铜绿假单胞菌蛋白酶IV：一种低免疫原性的角膜毒力因子。

Pseudomonas aeruginosa protease IV: a corneal virulence factor of low immunogenicity.

作者信息

Thibodeaux Brett A, Caballero Armando R, Dajcs Joseph J, Marquart Mary E, Engel Lee S, O'Callaghan Richard J

机构信息

Department of Microbiology, Louisiana State University Health Sciences Center, New Orleans, 70112, USA.

出版信息

Ocul Immunol Inflamm. 2005 Apr-Jun;13(2-3):169-82. doi: 10.1080/09273940490518937.

DOI:10.1080/09273940490518937

PMID:16019676

Abstract

PURPOSE

To study antibody production to Pseudomonas aeruginosa protease IV (PIV) for immunoassay development and to assess the possible role of antibody in arresting corneal damage.

METHODS

Rabbits were immunized with PIV, urea-soluble recombinant PIV (rPIV), or precipitated rPIV. Antibody was analyzed by ELISA and Western blotting. Antibody-mediated inhibition of PIV activity was tested by colorimetric assay and during keratitis by slit-lamp examination of infected eyes.

RESULTS

Antibody was not produced after PIV immunization but was induced by rPIV. Rabbits immunized first with soluble and then precipitated rPIV produced high titers (log(10)) to rPIV (4.28 +/- 0.09) and significantly higher titers to PIV (3.90 +/- 0.06) compared to the other immunized groups. Antibody to rPIV reacted with PIV, but neither neutralized enzyme activity in vitro nor protected infected rabbits in vivo.

CONCLUSIONS

The present study demonstrates that PIV is a virulence factor which can escape a protective immune response.

摘要

目的

研究针对铜绿假单胞菌蛋白酶IV（PIV）的抗体产生情况，以开发免疫测定方法，并评估抗体在阻止角膜损伤中可能发挥的作用。

方法

用PIV、尿素可溶性重组PIV（rPIV）或沉淀的rPIV免疫兔子。通过酶联免疫吸附测定（ELISA）和蛋白质印迹法分析抗体。通过比色法以及在角膜炎期间通过对感染眼睛进行裂隙灯检查来测试抗体介导的PIV活性抑制作用。

结果

用PIV免疫后未产生抗体，但rPIV可诱导产生抗体。先接种可溶性rPIV然后接种沉淀rPIV的兔子对rPIV产生高滴度（log(10)）（4.28±0.09），与其他免疫组相比，对PIV的滴度显著更高（3.90±0.06）。针对rPIV的抗体与PIV反应，但在体外既不中和酶活性，在体内也不保护受感染的兔子。