PMID- 9875321
OWN - NLM
STAT- MEDLINE
DCOM- 19990120
LR  - 20061115
IS  - 0042-6822 (Print)
IS  - 0042-6822 (Linking)
VI  - 252
IP  - 1
DP  - 1998 Dec 5
TI  - Localization and fine mapping of antigenic sites on the nucleocapsid protein N of 
      porcine reproductive and respiratory syndrome virus with monoclonal antibodies.
PG  - 106-14
AB  - The purpose of this study was to analyze the antigenic structure of the 
      nucleocapsid protein N of the Lelystad virus isolate of porcine reproductive and 
      respiratory syndrome virus (PRRSV) and to identify antigenic differences between 
      this prototype European isolate and other North American isolates. To do this, we 
      generated a panel of monoclonal antibodies (mAbs) directed against the N protein 
      of Lelystad virus and tested them in competition assays with other N-specific 
      mAbs described previously (Drew et al., 1995; Nelson et al., 1993; van Nieuwstadt 
      et al., 1996). Four different competition groups of mAbs were identified. Pepscan 
      analysis with solid-phase dodecapeptides was used to identify specific antigenic 
      regions in the N protein that were bound by the mAbs. In this pepscan analysis, 
      we found that the mAb of the first competition group reacted with linear peptides 
      whose core sequences consisted of amino acids 2-12 (site A), the mAbs of the 
      second group reacted with peptides whose core sequences consisted of amino acids 
      25-30 (site B), and the mAb of the third group reacted with peptides whose core 
      sequences consisted of amino acids 40-46 (site C). However, the fourth group of 
      mAbs binding to an antigenic region, provisionally designated as domain D, 
      reacted very weakly or did not react at all with solid-phase dodecapeptides. To 
      further characterize the structure of the epitopes in domain D, we produced 
      chimeric constructs composed of the N protein sequences of Lelystad virus and 
      another arterivirus lactate dehydrogenase-elevating virus, which was used because 
      its N protein has similarity in amino acid sequence and hydropathicity profile 
      but does not react with our mAbs. When the mAbs specific to domain D were tested 
      for binding to the chimeric N proteins expressed by Semliki Forest virus, we 
      found that the regions between amino acids 51-67 and amino acids 80-90 are 
      involved in the formation or are part of the epitopes in domain D. Therefore, we 
      conclude that the N protein contains four distinct antigenic regions. The 
      epitopes mapped to sites A-C are linear, whereas the epitopes mapped to domain D 
      are more conformation dependent or discontinuous. Sites A and C contain epitopes 
      that are conserved in European but not in North American isolates; site B 
      contains epitopes that are conserved in European and North American isolates; and 
      site D contains epitopes that are either conserved or not conserved in European 
      and North American isolates. The antigenic regions identified here might be 
      important for the development of diagnostic test for PRRSV in particular tests 
      that discriminate between different antigenic types of PRRSV.
FAU - Meulenberg, J J
AU  - Meulenberg JJ
AD  - Institute for Animal Science and Health, Lelystad, The Netherlands. 
      J.J.M.Meulenberg@id.dlo.nl
FAU - van Nieuwstadt, A P
AU  - van Nieuwstadt AP
FAU - van Essen-Zandbergen, A
AU  - van Essen-Zandbergen A
FAU - Bos-de Ruijter, J N
AU  - Bos-de Ruijter JN
FAU - Langeveld, J P
AU  - Langeveld JP
FAU - Meloen, R H
AU  - Meloen RH
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Virology
JT  - Virology
JID - 0110674
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (DNA, Viral)
RN  - 0 (Nucleocapsid Proteins)
RN  - 0 (Recombinant Fusion Proteins)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Antibodies, Monoclonal/*immunology
MH  - Antibody Specificity
MH  - Binding Sites
MH  - Binding, Competitive
MH  - Cell Line
MH  - Cloning, Molecular
MH  - Cricetinae
MH  - DNA, Viral/chemistry
MH  - Epitope Mapping/methods
MH  - Mice
MH  - Molecular Sequence Data
MH  - Nucleocapsid Proteins/chemistry/genetics/*immunology
MH  - Peptide Mapping/methods
MH  - Porcine respiratory and reproductive syndrome 
      virus/chemistry/genetics/*immunology
MH  - Recombinant Fusion Proteins/chemistry
EDAT- 1999/01/06 00:00
MHDA- 1999/01/06 00:01
CRDT- 1999/01/06 00:00
PHST- 1999/01/06 00:00 [pubmed]
PHST- 1999/01/06 00:01 [medline]
PHST- 1999/01/06 00:00 [entrez]
AID - S0042-6822(98)99436-3 [pii]
AID - 10.1006/viro.1998.9436 [doi]
PST - ppublish
SO  - Virology. 1998 Dec 5;252(1):106-14. doi: 10.1006/viro.1998.9436.