PMID- 15507310
OWN - NLM
STAT- MEDLINE
DCOM- 20041229
LR  - 20061115
IS  - 0165-2427 (Print)
IS  - 0165-2427 (Linking)
VI  - 102
IP  - 3
DP  - 2004 Dec 8
TI  - The primary GP5 neutralization epitope of North American isolates of porcine 
      reproductive and respiratory syndrome virus.
PG  - 263-75
AB  - I have used indirect ELISA with overlapping synthetic peptides representing the 
      GP5 ectodomain to study the generation and specificity of peptide-binding Abs in 
      pigs that were infected in utero with porcine reproductive and respiratory 
      syndrome virus (PRRSV) strain VR2332 and in North American field sera submitted 
      for PRRSV infection diagnosis. Peptide-binding Abs appeared in sera of the 
      VR2332-infected pigs within about 30 days post-farrowing (dpf), reaching maximum 
      titers 100-200 dpf and then decreasing slowly to about half of maximum titer by 
      about 400 dpf. The formation of peptide-binding Abs and of virus neutralizing Abs 
      correlated and their initial appearance coincided with disappearance of virus 
      from the circulation. The Abs were specific for VR2332-specific peptides. In 
      contrast, anti-N-protein Abs as measured by HerdCheck ELISA appeared within 7 
      dpf, reached maximum levels at about 100 dpf and had decreased below detectable 
      levels by about 200 dpf. Twenty-seven field serum samples with virus neutralizing 
      activity all possessed high levels of peptide binding Abs, but the Abs bound 
      about equally to VR2332 and strain Lelystad virus (LV)-specific peptides. The 
      indirect ELISA results using various large peptides and competition ELISA using 
      small peptides (8 or 9 amino acids long) confirmed that the epitope recognized by 
      the Abs is located in the GP5 ectodomain sequence 37SHLQLIYNL of VR2332. Use of 
      mutated peptides in the competition ELISA showed that 42I to T and 38HL to TY 
      substitutions blocked Ab recognition, whereas deletion of 41L had no effect. In 
      addition, 26 serum samples submitted by two farms for diagnostic tests were found 
      to possess low levels of Abs that bound to GP5 ectodomain peptides, even though 
      the sera were sero-negative in the HerdChek ELISA and lacked neutralizing 
      activity. Competition ELISA showed that the Abs recognized one or more epitopes 
      located downstream of the PRRSV neutralization epitope. An epitope(s) located in 
      the same area was recognized by Abs generated in mice by immunization with a GP5 
      ectodomain peptide conjugated to BSA. These Abs also lacked neutralizing 
      activity.
FAU - Plagemann, Peter G W
AU  - Plagemann PG
AD  - Department of Microbiology, University of Minnesota, MMC 196, Minneapolis, MN 
      55455, USA. plage001@umn.edu
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - Vet Immunol Immunopathol
JT  - Veterinary immunology and immunopathology
JID - 8002006
RN  - 0 (Antibodies, Viral)
RN  - 0 (Antigens, Viral)
RN  - 0 (Epitopes)
RN  - 0 (Viral Envelope Proteins)
RN  - 0 (glycoprotein 5, PRRSV)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Antibodies, Viral/biosynthesis/blood
MH  - Antigens, Viral/*immunology
MH  - Enzyme-Linked Immunosorbent Assay/veterinary
MH  - Epitopes/*chemistry/*immunology
MH  - Female
MH  - Mice
MH  - Molecular Sequence Data
MH  - Neutralization Tests/veterinary
MH  - North America
MH  - Porcine Reproductive and Respiratory Syndrome/immunology/virology
MH  - Porcine respiratory and reproductive syndrome virus/*immunology
MH  - Pregnancy
MH  - Pregnancy Complications, Infectious/immunology/veterinary/virology
MH  - Swine/virology
MH  - Time Factors
MH  - Viral Envelope Proteins/*immunology
EDAT- 2004/10/28 09:00
MHDA- 2004/12/30 09:00
CRDT- 2004/10/28 09:00
PHST- 2004/10/28 09:00 [pubmed]
PHST- 2004/12/30 09:00 [medline]
PHST- 2004/10/28 09:00 [entrez]
AID - S0165-2427(04)00260-0 [pii]
AID - 10.1016/j.vetimm.2004.09.011 [doi]
PST - ppublish
SO  - Vet Immunol Immunopathol. 2004 Dec 8;102(3):263-75. doi: 
      10.1016/j.vetimm.2004.09.011.