A study of anti-group A streptococcal monoclonal antibodies cross-reactive with myosin.
A study of anti-group A streptococcal monoclonal antibodies cross-reactive with myosin.
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DOI：
10.4049/jimmunol.136.1.293
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发表时间：
1986-01
影响因子：
4.4
通讯作者：
M. Cunningham;N. K. Hall;K. Krisher;A. M. Spanier
中科院分区：
文献类型：
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作者：
M. Cunningham;N. K. Hall;K. Krisher;A. M. Spanier
用纯化的M型5化脓性链球菌膜免疫BALB c/BYJ小鼠，获得抗A组链球菌单克隆抗体。两种抗链球菌单克隆抗体先前显示与肌球蛋白交叉反应。在本研究中，单克隆抗体与正常人心脏和骨骼肌组织切片反应。通过间接免疫荧光和免疫过氧化物酶技术估计抗体结合。本报告研究的两种单克隆抗体（36.2.2和54.2.8）均与心脏和/或骨骼肌切片反应。免疫荧光检测显示，单克隆抗体54.2.8可与心肌横纹肌细胞外周划界，并与肌上皮下成分发生较小程度的反应。单克隆抗体36.2.2不能与心脏切片反应，但两种单克隆抗体与骨骼肌切片反应强烈。用免疫过氧化物酶技术获得了与间接免疫荧光相似的结果。通过Western免疫印迹和竞争抑制实验，发现单克隆抗体36.2.2和54.2.8均与骨骼肌肌球蛋白重链反应。而与心肌肌球蛋白重链反应的只有54.2.8。单克隆抗体对骨骼肌肌球蛋白亚片段的特异性表明，单克隆抗体36.2.2对轻肌球蛋白片段具有特异性，而单克隆抗体54.2.8对重肌球蛋白和轻肌球蛋白都有特异性。数据表明，两种抗链球菌单克隆抗体对骨骼肌和/或心脏肌球蛋白以及肌球蛋白分子亚片段具有特异性。单克隆抗体与人组织切片的反应与单克隆抗体与变性肌球蛋白和天然肌球蛋白的免疫化学反应一致。
Anti-group A streptococcal monoclonal antibodies were obtained from BALB c/BYJ mice immunized with purified membranes from M type 5 Streptococcus pyogenes. Two of the anti-streptococcal monoclonal antibodies were previously shown to cross-react with muscle myosin. In this study the monoclonal antibodies were reacted with tissue sections of normal human heart and skeletal muscle. Antibody binding was estimated by indirect immunofluorescence and immunoperoxidase techniques. Both of the monoclonal antibodies (36.2.2 and 54.2.8) investigated in this report reacted with heart and/or skeletal muscle sections. When evaluated by immunofluorescence, monoclonal antibody 54.2.8 demarcated the periphery of cardiac striated muscle cells and reacted to a lesser degree with subsarcolemmal components. Monoclonal antibody 36.2.2 failed to react with heart sections, but both of the monoclonal antibodies reacted strongly with skeletal muscle sections. Results similar to those observed with indirect immunofluorescence were obtained with the immunoperoxidase technique. By Western immunoblotting and competitive inhibition assays, monoclonal antibodies 36.2.2 and 54.2.8 both were found to react with the heavy chain of skeletal muscle myosin. However, only 54.2.8 reacted with the heavy chain of cardiac myosin. The specificity of the monoclonal antibodies for subfragments of skeletal muscle myosin indicated that monoclonal antibody 36.2.2 was specific for light meromyosin fragments, whereas 54.2.8 reacted with both heavy and light meromyosin. The data demonstrated that two monoclonal antibodies against streptococci were specific for skeletal muscle and/or cardiac myosin and for subfragments of the myosin molecule. The reactions of the monoclonal antibodies with human tissue sections were consistent with the immunochemical reactions of the monoclonal antibodies with both denatured and native myosin.