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福氏 2a 志贺菌外膜蛋白 A（OmpA）在小鼠模型中诱导保护性免疫应答。

Outer membrane protein A (OmpA) of Shigella flexneri 2a, induces protective immune response in a mouse model.

机构信息

Division of Pathophysiology, National Institute of Cholera and Enteric Diseases, Beliaghata, Kolkata, West Bengal, India.

出版信息

PLoS One. 2011;6(7):e22663. doi: 10.1371/journal.pone.0022663. Epub 2011 Jul 26.

DOI:10.1371/journal.pone.0022663
PMID:21818362
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3144247/
Abstract

BACKGROUND

In our earlier studies 34 kDa outer membrane protein (OMP) of Shigella flexneri 2a has been identified as an efficient immunostimulant.

KEY RESULTS

In the present study MALDI-TOF MS analysis of the purified 34 kDa OMP of Shigella flexneri 2a shows considerable sequence homology (Identity 65%) with the OmpA of S. flexneri 2a. By using the specific primers, the gene of interest has been amplified from S. flexneri 2a (N.Y-962/92) genomic DNA, cloned in pET100/D-TOPO® vector and expressed using induction with isopropyl thiogalactoside (IPTG) for the first time. Immunogenicity and protective efficacy of the recombinant OmpA has been evaluated in an intranasally immunized murine pulmonary model. The recombinant protein induces significantly enhanced protein specific IgG and IgA Abs in both mucosal and systemic compartments and IgA secreting cells in the systemic compartment (spleen). The mice immunized with OmpA have been protected completely from systemic challenge with a lethal dose of virulent S. flexneri 2a. Immunization with the protein causes mild polymorphonuclear neutrophil infiltration in the lung, without inducing the release of large amounts of proinflammatory cytokines.

CONCLUSION

These results suggest that the OmpA of S. flexneri 2a can be an efficacious mucosal immunogen inducing protective immune responses. Our findings also demonstrate that antibodies and Th1 immune response may be associated with the marked protective efficacy of immunized mice after intranasal shigellae infection.

摘要

背景

在我们早期的研究中，已经鉴定出福氏 2a 志贺氏菌的 34kDa 外膜蛋白（OMP）是一种有效的免疫刺激剂。

主要结果

在本研究中，MALDI-TOF MS 分析纯化的福氏 2a 志贺氏菌 34kDa OMP 显示与福氏 2a 志贺氏菌的 OmpA 具有相当的序列同源性（同一性 65%）。使用特异性引物，从福氏 2a（N.Y-962/92）基因组 DNA 中扩增了感兴趣的基因，克隆到 pET100/D-TOPO®载体中，并首次使用异丙基硫代半乳糖苷（IPTG）诱导表达。在鼻内免疫的鼠肺模型中评估了重组 OmpA 的免疫原性和保护效力。重组蛋白在粘膜和全身部位以及全身部位（脾脏）的 IgA 分泌细胞中诱导显著增强的蛋白质特异性 IgG 和 IgA Abs。用 OmpA 免疫的小鼠完全免受致命剂量的强毒福氏 2a 的全身攻击。蛋白免疫引起肺部轻度多形核中性粒细胞浸润，而不会诱导大量促炎细胞因子的释放。

结论

这些结果表明，福氏 2a 志贺氏菌的 OmpA 可以作为有效的粘膜免疫原，诱导保护性免疫反应。我们的研究结果还表明，抗体和 Th1 免疫反应可能与免疫接种小鼠在鼻内感染志贺氏菌后具有显著的保护效力有关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3a3a/3144247/bfca2c7ccb6c/pone.0022663.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3a3a/3144247/b84933aa5df8/pone.0022663.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3a3a/3144247/5519cca35408/pone.0022663.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3a3a/3144247/fa14418e4aae/pone.0022663.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3a3a/3144247/0ff5cbda8c0e/pone.0022663.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3a3a/3144247/7fc2ef10656d/pone.0022663.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3a3a/3144247/bfca2c7ccb6c/pone.0022663.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3a3a/3144247/b84933aa5df8/pone.0022663.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3a3a/3144247/5519cca35408/pone.0022663.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3a3a/3144247/fa14418e4aae/pone.0022663.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3a3a/3144247/0ff5cbda8c0e/pone.0022663.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3a3a/3144247/7fc2ef10656d/pone.0022663.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3a3a/3144247/bfca2c7ccb6c/pone.0022663.g006.jpg

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