title: Egg yolk IgY against RHDV capsid protein VP60 promotes rabbit defense against RHDV infection.
abstract: VP60 capsid protein is the major structural and immunogenicity protein of RHDV (Rabbit hemorrhagic disease virus, RHDV), and has been implicated as a main protein antigen in RHDV diagnosis and vaccine design. In this report, egg yolk antibody (IgY) against N-terminal of VP60 was evaluated and developed as a new strategy for RHDV therapy. Briefly, N-terminal of VP60 (∼250aa) fragment was cloned and inserted into pET28a expression vector, and then the resultant plasmid, pET28a/VP60-N, was transformed into E. coli BL21(DE3) for recombinant VP60-N protein (rVP60-N) expression. Next, the rVP60-N was purified by Ni(+)-affinity purification chromatography and identified by Western blotting with RHDV antiserum. After immunizing the chickens with rVP60-N, the anti-rVP60-N IgY was isolated, and the activity and specificity of the IgY antibody were analyzed by ELISA and Western blotting. In our results, the rVP60-N could be expressed in E. coli as soluble fraction, and the isolated anti-rVP60-N IgY demonstrated a high specificity and titer (1:22,000) against rVP60-N antigen. For further evaluation of the IgY efficacy in vivo, rabbits were grouped randomly and challenged with RHDV, and the results showed that anti-rVP60-N IgY could significantly protect rabbits from virus infection and promote the host survival after a sustained treatment with anti-rVP60-N IgY for 5 days. Taken together, our study demonstrates evidence that production of IgY against VP60 could be as a novel strategy for the RHDV therapy.
authors.affiliation: Key Laboratory of Pharmaceutical Engineering Technology and Application, School of Chemical and Pharmaceutical Engineering, Sichuan University of Science and Engineering, Zigong 643000, China.
authors.forename: Zai Xin
authors.initials: ZX
authors.lastname: Li
authors.affiliation: Key Laboratory of Pharmaceutical Engineering Technology and Application, School of Chemical and Pharmaceutical Engineering, Sichuan University of Science and Engineering, Zigong 643000, China.
authors.forename: Wei Dong
authors.initials: WD
authors.lastname: Hu
authors.affiliation: Key Laboratory of Pharmaceutical Engineering Technology and Application, School of Chemical and Pharmaceutical Engineering, Sichuan University of Science and Engineering, Zigong 643000, China.
authors.forename: Bing Chao
authors.initials: BC
authors.lastname: Li
authors.affiliation: Key Laboratory of Pharmaceutical Engineering Technology and Application, School of Chemical and Pharmaceutical Engineering, Sichuan University of Science and Engineering, Zigong 643000, China.
authors.forename: Tian You
authors.initials: TY
authors.lastname: Li
authors.affiliation: Key Laboratory of Pharmaceutical Engineering Technology and Application, School of Chemical and Pharmaceutical Engineering, Sichuan University of Science and Engineering, Zigong 643000, China.
authors.forename: Xiao Yang
authors.initials: XY
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Pharmaceutical Engineering Technology and Application, School of Chemical and Pharmaceutical Engineering, Sichuan University of Science and Engineering, Zigong 643000, China. Electronic address: zhangzhi02@gmail.com.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D015153:Blotting, Western / Q000662:veterinary; D017250:Caliciviridae Infections / Q000276:immunology / Q000517:prevention & control / Q000662:veterinary* / Q000821:virology; D002645:Chickens; D004530:Egg Yolk / Q000276:immunology*; D005260:Female; D017929:Hemorrhagic Disease Virus, Rabbit / Q000235:genetics / Q000276:immunology*; D007114:Immunization / Q000379:methods / Q000662:veterinary; D007136:Immunoglobulins / Q000235:genetics / Q000276:immunology*; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics; D011817:Rabbits / Q000276:immunology / Q000821:virology*; D011897:Random Allocation; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D015678:Viral Structural Proteins / Q000235:genetics / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; C016512:IgY; D007136:Immunoglobulins; D012367:RNA, Viral; D015678:Viral Structural Proteins; C081149:viral protein 60, rabbit hemorrhagic disease virus
keywords: Egg yolk antibody (IgY); Rabbit hemorrhagic disease virus (RHDV); VP60 capsid protein