Blanchard-Channell M, Stott J L
Department of Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis 95616.
Infect Immun. 1991 Aug;59(8):2790-8. doi: 10.1128/iai.59.8.2790-2798.1991.
Three monoclonal antibodies (F6F3, F6B11, and F6B3) were developed against Borrelia coriaceae antigens. All three antibodies appeared to be specific for this species and did not cross-react with Borrelia burgdorferi (strains B31 and IRS), Borrelia hermsii, Borrelia anserina, Leptospira interrogans serovar hardjo, or Treponema hyodysenteriae, as determined by indirect fluorescent antibody staining, enzyme-linked immunosorbent assay, and Western immunoblot analysis. Only one of these antibodies, F6B3, bound to spirochetes present in organ smears from the argasid tick, Ornithodoros coriaceus. The antigens recognized by F6F3, F6B11, and F6B3 have apparent molecular weights of ca. 37,000, 35,000, and 16,000, respectively, as determined by Western blot analysis. Antigens were analyzed by immune electron microscopy as well as Western blot and indirect fluorescent antibody staining analysis of spirochetes after enzyme (trypsin and protease K) and detergent (Triton X-100) treatments. These studies suggest that all three antigens are integral membrane proteins. The characteristics of the 37K and 35K proteins are consistent with the outer surface proteins of B. burgdorferi (OSP A and OSP B) described by Barbour et al. (A. G. Barbour, S. L. Tessier, and S. F. Hayes, Infect. Immun. 45:94-100, 1984), while data regarding the 16K protein are less conclusive but may suggest a cytoplasmic membrane location. We suggest that the 37K, 35K, and 16K antigens be designated integral membrane proteins A, B, and C, respectively, as a result of these studies.
制备了三种抗科氏疏螺旋体抗原的单克隆抗体(F6F3、F6B11和F6B3)。通过间接荧光抗体染色、酶联免疫吸附测定和Western免疫印迹分析确定,这三种抗体似乎都对该物种具有特异性,并且不与伯氏疏螺旋体(菌株B31和IRS)、赫氏疏螺旋体、鹅疏螺旋体、问号钩端螺旋体哈焦血清型或猪痢疾密螺旋体发生交叉反应。这些抗体中只有一种,即F6B3,与钝缘蜱科氏钝缘蜱器官涂片中存在的螺旋体结合。通过Western印迹分析确定,F6F3、F6B11和F6B3识别的抗原的表观分子量分别约为37,000、35,000和16,000。通过免疫电子显微镜以及对螺旋体进行酶(胰蛋白酶和蛋白酶K)和去污剂(Triton X-100)处理后的Western印迹和间接荧光抗体染色分析对抗原进行了分析。这些研究表明,所有三种抗原都是整合膜蛋白。37K和35K蛋白的特征与Barbour等人(A.G.Barbour、S.L.Tessier和S.F.Hayes,《感染与免疫》45:94-100,1984)描述的伯氏疏螺旋体的外表面蛋白(OSP A和OSP B)一致,而关于16K蛋白的数据不太确凿,但可能表明其位于细胞质膜。由于这些研究,我们建议将37K、35K和16K抗原分别命名为整合膜蛋白A、B和C。