Serologic associations of anti-cytoplasmic antibodies identified during anti-nuclear antibody testing.

BACKGROUND: There are currently no guidelines concerning additional laboratory testing for specific autoantibodies among anti-nuclear antibody-negative sera with an anti-cytoplasmic staining pattern identified by indirect immunofluorescence assay. Moreover, few data are available that address this laboratory situation. METHODS: We performed specific autoantibody assays in 200 sera with an anti-nuclear antibody titer < or =1:32 and a cytoplasmic titer (undefined staining pattern) of > or =1:64, identified sequentially in the course of routine anti-nuclear antibody testing. RESULTS: A total of 85 sera (42.5%) were positive in one (n=57) or more (n=28) of the specific autoantibody tests performed. Autoantibodies identified were antimitochondrial (15%), antimicrosomal (13%), anti-neutrophil cytoplasmic (10%), anti-smooth muscle (6%), anti-parietal cell (4%), and extractable nuclear antigen (8.5%, including histones, SSA, SSB, Sm, Jo-1 or Scl-70). A positive result in one or more of these assays was more frequent at anti-cytoplasmic titers > or =1:1024 (77.8%) than at titers of 1:64-1:128 (7%) (chi2=25.3, p<0.001). CONCLUSIONS: The present data demonstrate that undefined anti-cytoplasmic staining in anti-nuclear antibody-negative sera is associated with, although not necessarily caused by, a high frequency and wide range of specific autoantibodies. Further work is needed before specific recommendations can be made concerning follow-up in subjects with this laboratory finding.     

The clinical significance of autoantibodies in gastrointestinal malignancies: an overview

Autoimmunity can be associated with cancer and one of the forms of its expression is the development of antibodies to autologous cellular antigens. The types of cellular proteins which induce autoantibody responses in gastrointestinal malignancies are quite varied and include cellular proteins encoded by mutated normal genes (p53), cellular proteins that are overexpressed and/or aberrantly expressed in malignant tissues (carcinoembryonic antigen), inhibitors of apoptosis (survivin and livin), major components of mucus (mucins), surface receptors of apoptosis (Fas) and nuclear-restricted proteins (double-stranded DNA, single-stranded DNA and Sm family proteins). In the past few years, due to the great clinical interest and the advancement in detection techniques, the above list has grown significantly and a large number of cancer-related antigens, which trigger a specific humoral immune response to the host, have also been identified. The authors review the autoantibodies associated with gastrointestinal malignancies and their clinical implications.     

Implications of anti-Ro/Sjögren''s syndrome A antigen autoantibody in normal sera for autoimmunity.

We have applied a sensitive assay to analyze lupus and Sjögren''s syndrome autoantibodies in 40 normal sera. Seven of these bound Ro/Sjögren''s syndrome A antigen (SSA). Although this binding was 1,000-fold lower than the highest anti-Ro/SSA level measured from patients, it was inhibited by human Ro/SSA. Positive normal serum-bound Ro/SSA in Western immunoblots and binding activity was demonstrated in the F(ab'')2 fragment of IgG. Affinity purification of normal anti-Ro/SSA IgG increased the specific anti-Ro/SSA binding by greater than 17-fold. This purified antibody formed a Ro/SSA precipitin and had a relative affinity for Ro/SSA identical to that of Ro/SSA precipitin-positive patients. These data demonstrate that the anti-Ro/SSA present in healthy normal donors is true autoantibody. Anti-La/Sjögren''s syndrome B antigen (SSB) autoantibodies were found in 3 of the 40 normal sera, while none bound nuclear ribonucleoprotein (Sm). Finding low levels of anti-Ro/SSA and anti-La/SSB among normals may indicate that anti-Ro/SSA and anti-La/SSB occur in disease by enhancement of a preexisting immune response.     

Lupus/Sjögren''s autoantibody specificities in sera with paraproteins.

Antinuclear antibody and anti-RNA-protein autoantibodies were determined in 143 sera containing paraproteins and 39 control sera. Antinuclear antibodies were commonly present in the paraprotein sera by indirect immunofluorescence. 19 of 143 sera (13%) had elevated anti-Ro/SSA activity in a solid phase Ro/SSA binding assay, and 5 (3.5%) had Ro/SSA precipitating autoantibody. Eighteen sera had La/SSB binding autoantibodies (12%) but only one had an anti-La/SSB precipitin. Anti-nRNP(Sm) was not detected in any of these sera. The solid phase anti-RNA protein assays were repeated using anti-lambda and anti-kappa conjugates. Both lambda and kappa light chain autoantibodies were found in all positive sera consistent with polyclonal anti-Ro/SSA and anti-La/SSB responses. Paraprotein sera containing Ro/SSA precipitins were analyzed by isoelectric focusing followed by exposure to 125I-labeled Ro/SSA and autoradiography. All sera with anti-Ro/SSA binding paraproteins also contained polyclonal anti-Ro/SSA. Our data are consistent with the hypothesis that anti-Ro/SSA paraproteins are common and arise from a previously present polyclonal anti-Ro/SSA response.     

利用重组抗原检测抗SSA＼Ro—52kD自身抗体及其临床意义

目的 重组表达ＳＳＡ／Ｒｏ－５２ｋＤ融合蛋白,并用于检测自身抗体。方法 应用ＤＮＡ重组技术构建ＳＳＡ／Ｒｏ－５２ｋＤ工程菌,并表达ＳＳＡ／Ｒｏ－５２ｋＤ融合蛋白,经谷胱基肽巯基转移酶（ＧＳＴ）柱层析纯化后,作为抗原,分别用免疫印迹法（ＩＢＴ）和酶联免疫吸附试验（ＥＬＩＳＡ）检测２０份ＳＳ患者血清、６０份ＳＬＥ患者血清和３０份正常人血清。结果 重组抗原检测抗 ＳＳＡ／Ｒｏ－５２ｋＤ自身抗体敏感性较高     

免疫斑点法测抗钙调素自身抗体及其与其他8种自身抗体的关系

目的 探讨抗钙调素（ＣａＭ）自身抗体与自身免疫病及其他自身抗体的相互关系。方法 采用ＳＰＡ－免疫斑点法（ＳＰＡ－ＩＤＡ）对３４０例自身免疫病患者,４８例肿瘤患者,５２例正常人血清进行抗ＣａＭ抗体测定,并随机取自身免疫病患者血清同时检测其他８种自身抗体。结果 各类延寿央免疫病患者体内不同程度地存在着抗ＣａＭ自身抗体,４８例肿瘤患者血清中仅１例阳性,５２例正常人血清中无１例阳性。经统计学分析,抗ＣａＭ     

自身免疫病患者可抽提核抗体谱与体液免疫的关系

目的　探讨可抽提核抗原（ Ｅ Ｎ Ａ） 抗体谱的临床意义及与其他免疫指标的关系。方法　对１５６ 份自身免疫病患者血清样本,采用免疫印迹法检测抗 Ｅ Ｎ Ａ 抗体,经免疫浊度法检测 Ｉｇ 和补体。结果　（１） 抗 Ｅ Ｎ Ａ 抗体在自身免疫病有较高的检出率,其中抗平滑肌（ Ｓｍ） 抗体、抗 Ｕ１ 核糖核蛋白抗体的阳性率在系统性红斑狼疮（ Ｓ Ｌ Ｅ） 中分别为３６ ．３ ％ 、２８ ．８ ％ ; 抗 Ｓ Ｓ Ａ 和抗 Ｓ Ｓ Ｂ 抗体的出现在干燥综合征（ Ｓ Ｓ） 患者中分别为５／１６ 和３／１６ 。（２） 与正常对照和抗体阴性组比较,抗 Ｅ Ｎ Ａ 阳性血清中 Ｉｇ Ｇ、 Ｉｇ Ｍ、 Ｉｇ Ａ 增高,分别为（２１ ．６ ±１５ ．０） 、（１ ．７ ±０ ．８） 、（２ ．２ ±１ ．２）ｇ／ Ｌ, 而 Ｃ３ 、 Ｃ４ 和总补体溶血活性（ Ｃ Ｈ５０）水平下降分别为（１ ．０８ ±０ ．３７） 、（０ ．２１ ±０ ．１２） 、（９８ ．±２９） ｇ／ Ｌ;（３） 随着抗 Ｅ Ｎ Ａ 抗体免疫印迹条带数的增多, Ｉｇ Ｇ 增高,而 Ｉｇ Ｍ、 Ｃ３ 、 Ｃ４ 及 Ｃ Ｈ５０ 与之相反,下降很明显;（４） Ｃ３ 、 Ｃ４ 、 Ｃ Ｈ５０ 与疾病活动性有相关关系。结论　抗 Ｅ Ｎ Ａ 抗体阳性者存在体液免疫应答增加,且与抗     

A new assay for thyroglobulin concentration in serum using monoclonal antibodies against synthetic peptides

The concentration of thyroglobulin (Tg) measured by radioimmunoassay (RIA) or enzyme-linked immunosorbent assay (ELISA) is greatly affected by the presence of anti-Tg autoantibodies in sera. We developed a new assay for detecting Tg in the presence of high concentrations of anti-Tg autoantibodies. A 48-kDa fragment was purified from Tg after treatment with V8 protease. This fragment did not appear to bind to two types of monoclonal antibodies (57Ab and 28D3) against a peptide in the C-terminus (amino acids 2735-2748) of Tg and intact Tg, respectively, by ELISA and Western blot analysis. In contrast, anti-Tg autoantibody or anti-Tg polyclonal antibody reacted well with this fragment. Our new ELISA used 57Ab as a solid phase antibody and 28D3 as a antibody conjugated to horseradish peroxidase. Buffer containing purified 48-kDa fragment was used to neutralize autoantibodies against Tg. With this assay, the recovery of Tg was 84.0-89.6% in normal healthy donors (n=5) in the presence of immunoglobulin G (IgG) purified from sera positive for anti-Tg autoantibody, and 76.2-104.4% in patient sera Grave's disease (n=15). Furthermore, the Tg concentrations in sera from patients with Grave's disease (n=20) ranged from 25 to 526 ng/ml, even though the Tg concentration, as measured by a commercial RIA did not exceed 55 ng/ml. There was good agreement between Tg concentrations measured by new Tg-ELISA and commercial Tg-RIA in sera that were negative for anti-Tg autoantibody. Overall, our new ELISA containing a Tg fragment to neutralize the presence of autoantibodies, showed good sensitivity and precision, and may be useful for routine use. Further investigations with the new assay should allow wider assessment of the prevalence and pattern of thyroid autoimmunity or thyroid neoplasms.     

A shared idiotype among human anti-Ro/SSA autoantibodies [published erratum appears in J Exp Med 1990 Feb 1;171(2):591]

Idiotypes and antiidiotypes are thought to be important immune regulators and have provided clues for the origin and pathogenicity of autoantibodies. Many lupus and Sj?gren's syndrome patients, as well as most neonatal lupus infants with congenital heart block or dermatitis, have antibodies to the ribonucleoprotein Ro/SSA, which is one of a group of RNA-protein autoantigens commonly found in human lupus sera. To characterize the fine specificity of anti-Ro/SSA antibodies, a rabbit antidiotypic serum was prepared against polyclonal affinity purified anti-Ro/SSA F(ab')2. The resulting antiidiotype, anti-Id-Rol, is specific for the F(ab')2 fraction of the anti-Ro/SSA immunogen and its binding to anti-Ro/SSA is inhibited by purified Ro/SSA. These data indicate that the Id-Rol epitope on anti-Ro/SSA is associated with the antigen binding site of these same antibodies. The Id-Rol idiotype was present by ELISA in 3 of 12 additional anti-Ro/SSA preparations from precipitin-positive donor sera and in anti-Ro/SSA from one normal donor with low level antibody. This is the first shared idiotype to be found in the human autoantibodies binding to this RNA-protein antigen. Idiotypic differences between anti-Ro/SSA autoantibodies have the potential to explain the variation in pathologic associations found in individuals who develop this autoantibody specificity.     

Enhanced membrane binding of autoantibodies to cultured keratinocytes of systemic lupus erythematosus patients after ultraviolet B/ultraviolet A irradiation.

Although sunlight is known to induce skin lesions in patients with systemic lupus erythematosus (SLE) and to exacerbate systemic manifestations, the underlying mechanisms remain obscure. We report experiments that show enhanced binding of IgG autoantibodies to the cell surface membrane of ultraviolet-B (UVB) irradiated (200-1,600 J/m2) cultured SLE keratinocytes in 10 out of 12 such cell strains. The autoantibody probes showing increased binding were directed against the soluble intracellular antigens, Sm, RNP, SSA/Ro, SSB/La, whereas serum with anti-dsDNA activity did not demonstrate such binding. Control keratinocytes from several sources shared low level binding of autoantibodies after ultraviolet light exposure. In addition, 4/6 UVB-sensitive SLE strains showed increased autoantibody binding to the surface of SLE keratinocytes after UVA exposure (50-150 kJ/m2), but of lower magnitude. When UVB-sensitive nonirradiated SLE strains were exposed to autologous serum, 3/8 sera demonstrated a striking increase in IgG binding, which increased further after UVB exposure. Enhanced expression of saline-soluble intracellular antigens on the cell surface membrane of patient, but not control, keratinocytes may, in part, explain the photosensitivity of patients with SLE.     

Rapid and sensitive detection of autoantibody in rheumatoid arthritis patients by heat-mediated ELISA

OBJECTIVE: Heat-mediated ELISA (HELISA) technique has been reported to shorten ELISA timings and has major clinical implications in diagnostic field [Bora U, Kannan K, Nahar P. Heat mediated enzyme linked immunosorbent assay. J Immunol Methods 2004; 293: 43-50]. Objective of this study is to find out whether anti-MBL autoantibody can be detected rapidly by HELISA technique. DESIGN AND METHODS: Activated polystyrene microtiter plate was prepared by a photolinker in a photochemical reaction carried out over a period of 10 min. Lectin was covalently immobilized onto the activated plate at 50 degrees C in 45 min. Antigenicity of the immobilized lectin was checked by HELISA carried out at elevated temperature (50 degrees C). The result was further compared with that obtained by conventional ELISA method carried out on an untreated plate over an incubation period of 18 h. RESULTS: Autoantibody detection carried out by HELISA method (intra- and inter-assay CVs were <9.8%) in 2 h 45 min gives absorbance value comparable to that of conventional ELISA (intra- and inter-assay CVs were <12%) carried out in 18 h in RA patients and healthy controls (n=100). HELISA on a photoactivated surface showed 1.5-fold higher absorbance than those obtained on untreated surface (p=0.00019). The HELISA method is more sensitive (AUC=0.967, 95% CI=0.948-0.987) and can be used to detect autoantibody even at higher dilution than by conventional assay. Excellent correlation was observed when autoantibodies were detected by HELISA and conventional ELISA (R=0.9706). CONCLUSION: The present method provides rapid and sensitive detection of autoantibody in rheumatoid arthritis patients. The method is precise and reliable similar to method currently used in diagnostics and could be potentially useful for other immunoassays.     

3种检测方法在诊断自身免疫性疾病中的价值

目的通过对抗核抗体、可提取性核抗原(ENA)及抗核抗体谱3检测方法的分析比较,探讨有益于临床诊断的检测组合。方法采用人喉癌上皮细胞及猴肝作为基质的间接免疫荧光法检测抗核抗体;应用欧蒙斑点法检测可提取性核抗原中的如下抗原:nRNP/Sm、Sm、SSA、SSB、Scl-70、Jo-1;以绿蝇短膜虫为基质,应用间接免疫荧光法检测抗双链DNA抗体;通过欧蒙印迹法检测15种自身抗体。结果将3种稀释度(1:10、1:80、1:100)抗核抗体检测的阳性率进行比较,1:10及1:80稀释的血清抗核抗体阳性率差异有统计学意义(P0.01),其余2种比较差异无统计学意义;在核点型、胞浆颗粒型及均质型等多种核型的检测中抗核抗体谱3的阳性检出率高于ENA谱;在抗双链DNA抗体的检测中间接免疫荧光法的阳性检出率略高于欧蒙印迹法。结论在应用间接免疫荧光法的抗核抗体检测中,使用1:80的稀释度更有利于提高检出率,而且涵盖15种抗原的抗核抗体谱3也为临床诊断提供了有利的依据。     

Autoantibody formation in burn patients after inhibition of suppressor T cell activity with polymyxin B

After thermal injury, treatment with polymyxin B blocks suppressor T cell activity by uncoupling endotoxin-mediated T cell activation, but the effect on autoantibody formation is unknown. We examined the presence of antinuclear antibodies to native DNA; to soluble antigens Ro/SSA, La/SSB, Sm, nRNP; and to antiepithelial antibodies in 12 burn patients before and after treatment with polymyxin B and in 24 samples from control burn patients. Low titer antinuclear antibody activity was detected in 25% of pretreatment and 78% of posttreatment samples (p less than 0.01) and in 16.7% of control patients. One polymyxin B-treated patient had a significant antinuclear antibody titer both before and after treatment. Antiepithelial antibodies were detected in 16.7% of early polymyxin B-treated samples and 11.1% of late samples (p less than 0.05) but were also present in 20.8% of controls. Antibodies to native DNA, Ro/SSA, La/SSB, Sm, and nRNP were not detected in any sera.     

Sensitive non-isotopic assays for autoantibodies to IA-2 and to a combination of both IA-2 and GAD65

BACKGROUND: A sensitive ELISA for measurement of IA-2 autoantibodies has been developed and assessed. Also, a combination ELISA for detection of both GAD65 autoantibodies and IA-2 autoantibodies is described. METHODS: The IA-2 autoantibody assay is based on the ability of IA-2 autoantibodies to form a bridge between IA-2 intracellular fragment coated onto ELISA plate wells and liquid-phase IA-2 labelled with biotin. The combination ELISA uses plates coated with both IA-2 and GAD65 and a mixture of IA-2-biotin and GAD65-biotin. Assay sensitivity was assessed using the WHO reference (NIBSC 97/550) for islet cell antibodies. IA-2 autoantibody measurements by ELISA were compared with measurements in immunoprecipitation assays (IPAs) based on 125I or 35S labelled IA-2. Combination ELISA results were compared with results obtained for individual autoantibodies. RESULTS: As little as 15 units/mL of NIBSC 97/550 was detectable in the IA-2 autoantibody ELISA compared to 125 units/mL by 125I-IA-2 IPA. 110/216(51%) sera from patients with type 1 DM were positive in the IA-2 autoantibody ELISA while 97/216 (45%) and 91/216 (42%) were positive in the 125I-IA2 and 35S-IA-2 IPAs, respectively. The IA-2 autoantibody ELISA showed 100% specificity for type 1 DM. The combination ELISA was able to detect GAD65 and/or IA-2 autoantibodies in 183/216 (85%) diabetic sera and 183/216 (85%) were also found positive for autoantibodies to IA-2 and/or to GAD65 in the assays for individual antibodies. Autoantibody measurements in the individual autoantibody assays and in the combination ELISA showed good agreement by Pearson correlation (r=0.92, n=216, p<0.001) and by Bland and Altman analysis. CONCLUSIONS: Sensitive and specific ELISAs for measurement of autoantibodies to IA-2 and to a combination of IA-2 and GAD65 have been developed. These assays are suitable for screening large numbers of samples in diabetes prediction and prevention trials.     

Screening autoantibody profiles in systemic rheumatic disease with a diagnostic protein microarray that uses a filtration-assisted nanodot array luminometric immunoassay (NALIA)

BACKGROUND: We developed a cost-efficient modular system for multiplex analysis of the multiple autoantibodies that characterize systemic rheumatoid diseases. METHODS: The nanodot array luminometric immunoassay (NALIA) system consists of conventional 96-well membrane-bottomed plates in which antigens or antibodies are adsorbed onto the underside of the membrane. Current arrays use a 5 x 5 format (25 dots/well), which allows 10 analytes to be measured in duplicate: double-stranded DNA (dsDNA), centromere protein B (CENP-B), PCNA, Sm, Sm ribonucleoprotein (Sm-RNP), U1-snRNP, Scl70, SSA/Ro, SSB/La, Jo-1, and controls. The test fluid, control sera, and subsequent reagents are drawn through the membrane. The captured analytes are quantified by monitoring chemiluminescence with a charge-coupled device (CCD) and analyzed with commercial array software. RESULTS: The assay can detect <20 x 10(3) IU/L of anti-dsDNA. The interwell CV was 10%-14%. There was an 83% concordance (kappa = 0.56) between the NALIA results obtained for anti-dsDNA assayed by beta-testing in a routine immunology diagnostic laboratory and the results obtained with a conventional ELISA reagent set. The concordance values for Ro, La, Sm, and RNP were 98% (kappa, 0.92), 93% (kappa, 0.41), 97% (kappa, 0.62), and 97% (kappa, 0.73), respectively. CONCLUSION: The NALIA approach promises to provide a highly economical platform for a wide range of applications that require assays of multiple analytes. The degree of concordance of our results with a conventional reagent set was no less than that occurring between different commercial products. A sample of serum from a finger stick provides a volume sufficient to perform the array assay.     

Protein microarray analysis reveals BAFF-binding autoantibodies in systemic lupus erythematosus

Autoantibodies against cytokines, chemokines, and growth factors inhibit normal immunity and are implicated in inflammatory autoimmune disease and diseases of immune deficiency. In an effort to evaluate serum from autoimmune and immunodeficient patients for Abs against cytokines, chemokines, and growth factors in a high-throughput and unbiased manner, we constructed a multiplex protein microarray for detection of serum factor–binding Abs and used the microarray to detect autoantibody targets in SLE. We designed a nitrocellulose-surface microarray containing human cytokines, chemokines, and other circulating proteins and demonstrated that the array permitted specific detection of serum factor–binding probes. We used the arrays to detect previously described autoantibodies against cytokines in samples from individuals with autoimmune polyendocrine syndrome type 1 and chronic mycobacterial infection. Serum profiling from individuals with SLE revealed that among several targets, elevated IgG autoantibody reactivity to B cell–activating factor (BAFF) was associated with SLE compared with control samples. BAFF reactivity correlated with the severity of disease-associated features, including IFN-α–driven SLE pathology. Our results showed that serum factor protein microarrays facilitate detection of autoantibody reactivity to serum factors in human samples and that BAFF-reactive autoantibodies may be associated with an elevated inflammatory disease state within the spectrum of SLE.     

重组抗原检测抗干燥综合征A 抗原的自身抗体

目的 建立便捷的检测抗干燥综合征（ＳＳ）Ａ抗原（分子量为６００００的多肽成分）的自身抗体（抗ＳＳＡ）的方法,以利用疾病的早期诊断和病程监控。方法 构建基因工程菌,表达ＳＳＡ－６００００融合蛋白。经ＧＳＴ柱层析法纯化后,分别经免疫印迹（ＩＢＴ）法和酶联免疫吸附法（ＥＬＩＳＡ）检测２０份ＳＳ患者血清、６０份系统性红斑狼疮（ＳＬＥ）患者血清和３０份正常人血清。结果 利用重组抗原检测抗ＳＳＡ自身抗体敏感性     

Analysis of speckled fluorescent antinuclear antibody test antisera using electrofocused nuclear antigens.

Antibodies to different components of the extractable nuclear antigen (ENA) have been thought to be serological markers for clinical subsets of rheumatic diseases. However, incomplete characterization and standardization of antigenic components such as ribonucleoprotein (RNP), Sm, and SS-B (Ha), and the multiplicity of autoantibodies produced by different patients have confounded correlations between autoantibody specificity and disease subsets. This study describes the preparative separation of the antigens Sm, RNP, and Ss-B (Ha) by electrofocusing and their use in a rocket electrophoretic assay that in one step identifies and quantifies the multiple reactivities of patient sera exhibiting the speckled FANA pattern. Preparative electrofocusing generates milligram quantities of these antigens with retention of their immunologic and biochemical characteristics, facilitating further study of their biological properties and relationships to disease subsets.     

SS-56, a novel cellular target of autoantibody responses in Sjögren syndrome and systemic lupus erythematosus

Certain autoimmune disorders, including Sj?gren syndrome (SS) and systemic lupus erythematosus (SLE), are characterized by autoantibodies against the Ro/SSA and La/SSB cellular antigens. Although the implication of these autoantibodies in disease pathogenesis is still unclear, it is believed that the aberrant responses against autoantigens may extend to other proteins that are not yet well defined. In an attempt to analyze the regulated gene expression in lymphocytes by an HIV-suppressive immunomodulator, we have identified and cloned a novel gene encoding a 56-kDa protein, named SS-56, which is structurally related to the 52-kDa Ro/SSA antigen. The new protein showed primarily perinuclear cytoplasmic localization, and recombinant SS-56 was found to react in ELISA with sera from most patients with SS or SLE. Western blot analysis confirmed the autoantigenic nature of native SS-56 in extracts from HeLa cells. Interestingly, the incidence of antibodies to SS-56 was associated with visceral complications in SLE, and roughly half of the 17 SS or SLE patients with no detectable antibodies to SSA and SSB antigens presented measurable antibodies against recombinant SS-56. Thus, SS-56 represents a new member of the SS family of autoantigens and could become an additional and important diagnostic marker for SS and SLE.     

基于免疫金银染色的蛋白质与抗原分子微阵列技术

目的　研制适于自身抗体检测的蛋白质与抗原分子微阵列。方法　利用氧化型琼脂糖凝胶膜结合戊二醛分子衍生的活性表面 ,制作蛋白质与抗原分子微阵列 ,建立了检测血清自身抗体的胶体金免疫金银染色方法 (IGSS)。结果　在未经选择的风湿病患者中 (包括SLE ,RA和MCTD) ,检出多种自身抗体阳性 ,其中又以子宫内膜抗体 (EmAb)、抗ssDNA抗体、抗TG抗体、抗TPO抗体和RF等的检出频率较高 ;但心磷脂抗体 (ACA)、抗LSP和抗精子抗体 (AsAb)均为阴性。经初步方法学对比考证 ,实验结果与ELISA有较好的一致性 (达 90 % )。结论　本法具有方法敏感 ,结果直观和实验条件易于控制等特点 ,为发展可视化阵列芯片提供了一种很好的模式。