Ribi E, Larson C, Wicht W, List R, Goode G
J Bacteriol. 1966 Mar;91(3):975-83. doi: 10.1128/jb.91.3.975-983.1966.
Ribi, Edgar (Rocky Mountain Laboratory, Hamilton, Mont.), Carl Larson, William Wicht, Robert List, and Granville Goode. Effective nonliving vaccine against experimental tuberculosis in mice. J. Bacteriol. 91:975-983. 1966.-Antituberculosis vaccines were prepared in one of three manners: lyophilized BCG suspended in light mineral oil was disrupted in a Sorvall pressure cell and the "oil disruption product" was collected by centrifugation; BCG was disrupted in water, lyophilized, and worked into a paste with a small amount of oil (about 0.16 ml per 50 mg); BCG was disrupted in water, and the cell wall fraction was isolated, lyophilized, and prepared in an oil paste. These vaccines were suspended in Tween-saline to a concentration of 5 mg/ml and heated at 65 C for 30 min. In protection tests based on pulmonary infection with Mycobacterium tuberculosis H37Rv, the median number of virulent organisms in lung tissue of mice immunized with a few hundred micrograms of these three vaccines was 3 to 4 logs lower than in unvaccinated control mice. A similar dose of viable BCG standard vaccine reduced the lung count 1 to 2 logs below the controls. Protection afforded by nonviable, whole BCG, with or without oil, was of only borderline significance. Since oil-treated fractions containing cell walls produced effective immunity, while the oil-treated protoplasm or whole cells were not active, the protective antigen appeared to be an inner component of the cell wall, exposed when the cell was disrupted, and activated by oil. Extraction of oil from immunogenic disruption products resulted in loss of ability of the products to confer protection against the aerosol challenge, whereas high protection against the conventional challenge by intravenous infection with up to 1.4 x 10(8) cells of M. tuberculosis H37Rv was retained. Retreatment with oil of these nonimmunogenic products restored the immunogenicity if the oil was applied to dried products. The consistent finding that moisture interferes with the enhancement of the vaccine potency by oil suggested that such enhancement may not be the same as that ordinarily produced by water-in-oil emulsions.
里比，埃德加（落基山实验室，蒙大拿州汉密尔顿），卡尔·拉尔森，威廉·维希特，罗伯特·利斯特，以及格兰维尔·古德。针对小鼠实验性结核病的有效非活性疫苗。《细菌学杂志》91:975 - 983。1966年。——抗结核疫苗通过以下三种方式之一制备：将冻干卡介苗悬浮于轻质矿物油中，在索瓦压力细胞中破碎，通过离心收集“油破碎产物”；卡介苗在水中破碎，冻干，与少量油（每50毫克约0.16毫升）制成糊剂；卡介苗在水中破碎，分离细胞壁部分，冻干，并制成油糊剂。这些疫苗悬浮于吐温 - 盐溶液中，浓度为5毫克/毫升，并在65℃加热30分钟。在基于结核分枝杆菌H37Rv肺部感染的保护试验中，用几百微克这三种疫苗免疫的小鼠肺组织中致病生物体的中位数数量比未接种疫苗的对照小鼠低3至4个对数级。类似剂量的活卡介苗标准疫苗使肺内菌数比对照低1至2个对数级。含细胞壁的经油处理部分提供的保护仅具有临界意义。含细胞壁的经油处理部分产生有效免疫力，而经油处理的原生质或全细胞无活性，保护性抗原似乎是细胞壁的内部成分，在细胞破碎时暴露，并被油激活。从免疫原性破碎产物中提取油导致产物丧失针对气溶胶攻击的保护能力，而对高达1.4×10⁸个结核分枝杆菌H37Rv细胞静脉感染的传统攻击仍具有高保护作用。如果将油应用于干燥产物，这些非免疫原性产物用油再处理可恢复免疫原性。始终发现水分会干扰油对疫苗效力的增强作用，这表明这种增强作用可能与通常由油包水乳液产生的增强作用不同。
