Diagnostic potential of parechovirus capsid proteins.
To study humoral and cellular immunity against human parechovirus type 1 (HPEV1), the viral capsid proteins VP0, VP1, and VP3 were expressed and purified as glutathione S-transferase (GST)-tagged recombinant proteins. The fusion proteins were used to raise antisera in rabbits. VP0 and VP1 antisera specifically detected HPEV1-infected cells in culture by immunoperoxidase staining and immunofluorescence. Furthermore, antisera against the VP0 and VP1 proteins had neutralizing effects against HPEV1 infection. When the HPEV1 antibody titers of 20 adults and 55 children were determined by a microneutralization test, the prevalence of HPEV1 antibodies in the adult population was 96%, while 50% of children were seropositive. Selected sera were used to evaluate HPEV1 fusion proteins as antigens in an enzyme immunoassay. The VP3 capsid protein appeared to be suitable for the purpose, with specificity of 100% and sensitivity of 96% compared to the neutralization test. Furthermore, T-cell responses to the purified HPEV1 and HPEV1 capsid fusion proteins were studied in 20 adults. Sixty percent of the subjects had T-cell proliferation responses to purified HPEV1, and 90% of the subjects also had positive T-cell responses to at least one of the GST capsid proteins.
Alho
Annu
A
Department of Virology, Haartman Institute, University of Helsinki, FIN-00014 Helsinki, Finland. annu.alho@helsinki.fi
Marttila
Jane
J
Ilonen
Jorma
J
Hyypiä
Timo
T
D000328:Adult; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D005455:Fluorescent Antibody Technique; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007124:Immunoenzyme Techniques; D008213:Lymphocyte Activation; D008297:Male; D008875:Middle Aged; D030061:Parechovirus / Q000276:immunology*; D010850:Picornaviridae Infections / Q000175:diagnosis* / Q000821:virology; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology
D000914:Antibodies, Viral; D036022:Capsid Proteins; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase