Baker's yeast expressing the Japanese encephalitis virus envelope protein on its cell surface: induction of an antigen-specific but non-neutralizing antibody response.
Live recombinant Saccharomyces cerevisiae yeast expressing the envelope antigen of Japanese encephalitis virus (JEV) on the outer mannoprotein layer of the cell wall were examined for their ability to induce antigen-specific antibody responses in mice. When used as a model antigen, parenteral immunization of mice with surface-expressing GFP yeast induced a strong anti-GFP antibody response in the absence of adjuvants. This antigen delivery approach was then used for a more stringent system, such as the envelope protein of JEV, which is a neurotropic virus requiring neutralizing antibodies for protection. Although 70% of cells were detected to express the total envelope protein on the surface by antibodies raised to the bacterially expressed protein, polyclonal anti-JEV antibodies failed to react with them. In marked contrast, yeast expressing the envelope fragments 238-398, 373-399 and 373-500 in front of a Gly-Ser linker were detected by anti-JEV antibodies as well as a monoclonal antibody but not by antibodies raised to the bacterially expressed protein. Immunization of mice with these surface-expressing recombinants resulted in a strong antibody response. However, the antibodies failed to neutralize the virus, although the fragments were selected based on neutralizing determinants.
Upadhyaya
Bhaskar
B
Department of Biochemistry, Indian Institute of Science, Bangalore 560012, India. upadhyayab@niaid.nih.gov
Manjunath
Ramanathapuram
R
D000284:Administration, Oral; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D004664:Encephalitis Virus, Japanese / Q000235:genetics / Q000276:immunology*; D005260:Female; D005822:Genetic Vectors; D022321:Japanese Encephalitis Vaccines / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014759:Viral Envelope Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*
D000914:Antibodies, Viral; D022321:Japanese Encephalitis Vaccines; D014759:Viral Envelope Proteins