T lymphocyte epitope mapping of porcine circovirus type 2.
Immunoreactive T lymphocyte epitopes within the ORF1, ORF2, and ORF 3 products of porcine circovirus type 2 (PCV2) were mapped. For this, overlapping linear 20-mer peptides were synthesized and tested for their ability to induce T lymphocyte proliferation in porcine peripheral blood mononuclear cells (PBMCs) isolated from experimentally PCV2-infected pigs. After a preliminary screening of 31 (ORF1), 23 (ORF2), and 10 (ORF3) peptides using PBMCs from 4 PCV2-infected pigs, none of the peptides appeared to be immunoreactive (stimulation index [SI] : 2) in all four pigs. Only 14 peptides appeared to be immunoreactive in 3 of the 4 pigs. These peptides were designated as immunodominant in the preliminary screening and selected for further analysis. The immunodominant peptides were resynthesized and purified by high-performance liquid chromatography and tested for their ability to induce T lymphocyte proliferation in PBMCs from another three PCV2-infected pigs. None of the immunodominant peptides appeared to be immunoreactive in all three pigs of the second screening. Only three peptides appeared to be immunoreactive in two of three pigs, two encoded by PCV2 ORF1 (amino acid residues 81-100 and 201-220) and one encoded by PCV2 ORF3 (amino acid residues 31-50), and were therefore considered to be immunodominant in both screenings. Although peptides encoded by ORF2 appeared to show the highest immunoreactivity in some pigs, none of these peptides displayed immunodominance in both screenings. In summary, the present study indicates that the T lymphocyte responses to PCV2 are primarily directed toward epitopes of the nonstructural proteins of ORF1 and ORF3.
Stevenson
Leanne S
LS
Department of Veterinary Science, Queen's University Belfast, Belfast BT4 3SD, United Kingdom. s.stevenson@qub.ac.uk
Gilpin
Deirdre F
DF
Douglas
Alastair
A
McNeilly
Francis
F
McNair
Irene
I
Adair
Brian M
BM
Allan
Gordon M
GM
D000818:Animals; D049109:Cell Proliferation; D002478:Cells, Cultured; D017925:Circovirus / Q000276:immunology*; D018604:Epitope Mapping; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D016056:Immunodominant Epitopes / Q000276:immunology; D007963:Leukocytes, Mononuclear; D008213:Lymphocyte Activation; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology; D013552:Swine; D014764:Viral Proteins / Q000276:immunology
D018984:Epitopes, T-Lymphocyte; D016056:Immunodominant Epitopes; D010455:Peptides; D014764:Viral Proteins