Lyons J, Sinos C, Destree A, Caiazzo T, Havican K, McKenzie S, Panicali D, Mahr A
Applied bioTechnology, Cambridge, Massachusetts 02142.
Infect Immun. 1990 Dec;58(12):4089-98. doi: 10.1128/iai.58.12.4089-4098.1990.
Eight Mycobacterium tuberculosis and M. leprae genes were inserted into the vaccinia virus genome by in vivo recombination. The resulting virus recombinants were shown to express five different M. tuberculosis proteins (71, 65, 35, 19, and 12 kDa) and three M. leprae proteins (65 and 18 kDa and a biotin-binding protein) by Western immunoblot analysis, radioimmunoprecipitation, or black-plaque assay. When injected into BALB/c mice, the recombinants expressing the M. tuberculosis 71-, 65-, or 35-kDa protein and the M. leprae 65-kDa protein or the biotin-binding protein elicited antibodies against the appropriate M. tuberculosis or M. leprae protein. These vaccinia virus recombinants are being tested for the ability to elicit immune protection against M. tuberculosis or M. leprae challenge in animal model systems. The recombinants are also useful in generating target cells for assays aimed at elucidating the cellular immune responses to mycobacterial proteins in leprosy and tuberculosis. Furthermore, the M. tuberculosis 65-kDa protein and four of the other mycobacterial proteins share homology with known eucaryotic and procaryotic stress proteins, some of which may play a role in autoimmunity.
通过体内重组将8个结核分枝杆菌和麻风分枝杆菌基因插入痘苗病毒基因组。通过蛋白质免疫印迹分析、放射免疫沉淀或黑斑试验表明，所得病毒重组体表达5种不同的结核分枝杆菌蛋白（71、65、35、19和12 kDa）和3种麻风分枝杆菌蛋白（65和18 kDa以及一种生物素结合蛋白）。当将这些重组体注射到BALB/c小鼠体内时，表达结核分枝杆菌71 kDa、65 kDa或35 kDa蛋白以及麻风分枝杆菌65 kDa蛋白或生物素结合蛋白的重组体可引发针对相应结核分枝杆菌或麻风分枝杆菌蛋白的抗体。目前正在动物模型系统中测试这些痘苗病毒重组体引发针对结核分枝杆菌或麻风分枝杆菌攻击的免疫保护的能力。这些重组体也可用于生成靶细胞，用于旨在阐明麻风病和结核病中针对分枝杆菌蛋白的细胞免疫反应的检测。此外，结核分枝杆菌65 kDa蛋白和其他4种分枝杆菌蛋白与已知的真核和原核应激蛋白具有同源性，其中一些可能在自身免疫中起作用。
