表达猪细小病毒VP2蛋白的重组干酪乳杆菌诱导小鼠产生特异性抗体

doi:10.3864/j.issn.0578-1752.2008.03.030

中国农业科学 ›› 2008, Vol. 41 ›› Issue (3): 846-851 .doi: 10.3864/j.issn.0578-1752.2008.03.030

表达猪细小病毒VP2蛋白的重组干酪乳杆菌诱导小鼠产生特异性抗体

徐义刚,崔丽春,葛俊伟,唐丽杰,赵丽丽,李一经

东北农业大学动物医学院

收稿日期:2006-12-29 修回日期:1900-01-01 出版日期:2008-03-10 发布日期:2008-03-10
通讯作者: 李一经

The Determination of Specific Antibodies in Mice orally Immuned Lactobacillus casei expressing Porcine Parvovirus VP2 protein on cell surface

东北农业大学动物医学院

Received:2006-12-29 Revised:1900-01-01 Online:2008-03-10 Published:2008-03-10

摘要/Abstract

摘要： 【目的】研究以干酪乳杆菌作为传递抗原活载体表达猪细小病毒主要免疫保护性抗原VP2蛋白的免疫特性。【方法】将构建的重组猪细小病毒（PPV）VP2基因的细胞表面表达型载体pPG-VP2电转化干酪乳杆菌L.casei 393，获得阳性重组菌pPG-VP2/L.casei 393。重组菌以2%乳糖为诱导物，在MRS培养基中进行诱导，经SDS-PAGE、Western blot及间接免疫荧光分析，目的蛋白获得表达。将重组菌及空质粒菌株分别口服接种Balb/c小鼠，收集粪便及肠黏液样品测定小鼠产生抗VP2的特异性sIgA，采集血液样品测定小鼠产生抗VP2的特异性IgG，并对获得的抗体进行PPV中和活性的测定。【结果】重组干酪乳杆菌pPG-VP2/L.casei393免疫小鼠能够产生明显的抗PPV的sIgA和IgG抗体水平，其对PPV的中和效价分别为1﹕24和1﹕128。【结论】为PPV重组乳酸菌口服疫苗的研制提供了重要的物质基础。

关键词: 猪细小病毒, VP2蛋白, 干酪乳杆菌, 黏膜免疫

Abstract: Lactobacillus casei strain 393 was selected as oral vaccine carrier for the expression of Porcine Parvovirus (PPV) main immune protective antigen VP2 protein. The constructed pPG-VP2 plasmid expressing PPV VP2 protein on cell surface was electroporated into Lactobacillus casei 393 generating recombinant strain pPG-VP2/L.casei393.The interest protein was detected by SDS-PAGE、Western-blot and indirect immunofluorescence after induced by 2% Lactose in MRS broth. Oral immunization of BALB/c mice were received with recombinant strain harboring pPG-VP2 and L.casei393 harboring pPG. Specific anti-PPV VP2 secret immunoglobulin A (sIgA) antibody was detected by indirect enzyme linked immunosorbent assay (ELISA) in the feces and intestines mucus after orogastric immunization, and anti-PPV VP2 immunoglobulin G (IgG) antibody was detected by indirect ELISA in the serum of immunized mice. The results indicated that the mice immunized with pPG-VP2/L.casei393 could produce clear anti-PPV antibody level. Our work establish important material basement for the development of PPV oral vaccine.

Key words: Porcine Parvovirus, VP2 protein, Lactobacillus casei 393, surface expression, sIgA, IgG

徐义刚,崔丽春,葛俊伟,唐丽杰,赵丽丽,李一经. 表达猪细小病毒VP2蛋白的重组干酪乳杆菌诱导小鼠产生特异性抗体[J]. 中国农业科学, 2008, 41(3): 846-851 .

. The Determination of Specific Antibodies in Mice orally Immuned Lactobacillus casei expressing Porcine Parvovirus VP2 protein on cell surface[J]. Scientia Agricultura Sinica, 2008, 41(3): 846-851 .

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表达猪细小病毒VP2蛋白的重组干酪乳杆菌诱导小鼠产生特异性抗体

The Determination of Specific Antibodies in Mice orally Immuned Lactobacillus casei expressing Porcine Parvovirus VP2 protein on cell surface

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