猪源粪肠球菌菌毛亚单位蛋白多抗对其生物被膜形成及黏附侵袭作用的影响

Effect of Ebp subunit protein polyclonal antibody of swine Enterococcus faecalis on its biofilm formation and adhesion and invasion

以猪源粪肠球菌N9、N41为受试菌株,探究了粪肠球菌N9、N41的生物膜形成能力和对猪肠上皮细胞(IPEC-J2)的黏附侵袭能力,然后与重组亚单位蛋白(Ebp)的多克隆抗血清及纯化IgG作用,分别测定其生物膜的形成量、对猪肠上皮细胞(IPEC-J2)的黏附侵袭能力。结果表明,多抗IgG EbpA1、EbpC1和EbpA3对生物被膜的形成有阻断作用(P<0.05),且EbpA1的阻断作用最强,其次是EbpC1。6个蛋白片段的多抗血清对细菌黏附、侵袭细胞均有一定的阻断作用,但血清EbpA1和EbpC1对菌株的黏附侵入阻断能力最强,且对N41菌株黏附、侵袭细胞的阻断能力强于N9菌株;6个纯化多抗IgG的黏附、侵入阻断作用基本同多抗血清的作用。这说明重组亚单位蛋白EbpA1和EbpC1可诱导试验动物机体产生相对应的特异性抗体,且在体外试验中具有明显的免疫保护作用。

Swine Enterococcus faecalis N9 and N41 were used as tested strains to explore the biofilm formation ability of Enterococcus faecalis N9 and N41 and the adhesion and invasion ability to porcine intestinal epithelial cells(IPEC-J2),and then with the polyclonal antiserum of the recombinant subunit protein(Ebp)and the purified IgG were used to determine the amount of biofilm formation and adhesion to the intestinal epithelial cells(IPEC-J2).The results showed that the polyclonal antibodies IgG EbpA1,EbpA3 and EbpC1 all displayed blocking effect on the formation of biofilm(P<0.05),with EbpA1 having the strongest blocking effect,followed by EbpC1.The multi-antiserum of the six protein fragments had certain blocking effect on the adhesion and invasion of the bacteria,but the serum EbpA1 and EbpC1 had the strongest adhesion and invasion ability to the strain,and the blocking ability of the adhesion and invasion cells of the N41 strain was higher than that of N9 strain.The adhesion and invasion blocking effects of the six polyclonal antibodies were basically the same as those of the polyclonal antiserum.The above test results indicate that the recombinant subunit proteins EbpA1 and EbpC1 can induce the corresponding specific antibodies in the tested animal body,and have obvious immunoprotective effects in the in vitro test.