Hubbard R D, Flory C M, Collins F M
Trudeau Institute, Inc., Saranac Lake, NY 12983.
Clin Exp Immunol. 1992 Jan;87(1):94-8. doi: 10.1111/j.1365-2249.1992.tb06419.x.
Culture filtrate proteins were obtained from Mycobacterium tuberculosis cultures after 7 days growth in Proskauer and Beck medium. The protein yield increased substantially to peak about the time the number of viable organisms reached its maximum level (day 8). Examination of the protein concentrate by SDS-PAGE revealed the presence of at least 12 separate protein bands varying from 10 to 90 kD. Mice were injected subcutaneously with 20 micrograms of M. tuberculosis culture filtrate (MTCF) protein suspended in saline or Freund's complete or incomplete adjuvant. The vaccinated mice were subjected to an aerogenic challenge with 10(3) colony-forming unit (CFU) M. tuberculosis Erdman and a significant reduction in the number of viable organisms was observed in the spleens and lungs determined over a 21-day period compared with age-matched normal controls. Mice immunized with the same culture filtrate proteins bound to nitrocellulose particles also showed some resistance to the virulent challenge, suggesting that individual antigens present in the culture filtrate were able to induce a protective T cell-mediated immune response in appropriately immunized mice.
在普罗斯考尔和贝克培养基中培养7天后，从结核分枝杆菌培养物中获得培养滤液蛋白。蛋白质产量在活菌数量达到最高水平（第8天）时大幅增加至峰值。通过SDS-PAGE对蛋白质浓缩物进行检测，发现存在至少12条单独的蛋白条带，分子量从10到90 kD不等。将20微克悬浮于生理盐水或弗氏完全或不完全佐剂中的结核分枝杆菌培养滤液（MTCF）蛋白皮下注射给小鼠。对接种疫苗的小鼠用10³菌落形成单位（CFU）的结核分枝杆菌埃尔德曼进行气溶胶攻击，与年龄匹配的正常对照相比，在21天的观察期内，脾脏和肺部的活菌数量显著减少。用结合到硝酸纤维素颗粒上的相同培养滤液蛋白免疫的小鼠也表现出对强毒攻击的一定抵抗力，这表明培养滤液中存在的单个抗原能够在适当免疫的小鼠中诱导保护性T细胞介导的免疫反应。
