Plant heat shock protein 70 as carrier for immunization against a plant-expressed reporter antigen
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ORIGINAL PAPER
Plant heat shock protein 70 as carrier for immunization against a plant-expressed reporter antigen Giampaolo Buriani • Camillo Mancini Eugenio Benvenuto • Selene Baschieri
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Received: 18 March 2010 / Accepted: 4 June 2010 / Published online: 18 June 2010 Ó Springer Science+Business Media B.V. 2010
Abstract Mammalian Heat Shock Proteins (HSP), have potent immune-stimulatory properties due to the natural capability to associate with polypeptides and bind receptors on antigen presenting cells. The present study was aimed to explore whether plant HSP, and in particular HSP70, share similar properties. We wanted in particular to evaluate if HSP70 extracted in association to naturally bound polypeptides from plant tissues expressing a recombinant ‘‘reporter’’ antigen, carry antigen-derived polypeptides and can be used to activate antigen-specific immune responses. This application of HSP70 has been very poorly investigated so far. The analysis started by structurally modeling the plant protein and defining the conditions that ensure maximal expression levels and optimal recovery from plant tissues. Afterwards, HSP70 was purified from Nicotiana benthamiana leaves transiently expressing a heterologous ‘‘reporter’’ protein. The purification was carried
out taking care to avoid the release from HSP70 of the polypeptides chaperoned within plant cells. The evaluation of antibody titers in mice sera subsequent to the subcutaneous delivery of the purified HSP70 demonstrated that it is highly effective in priming humoral immune responses specific to the plant expressed ‘‘reporter’’ protein. Overall results indicated that plantderived HSP70 shares structural and functional properties with the mammalian homologue. This study paves the way to further investigations targeted at determining the properties of HSP70 extracted from plants expressing foreign recombinant antigens as a readily available immunological carrier for the efficient delivery of polypeptides derived from these antigens. Keywords Plant biofactories HSP70 Subunit vaccines Recombinant antigen delivery
Introduction
Electronic supplementary material The online version of this article (doi:10.1007/s11248-010-9418-1) contains supplementary material, which is available to authorized users. G. Buriani C. Mancini E. Benvenuto S. Baschieri (&) Technical Unit Radiation Biology and Human Health, Biotechnologies Laboratory, ENEA C.R. Casaccia, Via Anguillarese 301, 00123 Rome, Italy e-mail: [email protected]
Heat shock proteins (HSP) are a heterogeneous group of ubiquitous proteins exerting a pivotal function in cell homeostasis by contributing to the folding and unfolding, translocation and stabilization of proteins (Hartl and Hayer-Hartl 2009). Each HSP family includes several highly homologous isoforms, associated to particular cell compartments and/or tissues and expressed in normal or stress conditions (Chang et al. 2007; Tang et al. 2007). Except for small HSP, which are exclusive to the plant kingdom, they are
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