Hou Xi-Lin, Yu Li-Yun, Liu Jianzhu, Wang Gui-Hua
College of Animal Science and Technology, Heilongjiang August First Land Reclamation University, Daqin 163319, China.
Vaccine. 2007 Dec 21;26(1):24-31. doi: 10.1016/j.vaccine.2007.10.065. Epub 2007 Nov 20.
In this report, for surface display of viral antigen on lactobacilli, we have developed a surface antigen display system using the poly-gamma-glutamate synthetase A protein (pgsA) of Bacillus subtilis as an anchoring matrix. Recombinant fusion proteins comprised of pgsA and neucleocapsid protein of PEDV were stably expressed in Lactobacillus casei. Surface location of fusion protein was verified by ELISA, immunofluoresence microscopy. Oral and intranasal inoculations of recombinant L. casei into pregnant sow and mice resulted in high levels of serum immunoglobuline G (IgG) and mucosal IgA, as demonstrated by rnELISA(recombinant N protein ELISA) using recombinant N protein. Absorbance of IgG in pregnant sow sera highly increased duration of the experiment. More importantly, the level of IgA in colostrum were increased significantly higher than that of IgG. The IgG levels of the piglets were increased after suckling colostrum secreted from sows previously inoculated recombinant L. casei. These results indicate that mucosal immunization with recombinant L. casei expressing PEDV N protein (neucleoprotein of PEDV) on its surface elicited high levels of mucosal IgA and circulation IgG immune responses against the antigen N of PEDV.
在本报告中，为了在乳酸杆菌表面展示病毒抗原，我们开发了一种表面抗原展示系统，该系统使用枯草芽孢杆菌的聚γ-谷氨酸合成酶A蛋白（pgsA）作为锚定基质。由pgsA和猪流行性腹泻病毒（PEDV）的核衣壳蛋白组成的重组融合蛋白在干酪乳杆菌中稳定表达。通过酶联免疫吸附测定（ELISA）、免疫荧光显微镜检查验证了融合蛋白的表面定位。将重组干酪乳杆菌经口服和鼻内接种到怀孕母猪和小鼠体内，使用重组N蛋白的重组N蛋白ELISA（rnELISA）结果显示，这导致了高水平的血清免疫球蛋白G（IgG）和粘膜IgA。在整个实验期间，怀孕母猪血清中IgG的吸光度大幅增加。更重要的是，初乳中IgA的水平显著高于IgG。仔猪在吸食先前接种重组干酪乳杆菌的母猪分泌的初乳后，其IgG水平升高。这些结果表明，用表面表达PEDV N蛋白（PEDV的核蛋白）的重组干酪乳杆菌进行粘膜免疫可引发针对PEDV抗原N的高水平粘膜IgA和循环IgG免疫反应。
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