Skip to main content
SpringerLink
Log in

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

  • Published:
Veterinary Research Communications Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

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

References

  • Allan, G.M., Meehan, B.M., Todd, D., Kennedy, S., McNeilly, F., Ellis, J., Clark, E.G., Harding, J., Espuna, E., Botner, A. and Charreyre, C., 1998a. Novel porcine circoviruses from pigs with wasting disease syndromes. Veterinary Record, 142, 467–468

    PubMed  CAS  Google Scholar 

  • Allan, G.M., McNeilly, F., Kennedy, S., Daft, B., Clark, E.G., Ellis, J.A., Haines, D.M., Meehan, B.M. and Adair, B.M., 1998b. Isolation of porcine circovirus-like viruses from pigs with a wasting disease in the USA and Europe. Journal of Veterinary Diagnostic Investigation, 10, 3–10

    PubMed  CAS  Google Scholar 

  • Allan, G.M. and Ellis, J.A., 2000. Porcine circovirus: a review. Journal of Veterinary Diagnostic Investigation, 12, 3–14

    PubMed  CAS  Google Scholar 

  • Allan, G.M., McNeilly, F., Ellis, J., Krakowka, S., Meehan, B., McNair, I. and Walker, I., Kennedy, S., 2000. Experimental infection of colostrum-deprived piglets with porcine circovirus 2 (PCV2) and porcine reproductive and respiratory syndrome virus (PRRSV) potentiates PCV2 replication. Archives of Virology, 145, 2421–2429

    Article  PubMed  CAS  Google Scholar 

  • Bevan, M.J., 1976a. Cross-priming for a secondary cytotoxic response to minor H antigens with H-2 congenic cells which do not cross-react in the cytotoxic assay. Journal of Experimental Medicine, 143(5), 1283–1288

    Article  CAS  Google Scholar 

  • Bevan, M.J., 1976b. Minor H antigens introduced on H-2 different stimulating cells cross-react at the cytotoxic T-cell level during in vivo priming. Journal of Immunology, 117(6), 2233–2238

    CAS  Google Scholar 

  • Blanchard, P., Mahe, D., Cariolet, R., Keranflec'h, A., Baudouard, M.A., Cordioli, P., Albina, E. and Jestin, A., 2003. Protection of swine against post-weaning multisystemic wasting syndrome (PMWS) by porcine circovirus type 2 (PCV2) proteins. Vaccine, 21, 4565–4575

    Article  PubMed  CAS  Google Scholar 

  • Boisgérault, F., Moron, G. and Leclerc, C., 2002. Virus-like particles: a new family of delivery systems. Expert Review of Vaccines, 1, 101–109

    Article  PubMed  Google Scholar 

  • Clark, E.G., 1997. Post-weaning multisystemic wasting syndrome. In: Proceedings of the American Associations of Swine Practitioners, 499–501

  • De Arriba, M.L., Carvajal, A., Pozo, J. and Rubio, P., 2002. Isotype-specific antibody-secreting cells in systemic and mucosal associated lymphoid tissues and antibody responses in serum of conventional pigs inoculated with PEDV. Veterinary Immunology and Immunopathology, 84, 1–16

    Article  PubMed  Google Scholar 

  • Ellis, J., Hassard, L., Clark, E., Harding, J., Allan, G., Willson, P., Strokappe, J., Martin, K., McNeilly, F., Meehan, B., Todd, D. and Haines, D., 1998. Isolation of circovirus from lesions of pigs with postweaning multisystemic wasting syndrome. Canadian Veterinary Journal, 39, 44–51

    CAS  Google Scholar 

  • Ellis, J.A., Bratanich, A., Clark, E.G., Allan, G., Meehan, B., Haines, D.M., Harding, J., Werb, K.H., Krakowka, S, Konoby, C., and Hassand, L., 2000. Coinfection by porcine circoviruses and porcine parvovirus in pigs with naturally acquired postweaning multisystemic wasting syndrome. Journal of Veterinary Diagnostic Investigation, 12, 21–27

    PubMed  CAS  Google Scholar 

  • Fenaux M., Oprissing, T., Halbur, P.G. and Meng, X.J., 2003. Immunogenicity and pathogenicity of chimeric infectious DNA clones of pathogenic porcine circovirus type 2 (PCV2) and nonpathogenic PCV1 in weaning pigs. Journal of Virology, 77(2), 11232–11243

    Article  PubMed  CAS  Google Scholar 

  • Fenaux, M., Oprissing, T., Halbur, P.G., Elvinger, F. and Meng, X.J., 2004. A chimeric procine circovirus (PCV2) with the immunogenic capsid gene of the pathogenic PCV2 type 2 (PCV2) cloned into the genomic backbone of the nonpathogenic PCV1 induces protective immunity against PCV2 infection in pigs. Journal of Virology, 78(12), 6297–6303

    Article  PubMed  CAS  Google Scholar 

  • Hamel, A.L., Lin, L.L. and Nayar, G.P., 1998. Nucleotide sequence of porcine circovirus associated with postweaning multisystemic wasting syndrome in pigs. Journal of Virology, 72: 5262–5267

    PubMed  CAS  Google Scholar 

  • Heermann, K.H. and Gerlich, W.H., 1991. In: A. McLachlan (ed.), Molecular Biology of Hepatitis B Virus, (CRC Press, Boca Raton, FL), 109

    Google Scholar 

  • Kennedy, S., Moffett, D., McNeilly, F., Meehan, B., Ellis, J., Krakowka, S. and Allan, G.M., 2000. Reproduction of lesion of postweaning multisystemic wasting syndrome by infection of conventional pigs with porcine circovirus type 2 alone or in combination with porcine parvovirus. Journal of Comparative Pathology, 122, 9–24

    Article  PubMed  CAS  Google Scholar 

  • Kim, Y., Kim, J., Kang, K. and Lyoo, Y.S., 2002. Characterization of the recombinant proteins of porcine circovirus type 2 field isolate expressed in the baculovirus system. Journal of Veterinary Science, 3(1), 19–23

    PubMed  Google Scholar 

  • Koch, G., van Roozelaar, D.J., Verschueren, C.A., Van der Eb, A.J. and Noteborn, M.H., 1995. Immunogenic and protective properties of chicken anemia virus proteins expressed by baculovirus. Vaccine, 13, 763–770

    Article  PubMed  CAS  Google Scholar 

  • Krakowka, S., Ellis, J.A., McNeilly, F., Rings, D.M. and Allan, G., 2001. Activation of the immune system is the pivotal event in the production of wasting disease in pigs infected with porcine circovirus-2 (PCV2). Veterinary Pathology, 38, 31–42

    Article  PubMed  CAS  Google Scholar 

  • Kuntz-Simon, G., Blanchard, P., Cherbonnel, M., Jestin, A. and Jestin, V., 2002. Baculovirus-expressed muscovy duck reovirus sigmaC protein induces serum neutralizing antibodies and protection against challenge. Vaccine, 20, 3113

    Article  PubMed  CAS  Google Scholar 

  • Liu, Q., Tikoo, S.K. and Babiuk, L.A., 2001. Nuclear localization of the ORF2 protein encoded by porcine circovirus Type 2. Virology, 285, 91–99

    Article  PubMed  CAS  Google Scholar 

  • Madec, F., Eveno, E., Morvan, P., Hamon, L., Blanchard, P., Cariolet, R., Amenna, N., Marvan, H., Truong, C., Mahe, D., Albina, E. and Jestin, A., 2000. Post-weaning multisystemic wasting syndrome (PMWS) in pigs in France: clinical observations from follow-up studies on affected farms. Livestock Production Science, 63, 223–233

    Article  Google Scholar 

  • Morozov, I., Sirinarumitr, T., Sorden, S.D., Halbur, P.G., Morgen, M.K., Yoon, K.J., and Paul, P.S., 1998. Detection of a novel strain of porcine circovirus in pigs with postweaning multisystemic wasting syndrome, Journal of Clinical Microbiology, 36, 2535–2541

    PubMed  CAS  Google Scholar 

  • Nawagitgul, P., Morozov, I., Bolin, S.R., Harms, P.A., Sorden, S.D. and Paul, P.S., 2000. Open reading frame 2 of porcine circovirus type 2 encodes a major capsid protein. Journal of General Virology, 81, 2281–2287

    PubMed  CAS  Google Scholar 

  • Nawagitgul, P., Harms, P.A., Morozov, I., Thacker, B.J., Sorden, S.D., Lekcharoensuk, C. and Paul, P.S., 2002. Modified indirect porcine circovirus (PCV) type 2-based and recombinant capsid protein (ORF2)-based enzyme-linked immunosorbent assays for detection of antibodies to PCV. Clinical and Diagnostic Laboratory Immunology, 9, 33–40

    Article  PubMed  CAS  Google Scholar 

  • O'Reilly, D.R., Miller, L.K. and Luckow, U.A., 1992. Baculovirus Expression Vectors: A Laboratory Manual (W.H. Freeman, New York), 109–134

    Google Scholar 

  • Rovira, A., Balasch, M., Segales, J., Garcia, L., Plana-Duran, J., Rosell, C., Ellerbrok, H., Mankertz, A. and Domingo, M., 2002. Experimental inoculation of conventional pigs with porcine reproductive and respiratory syndrome virus and porcine circovirus 2. Journal of Virology, 76, 3232–3239

    Article  PubMed  CAS  Google Scholar 

  • Todd, D., McNulty, M.S., Mankertz, A., et al., 2000. Family Circoviridae. In: F.A. Murphy, C.M. Fauquet, D.H.L. Bishop, et al. (eds.), Virus Taxonomy. Classification and Nomenclature of Viruses, Seventh Report of the International Committee of Taxonomy of Viruses (Academic Press, New York), 299–303

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratory of Animal Virology, College of Veterinary Medicine, and The National Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, China

    H. Fan, C. Ju, T. Tong, H. Huang, J. Lv & H. Chen

Authors
  1. H. Fan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. C. Ju
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. T. Tong
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. H. Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. J. Lv
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. H. Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to H. Chen.

Rights and permissions

Reprints and permissions

About this article

Cite this article

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

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11259-007-3469-7

Keywords

Search

Navigation