Detoxified  Pseudomonas Exotoxin A Enhanced the Immune Response of a Porcine Circovirus type 2 (PCV2) ORF2Recombinant Protein

本研究目的係將五個不同片段的豬環狀病毒第二型（PCV2）ORF2 之抗原區基因，進行選殖及以大腸桿菌表現大量的蛋白，並以大鼠評估各重組蛋白引發的免疫原性。結果顯示，PCV2 ORF2 F2 片段（氨基酸序列第78－156區塊）具有最佳的免疫原性。將去毒綠膿桿菌外毒素A（PE）基因及KDEL訊號序列基因與F2基因融和（稱PE－F2－KDEL重組蛋白），來評估應用PE的結合及轉位區塊作為攜帶PCV2 ORF2 F2免疫抗原的效果。本實驗分別將F2及PE－F2－KDEL重組蛋白進行小鼠的腹腔免疫注射。結果顯示，PE－F2－KDEL有顯著性地，比F2更能誘發較佳的抗PCV2血清IgG抗體力價。此外，與PCV2不活化病毒抗原比較，PE－F2－KDEL重組蛋白亦有顯著性地提高抗PCV2血清IgG的力價。本實驗證實，去毒綠膿桿菌外毒素A可誘發PCV2　ORF2重組蛋白較強的免疫反應。

關鍵字

豬環狀病毒第二型；綠膿桿菌外毒素A ； PCV2 ORF2重組蛋白

並列摘要

In this study, five different fragments of porcine circovirus type 2 (PCV2) ORF2 antigenic regions were cloned, over-expressed in E. coli. Rats were immunized with the fragments to evaluate the immunogenicity of the PCV2 ORF2 recombinant proteins. Results showed that the ORF2 F2 fragment (residues 78-156) was the most immunogenic in terms of the antibody response. Detoxified Pseudomonas exotoxin A (PE) and KDEL signal peptide were fused with F2 fragment at the N and C terminuses, respectively. This study evaluated the application of using the binding and translocation domains of PE as a vehicle for PCV2 F2 fragment, resulting in the construction of a PE-F2- KDEL recombinant protein. F2 and PE-F2-KDEL recombinant proteins were intraperitoneally injected to mice. Results showed that PE-F2-KDEL induced significantly higher levels of anti-PCV2 serum IgG antibodies than F2. Additionally, PE-F2-KDEL recombinant protein also stimulated significantly higher levels of PCV2-specific IgG compared to inactivated PCV2 whole virus antigen. These results showed that detoxified Pseudomonas exotoxin A could enhance the immune response of a PCV2 ORF2 recombinant protein.

並列關鍵字

PVC2 ORF2 recombinant protein ； porcine circovirus type 2 ； Pseudomonas aeruginosa exotoxin A

國際替代計量

Detoxified Pseudomonas Exotoxin A Enhanced the Immune Response of a Porcine Circovirus type 2 (PCV2) ORF2Recombinant Protein

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