Induction of toxin‐specific neutralizing immunity by molecularly uniform rice‐based oral cholera toxin B subunit vaccine without plant‐associated sugar modification

Induction of toxin‐specific neutralizing immunity by molecularly uniform rice‐based oral... Plants have been used as expression systems for a number of vaccines. However, the expression of vaccines in plants sometimes results in unexpected modification of the vaccines by N‐terminal blocking and sugar‐chain attachment. Although MucoRice‐CTB was thought to be the first cold‐chain‐free and unpurified oral vaccine, the molecular heterogeneity of MucoRice‐CTB, together with plant‐based sugar modifications of the CTB protein, has made it difficult to assess immunological activity of vaccine and yield from rice seed. Using a T‐DNA vector driven by a prolamin promoter and a signal peptide added to an overexpression vaccine cassette, we established MucoRice‐CTB/Q as a new generation oral cholera vaccine for humans use. We confirmed that MucoRice‐CTB/Q produces a single CTB monomer with an Asn to Gln substitution at the 4th glycosylation position. The complete amino acid sequence of MucoRice‐CTB/Q was determined by MS/MS analysis and the exact amount of expressed CTB was determined by SDS‐PAGE densitometric analysis to be an average of 2.35 mg of CTB/g of seed. To compare the immunogenicity of MucoRice‐CTB/Q, which has no plant‐based glycosylation modifications, with that of the original MucoRice‐CTB/N, which is modified with a plant N‐glycan, we orally immunized mice and macaques with the two preparations. Similar levels of CTB‐specific systemic IgG and mucosal IgA antibodies with toxin‐neutralizing activity were induced in mice and macaques orally immunized with MucoRice‐CTB/Q or MucoRice‐CTB/N. These results show that the molecular uniformed MucoRice‐CTB/Q vaccine without plant N‐glycan has potential as a safe and efficacious oral vaccine candidate for human use. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Plant Biotechnology Journal Wiley

Induction of toxin‐specific neutralizing immunity by molecularly uniform rice‐based oral cholera toxin B subunit vaccine without plant‐associated sugar modification

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Publisher
Wiley
Copyright
Copyright © 2013 Society for Experimental Biology, Association of Applied Biologists and John Wiley & Sons Ltd
ISSN
1467-7644
eISSN
1467-7652
D.O.I.
10.1111/pbi.12071
Publisher site
See Article on Publisher Site

Abstract

Plants have been used as expression systems for a number of vaccines. However, the expression of vaccines in plants sometimes results in unexpected modification of the vaccines by N‐terminal blocking and sugar‐chain attachment. Although MucoRice‐CTB was thought to be the first cold‐chain‐free and unpurified oral vaccine, the molecular heterogeneity of MucoRice‐CTB, together with plant‐based sugar modifications of the CTB protein, has made it difficult to assess immunological activity of vaccine and yield from rice seed. Using a T‐DNA vector driven by a prolamin promoter and a signal peptide added to an overexpression vaccine cassette, we established MucoRice‐CTB/Q as a new generation oral cholera vaccine for humans use. We confirmed that MucoRice‐CTB/Q produces a single CTB monomer with an Asn to Gln substitution at the 4th glycosylation position. The complete amino acid sequence of MucoRice‐CTB/Q was determined by MS/MS analysis and the exact amount of expressed CTB was determined by SDS‐PAGE densitometric analysis to be an average of 2.35 mg of CTB/g of seed. To compare the immunogenicity of MucoRice‐CTB/Q, which has no plant‐based glycosylation modifications, with that of the original MucoRice‐CTB/N, which is modified with a plant N‐glycan, we orally immunized mice and macaques with the two preparations. Similar levels of CTB‐specific systemic IgG and mucosal IgA antibodies with toxin‐neutralizing activity were induced in mice and macaques orally immunized with MucoRice‐CTB/Q or MucoRice‐CTB/N. These results show that the molecular uniformed MucoRice‐CTB/Q vaccine without plant N‐glycan has potential as a safe and efficacious oral vaccine candidate for human use.

Journal

Plant Biotechnology JournalWiley

Published: Jan 1, 2013

Keywords: ; ; ; ;

References

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