (except for two cases of nephrotic syndrome), nor in normal subjects. Thus detection of 7S IgM in urine may be helpful in the differential diagnosis of primary and secondary renal diseases.     

Characterization and antigenic specificity of chlorpromazine-induced antinuclear antibodies

Prolonged treatment with chlorpromazine is often associated with the development of antinuclear antibodies, an immunoglobulin M lupus anticoagulant, and polyclonal serum IgM elevation, but not with clinical features of systemic lupus erythematosus (SLE). Sera from 62 long-term psychiatric patients given treatment daily with 100 mg or more of chlorpromazine for at least 1 year were screened for antinuclear antibodies by indirect immunoperoxidase assay using HEp-2 cells. In 26 samples, antinuclear antibody titers greater than or equal to 1:40 with a homogeneous pattern were seen when anti-human IgM was used as the second antibody, three sera samples reacted with IgG, and four samples reacted with both IgG and IgM antisera. The antinuclear antibody antigenic reactivity was investigated by using histone and nonhistone nuclear antigens by enzyme-linked immunosorbent assay and passive hemagglutination techniques. Forty serum samples reacted with histone. Twenty-five samples reacted with deoxyribonucleoprotein (DNP), 28 with single-stranded DNA, and two with double-stranded DNA. No reaction was obtained with the extractable nuclear antigens RNP or Sm. These results indicate that chlorpromazine-induced antinuclear antibodies, like the antinuclear antibodies induced by hydralazine and procainamide, react mainly with histone nuclear antigens. Unlike the hydralazine and procainamide response, in which both IgG and IgM antibodies are demonstrated, the chlorpromazine-induced autoantibodies are predominantly of the IgM class.     

Antibody-dependent cell-mediated cytotoxicity in selected autoimmune diseases.

Antibody-dependent cell-mediated cytotoxicity mediated by peripheral blood lymphocytes was studied in patients with systemic lupus erythematosus, polyarteritis nodosa. Sjogren's syndrome, and rheumatoid arthritis. The target cells were chicken erythrocytes coated with rabbit anti-chicken erythrocyte antibody. Antibody-dependent cell-mediated cytotoxic activity was normal in Sjogren's syndrome and rheumatoid arthritis but significantly decreased (P is less than 0.001) in active systemic lupus erythematosus and in two patients with polyarteritis nodosa. A partial regeneration of antibody-dependent cell-mediated cytotoxic activity was obtained by treatment with pronase and DNase followed by overnight incubation. Sera from patients with systemic lupus erythematosus inhibited antibody-dependent cell-mediated cytotoxic activity of normal lymphocytes. The inhibitory activity was studied by specific immunoadsorption and sucrose density geadient ultracentrifugation. Removal of IgG but not IgM greatly reduced inhibition. Inhibitory factors were present in 7S and heavier fractions containing IgG. Five systemic lupus erythematosus patients were studied serially to determine if improvement in clinical status could be correlated with a decrease in serum inhibitory factors as studied by inhibition of normal antibody-dependent cell-mediated cytotoxicity. Indeed, a greater serum inhibitory capacity was found in each patient during periods of greater disease activity.     

Immunoglobulin in clinically uninvolved skin in systemic lupus erythematosus: association with renal disease

23 of 42, or 55%, of patients with systemic lupus erythematous had immunoglobulin deposits along the epidermal basement membrane of uninvolved skin (positive lupus band test [LBT]). In patients with low serum complement levels, 91% had a positive LBT), as compared with 15% in those with normal complement levels. The LBT was positive in 70% of patients with clinical and laboratory evidence of renal disease, but in only 31% of patients without renal disease. 81% of patients with the more severe histologic forms of lupus nephritis, i.e., proliferative glomerulonephritis and membranous glomerulonephritis, and positive tests, whereas only 23% with mesangial glomerulitis or normal histologic findings were positive. Immunoglobulins of the same class found in the skin were detected in the glomeruli of patients examined by renal biopsy. These results suggest that there is a relationship between the occurrence of immunoglobulin in the epidermal basement membrane and the presence of the more severe forms of lupus nephritis.     

Antibodies to native DNA: detection by immunoperoxidase assay and determination of their immunoglobulin classes.

Antinuclear antibodies are almost always found in sera of patients with systemic lupus erythematosus. To differentiate antinuclear antibodies from antibodies to DNA in the recently described Crithidia luciliae assay, we developed an immunoperoxidase technique for detecting antibodies to native, double-stranded DNA and compared results by it with those by the Farr assay. Smears of cultured Crithidia luciliae were incubated with human sera, peroxidase-labeled anti-human IgG serum, and diaminobenzidine. The peroxidase stain was examined by conventional light microscopy, which facilitated differentiation between the kinetoplast and the nuclear staining. The Crithidia assay appeared to be specific for double-stranded DNA antibodies, seemed to be more sensitive than the Farr assay, and allowed us to determine the immunoglobulin classes of antibodies to native DNA. Some patients with systemic lupus erythematosus had only IgM or IgA antibodies to DNA.     

Evaluation of four commercially available ELISA assays for the serologic diagnosis of Lyme disease

We evaluated four commercially available ELISAs for detection of antibody to Borrelia burgdorferi with 21 sera from patients with clinically diagnosed Lyme disease and 89 patient control sera. Patient control sera included 28 sera from patients with rheumatoid arthritis (RA), 17 sera from patients with systemic lupus erythematosus (SLE), and 44 sera containing antibodies reported to cross-react in some Lyme disease tests. The ELISAs tested (Cambridge Bioscience, Diamedix, 3M, and Zeus) detect antibodies (IgM and/or IgG) that bind Borrelia burgdorferi antigen attached to microtiter wells. Antibody reactivity in the sera from patients with clinically diagnosed Lyme disease was characterized by using Zeus immunoglobulin class-specific assays (IgM and IgG). Sensitivities in early and late Lyme disease were as follows: Cambridge and Diamedix, 57% and 100%; 3M, 57% and 93%; and Zeus, 71% and 86%. Reactivities within a patient control population were: Cambridge and Diamedix, 3%; 3M, 7%; and Zeus, 10%.     

系统性红斑狼疮患者的血清蛋白电泳谱及免疫球蛋白测定分析

目的探讨系统性红斑狼疮(systemic lupus erythematosus,SLE)患者的血清蛋白电泳谱、免疫球蛋白和补体的变化情况.方法 对64例系统性红斑狼疮患者与53例健康者血清进行琼脂糖凝胶电泳及免疫球蛋白和补体测定,并对其各测定结果进行分析.结果 清蛋白、α1-球蛋白、α2-球蛋白、γ-球蛋白、免疫球蛋白IgG、IgA、IgM高于对照组,而C3、C4比对照组低,β-球蛋白无差异.结论 血清蛋白电泳、免疫球蛋白和补体的联合检测对系统性红斑狼疮的诊断和疗效观查有重要意义.     

TLR9/MyD88 signaling is required for class switching to pathogenic IgG2a and 2b autoantibodies in SLE

Loss of tolerance in systemic lupus erythematosus (SLE) leads to the generation of autoantibodies, which accumulate in end-organs where they induce disease. Here we show that immunoglobulin (Ig)G2a and 2b autoantibodies are the pathogenic isotypes by recruiting FcgammaRIV expressing macrophages. Class switching, but not development, of IgM anti-self B cells to these pathogenic subclasses requires the innate immune receptor Toll-like receptor (TLR)9 and MyD88 signaling. In their absence, switching of autoreactive B cells to the IgG2a and 2b subclasses is blocked, resulting in reduced pathology and mortality. In contrast, switching of anti-self B cells to IgG1 is not perturbed and generation of nonautoreactive IgG2a and 2b antibodies is not impaired in TLR9-deficient mice. Thus, the TLR9 pathway is a potential target for therapeutic intervention in SLE.     

系统性红斑狼疮患者抗核小体抗体与疾病活动的关系

目的探讨系统性红斑狼疮(SLE)患者血清中抗核小体抗体的发生与补体含量及SLE疾病活动的关系。方法用ELISA法检测抗核小体抗体及抗双链DNA(ds鄄DNA)抗体,用全自动特种蛋白仪检测免疫球蛋白及补体。根据抗核小体抗体的含量将46例患者分成抗核小体抗体阳性组(A组)和抗核小体抗体阴性组(B组),同时分别对这两组进行各实验指标分析。结果两组结果显示,A组中抗ds鄄DNA抗体、C3、C4、IgG、SLE积分与B组相比较差异均有显著性(P<0.05),而红细胞沉降率(ESR)、Hb、IgA、IgM两组相比较差异无显著性(P>0.05)。抗核小体抗体含量与补体C3呈显著负相关(P<0.001),与补体C4呈显著负相关(P<0.05),与SLE积分呈显著正相关(P<0.05)。结论抗核小体抗体在SLE的发生、发展及诊断中起着重要作用,有望成为标志SLE活动的新指标。     

泼尼松联合甲氨喋啶治疗对系统性红斑狼疮患者炎症因子及免疫球蛋白表达水平的影响

[目的]探讨泼尼松联合甲氨喋啶治疗系统性红斑狼疮(SLE)患者对外周血清中的炎症因子白细胞介素-17(IL-17)和白细胞介素-23(IL-23)及免疫球蛋白表达水平的影响.[方法]在本院治疗的74例SLE患者,随机分为两组,各37例.对照组采用醋酸泼尼松治疗,观察组在对照组基础上联合甲氨喋啶治疗,3个月治疗后对患者治...     

抗核抗体、免疫球蛋白及补体联合检测对狼疮性肾炎的诊断意义

目的探讨抗核抗体(ANA)、免疫球蛋白和补体检测对狼疮性肾炎(LN)和不伴肾炎的系统性红斑狼疮(SLE)的诊断意义。方法对406例SLE患者和120例健康体检者采用间接免疫荧光法测定ANA,应用散射比浊法测定补体(C3、C4)及免疫球蛋白(IgG、IgA、IgM、IgE)。结果 406例SLE患者中,ANA阳性率94.49%,与对照组比较,差异有统计学意义(P<0.01)。SLE患者LN组与非LN组ANA核型均以核均质型和核颗粒型为主,LN组这2种核型占84.35%,非LN组占72.12%,两者差异有统计学意义(P<0.05);LN组胞浆颗粒型占11.31%,与非LN组的21.93%差异有统计学意义(P<0.05)。LN组与非LN组IgG、IgA、IgM、IgE水平均增高,而C3、C4水平均降低,与对照组比较差异有统计学意义(P<0.05);C3水平在LN组降低更明显,与非LN组比较差异有统计学意义(P=0.001)。结论 ANA、免疫球蛋白和补体的联合检测对提高SLE的临床诊断、预后判断、疗效观察等方面意义重大。     

Intrinsic B cell defects in NZB and NZW mice contribute to systemic lupus erythematosus in (NZB x NZW)F1 mice

We have previously shown that long-term in vitro proliferating fetal liver pre-B cell lines derived from autoimmune-prone (NZB x NZW)F1 (BW) mice, but not normal (B6 x DBA2)F1 mice, can differentiate in severe combined immunodeficient (SCID) mice to produce elevated levels of serum immunoglobulin (Ig) M and IgG, and high titers of antinuclear antibodies The contribution of parental NZB and NZW strains to B cell abnormalities of BW hybrid mice was investigated here by preparing pre- B cells and transferring them into immunodeficient SCID- and RAG-2- targeted mice. We show that transfer of NZB pre-B cells led to a marked IgM hypergammaglobulinemia and to the production of limited amounts of IgG2a. On the other hand, the transfer of NZW pre-B cell lines led to moderately elevated IgM levels and marked hypergammaglobulinemia of IgG2a. High IgM and low IgG anti-DNA titers are found in the recipients of NZB pre-B cells, whereas those receiving NZW pre-B cells contained lower levels of IgM and high titers of IgG anti-DNA. In marked contrast, essentially identical titers of antibodies directed against a non-self-antigen, DNP, are found in all group of pre-B cell recipients. Thus, B-lineage cells of both NZB and NZW parental strains manifest abnormalities associated with the development of this lupus-like disease. Therefore, the present study strongly suggests a complex inheritance of B cell abnormalities in autoimmune-prone (NZB x NZW)F1 mice and emphasizes the critical importance of intrinsic B cell defects in the development of murine systemic lupus erythematosus.     

Sjogren's Syndrome: 2. CLINICAL ASSOCIATIONS AND IMMUNOLOGICAL PHENOMENA

The diverse clinical associations and wide variety of immunologicalphenomena and their incidence occurring in a large series ofpatients with Sjøgren's syndrome is presented and discussed,with reference to individual cases where relevant. Althoughthere was an increased prevalence of organ-specific auto-antibodies,we were unable to show an association between organ-specificauto-immune diseases and Sjøgren's syndrome. The previouslyreported increased prevalence of mitochondrial antibody andliver disease have been confirmed, as has renal tubular dysfunction.Drug allergy and eosinophilia were commonly found. Serum IgG, IgM, and IgA levels were elevated, and serum secretory-IgAlevels were markedly raised. The increased incidence of non-organ-specificauto-antibodies has been confirmed, and sera negative for rheumatoidfactor by conventional tests were found to have a high prevalenceof antiglobulin factors. Three patients without other evidenceof systemic lupus erythematosus had raised serum native-DNA-bindingcapacities, and another eight patients had antibodies to heat-denaturedDNA.     

Role of interleukin 10 in the B lymphocyte hyperactivity and autoantibody production of human systemic lupus erythematosus

Interleukin-10 (IL-10) is produced at a high level by B lymphocytes and monocytes of patients with systemic lupus erythematosus (SLE). In the present work, we analyzed whether this increased production of IL-10 contributed to the abnormal production of immunoglobulins (Ig) and of autoantibodies in SLE. The role of IL-10 was compared with that of IL- 6, another cytokine suspected to play a role in these abnormalities. The spontaneous in vitro production of IgM, IgG, and IgA by peripheral blood mononuclear cells from SLE patients was weakly increased by recombinant IL (rIL)-6, but strongly by rIL-10. This production was not significantly affected by an anti-IL-6 mAb but was decreased by an anti- IL-10 mAb. We then tested the in vivo effect of these antibodies in severe combined immunodeficiency mice injected with PBMC from SLE patients. The anti-IL-6 mAb did not significantly affect the serum concentration of total human IgG and of anti-double-stranded DNA IgG in the mice. In contrast, the anti-IL-10 mAb strongly inhibited the production of autoantibodies, and, to a lesser extent, that of total human IgG. These results indicate that the Ig production by SLE B lymphocytes is largely IL-10 dependent, and that the increased production of IL-10 by SLE B lymphocytes and monocytes may represent a critical mechanism in the emergence of the autoimmune manifestations of the disease.     

Selective deposition of immunoglobulin A1 in immunoglobulin A nephropathy, anaphylactoid purpura nephritis, and systemic lupus erythematosus.

To further characterize the IgA deposits found in glomeruli of patients with IgA nephropathy, anaphylactoid purpura nephritis, and systemic lupus erythematosus, renal biopsies from patients with these disorders were stained by immunofluorescence with monoclonal anti-IgA subclass reagents, anti-light chain reagents and anti-J chain. The mesangium and peripheral capillary were brightly stained for IgA1 and were negative for IgA2. IgA1 and, to a lesser extent, IgA2 were contained in tubular casts. Both kappa and lambda light chains were found in all deposits. The intensity of J chain staining correlated with the intensity of IgM and not IgA staining. Biopsies brightly stained for IgA but negative for IgM were negative for J chain. These results indicate that glomerular IgA deposits in these disorders consist predominantly of monomers of IgA1.     

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。     

Synapsin I identified as a novel brain-specific autoantigen.

BACKGROUND: We report the identification and characterization of a novel 74-kd brain-specific autoantigen that is reactive with serum from a patient with discoid lupus erythematosus and chronic lymphocytic leukemia. METHODS: We determined the molecular weight, tissue distribution and subcellular distribution of the autoantigen and obtained limited amino acid sequence after purification by ion-exchange chromatography and trypsin digestion. RESULTS: We identified the 74-kd autoantigen as synapsin I on the basis of the following observations. First, the autoantigen has properties consistent with synapsin I: molecular weight of approximately equals 74 kd, brain-specific distribution, presence in cytosol and on synaptosomes, and association with taxol-stabilized microtubules. Second, limited amino acid sequence determination after trypsin digestion of the autoantigen shows identity with synapsin I. Third, the autoimmune serum immunoblots fusion proteins that incorporate rat synapsin Ia. The autoantibodies reactive to synapsin Ia are of immunoglobulin (Ig) G and IgM class. CONCLUSIONS: This is the first report of autoantibodies that are reactive to synapsin Ia. Autoantibodies that are reactive to synapsin Ia are not restricted to discoid lupus erythematosus patients, because we found identical reactivity in two of 18 sera from dsDNA-positive systemic lupus erythematosus patients and in two of 14 rheumatoid factor-positive sera. Whether autoantibodies to synapsin I are associated with neuropsychiatric manifestations is currently unknown.