title: Characterization of porcine circovirus type 2 (PCV2) capsid particle assembly and its application to virus-like particle vaccine development.
abstract: Porcine circovirus type 2 (PCV2) is the primary causative agent of porcine circovirus-associated diseases in pigs. The sole structural capsid protein of PCV2, Cap, consists of major antigenic domains, but little is known about the assembly of capsid particles. The purpose of this study is to produce a large amount of Cap protein using Escherichia coli expression system for further studying the essential sequences contributing to formation of particles. By using codon optimization of rare arginine codons near the 5'-end of the cap gene for E. coli, a full-length Cap without any fusion tag recombinant protein (Cap1-233) was expressed and proceeded to form virus-like particles (VLPs) in normal Cap appearance that resembled the authentic PCV2 capsid. The N-terminal deletion mutant (Cap51-233) deleted the nuclear localization signal (NLS) domain, while the internal deletion mutant (CapΔ51-103) deleted a likely dimerization domain that failed to form VLPs. The unique Cys108 substitution mutant (CapC/S) exhibited most irregular aggregates, and only few VLPs were formed. These results suggest that the N-terminal region within the residues 1 to 103 possessing the NLS and dimerization domains are essential for self-assembly of stable Cap VLPs, and the unique Cys108 plays an important role in the integrity of VLPs. The immunogenicity of PCV2 VLPs was further evaluated by immunization of pigs followed by challenge infection. The Cap1-233-immunized pigs demonstrated specific antibody immune responses and are prevented from PCV2 challenge, thus implying its potential use for a VLP-based PCV2 vaccine.
authors.affiliation: College of Veterinary Medicine, National Chung Hsing University, 250 Kuo Kuang Road, Taichung 40227 Taiwan, Republic of China.
authors.forename: Pei-Ching
authors.initials: PC
authors.lastname: Wu
authors.forename: Wei-Li
authors.initials: WL
authors.lastname: Lin
authors.forename: Chi-Ming
authors.initials: CM
authors.lastname: Wu
authors.forename: Jiun-Ni
authors.initials: JN
authors.lastname: Chi
authors.forename: Maw-Sheng
authors.initials: MS
authors.lastname: Chien
authors.forename: Chienjin
authors.initials: C
authors.lastname: Huang
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D036022:Capsid Proteins / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D018173:Circoviridae Infections / Q000276:immunology / Q000517:prevention & control / Q000662:veterinary* / Q000821:virology; D017925:Circovirus / Q000235:genetics / Q000276:immunology* / Q000502:physiology; D013552:Swine; D013553:Swine Diseases / Q000276:immunology* / Q000517:prevention & control / Q000821:virology; D058425:Vaccines, Virus-Like Particle / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D014765:Viral Vaccines / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D019065:Virus Assembly
chemical_list: D000914:Antibodies, Viral; D036022:Capsid Proteins; D058425:Vaccines, Virus-Like Particle; D014765:Viral Vaccines