Kemeny E, Husby G, Williams R C, Zabriskie J B
Department of Medicine, University of Florida, Gainesville 32610.
Clin Immunol Immunopathol. 1994 Jul;72(1):35-43. doi: 10.1006/clin.1994.1104.
Monoclonal antibody D8/17 originally prepared by immunization of mice with B cells from a patient with rheumatic heart disease (RHD) reacts with epitopes expressed on significantly elevated proportions of B cells (10-35%) from all patients with acute rheumatic fever or rheumatic heart disease. This B cell marker does not segregate with any known HLA or DR phenotype. We have examined the reactivity of D8/17 with a broad assortment of human tissues, using indirect immunofluorescence. Strong positive reactivity of D8/17 was observed with cardiac muscle, skeletal muscle, and smooth muscle of blood vessels. Positive fluorescent staining was also noted in cell membranes of hepatocytes and cells lining bile canaliculi as well as in epithelial cells of skin, esophagus, and cervix. D8/17 mAb binding to cardiac muscle was markedly diminished by preincubation of mAb with KH B cell line originally established from an RHD patient. D8/17 mAb binding to human heart was also inhibited by preincubation with myosin and tropomyosin but not by actin. Using the D8/17 mAb in immunoblots, positive binding was noted by the antibody to recombinant type M6 protein, vimentin, and myosin. Our findings indicate that the B cell antigen reacting with mAb D8/17 may be related to contractile proteins present in heart, skeletal and smooth muscle, and may also share epitopes with some components of group A streptococci.
最初通过用风湿性心脏病(RHD)患者的B细胞免疫小鼠制备的单克隆抗体D8/17,可与所有急性风湿热或风湿性心脏病患者中比例显著升高(10%-35%)的B细胞上表达的表位发生反应。这种B细胞标志物与任何已知的HLA或DR表型均无关联。我们使用间接免疫荧光法检测了D8/17与多种人体组织的反应性。观察到D8/17在心肌、骨骼肌和血管平滑肌上有强阳性反应。在肝细胞和胆小管内衬细胞的细胞膜以及皮肤、食管和宫颈的上皮细胞中也发现了阳性荧光染色。用最初从一名RHD患者建立的KH B细胞系预先孵育单克隆抗体后,D8/17单克隆抗体与心肌的结合明显减弱。用肌球蛋白和原肌球蛋白预先孵育也可抑制D8/17单克隆抗体与人心脏的结合,但肌动蛋白无此作用。在免疫印迹中使用D8/17单克隆抗体时,该抗体与重组M6型蛋白、波形蛋白和肌球蛋白有阳性结合。我们的研究结果表明,与单克隆抗体D8/17发生反应的B细胞抗原可能与心脏、骨骼肌和平滑肌中存在的收缩蛋白有关,并且可能与A组链球菌的某些成分共享表位。