PMID- 19427138
OWN - NLM
STAT- MEDLINE
DCOM- 20091026
LR  - 20090612
IS  - 1873-2542 (Electronic)
IS  - 0378-1135 (Linking)
VI  - 138
IP  - 1-2
DP  - 2009 Jul 2
TI  - Monoclonal antibodies and conserved antigenic epitopes in the C terminus of GP5 
      protein of the North American type porcine reproductive and respiratory syndrome 
      virus.
PG  - 1-10
LID - 10.1016/j.vetmic.2009.01.041 [doi]
AB  - Glycoprotein 5 (GP5) is the major glycoprotein of porcine reproductive and 
      respiratory syndrome virus (PRRSV). In this study, the gene encoding rtGP5, 
      lacking signal peptide sequence, was expressed as GST-fusion protein in E. coli. 
      Fifteen monoclonal antibodies (MAbs) against rtGP5 were developed and used to 
      probe a series of GP5 peptides by ELISA, in which two MAbs specifically 
      recognized the epitope GP5EP3 (146-156aa), four recognized GP5EP5 (164-180aa) and 
      nine recognized GP5EP7 (192-200aa). After precise analysis by sequential deletion 
      of the terminal amino acid residues, the three minimal epitopes 
      (R(152)LYRWR(156), E(169)GHLIDLKRV(178) and Q(196)WGRL(200)) were determined, 
      which were highly conserved among the North American type isolates, with the 
      exception of one amino acid mutation (L(200) to P(200)). Mutational analysis 
      showed that the mutant (Q(196)WGRP(200)) could be recognized by four of nine 
      anti-GP5EP7 MAbs, indicating Q(196)WGRP(200) was also one minimal epitope. 
      Western blot analysis showed that GP5EP5 and GP5EP7 (L(200) or P(200)) could be 
      recognized by PRRSV-positive sera of CH-1a and/or BJ-4, suggesting GP5EP5 and 
      GP5EP7 (L(200) or P(200)) were antigenic epitopes in the PRRSV-infected pigs. 
      MAbs against GP5EP3, GP5EP5, and GP5EP7 could react with MARC-145 cells infected 
      with the North American type isolates from China in IFA. However, very 
      interestingly, when the highly pathogenic PRRSV, represented by HUN4, was 
      passaged in MARC-145 cells, MAbs against GP5EP7 did not react with 
      HUN4-F20-HUN4-F112 (20-112th passage virus), where Q(196)WGRL(200) had mutated to 
      R(196)WGRL(200). Due to no mutations observed in GP5EP3 and GP5EP5, MAbs against 
      GP5EP3 and GP5EP5 could recognize HUN4-F20-HUN4-F112. All the results herein 
      might deepen the understanding of the antigen structure of in the C terminus of 
      GP5 and facilitate the development of diagnostic antigens of the North American 
      type PRRSV in China.
FAU - Zhou, Yan-Jun
AU  - Zhou YJ
AD  - Division of Swine Infectious Diseases, Shanghai Veterinary Research Institute, 
      Chinese Academy of Agricultural Sciences, No. 518, Ziyue Road, Minhang District, 
      Shanghai 200241, China.
FAU - Yu, Hai
AU  - Yu H
FAU - Tian, Zhi-Jun
AU  - Tian ZJ
FAU - Liu, Jin-Xia
AU  - Liu JX
FAU - An, Tong-Qing
AU  - An TQ
FAU - Peng, Jin-Mei
AU  - Peng JM
FAU - Li, Guo-Xin
AU  - Li GX
FAU - Jiang, Yi-Feng
AU  - Jiang YF
FAU - Cai, Xue-Hui
AU  - Cai XH
FAU - Xue, Qiang
AU  - Xue Q
FAU - Wang, Mei
AU  - Wang M
FAU - Wang, Yun-Feng
AU  - Wang YF
FAU - Tong, Guang-Zhi
AU  - Tong GZ
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20090206
PL  - Netherlands
TA  - Vet Microbiol
JT  - Veterinary microbiology
JID - 7705469
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Antigens, Viral)
RN  - 0 (DNA Primers)
RN  - 0 (Epitopes)
RN  - 0 (Peptide Fragments)
RN  - 0 (Viral Envelope Proteins)
RN  - 0 (glycoprotein 5, PRRSV)
SB  - IM
MH  - Amino Acid Sequence
MH  - Amino Acid Substitution
MH  - Animals
MH  - Antibodies, Monoclonal/*genetics/immunology
MH  - Antibody Specificity
MH  - Antigens, Viral/genetics
MH  - Conserved Sequence
MH  - DNA Primers
MH  - Epitopes/*genetics/immunology
MH  - Escherichia coli/genetics
MH  - Genetic Vectors
MH  - Open Reading Frames/genetics
MH  - Peptide Fragments/chemistry
MH  - Plasmids/genetics
MH  - Porcine Reproductive and Respiratory Syndrome/*immunology
MH  - Porcine respiratory and reproductive syndrome virus/*genetics/*immunology
MH  - Restriction Mapping
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Swine
MH  - Viral Envelope Proteins/*genetics
EDAT- 2009/05/12 09:00
MHDA- 2009/10/27 06:00
CRDT- 2009/05/12 09:00
PHST- 2008/11/09 00:00 [received]
PHST- 2009/01/05 00:00 [revised]
PHST- 2009/01/12 00:00 [accepted]
PHST- 2009/05/12 09:00 [entrez]
PHST- 2009/05/12 09:00 [pubmed]
PHST- 2009/10/27 06:00 [medline]
AID - S0378-1135(09)00039-X [pii]
AID - 10.1016/j.vetmic.2009.01.041 [doi]
PST - ppublish
SO  - Vet Microbiol. 2009 Jul 2;138(1-2):1-10. doi: 10.1016/j.vetmic.2009.01.041. Epub 
      2009 Feb 6.