Cabiedes Javier, Cabral Antonio R, López-Mendoza A Tamara, Cordero-Esperón Hugo A, Huerta M Teresa, Alarcón-Segovia Donato
Department of Immunology and Rheumatology, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Mexico City, México.
J Autoimmun. 2002 Mar;18(2):181-90. doi: 10.1006/jaut.2001.0575.
In 1990 our group reported a patient with autoimmune hemolytic anemia and high titers of IgM anticardiolipin antibodies that cross-reacted with phosphatidylcholine (PTC). These autoantibodies also recognized bromelain-treated erythrocytes (BrE) and in vitro aged erythrocytes. The epitope exposed with this treatment is PTC. To detect and characterize antiphosphatidylcholine antibodies (anti-PTC) in a normal human population, we studied by ELISA the presence of serum anti-PTC (IgG and IgM) in clinically healthy human subjects. The most representative samples were also studied for IgG or IgM activity against BrE by flow cytometry, rheumatoid factor activity, anti-dsDNA, anti-ssDNA by ELISA and by indirect immunofluorecence (IIF) using HEp-2 line and a healthy human fibroblast strain as substratum. Eighty five percent of sera had IgM anti-PTC and none had IgG. IgM antibodies against BrE were inhibited by PTC micelles (mPTC). Anti-PTC were also inhibited by phosphorylcholine and phosphatidic acid. Aggregated gammaglobulin (AGG) reactivity was inhibited by dsDNA and mPTC. The IgM anti-dsDNA activity was inhibited by soluble dsDNA, AGG and mPTC. All sera gave intermediate filaments pattern by IIF and reacted against purified vimentin by dot blot and Western blot.Our study shows hemolytic IgM anti-PTC present in normal human serum. The main epitope recognized by these autoantibodies is phosphorylcholine. The physicochemical characteristics, crossreactivity with self-antigens and functional properties are typical features of natural autoantibodies.
1990年,我们小组报告了一名患有自身免疫性溶血性贫血且IgM抗心磷脂抗体滴度高的患者,该抗体与磷脂酰胆碱(PTC)发生交叉反应。这些自身抗体还识别菠萝蛋白酶处理的红细胞(BrE)和体外老化的红细胞。这种处理后暴露的表位是PTC。为了检测和表征正常人群中的抗磷脂酰胆碱抗体(抗PTC),我们通过ELISA研究了临床健康人类受试者血清中抗PTC(IgG和IgM)的存在情况。还通过流式细胞术研究了最具代表性样本针对BrE的IgG或IgM活性、类风湿因子活性、通过ELISA检测抗双链DNA、抗单链DNA以及使用HEp-2细胞系和健康人成纤维细胞系作为基质通过间接免疫荧光法(IIF)进行检测。85%的血清含有IgM抗PTC,无一含有IgG。PTC微团(mPTC)可抑制针对BrE的IgM抗体。抗PTC也可被磷酸胆碱和磷脂酸抑制。聚集的丙种球蛋白(AGG)反应性可被双链DNA和mPTC抑制。IgM抗双链DNA活性可被可溶性双链DNA、AGG和mPTC抑制。所有血清通过IIF呈现中间丝模式,并通过斑点印迹和蛋白质印迹法与纯化的波形蛋白发生反应。我们的研究表明正常人血清中存在溶血性IgM抗PTC。这些自身抗体识别的主要表位是磷酸胆碱。其物理化学特性、与自身抗原的交叉反应性和功能特性是天然自身抗体的典型特征。