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

• Published in: Viruses Vol. 6; no. 2; pp. 727 - 751
• Main Authors: Gnanandarajah, Josephine S, Gillis, Peter A, Hernandez-Alvarado, Nelmary, Higgins, LeeAnn, Markowski, Todd W, Sung, Heungsup, Lumley, Sheila, Schleiss, Mark R
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 13.02.2014; MDPI
• Subjects: Animals; Antibodies; Antibodies, Viral - blood; Antigens, Viral - genetics; Baculovirus; Blotting, Western; Clinical trials; congenital cytomegalovirus infection; Congenital diseases; Cytomegalovirus; cytomegalovirus vaccine; Fetuses; Fibroblasts; Glycoproteins; Guinea pig cytomegalovirus; Guinea Pigs; Human cytomegalovirus; Infections; Macromolecular Substances - chemistry; Mass Spectrometry; pentameric complex; Proteins; Public health; Recombinant Proteins - genetics; Roseolovirus - chemistry; Scientific imaging; Vaccines; Viral Structural Proteins - analysis; Viral Structural Proteins - genetics; Viral Structural Proteins - immunology; Viruses; Minneapolis Minnesota; United States > US
• ISSN: 1999-4915; 1999-4915
• DOI: 10.3390/v6020727

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.