Youmans A S, Youmans G P
J Bacteriol. 1966 Jun;91(6):2139-45. doi: 10.1128/jb.91.6.2139-2145.1966.
Youmans, Anne S. (Northwestern University Medical School, Chicago, Ill.), and Guy P. Youmans. Preparation of highly immunogenic ribosomal fractions of Mycobacterium tuberculosis by use of sodium dodecyl sulfate. J. Bacteriol. 91:2139-2145. 1966.-Ribosomal fractions of Mycobacterium tuberculosis, strain H37Ra, were prepared by treatment of the intracellular particulate fraction with 0.25 or 0.5% sodium dodecylsulfate (SDS) followed by centrifugation at 144,700 x g for 3 hr. This procedure has greatly simplified the preparation of ribosomal fractions and has given fractions composed of approximately 50% ribonucleic acid (RNA) and 15 to 20% protein. When incorporated into Freund's incomplete adjuvant and injected intraperitoneally into CF-1 mice, the SDS ribosomal fractions were more immunogenic than the particulate fractions from which they were prepared. They were as much as 100 times more immunogenic than ribosomal fractions prepared by differential centrifugation, 1 mug (dry weight) per mouse being sufficient for the induction of some immunity. However, none of these ribosomal preparations, in comparable doses, was as immunogenic as the living cells from which they were prepared. It was also shown that the addition of 10(-4)m MgCl(2) to the final diluent increased immunogenic activity, whereas larger concentrations (10(-3)m) reduced immunogenic activity. Preparation of the ribosomal fraction from ruptured cells in one continuous process during the course of 1 day increased the activity. Two-week-old H37Ra cells contained more RNA and were more immunogenic than the older cultures which have been used in the past.
尤曼斯，安妮·S.（伊利诺伊州芝加哥西北大学医学院），以及盖伊·P.尤曼斯。使用十二烷基硫酸钠制备结核分枝杆菌的高免疫原性核糖体组分。《细菌学杂志》91:2139 - 2145。1966年。——通过用0.25%或0.5%的十二烷基硫酸钠（SDS）处理细胞内颗粒组分，然后在144,700×g下离心3小时，制备结核分枝杆菌H37Ra菌株的核糖体组分。该方法极大地简化了核糖体组分的制备，并得到了由约50%核糖核酸（RNA）和15%至20%蛋白质组成的组分。当将SDS核糖体组分掺入弗氏不完全佐剂并腹腔注射到CF - 1小鼠体内时，它们比制备它们的颗粒组分更具免疫原性。它们的免疫原性比通过差速离心制备的核糖体组分高多达100倍，每只小鼠1微克（干重）就足以诱导一定的免疫力。然而，这些核糖体制剂在相当剂量下，都不如制备它们的活细胞具有免疫原性。还表明，在最终稀释剂中添加10⁻⁴m氯化镁会增加免疫原活性，而更高浓度（10⁻³m）则会降低免疫原活性。在一天的过程中通过一个连续过程从破裂细胞制备核糖体组分可提高活性。两周大的H37Ra细胞比过去使用的较老培养物含有更多的RNA且更具免疫原性。
