Xác định khả năng kích thích tạo kháng thể đặc hiệu của kháng nguyên tái tổ hợp GP5-ELB của virus PRRS gây hội chứng rối loạn sinh sản và hô hấp ở lợn trên động vật thí nghiệm=Possibility in creating specific antibody of GP5-ELP recombinant antigen of Porcine reproductive and respiratory syndrome virus (PRRSV) on experimental animals

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Tác giả: Chu Hoàng Hà, Hồ Thị Hương, Minh Hằng Nguyễn Thị, Nam Nguyễn Trung, Phạm Bích Ngọc

Ngôn ngữ: vie

Ký hiệu phân loại:

Thông tin xuất bản: Tạp chí Khoa học kỹ thuật Thú y, 2018

Mô tả vật lý: tr.35

Bộ sưu tập: Metadata

ID: 545005

Thêm vào giỏ

Hội chứng rối loạn sinh sản và hô hấp ở lợn (Porcine reproductive and respiratory syndrome -PRRS) do virus PRRS thuộc chi Arterivirus, họ Arteriviridae, thuộc bộ Nidovirales gây ra đã và đanggây thiệt hại lớn về kinh tế cho ngành chăn nuôi ở nhiều quốc gia trên khắp thế giới. Glycoprotein 5(GP5) là một protein vỏ quan trọng của virus PRRS, có khối lượng phân tử là 24-25 kDa, được biếtđến như yếu tố kích thích việc sản sinh kháng thể trung hòa ở lợn. GP5 được nghiên cứu để thiết kếvaccine tiểu đơn vị phòng chống PRRS. Để có cơ sở khoa học cho việc nghiên cứu phát triển vaccinetiểu đơn vị có nguồn gốc thực vật phòng chống PRRS, GP5 đã được dung hợp với ELP tạo proteintái tổ hợp và tích luỹ trong cây thuốc lá bằng phương pháp biểu hiện gen tạm thời. Protein GP5-ELPtừ mô lá thuốc lá N. benthamiana được tách chiết, tinh sạch bằng phương pháp mITC và thu hồiprotein mục tiêu. Hiệu suất thu hồi protein GP5-ELP là 98%, mức độ tinh sạch của protein GP5-ELPlà 81,1%. Protein tinh sạch được sử dụng để gây miễn dịch trên chuột bạch, chủng BALB/C 4-6tuần tuổi và thu huyết thanh từ máu chuột ở các thời điểm: Trước khi tiêm kháng nguyên, ngày thứ21 và 35 sau lần tiêm kháng nguyên đầu tiên. Kháng nguyên tinh sạch GP5-ELP được tạo ra có khảnăng kích thích sản sinh kháng thể IgG đặc hiệu kháng GP5-ELP trên chuột. Kháng thể IgG đặc hiệukháng GP5-ELP đã tăng lên hơn 2,4 lần sau ba lần tiêm so với sau hai lần tiêm. Như vậy, có thể khẳngđịnh rằng kháng nguyên GP5-ELP được tổng hợp trong mô lá thuốc lá N. benthamiana bằng phươngpháp biểu hiện gen tạm thời trong thực vật có khả năng gây ra đáp ứng miễn dịch trên động vật thínghiệm. Đây là căn cứ khoa học quan trọng để nghiên cứu sử dụng GP5-ELP như một ứng viên trongviệc phát triển vaccine tiểu đơn vị phòng chống PRRSPorcine reproductive and respiratory syndrome (PRRS) caused by PRRS virus has resultedin huge economic losses to the livestock sector in many countries around the world. PRRSVbelongs to Arterivirus, Arteriviridae and Nidovirales. GP5 is an important PRRSV envelopeglycoprotein with molecular weight of 24-25 kDa, known as the stimulant for the production of neutralizing antibodies in pigs, consequently to design the PRRS subunit vaccine. In order to havea scientific evidence for the development of a PRRS-derived plant-derived subunit vaccine, GP5was fused with ELP to create recombinant protein and accumulated in tobacco tree by transientgene expression. The GP5-ELP protein from N. benthamiana leaf extract was purified by mITC andrecovered target protein, with 98% GP5-ELP protein recovery efficiency and 81.1% GP5-ELP purityprotein. Pure protein was used to induce immunity in 4-6 week BALB/C mice and blood sampleswere collected at the times: before antigen injection and day 21, 35 after the first antigen injection.IgG antigen-specific antibody to GP5-ELP antigen in mouse serum was detected by ELISA andWestern blot analysis. The studied results showed that GP5-ELP purified antigen was capable ofstimulating production of GP5-ELP-specific IgG antibody in mice. GP5-ELP-specific IgG productionwas increased more than 2.4 times after three injections compared to two injections. Thus, it can beconcluded that GP5-ELP antigen synthesized in tobacco leaf tissue by transient gene expressionwas capable of inducing immune responses in mice. This is an important scientific evidence for usingGP5-ELP as a candidate to develop a PRRS subunit vaccine.

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