Kersten G F, Teerlink T, Derks H J, Verkleij A J, van Wezel T L, Crommelin D J, Beuvery E C
Department of Inactivated Viral Vaccines, National Institute for Public Health and Environmental Hygiene, Bilthoven, The Netherlands.
Infect Immun. 1988 Feb;56(2):432-8. doi: 10.1128/iai.56.2.432-438.1988.
We incorporated the major outer membrane protein (PI) of Neisseria gonorrhoeae into immunostimulating complexes (iscoms) and examined some analytical, physicochemical, and immunological properties of these structures. The immunogenicity was compared with that of three other PI-containing structures, i.e., liposomes, outer membrane complexes produced by the bacterium, and protein-detergent-adjuvant complexes. AIPO4 and dioctadecyldimethylammonium bromide were used as adjuvants. Our results show that iscoms are much more immunogenic than liposomes and protein-detergent complexes but are also much more toxic. The localization of PI in iscoms was investigated. Therefore, the chymotrypsin susceptibility of PI in iscoms was tested, and the incorporation of fragments of PI was determined. Amphiphilic fragments of PI were incorporated in iscoms, but hydrophilic and hydrophobic fragments were not. Chymotrypsin degradation of PI in iscoms indicated that the protein is exposed to the environment in a similar manner as PI in outer membrane complexes, i.e., with both termini anchored in the iscom.
我们将淋病奈瑟菌的主要外膜蛋白(PI)整合到免疫刺激复合物(iscoms)中,并研究了这些结构的一些分析、物理化学和免疫学特性。将其免疫原性与其他三种含PI的结构进行了比较,即脂质体、细菌产生的外膜复合物以及蛋白质-去污剂-佐剂复合物。使用磷酸铝(AIPO4)和二辛基二甲基溴化铵作为佐剂。我们的结果表明,iscoms的免疫原性比脂质体和蛋白质-去污剂复合物强得多,但毒性也大得多。研究了PI在iscoms中的定位。因此,测试了iscoms中PI对胰凝乳蛋白酶的敏感性,并确定了PI片段的掺入情况。PI的两亲性片段被整合到iscoms中,但亲水性和疏水性片段未被整合。iscoms中PI的胰凝乳蛋白酶降解表明,该蛋白在外膜复合物中与PI一样以类似方式暴露于环境中,即两端都锚定在iscom中。