PMID- 11238854
OWN - NLM
STAT- MEDLINE
DCOM- 20010405
LR  - 20201208
IS  - 0022-538X (Print)
IS  - 1098-5514 (Electronic)
IS  - 0022-538X (Linking)
VI  - 75
IP  - 7
DP  - 2001 Apr
TI  - Epitope mapping porcine reproductive and respiratory syndrome virus by phage 
      display: the nsp2 fragment of the replicase polyprotein contains a cluster of 
      B-cell epitopes.
PG  - 3277-90
AB  - We screened phage display libraries of porcine reproductive and respiratory 
      syndrome virus (PRRSV) protein fragments with sera from experimentally infected 
      pigs to identify linear B-cell epitopes that are commonly recognized during 
      infection in vivo. We identified 10 linear epitope sites (ES) 11 to 53 amino 
      acids in length. In the replicase polyprotein, a total of eight ES were 
      identified, six of which localized to the Nsp2 replicase polyprotein processing 
      end product. In the structural proteins, a total of two ES were identified, in 
      the ORF3 and ORF4 minor envelope glycoproteins. The ORF4 ES was previously 
      identified by monoclonal antibody mapping (J. J. M. Meulenberg, A. P. van 
      Nieuwstadt, A. van Essen-Zandenbergen, and J. P. M. Langeveld, J. Virol. 
      71:6061-6067, 1997), but its immunogenicity had not been examined in pigs. We 
      found that six experimentally PRRSV-infected pigs consistently had very high 
      antibody titers against the ORF4 ES. In some animals, sera diluted 1:62,500 still 
      gave weak positive enzyme immunoassay reactivity against the ORF4 ES. This 
      hitherto unrecognized immunodominance likely caused phages displaying the ORF4 ES 
      to outcompete phages displaying other ES during library screening with porcine 
      sera and accounted for our failure to identify more than two ES in the structural 
      genes of PRRSV. Genetic analysis showed that variable ES were also the most 
      immunogenic in vivo. Serological analysis indicated differences in the 
      immunoglobulin A responses between short-term and longer-term viremic pigs 
      towards some ES. The implications of these findings for PRRSV diagnostics and 
      immunopathogenesis are discussed.
FAU - Oleksiewicz, M B
AU  - Oleksiewicz MB
AD  - Danish Veterinary Institute for Virus Research, Lindholm, 4771 Kalvehave, 
      Denmark.
FAU - Botner, A
AU  - Botner A
FAU - Toft, P
AU  - Toft P
FAU - Normann, P
AU  - Normann P
FAU - Storgaard, T
AU  - Storgaard T
LA  - eng
SI  - GENBANK/M96262
PT  - Journal Article
PL  - United States
TA  - J Virol
JT  - Journal of virology
JID - 0113724
RN  - 0 (Antibodies, Viral)
RN  - 0 (Epitopes, B-Lymphocyte)
RN  - 0 (Immune Sera)
RN  - 0 (Peptide Library)
RN  - 0 (Viral Nonstructural Proteins)
RN  - 0 (Viral Structural Proteins)
RN  - EC 2.7.7.48 (RNA-Dependent RNA Polymerase)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Antibodies, Viral/blood
MH  - Enzyme-Linked Immunosorbent Assay
MH  - *Epitope Mapping
MH  - *Epitopes, B-Lymphocyte
MH  - Genes, Viral
MH  - Genetic Variation
MH  - Immune Sera/immunology
MH  - Molecular Sequence Data
MH  - Open Reading Frames
MH  - *Peptide Library
MH  - Porcine respiratory and reproductive syndrome 
      virus/genetics/*immunology/pathogenicity
MH  - RNA-Dependent RNA Polymerase/*immunology
MH  - Swine
MH  - Viral Nonstructural Proteins/*immunology
MH  - Viral Structural Proteins/genetics
MH  - Viremia/virology
PMC - PMC114121
EDAT- 2001/03/10 10:00
MHDA- 2001/04/06 10:01
PMCR- 2001/04/01
CRDT- 2001/03/10 10:00
PHST- 2001/03/10 10:00 [pubmed]
PHST- 2001/04/06 10:01 [medline]
PHST- 2001/03/10 10:00 [entrez]
PHST- 2001/04/01 00:00 [pmc-release]
AID - 1739 [pii]
AID - 10.1128/JVI.75.7.3277-3290.2001 [doi]
PST - ppublish
SO  - J Virol. 2001 Apr;75(7):3277-90. doi: 10.1128/JVI.75.7.3277-3290.2001.