USE OF A PURIFIED OUTER-MEMBRANE PROTEIN-F (PORIN) PREPARATION OF PSEUDOMONAS-AERUGINOSA AS A PROTECTIVE VACCINE IN MICE
USE OF A PURIFIED OUTER-MEMBRANE PROTEIN-F (PORIN) PREPARATION OF PSEUDOMONAS-AERUGINOSA AS A PROTECTIVE VACCINE IN MICE
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DOI：
10.1128/iai.44.1.49-54.1984
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发表时间：
1984-01-01
影响因子：
3.1
通讯作者：
BERG, RD
中科院分区：
文献类型：
--
作者：
GILLELAND, HE;PARKER, MG;BERG, RD
采用2种方法对铜绿假单胞菌PAO 1菌株的外膜蛋白F（孔蛋白）进行了纯化。一个程序涉及分离柱层析分离的蛋白质提取分离的外膜;其他涉及提取凝胶后，平板聚丙烯酰胺凝胶电泳的蛋白质提取细胞包膜。这两种方法都产生了蛋白F制剂，其成功地免疫了随后用PAO 1菌株攻击的小鼠。蛋白F制剂含有少量脂多糖（LPS）。这种水平的LPS污染保护免疫小鼠免受同源LPS血清型菌株的攻击。用来自PAO 1菌株的蛋白F制剂免疫小鼠也提供了针对2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。显然，当作为疫苗给予时，蛋白F具有保护能力，这不是由于LPS污染。用蛋白F制剂免疫后，通过酶联免疫吸附测定，小鼠显示对纯化的蛋白F制剂的抗体滴度增加。通过给予针对蛋白F制备物产生的兔抗血清来被动保护小鼠。蛋白F制剂在免疫动物中激发特异性体液抗体应答。纯化的蛋白F可能具有作为铜绿假单胞菌的有效疫苗的潜力。
The outer membrane protein F (porin) from the PAO1 strain of P. aeruginosa was purified by 2 methods. One procedure involved separation by column chromatography of proteins extracted from isolated outer membranes; 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. Immunization of mice with protein F preparations from the PAO1 strain also afforded protection against challenge with 2 different LPS serotype strains. This protective ability was lost when the protein F preparation was treated with papain before use as a vaccine. Evidently, 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. The protein F preparation elicits a specific humoral antibody response in immunized animals. Purified protein F may have potential as an effective vaccine for P. aeruginosa.