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首页> 外文期刊>Infection and immunity >Production of a monoclonal antibody specific for Mycobacterium avium and immunological activity of the affinity-purified antigen.
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Production of a monoclonal antibody specific for Mycobacterium avium and immunological activity of the affinity-purified antigen.

机译：对鸟分枝杆菌有特异性的单克隆抗体的生产和亲和纯化抗原的免疫活性。

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Three hybridomas which secrete antibodies to Mycobacterium avium were obtained by the fusion of p3u1 myeloma cells with spleen cells of mice immunized with M. avium culture sonicate. The reactivity of these monoclonal antibodies was determined in 16 species of mycobacteria by an enzyme-linked immunosorbent assay. An antibody, designated Avi-3, reacted only with M. avium and not with the reference strains of the other 15 species of mycobacteria tested, including M. intracellulare, M. paratuberculosis, and M. lepraemurium. Specificity of the antibody was confirmed by assay, using a specific DNA probe of M. avium complex in 29 M. avium complex isolates. An antigen was purified from M. avium culture sonicate on a monoclonal antibody Avi-3-coupled affinity column. The purified antigen gave a single band (molecular size, about 27 kilodaltons) upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The antigen Avi-3 showed strong skin test activity in guinea pig preimmunized with heat-killed M. avium but not in those sensitized with heat-killed M. intracellulare or M. bovis BCG. Purified protein derivative elicited positive skin reactions in all the immunized guinea pigs. When heat-killed M. avium was used as the immunogen, strong lymphoproliferative responses were observed in cultures stimulated with the antigen Avi-3. These results suggest that M. avium-specific antigen Avi-3 may facilitate the diagnosis of mycobacterial infections.
机译：通过将p3u1骨髓瘤细胞与用鸟分枝杆菌培养物超声处理的小鼠的脾细胞融合，获得了三种分泌鸟分枝杆菌抗体的杂交瘤。这些单克隆抗体的反应性通过酶联免疫吸附测定法在16种分枝杆菌中测定。命名为Avi-3的抗体仅与鸟分枝杆菌发生反应，而与其他15种分枝杆菌的参考菌株（包括胞内分枝杆菌，副结核分枝杆菌和麻风分枝杆菌）没有反应。通过使用29个鸟形支原体复合体分离物中的鸟形支原体复合体的特异性DNA探针，通过测定证实了抗体的特异性。在单克隆抗体Avi-3偶联的亲和柱上从鸟分枝杆菌培养物中超声纯化抗原。经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳后，纯化的抗原产生一条单一的条带（分子大小，约27道尔顿）。抗原Avi-3在用热灭活的鸟分枝杆菌预免疫的豚鼠中显示出强大的皮肤测试活性，但在用热灭活的胞内分枝杆菌或牛分枝杆菌BCG致敏的豚鼠中却没有。纯化的蛋白衍生物在所有免疫的豚鼠中引起阳性皮肤反应。当使用热杀死的鸟分枝杆菌作为免疫原时，在用抗原Avi-3刺激的培养物中观察到了强烈的淋巴增生反应。这些结果表明，鸟分枝杆菌特异性抗原Avi-3可能有助于分枝杆菌感染的诊断。

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