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Immunogenicity of Empty Capsids of Porcine Circovius Type 2 Produced in Insect Cells

Abstract

Porcine circovirus type 2 (PCV2), a single-stranded DNA virus, is associated with postweaning multisystemic wasting syndrome (PMWS). ORF2 protein (capsid) of PCV2 was recently demonstrated to be a major immunogenable to induce protection in pigs with a prime–boost protocol. In this study, the ORF2 gene of PCV2 was expressed in insect cells. The product self-assembled into particles that were structurally and antigenically indistinguishable from regular PCV2 capsids. To evaluated the immunogenicity of these virus-like particles, PCV2-free piglets were vaccinated with the crude lysate from recombinant baculovirus (Ac.ORF2)-infected insect cells, at doses of 0.1 ml (106 cells), 0.5 ml (5 × 106 cells) or 1.0 ml (107 cells). The immune response was monitored by an indirect enzyme-linked immunosorbent assay (ELISA) for PCV2 antibody and lymphocyte proliferation assay. The ELISA results indicated that primary immune response was elicited with 0.5 ml or 1.0 ml of crude lysate from Ac.ORF2. After boost immunization, relatively higher levels of PCV2 antibody were elicited in 0.5-ml or 1.0-ml vaccinated groups, compared to the 0.1-ml group. In addition, higher PCV2 specific lymphocyte proliferation response was developed in piglets vaccinated with 0.5 ml or 1.0 ml of crude lysate, especially in those vaccinated with with 1.0 ml of crude lysate. Thus, the expressed ORF2 protein has significant potential as a subunit vaccine against PCV2 infection.

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Abbreviations

AcNPV:

Autographa california nuclear ployhedrosis virus

Ac.ORF2:

recombinant AcNPV expressing ORF2 protein

CTL:

cytotoxic lymphocyte

ELISA:

enzyme-linked immunosorbent assay

HRP:

horseradish peroxidase

MHC:

major histocompatibility complex

PBMC:

peripheral blood mononuclear cell

PCR:

polymerase chain reaction

PCV:

porcine circovirus

pfu:

plaque-forming units

p.i.:

post infection

PMWS:

post-weaning multisystemic wasting syndrome

PTA:

phosphotungstic acid

Sf9:

Spodoptera frugiperda

SI:

stimulation index

VLP:

virus-like particle

wt. AcNPV:

wild-type Autographa california nuclear polyhedrosis virus

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Correspondence to H. Chen.

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Fan, H., Ju, C., Tong, T. et al. Immunogenicity of Empty Capsids of Porcine Circovius Type 2 Produced in Insect Cells. Vet Res Commun 31, 487–496 (2007). https://doi.org/10.1007/s11259-007-3469-7

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Keywords

  • ORF2
  • porcine circovirus type 2 (PCV2)
  • postweaning multisystemic wasting syndrome
  • PMWS subunit vaccine