- 學位論文
豬鏈球菌之溶血素及細胞外因素重組蛋白之抗原性分析
Evaluation on the Antigenicity of Recombinant Suilysin and Extracellular Factor of Streptococcus suis
摘要
豬鏈球菌（Streptococcus suis, S. suis）第二血清型係一重要豬隻病原，它能造成諸如腦膜炎、敗血症、心內膜炎、關節炎及敗血性休克等症狀,死亡率高。本病原屬人畜共通，以密集與豬隻接觸之人員風險較高。豬鏈球菌共有35種血清型，其中部分致病因子110KDa大小的細胞外因子（extracellular factor, EF）及54~65 KDa的溶血素（suilysin, Sly）可於部分血清型中表現，分別由epf及sly基因表現出。而EF和SLY蛋白表現與否往往與高毒力第二血清型菌株有關，但它們的功能目前仍不明。本篇研究目的為利用原核表現系統表現重組細胞外因子(rEFa)及溶血素(rSly)蛋白，以作為抗原製作次單位疫苗。由結果可知單獨以rEFa免疫,經過2至4週仍無法誘發抗原特異性抗體；然而，如再搭配全菌，rEFa則能增強誘發抗體。另一方面，單獨以rSly免疫，經過2至4週能增加抗原特異性抗體產生。當rEFa及rSly搭配全菌，抗體的增加將是可預期的。rEFa雖無極佳免疫原性，仍可做為評估商品化ISA VG系列佐劑效力之抗原。結果顯示混合商品化佐劑 ISA 201 VG組別可產生較高的特異性rEFa抗體力價，較其他組別具顯著性差異(P <0.05)。利用血清型第一型豬鏈球菌選殖rEFa 及rEFb 的重組蛋白混合ISA 201 VG，結果顯示在單單施打rEFa或rEFb疫苗在施打後2及4週，可促使抗體產生，並且將rEFa混合rEFb免疫結果顯示可引起更高的抗體產生。
並列摘要
Streptococcus suis (S. suis) serotype 2 is an important swine pathogen responsible for diverse infections, including meningitis, septicaemia, endocarditis, arthritis, and septic shock, and the mortality is high. It can be transmitted to human beings through close contact with sick pigs or carriers. Totally, thirty-five serotypes have been identified. The 110 kDa extracellular factor (EF) and the 54 to 65 kDa Suilysin (Sly) are expressed by some S. suis strains. EF and Sly are frequently associated with the highly virulent serotype 2 strains, but their functions are still unclear. The objective of this research is using recombinant EF (rEFa from serotype 2, P2), (rEFa, rEFb from serotype 1, P1), and Sly (rSly from 14750, standard strain from BCRC) proteins, expressed by a prokaryotic expression system, as the antigen to produce subunit vaccines. The results showed that immunization with rEFa alone in mice failed to trigger the production of antigen-specific antibody 2 and 4 weeks after the primary vaccination. However, when combined with the whole cell S. suis, rEFa was able to enhance the production of antigen-specific antibody. On the other hand, immunization with, rSly in mice was able to increase the production of antigen-specific antibody 2 and 4 weeks after the primary vaccination. When rSly and rEFa were combined with the whole cell S. suis the production of antigen-specific antibody was further augmented. Since the immunogenicity of rEFa is low, it was used as the antigen to evaluate the efficacy of various commercial ISA VG adjuvants. Results indicated that some of these products significantly (P <0.05) enhanced the production of rEFa - specific antibodies. Among them, ISA 201 VG adjuvants showed the best potential in terms of augmenting antibody production. As a consequence, the full range of rEF sequences including, (rEFa, rEFb) from serotype 1 plus commercial ISA 201 VG adjuvants were carried out. The results showed that rEFa and rEFb alone were able to trigger the production of antigen-specific antibody 2 and 4 weeks after the primary vaccination. Moreover, when rEFa and rEFb were combined, the production of antigen-specific antibody was further enhanced.
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