TY - JOUR
T1 - Identification by mass spectrometry and immune response analysis of guinea pig cytomegalovirus (GPCMV) pentameric complex proteins GP129, 131 and 133
AU - Gnanandarajah, Josephine S.
AU - Gillis, Peter A.
AU - Hernandez-Alvarado, Nelmary
AU - Higgins, LeeAnn
AU - Markowski, Todd W.
AU - Sung, Heungsup
AU - Lumley, Sheila
AU - Schleiss, Mark R
PY - 2014/2/13
Y1 - 2014/2/13
N2 - Development of a vaccine against congenital infection with human cytomegalovirus (HCMV) is a major public health priority. A potential vaccine target receiving considerable recent attention is the pentameric complex (PC) of HCMV proteins consisting of gL, gH, UL128, UL130, and UL131, since some antibodies against these target proteins are capable of potently neutralizing virus at epithelial and endothelial cell surfaces. Recently, homologous proteins have been described for guinea pig cytomegalovirus (GPCMV), consisting of gH, gL, and the GPCMV proteins GP129, GP131, and GP133. To investigate these proteins as potential vaccine targets, expression of GP129-GP133 transcripts was confirmed by reverse-transcriptase PCR. Mass spectrometry combined with western blot assays demonstrated the presence of GP129, GP131, and GP133 proteins in virus particles. Recombinant proteins corresponding to these PC proteins were generated in baculovirus, and as GST fusion proteins. Recombinant proteins were noted to be immunoreactive with convalescent sera from infected animals, suggesting that these proteins are recognized in the humoral immune response to GPCMV infection. These analyses support the study of PC-based recombinant vaccines in the GPCMV congenital infection model.
AB - Development of a vaccine against congenital infection with human cytomegalovirus (HCMV) is a major public health priority. A potential vaccine target receiving considerable recent attention is the pentameric complex (PC) of HCMV proteins consisting of gL, gH, UL128, UL130, and UL131, since some antibodies against these target proteins are capable of potently neutralizing virus at epithelial and endothelial cell surfaces. Recently, homologous proteins have been described for guinea pig cytomegalovirus (GPCMV), consisting of gH, gL, and the GPCMV proteins GP129, GP131, and GP133. To investigate these proteins as potential vaccine targets, expression of GP129-GP133 transcripts was confirmed by reverse-transcriptase PCR. Mass spectrometry combined with western blot assays demonstrated the presence of GP129, GP131, and GP133 proteins in virus particles. Recombinant proteins corresponding to these PC proteins were generated in baculovirus, and as GST fusion proteins. Recombinant proteins were noted to be immunoreactive with convalescent sera from infected animals, suggesting that these proteins are recognized in the humoral immune response to GPCMV infection. These analyses support the study of PC-based recombinant vaccines in the GPCMV congenital infection model.
KW - Baculovirus
KW - Congenital cytomegalovirus infection
KW - Cytomegalovirus vaccine
KW - Guinea pig cytomegalovirus
KW - Pentameric complex
UR - http://www.scopus.com/inward/record.url?scp=84895107518&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84895107518&partnerID=8YFLogxK
U2 - 10.3390/v6020727
DO - 10.3390/v6020727
M3 - Article
C2 - 24531333
AN - SCOPUS:84895107518
SN - 1999-4915
VL - 6
SP - 727
EP - 751
JO - Viruses
JF - Viruses
IS - 2
ER -