Setting up a platform for plant-based influenza virus vaccine production in South Africa

Elizabeth Mortimer,Sandiswa Mbewana,Anna-Lise Williamson,Edward P Rybicki,James M Maclean,Amelia Buys,Inga I Hitzeroth

doi:10.1186/1472-6750-12-14

Abstract

BackgroundDuring a global influenza pandemic, the vaccine requirements of developing countries can surpass their supply capabilities, if these exist at all, compelling them to rely on developed countries for stocks that may not be available in time. There is thus a need for developing countries in general to produce their own pandemic and possibly seasonal influenza vaccines. Here we describe the development of a plant-based platform for producing influenza vaccines locally, in South Africa. Plant-produced influenza vaccine candidates are quicker to develop and potentially cheaper than egg-produced influenza vaccines, and their production can be rapidly upscaled. In this study, we investigated the feasibility of producing a vaccine to the highly pathogenic avian influenza A subtype H5N1 virus, the most generally virulent influenza virus identified to date. Two variants of the haemagglutinin (HA) surface glycoprotein gene were synthesised for optimum expression in plants: these were the full-length HA gene (H5) and a truncated form lacking the transmembrane domain (H5tr). The genes were cloned into a panel of Agrobacterium tumefaciens binary plant expression vectors in order to test HA accumulation in different cell compartments. The constructs were transiently expressed in tobacco by means of agroinfiltration. Stable transgenic tobacco plants were also generated to provide seed for stable storage of the material as a pre-pandemic strategy.ResultsFor both transient and transgenic expression systems the highest accumulation of full-length H5 protein occurred in the apoplastic spaces, while the highest accumulation of H5tr was in the endoplasmic reticulum. The H5 proteins were produced at relatively high concentrations in both systems. Following partial purification, haemagglutination and haemagglutination inhibition tests indicated that the conformation of the plant-produced HA variants was correct and the proteins were functional. The immunisation of chickens and mice with the candidate vaccines elicited HA-specific antibody responses.ConclusionsWe managed, after synthesis of two versions of a single gene, to produce by transient and transgenic expression in plants, two variants of a highly pathogenic avian influenza virus HA protein which could have vaccine potential. This is a proof of principle of the potential of plant-produced influenza vaccines as a feasible pandemic response strategy for South Africa and other developing countries.

Highlights

During a global influenza pandemic, the vaccine requirements of developing countries can surpass their supply capabilities, if these exist at all, compelling them to rely on developed countries for stocks that may not be available in time
Transient expression of H5 in tobacco plants We compared the transient expression of eight H5N1 HA constructs in N. benthamiana plants
The yields of H5 and H5tr varied with experiment and by transgenic plant line, but in general were higher than those obtained by others: this may be due to the vector system used, as well as to the human codon optimisation of the genes

Summary

Introduction

During a global influenza pandemic, the vaccine requirements of developing countries can surpass their supply capabilities, if these exist at all, compelling them to rely on developed countries for stocks that may not be available in time. There is a need for developing countries in general to produce their own pandemic and possibly seasonal influenza vaccines. We describe the development of a plant-based platform for producing influenza vaccines locally, in South Africa. Stable transgenic tobacco plants were generated to provide seed for stable storage of the material as a pre-pandemic strategy. During 2009, South Africa was faced with the novel (“swine”) influenza A H1N1 virus pandemic. As the country does not have the ability to produce influenza vaccine stocks, we had to rely on the World Health Organisation (WHO) and developed countries – and as expected, even developed countries did not have adequate vaccine stocks to meet their own demands. South Africa should be self-sufficient; producing its own influenza vaccines to reduce morbidity and mortality in its large and relatively poor population

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