高应答患者牙龈卟啉单胞菌381免疫优势抗原分析。

E A Boutsi
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引用次数: 0

摘要

本研究的目的是测定牙龈假单胞菌381的免疫优势抗原,并检测其组成和分子量。选择对牙龈假单胞菌381超声提取物具有高抗体滴度的牙周患者14例。以牙龈假单胞菌381的全细胞部分、超声提取液和外膜蛋白为抗原,分别进行了未处理和加热后的木瓜蛋白酶处理。测定三种抗原中糖、蛋白质和脂多糖(LPS)的总体积。通过酶联免疫吸附试验(ELISA)评估热和木瓜蛋白酶处理前后抗体对三种抗原的结合能力。此外,进行免疫印迹分析。定量分析表明,全细胞部分的脂多糖含量是另外两种制剂的10倍左右,外膜蛋白的碳水化合物含量是另外两种制剂的2倍左右。根据热和木瓜蛋白酶处理下抗体结合降低率将14份血清分为3组。在热处理方面,大多数血清在与超声提取液反应时显示出抗体结合的高度降低。然而,三种血清经热处理后,抗体与外膜蛋白抗原的结合几乎没有降低。同时,这三种血清在热处理下对全细胞抗原表现出几乎相同的反应。在木瓜蛋白酶处理下,几乎所有血清与超声提取液和全细胞部分反应时,抗体结合率均有中等程度的降低,而与外膜蛋白反应时,抗体结合率则有较低的降低。从本研究可以看出,牙龈假单胞菌的主要蛋白质抗原被大多数患者识别,提示蛋白质部分是抗原的重要组成部分,而一些患者似乎将LPS或碳水化合物识别为抗原决定因素。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Analysis of immunodominant antigens of Porphyromonas gingivalis 381 in high responder patients.

The purpose of this study was to determine the immunodominant antigens of P. gingivalis 381 and to examine their composition and molecular weight. Fourteen periodontal patients, with high antibody titers to P. gingivalis 381 sonicated extract, were selected. Whole cell fraction, sonicated extract and outer membrane protein of P. gingivalis 381 were used as antigens in the untreated form as well as in the heated form and treated with papain. Total volume of sugar, protein and lipopolysaccharide (LPS) was estimated in each one of the three antigens. Antibody binding capacity to the three antigens was evaluated, before and after heat and papain treatment, by an enzyme-linked immunosorbent assay (ELISA). In addition, the immunoblotting analysis was performed. The quantitative assays showed that the whole cell fraction contained about ten times more LPS than the other two preparations while the outer membrane protein contained twice the amount of carbohydrates than the other two preparations. The 14 sera were classified into three groups according to the rate of reduction of antibody binding under heat and papain treatment. Concerning heat treatment, most of the sera showed a high reduction of antibody binding when reacting with the sonicated extract. However, antibody binding to the outer membrane protein antigen was hardly decreased by heat treatment in three sera. Also, these three sera showed almost the same response to the whole cell fraction antigen under heat treatment. Under papain treatment, almost all sera showed a moderate reduction of antibody binding when they reacted with the sonicated extract and whole cell fraction while they showed a low reduction of antibody binding when they reacted with the outer membrane protein. From the present study it could be concluded that a main proteinaceous antigen of P. gingivalis was recognized by the majority of the patients suggesting that the proteinaceous portion is an important part of the antigen, while some patients seemed to recognize the LPS or carbohydrate as the antigenic determinant.

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