豬多價次單位疫苗之效力評估

豬巴氏桿菌 (Pasteurella multocida, Pm) 感染8-12週齡仔豬，引起豬萎縮性鼻炎，其中巴氏桿菌毒素 (Pasteurella multocida toxin, PMT) 是很重要的毒力因子；豬鏈球菌(Streptococcus suis, S.suis) 常感染4-12週齡仔豬造成腦膜炎、關節炎、敗血症，而其表面抗原蛋白 (Surface antigen one, Sao ) 為一種細胞壁表面相關蛋白，存在很多不同血清型中，且能夠引起很好免疫反應；豬丹毒 (Erysipelothrix rhusiopathiae, E.rhusiopathiae) 能感染全年齡豬造成急性敗血症，其中表面保護性蛋白 (Surface protective antigen A , SpaA) 可以誘導出良好的保護性抗體反應，並對其他血清型具有交叉保護力。我們利用原核系統成功表現出rSao-tm、rSpaA-N及rPMT-C重組蛋白，經抗原純化後，並添加不同佐劑評估其免疫效力。初步結果顯示，在豬隻實驗中，w/o/w + CpG ODN在免疫後的第4週Sao抗體反應相較於其他組具顯著性差異 (p < 0.05)，w/o/w + CpG ODN在免疫後的第6週PMT-C及SpaA-N抗體反應相較於其他組具顯著性差異 (p < 0.05)；在細胞激素方面，透過豬鏈球菌刺激下，w/o/w + CpG ODN引起IFN-γ、IL-2、IL-12、IL-4高於w/o/w (p < 0.05) ;在豬丹毒桿菌刺激下，w/o/w引起IFN-γ、IL-2、IL-12高於 + CpG ODN (p < 0.05)，但在IL-4、IL-6表現量兩組之間並無顯著性差異。綜合上述結果，w/o/w + CpG ODN能夠提供多價次單位疫苗更快速的體液性免疫反應及更好的細胞性免疫反應。

關鍵字

豬巴氏桿菌；豬鏈球菌；豬丹毒；多價疫苗；次單位

並列摘要

Pasteurella multocida (P. multocida) can infect 8-12-week piglet and mainly causes atrophic rhinitis (AR). Pasteurella multocida toxin (PMT) is an important virulence factor. Streptococcus suis (S. suis) causes meningitis, arthritis and septicemia in 4-12-week piglet. The Surface antigen one (Sao), which is a cell wall surface-associated protein and found in all serotypes of S.suis, can induce good immune responses. Erysipelothrix rhusiopathiae (E. rhusiopathiae) can infect pigs of all age and cause acute septicemia and chronic diseases. The Surface protective antigen A (SpaA) not only induces protective antibody responses, but also provides cross protection against other serotypes. We produced and puriified rSao-tm, rSpaA-N and rPMT-C recombinant proteins using a prokaryotic expression system. We combined recombinant proteins with various adjuvants to evaluate the immune responses. Our resulst show that the Sao antibody titer of w/o/w + CpG ODN in the swine test was significantly higher than that of other groups at 4 weeks after immunization. PMT-C and SpaA-N antibody titer of w/o/w + CpG ODN in the swine test was significantly higher than that of other groups at 6 weeks after immunization (p < 0.05). In terms of cytokines, in response to Streptococcus suis antigen, w/o/w + CpG ODN induced higher level of IFN-γ, IL-2, IL-12 and IL-4 than the w/o/w alone (p < 0.05). Moreover, in response to Erysipelothrix rhusiopathiae antigen, w/o/w induced higher level of IFN-γ, IL-2 and IL-12 than the w/o/w + CpG ODN (p < 0.05). However, IL-4 and IL-6 showed no significant difference between two groups. In conclusion, w/o/w + CpG ODN can provide a more rapid humoral immune response and better cellular immune response in multivalent subunit vaccines.