title: Identification by mass spectrometry and immune response analysis of guinea pig cytomegalovirus (GPCMV) pentameric complex proteins GP129, 131 and 133.
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.
authors: Departments of Pediatrics, University of Minnesota Medical School, 2001 6th Street SE, Minneapolis, MN 55455, USA. gnan0007@umn.edu.
authors: Josephine S
authors: JS
authors: Gnanandarajah
authors: Departments of Pediatrics, University of Minnesota Medical School, 2001 6th Street SE, Minneapolis, MN 55455, USA. gill0221@umn.edu.
authors: Peter A
authors: PA
authors: Gillis
authors: Departments of Pediatrics, University of Minnesota Medical School, 2001 6th Street SE, Minneapolis, MN 55455, USA. hernande@umn.edu.
authors: Nelmary
authors: N
authors: Hernandez-Alvarado
authors: Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, 321 Church Street SE, Minneapolis, MN 55455, USA. higgi022@umn.edu.
authors: LeeAnn
authors: L
authors: Higgins
authors: Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, 321 Church Street SE, Minneapolis, MN 55455, USA. marko025@umn.edu.
authors: Todd W
authors: TW
authors: Markowski
authors: Departments of Pediatrics, University of Minnesota Medical School, 2001 6th Street SE, Minneapolis, MN 55455, USA. sung@amc.seoul.kr.
authors: Heungsup
authors: H
authors: Sung
authors: Departments of Pediatrics, University of Minnesota Medical School, 2001 6th Street SE, Minneapolis, MN 55455, USA. sheilalumley@gmail.com.
authors: Sheila
authors: S
authors: Lumley
authors: Departments of Pediatrics, University of Minnesota Medical School, 2001 6th Street SE, Minneapolis, MN 55455, USA. schleiss@umn.edu.
authors: Mark R
authors: MR
authors: Schleiss
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D000956:Antigens, Viral / Q000235:genetics; D015153:Blotting, Western; D006168:Guinea Pigs; D046911:Macromolecular Substances / Q000737:chemistry*; D013058:Mass Spectrometry; D011994:Recombinant Proteins / Q000235:genetics; D019211:Roseolovirus / Q000737:chemistry*; D015678:Viral Structural Proteins / Q000032:analysis* / Q000235:genetics / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D046911:Macromolecular Substances; D011994:Recombinant Proteins; D015678:Viral Structural Proteins