Expression of an immunogenic F1-V fusion protein in lettuce as a plant-based vaccine against plague

Abstract

Yersinia pestis is a pathogenic agent that causes the bubonic and pneumonic plague. The development of an efficient and low-cost oral vaccine against these diseases is highly desirable. In this study, the immunogenic fusion protein F1-V from Y. pestis was introduced into lettuce via Agrobacterium-mediated transformation, and putative transgenic lines were developed. The presence of the transgene in these putative transgenic lines was determined using polymerase chain reaction (PCR), and transgene integration and transgene copy number were confirmed following Southern blot analysis. The presence of specific F1-V transcripts was confirmed by reverse-transcriptase (RT)-PCR. Using monoclonal antibodies, ELISA and western blot analysis revealed that the expected antigenic F1-V protein was successfully expressed in transgenic lines. Mice immunized subcutaneously with lettuce expressing the F1-V antigen developed systemic humoral responses as 'proof of concept' of using lettuce as a production platform for the F1-V immunogen that could be used as a candidate plant-based vaccine against plague.

Transgenic carrot tap roots expressing an immunogenic F1–V fusion protein from Yersinia pestis are immunogenic in mice

“…An estimate of the accumulation levels of the F1-V protein using a pure V protein as a standard revealed up to 0.12-0.3% of TSP in these transgenic carrot lines. This is certainly higher than F1-V protein levels previously expressed in lettuce (0.08% TSP) (Rosales-Mendoza et al, 2010). Moreover, this is also deemed a relatively high level of expression as native genes of bacterial antigens are often poorly expressed in plants systems (∼0.001% TSP) (Fischer and Emans, 2000).…”

“…Although F1-V proteins have been previously expressed in other fresh edible plants, tomato (Alvarez et al, 2006) and lettuce (Rosales-Mendoza et al, 2010), it is important explore the expression of such antigenic proteins in a variety of plant systems as levels of antigens in tomato for example, such as the F protein for the Respiratory Syncytial Virus (RSV), have been reported to decrease in advanced generations (Lau and Korban, 2010). Moreover, neither tomato nor lettuce has long shelf-lives.…”

Molecular farming on rescue of pharma industry for next generations

“…Recently, lettuce was tested to produce a multiepitopic HIV protein (C4(V3)6) that has shown high immunogenic potential compared with Escherichia coliderived C4(V3)6 (Govea- Alonso et al, 2013). Lettuce was also used to produce proinsulin (Boyhan & Daniell, 2011), immunogenic F1-V fusion protein as a vaccine against plague (Rosales-Mendoza et al, 2010), cholera toxin B subunit (Huy et al, 2011) and cholera toxin B-proinsulin fusion (Ruhlman et al, 2007).…”

Recent Advancement in the Development of Vaccines Against Y. pestis - A Potential Agent of Bioterrorism

“…Recently, an F1-V fusion protein expressed in carrot tap roots and lettuce was found to be stable and immunogenic for mice. (Rosales-Mendoza et al, 2010a;Rosales-Mendoza et al, 2010b).…”