The immunogenicity of VP7, a rotavirus antigen resident in the endoplasmic reticulum, is enhanced by cell surface expression.
The glycoprotein VP7, the major serotype antigen of rotaviruses, is localized to the endoplasmic reticulum (ER) of the cell, where it is retained as a membrane-associated protein before assembly into mature virus particles. Wild-type VP7 expressed by a recombinant vaccinia virus was also located internally and was poorly antigenic. Using recombinant techniques, a correctly processed, secreted form of VP7 (S.C. Stirzaker and G.W. Both, Cell 56:741-747, 1989) was modified by addition to its C terminus of the membrane anchor and cytoplasmic domains from the influenza virus hemagglutinin. The hybrid protein was directed to the surface of cells, where it was anchored in the plasma membrane. When expressed in mice and rabbits by a recombinant vaccinia virus, the surface-anchored antigen stimulated a level of rotavirus-specific antibodies that was greater than 100-fold above the level induced by wild-type VP7. T-cell responses to the novel antigen were also elevated in comparison with the wild-type, intracellular protein. Cell surface anchoring may provide a strategy to increase the immunogenicity of intracellular antigens from other parasites and viruses.
Andrew
M E
ME
CSIRO Division of Biomolecular Engineering, North Ryde, New South Wales, Australia.
Boyle
D B
DB
Whitfeld
P L
PL
Lockett
L J
LJ
Anthony
I D
ID
Bellamy
A R
AR
Both
G W
GW
D000595:Amino Acid Sequence; D000818:Animals; D000917:Antibody Formation; D000956:Antigens, Viral / Q000276:immunology*; D002213:Capsid / Q000235:genetics / Q000276:immunology*; D036022:Capsid Proteins; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology; D004721:Endoplasmic Reticulum / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005814:Genes, Viral; D006801:Humans; D009980:Influenza A virus / Q000235:genetics; D051379:Mice; D008808:Mice, Inbred CBA; D008969:Molecular Sequence Data; D009154:Mutation; D009500:Neutralization Tests; D021382:Protein Sorting Signals / Q000235:genetics; D011817:Rabbits / Q000276:immunology; D012401:Rotavirus / Q000235:genetics* / Q000276:immunology; D014616:Vaccinia virus / Q000235:genetics
D000956:Antigens, Viral; D036022:Capsid Proteins; D021382:Protein Sorting Signals; C050779:VP7 protein, Rotavirus