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使用铜绿假单胞菌纯化的外膜蛋白F(孔蛋白)制剂作为小鼠的保护性疫苗。

Use of a purified outer membrane protein F (porin) preparation of Pseudomonas aeruginosa as a protective vaccine in mice.

作者信息

Gilleland H E, Parker M G, Matthews J M, Berg R D

出版信息

Infect Immun. 1984 Apr;44(1):49-54. doi: 10.1128/iai.44.1.49-54.1984.

Abstract

The outer membrane protein F (porin) from the PAO1 strain of Pseudomonas aeruginosa was purified by two different methods. One procedure involved separation by column chromatography of proteins extracted from isolated outer membranes, whereas the other involved extraction from gels after slab polyacrylamide gel electrophoresis of proteins extracted from cell envelopes. Both procedures yielded protein F preparations which successfully immunized mice from subsequent challenge with the PAO1 strain. The protein F preparations contained small quantities of lipopolysaccharide (LPS). This level of LPS contamination protected immunized mice from challenge with the homologous LPS serotype strain. However, immunization of mice with protein F preparations from the PAO1 strain also afforded protection against challenge with two different LPS serotype strains. This protective ability was lost when the protein F preparation was treated with papain before use as a vaccine. These observations support the conclusion that protein F has protective ability, which is not due to LPS contamination, when given as a vaccine. After immunization with the protein F preparation, mice showed an increase in antibody titer to the purified protein F preparation by enzyme-linked immunosorbent assay. Mice were protected passively by administration of rabbit antisera raised to the protein F preparation. These results indicate that the protein F preparation elicits a specific humoral antibody response in immunized animals. Our results suggest that purified protein F has potential as an effective vaccine for P. aeruginosa.

摘要

通过两种不同方法纯化了铜绿假单胞菌PAO1菌株的外膜蛋白F(孔蛋白)。一种方法是通过柱色谱法分离从分离的外膜中提取的蛋白质,而另一种方法是在对从细胞包膜中提取的蛋白质进行平板聚丙烯酰胺凝胶电泳后从凝胶中提取。两种方法都得到了蛋白F制剂,这些制剂成功地使小鼠免受PAO1菌株的后续攻击。蛋白F制剂含有少量脂多糖(LPS)。这种LPS污染水平保护免疫小鼠免受同源LPS血清型菌株的攻击。然而,用PAO1菌株的蛋白F制剂免疫小鼠也能提供针对两种不同LPS血清型菌株攻击的保护。当蛋白F制剂在用作疫苗之前用木瓜蛋白酶处理时,这种保护能力丧失。这些观察结果支持这样的结论,即蛋白F作为疫苗使用时具有保护能力,且这种能力并非由于LPS污染。用蛋白F制剂免疫后,通过酶联免疫吸附测定法,小鼠对纯化蛋白F制剂的抗体滴度有所增加。通过给予针对蛋白F制剂产生的兔抗血清,小鼠获得了被动保护。这些结果表明,蛋白F制剂在免疫动物中引发了特异性体液抗体反应。我们的结果表明,纯化的蛋白F有潜力成为一种有效的铜绿假单胞菌疫苗。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f1a4/263467/255af4f4c4a7/iai00127-0058-a.jpg

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