Generation of a recombinant cytomegalovirus for expression of a hantavirus glycoprotein.
A cytomegalovirus (CMV) was isolated from its natural host, Peromyscus maniculatus, and was designated Peromyscus CMV (PCMV). A recombinant PCMV was constructed that contained Sin Nombre virus glycoprotein G1 (SNV-G1) fused in frame to the enhanced green fluorescent protein (EGFP) gene inserted into a site homologous to the human CMV UL33 (P33) gene. The recombinant CMV was used for expression and immunization of deer mice against SNV-G1. The results of the study indicate that P. maniculatus could be infected with as few as 10 virus particles of recombinant virus. Challenge of P. maniculatus with either recombinant or wild-type PCMV produced no overt pathology in infected animals. P. maniculatus immunized with recombinant virus developed an antibody response to SNV and EGFP. When rechallenged with recombinant virus, animals exhibited an anamnestic response against SNV. Interestingly, a preexisting immune response against PCMV did not prevent reinfection with recombinant PCMV.
Rizvanov
Albert A
AA
Department of Microbiology, University of Nevada at Reno, Reno, Nevada 89557, USA.
van Geelen
Albert G M
AG
Morzunov
Sergey
S
Otteson
Elmer W
EW
Bohlman
Charlotte
C
Pari
Gregory S
GS
St  Jeor
Stephen C
SC
D000595:Amino Acid Sequence; D000818:Animals; D003587:Cytomegalovirus / Q000235:genetics* / Q000276:immunology; D004259:DNA-Directed DNA Polymerase / Q000032:analysis; D006023:Glycoproteins / Q000096:biosynthesis*; D007114:Immunization; D008969:Molecular Sequence Data; D010542:Peromyscus / Q000821:virology*; D019707:Receptors, Chemokine / Q000032:analysis; D011994:Recombinant Proteins / Q000096:biosynthesis*; D029261:Sin Nombre virus / Q000737:chemistry / Q000276:immunology*; D014614:Vaccines, Synthetic / Q000276:immunology*; D014764:Viral Proteins / Q000032:analysis / Q000096:biosynthesis*; D014765:Viral Vaccines / Q000276:immunology*
D006023:Glycoproteins; D019707:Receptors, Chemokine; D011994:Recombinant Proteins; C461643:UL33 protein, human cytomegalovirus; D014614:Vaccines, Synthetic; D014764:Viral Proteins; D014765:Viral Vaccines; D004259:DNA-Directed DNA Polymerase