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用表达痢疾志贺菌1型外膜蛋白A的重组乳酸乳球菌进行免疫:口服和鼻内给药途径的评估。

Immunization with r-Lactococcus lactis expressing outer membrane protein A of Shigella dysenteriae type-1: evaluation of oral and intranasal route of administration.

作者信息

Yagnik B, Sharma D, Padh H, Desai P

机构信息

Department of Cell and Molecular Biology, B. V. Patel Pharmaceutical Education and Research Development (PERD) Centre, Ahmedabad, 380054, Gujarat, India.

B. R. D School of Biosciences, Sardar Patel University, Vallabh Vidhyanagar, 388120, Gujarat, India.

出版信息

J Appl Microbiol. 2017 Feb;122(2):493-505. doi: 10.1111/jam.13353. Epub 2016 Dec 29.

DOI:10.1111/jam.13353
PMID:27860045
Abstract

AIMS

To evaluate the comparative immunogenic potential of food grade Lactococcus lactis expressing outer membrane protein A (OmpA) of Shigella dysenteriae type-1 (SD-1) when administered either orally or intranasally.

METHODS AND RESULTS

OmpA of SD-1 was cloned and expressed first in Escherichia coli and then in L. lactis. Presence of recombinant gene was confirmed by restriction enzyme digestion and immunoblot analysis. Using immobilized metal affinity chromatography, OmpA was purified from recombinant E. coliBL21 (DE3) and subcutaneously administered in BALB/c mice. Detection of OmpA-specific IgG antibodies by enzyme-linked immunosorbent assay (ELISA) confirmed the immunogenicity of OmpA. In order to establish r-L. lactis as a mucosal delivery vehicle, it was administered orally and nasally in BALB/c mice. Serum IgG and faecal IgA were assessed through ELISA to compare the relative potential of immunization routes and immunogenic potential of r-L. lactis. Immunization via the oral route proved superior to intranasal exposure.

CONCLUSION

Recombinant L. lactis expressing OmpA of SD-1 was found to be immunogenic. Oral administration of r-L. lactis elicited higher systemic and mucosal immune response when compared with the nasal route.

SIGNIFICANCE AND IMPACT OF THE STUDY

Using food grade recombinant L. lactis has implications in the development of a prophylactic against multidrug-resistant Shigella, which can be used as a prospective vaccine candidate. Evaluating mucosal routes of immunization demonstrated that the oral route of administration elicited better immune response against OmpA of Shigella.

摘要

目的

评估表达1型痢疾志贺菌(SD-1)外膜蛋白A(OmpA)的食品级乳酸乳球菌经口服或鼻内给药后的相对免疫原性潜力。

方法与结果

首先在大肠杆菌中克隆并表达SD-1的OmpA,随后在乳酸乳球菌中表达。通过限制性内切酶消化和免疫印迹分析确认重组基因的存在。利用固定化金属亲和层析从重组大肠杆菌BL21(DE3)中纯化OmpA,并皮下注射到BALB/c小鼠体内。通过酶联免疫吸附测定(ELISA)检测OmpA特异性IgG抗体,证实了OmpA的免疫原性。为了将重组乳酸乳球菌确立为黏膜递送载体,将其经口服和鼻内给药于BALB/c小鼠。通过ELISA评估血清IgG和粪便IgA,以比较免疫途径的相对潜力和重组乳酸乳球菌的免疫原性潜力。经口免疫证明优于鼻内暴露。

结论

发现表达SD-1的OmpA的重组乳酸乳球菌具有免疫原性。与鼻内途径相比,口服重组乳酸乳球菌可引发更高的全身和黏膜免疫反应。

研究的意义和影响

使用食品级重组乳酸乳球菌对开发针对多重耐药志贺菌的预防性疫苗具有重要意义,该疫苗可作为一种有前景的候选疫苗。评估黏膜免疫途径表明,口服给药途径对志贺菌OmpA引发的免疫反应更好。

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