Molecular cloning and expression of major structural protein VP1 of the human polyomavirus JC virus: formation of virus-like particles useful for immunological and therapeutic studies.
The major structural viral protein, VP1, of the human polyomavirus JC virus (JCV), the causative agent of progressive multifocal leukoencephalopathy (PML), was expressed by using recombinant baculoviruses. Recombinant VP1 formed virus-like particles (VLP) with the typical morphology of empty JCV capsids. Purified VP1 VLP bind to SVG, B, and T cells, as well as to monkey kidney cells. After binding, VP1 VLP were also internalized with high efficiency and transported to the nucleus. Immunization studies revealed these particles as highly immunogenic when administered with adjuvant, while immunization without adjuvant induced no immune response. VP1 VLP hyperimmune serum inhibits binding to SVG cells and neutralizes natural JCV. Furthermore, the potential of VP1 VLP as an efficient transporter system for gene therapy was demonstrated. Exogenous DNA could be efficiently packaged into VP1 VLP, and the packaged DNA was transferred into COS-7 cells as shown by the expression of a marker gene. Thus, VP1 VLP are useful for PML vaccine development and represent a potential new transporter system for human gene therapy.
Goldmann
C
C
Department of Virology and Immunology, German Primate Centre, D-37077 Göttingen, Germany.
Petry
H
H
Frye
S
S
Ast
O
O
Ebitsch
S
S
Jentsch
K D
KD
Kaup
F J
FJ
Weber
F
F
Trebst
C
C
Nisslein
T
T
Hunsmann
G
G
Weber
T
T
Lüke
W
W
D000818:Animals; D019556:COS Cells; D002213:Capsid / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D036022:Capsid Proteins; D002460:Cell Line; D003001:Cloning, Molecular; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006801:Humans; D007577:JC Virus / Q000235:genetics / Q000276:immunology / Q000502:physiology* / Q000648:ultrastructure; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D018411:Spodoptera / Q000166:cytology; D014614:Vaccines, Synthetic; D014765:Viral Vaccines; D014771:Virion / Q000502:physiology / Q000648:ultrastructure; D019065:Virus Assembly
D036022:Capsid Proteins; D011993:Recombinant Fusion Proteins; C062481:VP1 protein, polyomavirus; D014614:Vaccines, Synthetic; D014765:Viral Vaccines