Complexity of the single linear neutralization epitope of the mouse arterivirus lactate dehydrogenase-elevating virus.
Results from indirect ELISAs using synthetic peptides of various length that represent segments of the ectodomain of the envelope glycoprotein, VP-3P, of lactate dehydrogenase-elevating virus (LDV) showed that the primary neutralization epitope of LDV is located in a short linear hydrophilic segment in the center of the ectodomain. The epitope becomes slightly altered by amino acid substitutions in the ectodomain and inactivation of virions by various treatments. Neutralizing anti-VP-3P antibodies (Abs) to the epitope interact with the synthetic peptides only if they possess a certain conformation. When the peptides were immobilized on ELISA plates, neutralizing mAbs elicited to inactivated LDV and neutralizing Abs from infected mice bound best to the peptides that consisted of the full-length, 30-amino-acid-long ectodomain. The Abs bound poorly, if at all, to most of the shorter peptides when immobilized, whether truncated at the N- or C-end, but when in solution the same peptides strongly inhibited the binding of the Abs to immobilized full-length peptides. Thus, a conformation of the epitope required for Ab binding and (or) its steric accessibility were lost upon immobilization of the shorter peptides on ELISA plates. Abs raised in mice to peptide-bovine serum albumin conjugates reacted only with immobilized peptides in the indirect ELISA and failed to neutralize LDV. The neutralization epitope of the common LDV quasispecies, LDV-P and LDV-vx, is flanked by N-glycans that block the immunogenicity of the epitope and the neutralization of these LDVs. Abs to a second weakly immunogenic and probably discontinuous epitope appear in LDV infected mice about 1 month postinfection.
Plagemann
P G
PG
Department of Microbiology, University of Minnesota, Box 196 UMHC, Minneapolis, Minnesota 55455, USA.
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D018174:Arterivirus Infections / Q000276:immunology*; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000276:immunology*; D007772:Lactate dehydrogenase-elevating virus / Q000276:immunology*; D008562:Membrane Glycoproteins / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D009842:Oligopeptides / Q000276:immunology* / Q000302:isolation & purification; D012996:Solutions; D013997:Time Factors; D014759:Viral Envelope Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D018985:Epitopes, B-Lymphocyte; D008562:Membrane Glycoproteins; D009842:Oligopeptides; D012996:Solutions; C096091:VP-3 protein, lactate dehydrogenase-elevating virus; D014759:Viral Envelope Proteins