TOXICITY AND IMMUNOGENICITY OF NEISSERIA-MENINGITIDIS LIPOPOLYSACCHARIDE INCORPORATED INTO LIPOSOMES
TOXICITY AND IMMUNOGENICITY OF NEISSERIA-MENINGITIDIS LIPOPOLYSACCHARIDE INCORPORATED INTO LIPOSOMES
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DOI：
10.1128/iai.60.9.3897-3903.1992
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发表时间：
1992-09-01
影响因子：
3.1
通讯作者：
DMITRIEV, BA
中科院分区：
文献类型：
--
作者：
PETROV, AB;SEMENOV, BF;DMITRIEV, BA
为了获得无毒且高免疫原性的脂多糖（LPS）用于免疫，我们将脑膜炎奈瑟氏菌 LPS 掺入脂质体中。通过囊泡脱水-再水合或长时间共超声处理的方法掺入天然 LPS 及其盐。通过囊泡脱水-复水的方法实现了LPS最完全地融入脂质体并降低了毒性。三种形式的LPS（H+形式、Mg2+盐和三乙醇胺盐）在水中表现出不同的溶解度，酸性形式的LPS具有最明显的疏水性，几乎能够与脂质体膜完全缔合。对脂质体 LPS 体外（通过鲎变形细胞试验）和体内（通过监测兔子的热原反应）活性的评估表明，内毒素活性降低了高达 1,000 倍。此外，脂质体LPS的热原活性与脑膜炎球菌多糖疫苗的热原活性相当。脂质体对 LPS 的免疫反应具有明显的辅助作用。因此，与用游离LPS免疫的小鼠相比，用脂质体LPS免疫的小鼠脾脏中抗LPS噬斑形成细胞的水平高1个数量级，并且观察的时间更长（直到第21天，即观察期）。通过酶联免疫吸附测定进行的一项研究也揭示了同样的规律。这项研究还确定，免疫诱导的抗体属于免疫球蛋白 M 和 G 类，能够延长循环时间。此外，脂质体 LPS 在 CBA/N 小鼠（LyB-5+ 亚群的 B 淋巴细胞有缺陷）中诱导了明显的免疫反应。后者的结果表明脂质体 LPS 的免疫原性作用发生在早期。
To obtain nontoxic and highly immunogenic lipopolysaccharide (LPS) for immunization, we incorporated Neisseria meningitidis LPS into liposomes. Native LPS and its salts were incorporated by the method of dehydration-rehydration of vesicles or prolonged cosonication. The most complete incorporation of LPS into liposomes and a decrease in toxicity were achieved by the method of dehydration-rehydration of vesicles. Three forms of LPS (H+ form, Mg2+ salt, and triethanolamine salt) showed different solubilities in water, the acidic form of LPS, with the most pronounced hydrophobic properties, being capable of practically complete association with liposomal membranes. An evaluation of the activity of liposomal LPS in vitro (by the Limulus amoebocyte test) and in vivo (by monitoring the pyrogenic reaction in rabbits) revealed a decrease in endotoxin activity of up to 1,000-fold. In addition, the pyrogenic activity of liposomal LPS was comparable to that of a meningococcal polysaccharide vaccine. Liposomes had a pronounced adjuvant effect on the immune response to LPS. Thus, the level of anti-LPS plaque-forming cells in the spleens of mice immunized with liposomal LPS was 1 order of magnitude higher and could be observed for a longer time (until day 21, i.e., the term of observation) than in mice immunized with free LPS. The same regularity was revealed in a study done with an enzyme-linked immunosorbent assay. This study also established that antibodies induced by immunization belonged to the immunoglobulin M and G classes, which are capable of prolonged circulation. Moreover, liposomal LPS induced a pronounced immune response in CBA/N mice (defective in B lymphocytes of the LyB-5+ subpopulation). The latter results indicate that the immunogenic action of liposomal LPS occurs at an early age.