Summary
To prevent vaccine‐associated paralytic poliomyelitis,
WHO
recommended withdrawal of Oral Polio Vaccine (Serotype‐2) and a single dose of Inactivated Poliovirus Vaccine (
IPV
).
IPV
however is expensive, requires cold chain, injections and offers limited intestinal mucosal immunity, essential to prevent polio reinfection in countries with open sewer system. To date, there is no virus‐free and cold chain‐free polio vaccine capable of inducing robust mucosal immunity. We report here a novel low‐cost, cold chain/poliovirus‐free, booster vaccine using poliovirus capsid protein (
VP
1, conserved in all serotypes) fused with cholera non‐toxic B subunit (
CTB
) expressed in lettuce chloroplasts.
PCR
using unique primer sets confirmed site‐specific integration of
CTB
‐
VP
1 transgene cassettes. Absence of the native chloroplast genome in Southern blots confirmed homoplasmy. Codon optimization of the
VP
1 coding sequence enhanced its expression 9–15‐fold in chloroplasts.
GM
1‐ganglioside receptor‐binding
ELISA
confirmed pentamer assembly of
CTB
‐
VP
1 fusion protein, fulfilling a key requirement for oral antigen delivery through gut epithelium. Transmission Electron Microscope images and hydrodynamic radius analysis confirmed
VP
1‐
VLP
s of 22.3 nm size. Mice primed with
IPV
and boosted three times with lyophilized plant cells expressing
CTB
‐
VP
1co, formulated with plant‐derived oral adjuvants, enhanced
VP
1‐specific IgG1,
VP
1‐IgA titres and neutralization (80%–100% seropositivity of Sabin‐1, 2, 3). In contrast,
IPV
single dose resulted in <50%
VP
1‐IgG1 and negligible
VP
1‐IgA titres, poor neutralization and seropositivity (<20%, <40% Sabin 1,2). Mice orally boosted with
CTB
‐
VP
1co, without
IPV
priming, failed to produce any protective neutralizing antibody. Because global population is receiving
IPV
single dose, booster vaccine free of poliovirus or cold chain offers a timely low‐cost solution to eradicate polio.