Cucumber mosaic virus as a presentation system for a double hepatitis C virus-derived epitope

Abstract

Chimeric plant viruses are emerging as promising vectors for use in innovative vaccination strategies. In this context, cucumber mosaic virus (CMV) has proven to be a suitable carrier of the hepatitis C virus (HCV)-derived R9 mimotope. In the present work, a new chimeric CMV, expressing on its surface the HCV-derived R10 mimotope, was produced but lost the insert after the first passage on tobacco. A comparative analysis between R10- and R9-CMV properties indicated that R9-CMV stability was related to structural features typical of the foreign insert. Thus, in order to combine high virus viability with strong immuno-stimulating activity, we doubled R9 copies on each of the 180 coat protein (CP) subunits of CMV. One of the chimeras produced by this approach (2R9-CMV) was shown to systemically infect the host, stably maintaining both inserts. Notably, it was strongly recognized by sera of HCV-infected patients and, as compared with R9-CMV, displayed an enhanced ability to stimulate lymphocyte IFN-gamma production. The high immunogen levels achievable in plants or fruits infected with 2R9-CMV suggest that this chimeric form of CMV may be useful in the development of oral vaccines against HCV.

Potato virus X displaying the E7 peptide derived from human papillomavirus type 16: a novel position for epitope presentation

“…One way to combine fast and cost-effective plant transient expression with the ability of plant viruses to serve as safe antigen displaying systems is to produce recombinant viruses carrying surfaced-exposed antigens in plants. As a platform for this purpose, Cowpea mosaic virus (icosahedral), Cucumber mosaic virus (icosahedral), Tobacco mosaic virus (rod-shaped), and Potato virus X (filamentous) have been used (Bendahmane et al 1999;Uhde et al 2005;Zhao and Hammond 2005;Smith et al 2006;Nuzzaci et al 2007;Sainsbury et al 2010).…”

Virus-like Particles in Vaccine Development

“…Moreover, only a few studies have been performed to assess the induction of the systemic and mucosal immune responses upon mucosal (i.e., intranasal) administration [32,33]. In particular, we have recently shown in nonhuman primates that HIV-VLPs administered by intranasal route do not elicit a measurable immune response but are able to prime the response at the mucosal level after subsequent intramuscular administration [52].…”

Plant-based vaccines against viruses

“…Use of rTMV to express R9 epitope of E2 protein as CTB fusion [30] Expression of chimaeric plant virus coat protein molecules containing R9 [31][32][33] R9-CMV CP VLPs orally immunogenic via feeding of lettuce leaves in rabbits [34] Mixed Th1/Th2 response in mice to rodlike PMV-E2 epitope chimaeric VLPs [36] Co-expression of whole E2 and calnexin and calreticulin increases E2 accumulation in plants [37] Influenza viruses…”

“…The plant-derived HVR1/CTB reacted with immune sera from individuals infected with four of the major genotypes of HCV, and mice immunised intranasally with crude plant extract produced anti-HVR1 antibodies which specifically bound HCV VLPs [30]. This was followed by a succession of reports on plant expression of chimaeric plant virus coat protein molecules containing the same (R9) epitope: these included the use of Cucumber mosaic virus (CMV) CP, with immunoreactivity of sera from chronically infected patients with the recombinant CP [31] leading to use of purified R9-CMV to elicit in vivo responses in rabbits, and in vitro cellular responses as measured by interferon gamma production for lymphocytes from human patients [32,33]. Subsequently, the same group showed that purified R9-CMV particles were stable under simulated gastric and intestinal conditions, and could elicit a humoral immune response in rabbits fed with R9-CMV infected lettuce plants [34].…”