Analyses of rat Pneumocystis carinii antigens recognized by human and rat antibodies by using western immunoblotting
Analyses of rat Pneumocystis carinii antigens recognized by human and rat antibodies by using western immunoblotting
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使用蛋白质免疫印迹法分析人和大鼠抗体识别的大鼠卡氏肺孢子虫抗原
DOI：
10.1128/iai.54.1.96-103.1986
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发表时间：
1986
影响因子：
3.1
通讯作者：
M. Ivey
中科院分区：
文献类型：
--
作者：
D. Graves;S. McNabb;M. Worley;T. Downs;M. Ivey
用免疫印迹技术鉴定了卡氏肺孢子虫感染宿主中诱导抗体应答的主要抗原。这些抗原的生化性质也被阐明。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳，然后进行蛋白质染色，发现分子量（MW）大于205，000的主要成分。当在β-巯基乙醇存在下进行电泳时，该主要组分消失，并出现约110，000至116，000 MW的新的主要蛋白质染色组分。高碘酸-希夫染色显示，这一主要成分含有碳水化合物部分。在55，000 - 60，000-MW区域中的主要组分在β-巯基乙醇存在下电泳后，用过碘酸-希夫染色可见，但用蛋白质染色不可见。从诊断为肺孢子虫病的人和从类固醇诱导的肺孢子虫病恢复的大鼠中检测的大多数血清与110，000 - 116，000和55，000 - 60，000-MW组分反应强烈。这些血清通常但不总是检测到MW约为170，000、125，000和30，000至32，000的抗原。这些数据表明，卡氏肺孢子虫的抗原组成相对复杂，大鼠和人卡氏肺孢子虫可能共享抗原决定簇。感染衍生的人类和大鼠抗血清的主要大鼠卡氏肺孢子虫组分之间的竞争性研究显示竞争;大鼠抗血清似乎比人类抗血清识别更大范围的抗原表位。蛋白酶处理的抗原成分，已被固定在硝酸纤维素纸破坏其与大鼠抗体的抗原反应性。用高碘酸钠处理降低了这种110，000至116，000 MW组分的反应性，并完全破坏了55，000至60，000 MW组分与大鼠抗体的反应性。
The major Pneumocystis carinii antigens inducing antibody responses in infected hosts were identified by Western immunoblotting techniques. The biochemical nature of these antigens was also elucidated. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by protein staining revealed a major component with a molecular weight (MW) of greater than 205,000. This major component disappeared and a new major protein staining component of approximately 110,000 to 116,000 MW appeared when electrophoresis was done in the presence of beta-mercaptoethanol. Periodic acid-Schiff staining revealed that this major component contains carbohydrate moieties. A major component in the 55,000- to 60,000-MW region was visible with periodic acid-Schiff stain, but not with a protein stain, after electrophoresis in the presence of beta-mercaptoethanol. The majority of sera tested from humans with diagnosed pneumocystosis and from rats allowed to recover from steroid-induced pneumocystosis reacted strongly with 110,000- to 116,000-, and 55,000- to 60,000-MW components. These sera often, but not always, detected antigens with MWs of approximately 170,000, 125,000, and 30,000 to 32,000. The data suggest that the antigenic composition of P. carinii is relatively complex and that rat and human P. carinii probably share antigenic determinants. Competitive studies between infection-derived human and rat antisera for the major rat P. carinii components revealed competition; rat antisera appeared to recognize a greater range of antigenic epitopes than did human antisera. Protease treatment of the antigenic components that had been immobilized on nitrocellulose paper destroyed their antigenic reactivity with rat antibody. Treatment with sodium periodate decreased reactivity of this 110,000- to 116,000-MW component and completely destroyed the reactivity of the 55,000- to 60,000-MW component with rat antibody.