Abstract
Transgene introgression is a major concern associated with transgenic plant-based vaccines. Agroinfiltration can be used to selectively transform nonreproductive organs and avoid introgression. Here, we introduce a new vaccine modality in which Staphylococcal enterotoxin B (SEB) genes are agroinfiltrated into radishes (Raphanw sativus L.), resulting in transient expression and accumulation of SEB in planta. This approach can simultaneously express multiple antigens in a single leaf. Furthermore, the potential of high-throughput vaccine production was demonstrated by simultaneously agroinfiltrating multiple radish leaves using a multichannel pipette. The expression of SEB was detectable in two leaf cell types (epidermal and guard cells) in agroinfiltrated leaves. ICR mice intranasally immunized with homogenized leaves agroinfiltrated with SEB elicited detectable antibody to SEB and displayed protection against SEB-induced interferon-gamma (IFN-γ) production. The concept of encapsulating antigens in leaves rather than purifying them for immunization may facilitate rapid vaccine production during an epidemic disease.
Highlights
Transgenic plants have emerged as a promising technology to generate recombinant biopharmaceutical proteins and vaccines [1, 2]
Agrobacterium has been utilized as a vector to deliver foreign DNA and induce transient expression of recombinant proteins in various plants [34]
Infiltration of radish, lettuce, and Arabidopsis leaves with A. tumefaciens harboring 35S::GUS constructs resulted in GUS expression in all three plants
Summary
Transgenic plants have emerged as a promising technology to generate recombinant biopharmaceutical proteins and vaccines [1, 2]. Transient expression of recombinant proteins in leaf tissue avoids transgene introgression and provides a fast platform for protein production without an effort-exhaustive process to generate stably transformed transgenic plants [9]. There are at least four approaches to transforming and inducing transient expression in plants: (1) delivery of “naked” DNA by particle bombardment [10], (2) infection with modified viral vectors [6, 10, 11], (3) agroinfiltration of plant tissues with Agrobacteria [10, 12], and (4) polyethylene glycol- (PEG-) mediated gene transfer and electroporation of protoplasts [13]. Agroinfiltration accommodates transforming plants with large genes encoding complex proteins, such
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