Epitope mapping of the major capsid protein of type 2 porcine circovirus (PCV2) by using chimeric PCV1 and PCV2

Porntippa Lekcharoensuk, Igor Morozov, Prem S. Paul, Nattarat Thangthumniyom, Worawidh Wajjawalku, X. J. Meng

Research output: Contribution to journalArticle

134 Citations (Scopus)

Abstract

Type 2 porcine circovirus (PCV2) is associated with postweaning multisystemic wasting syndrome in pigs, whereas the genetically related type 1 PCV (PCV1) is nonpathogenic. In this study, seven monoclonal antibodies (MAbs) against PCV2-ORF2 capsid protein were generated, biologically characterized, and subsequently used to map the antigenic sites of PCV2 capsid protein by using infectious PCV DNA clones containing PCV1/PCV2-ORF2 chimeras. The PCV1/PCV2-ORF2 chimeras were constructed by serial deletions of PCV2-ORF2 and replacement with the corresponding sequences of the PCVI-ORF2. The reactivities of chimeric PCV1/PCV2 ciones in transfected PK-15 cells with the seven MAbs were detected by an immunofluorescence assay (IFA). The chimera (r140) with a deletion of 47 amino acids at the N terminus of PCV2-ORF2 reacted strongly to all seven MAbs. Expanding the deletion of PCV2-ORF42 from residues, 17 to 57 (r175) abolished the recognition of MAb 3B7, 3C11, 4A10, 6H2, or 8F6 to the chimera. Further deletion of PCV2-ORF2 to 62 residues disrupted the binding of this chimera to all seven MAbs. IFA reactivities with all MAbs were absent when residues 165 to 233 at the C terminus of PCV2-ORF2 was replaced with that of PCV1-ORF2. Extending the sequence of PCV2-ORF2 from residues 165 (r464) to 185 (r526), 200 (r588), or 224 (r652) restored the ability of the three chimeras to react with MAbs 3C11, 6H2, 9H7, and 12G3 but not with 8F6, 3B7, or 4A10. When the four amino acids at the C terminus of r588 were replaced with that of PCV2-ORF2, the resulting chimera (r588F) reacted with all seven MAbs. The results from this study suggest that these seven MAbs recognized at least five different but overlapping conformational epitopes within residues 47 to 63 and 165 to 200 and the last four amino acids at the C terminus of the PCV2 capsid protein.

Original languageEnglish (US)
Pages (from-to)8135-8145
Number of pages11
JournalJournal of virology
Volume78
Issue number15
DOIs
StatePublished - Aug 1 2004

Fingerprint

Circovirus
Epitope Mapping
Porcine circovirus-2
Capsid Proteins
coat proteins
chimerism
monoclonal antibodies
Monoclonal Antibodies
Amino Acids
fluorescent antibody technique
amino acids
Fluorescent Antibody Technique
Wasting Syndrome
postweaning multisystemic wasting syndrome
epitope mapping
assays
epitopes
Epitopes
Swine
Clone Cells

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Lekcharoensuk, P., Morozov, I., Paul, P. S., Thangthumniyom, N., Wajjawalku, W., & Meng, X. J. (2004). Epitope mapping of the major capsid protein of type 2 porcine circovirus (PCV2) by using chimeric PCV1 and PCV2. Journal of virology, 78(15), 8135-8145. https://doi.org/10.1128/JVI.78.15.8135-8145.2004

Epitope mapping of the major capsid protein of type 2 porcine circovirus (PCV2) by using chimeric PCV1 and PCV2. / Lekcharoensuk, Porntippa; Morozov, Igor; Paul, Prem S.; Thangthumniyom, Nattarat; Wajjawalku, Worawidh; Meng, X. J.

In: Journal of virology, Vol. 78, No. 15, 01.08.2004, p. 8135-8145.

Research output: Contribution to journalArticle

Lekcharoensuk, P, Morozov, I, Paul, PS, Thangthumniyom, N, Wajjawalku, W & Meng, XJ 2004, 'Epitope mapping of the major capsid protein of type 2 porcine circovirus (PCV2) by using chimeric PCV1 and PCV2', Journal of virology, vol. 78, no. 15, pp. 8135-8145. https://doi.org/10.1128/JVI.78.15.8135-8145.2004
Lekcharoensuk, Porntippa ; Morozov, Igor ; Paul, Prem S. ; Thangthumniyom, Nattarat ; Wajjawalku, Worawidh ; Meng, X. J. / Epitope mapping of the major capsid protein of type 2 porcine circovirus (PCV2) by using chimeric PCV1 and PCV2. In: Journal of virology. 2004 ; Vol. 78, No. 15. pp. 8135-8145.
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