Identification by mass spectrometry and immune response analysis of guinea pig cytomegalovirus (GPCMV) pentameric complex proteins GP129, 131 and 133

Josephine S. Gnanandarajah, Peter A. Gillis, Nelmary Hernandez-Alvarado, LeeAnn Higgins, Todd W. Markowski, Heungsup Sung, Sheila Lumley, Mark R Schleiss

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Abstract

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.

Original languageEnglish (US)
Pages (from-to)727-751
Number of pages25
JournalViruses
Volume6
Issue number2
DOIs
StatePublished - Feb 13 2014

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Keywords

  • Baculovirus
  • Congenital cytomegalovirus infection
  • Cytomegalovirus vaccine
  • Guinea pig cytomegalovirus
  • Pentameric complex

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