Schirmbeck R, Böhm W, Reimann J
Department of Bacteriology, University of Ulm, FRG.
Eur J Immunol. 1994 Sep;24(9):2068-72. doi: 10.1002/eji.1830240920.
Complex adjuvant formulations have been used to introduce soluble protein antigens into the "endogenous" processing pathway and hence to elicit specific, major histocompatibility complex class I-restricted cytotoxic T lymphocyte (CTL) responses. We tested if simple modifications of a model protein antigen, i.e. ovalbumin (OVA), can render it immunogenic for murine class I-restricted CTL when injected into mice in soluble form. Injection of 1-100 micrograms native OVA into C57BL/6 (H-2b) mice did not stimulate a class I-restricted CTL response. In contrast, immunization of mice with 0.5 to 10 micrograms sodium dodecyl sulfate (SDS)- or deoxycholate (DOC)-denatured OVA efficiently primed CD8+ CTL specific for the well-characterized Kb-restricted OVA257-264 epitope. Gel-purified SDS-denatured OVA devoid of protein fragments and excess detergent efficiently stimulated a specific CTL response in vivo. OVA preparations denatured by heat or urea treatment were not immunogenic for murine CTL. Injection of non-treated or detergent-treated, antigenic OVA257-264 peptide into mice did not elicit a CTL response. Thus, denaturation of OVA by simple detergents such as SDS or DOC dramatically enhances its immunogenicity for class I-restricted CTL but not all modes of denaturation are equally effective.
复杂的佐剂配方已被用于将可溶性蛋白质抗原引入“内源性”加工途径，从而引发特异性的、主要组织相容性复合体I类限制性细胞毒性T淋巴细胞（CTL）反应。我们测试了对模型蛋白质抗原（即卵清蛋白，OVA）进行简单修饰，是否能使其以可溶性形式注入小鼠体内时，对小鼠I类限制性CTL具有免疫原性。将1 - 100微克天然OVA注射到C57BL/6（H-2b）小鼠体内，并未刺激I类限制性CTL反应。相比之下，用0.5至10微克十二烷基硫酸钠（SDS）或脱氧胆酸盐（DOC）变性的OVA免疫小鼠，能有效地启动针对特征明确的Kb限制性OVA257 - 264表位的CD8 + CTL。不含蛋白质片段和过量去污剂的凝胶纯化SDS变性OVA在体内能有效刺激特异性CTL反应。经加热或尿素处理变性的OVA制剂对小鼠CTL无免疫原性。将未处理或经去污剂处理的抗原性OVA257 - 264肽注射到小鼠体内，未引发CTL反应。因此，用SDS或DOC等简单去污剂使OVA变性，可显著增强其对I类限制性CTL的免疫原性，但并非所有变性方式都同样有效。
