Relocation of antigens to the cell surface membrane can enhance immune stimulation and protection.
The major outer capsid glycoprotein of rotaviruses, VP7, is normally synthesized and directed to the ER, where it is required for virus assembly. By substituting a foreign signal sequence for the VP7 signal peptide, a secreted form of VP7 with an authentic amino terminus was produced. Secreted VP7 was further modified by the addition of a transmembrane anchor and cytoplasmic domain to its C-terminus. When the novel chimeric protein was expressed in transfected cells it became anchored in the cell surface membrane. The antigenicity of the chimeric protein was compared with that of the intracellular form of VP7 using recombinant vaccinia viruses to deliver the antigens in vivo. The novel antigen produced enhanced stimulation of both B and T lymphocytes of the immune system, and in mice it was able to induce protection against rotavirus-induced diarrhoeal disease. Other secreted and intracellular antigens show a similar improved level of antigenicity as a result of their relocation to the cell surface. Surface localization may therefore have general utility in the development of recombinant subunit vaccines.
Both
G W
GW
CSIRO, Divisions of Biomolecular Engineering, North Ryde, NSW, Australia.
Andrew
M E
ME
Boyle
D B
DB
Coupar
B E
BE
Bellamy
A R
AR
D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis; D000954:Antigens, Surface / Q000276:immunology*; D000956:Antigens, Viral / Q000276:immunology*; D001692:Biological Transport; D002213:Capsid / Q000235:genetics / Q000276:immunology; D003967:Diarrhea / Q000235:genetics / Q000276:immunology / Q000382:microbiology; D008562:Membrane Glycoproteins / Q000276:immunology*; D051379:Mice; D008808:Mice, Inbred CBA; D011817:Rabbits; D012401:Rotavirus / Q000235:genetics / Q000276:immunology*; D012400:Rotavirus Infections / Q000235:genetics / Q000276:immunology / Q000382:microbiology
D000914:Antibodies, Viral; D000954:Antigens, Surface; D000956:Antigens, Viral; D008562:Membrane Glycoproteins