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Localization of Rabies Virus Glycoprotein into the Endoplasmic Reticulum Produces Immunoprotective Antigen

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Abstract

Rabies virus surface glycoprotein (rabies G-protein) with (G+RS) and without (G−RS) endoplasmic reticulum retrieval signal was expressed and characterized in tobacco plants. Transgenically expressed rabies G-protein was estimated at 0.015–0.38 % of total leaf protein. The relative migration of the rabies G-protein on SDS-PAGE was at the position, as anticipated for the viral coat protein (~66 kDa). Immunolocalization by confocal microscopy established that immunoprotective G+RS expressed in tobacco was primarily confined to ER. G+RS showed binding to Con A lectin and was susceptible to N-glycosidase F activity similar to native rabies G-protein. However, the G−RS transgenically expressed in tobacco leaves was glycosylated differently and was resitant to N-glycosidase F. Immunological studies and Rapid Fluorescent Foci Inhibition Test (RFFIT) showed that G+RS was immunogenic and immunoprotective, whereas G−RS was moderately immunogenic but non-protective against live virus challenge. Hence, plants can express the antigenic component of rabies virus with suitable glycosylation, which is important to give protection against rabies virus infection.

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Abbreviations

BSA:

Bovine serum albumin

Con A:

Concanavalin A

ER:

Endoplasmic reticulum

G+RS:

Rabies G-protein with endoplasmic retrieval signal

G−RS:

Rabies G-protein without endoplasmic retrieval signal

PBS:

Phosphate buffer saline

RFFIT:

Rapid Fluorescent Foci Inhibition Test

VIR:

Authentic rabies virus

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Acknowledgments

The work was carried out at National Botanical Research Institute (NBRI), Lucknow. Council of Scientific and Industrial Research, India, is gratefully acknowledged for financial support to DKY as Senior Research Fellow at NBRI. We thank Central Drug Research Institute, Lucknow for providing confocal microscopy facility and Indian Veterinary Research Institute, Izatnagar for conducting challenge virus standard challenge of immunized mice. We thank Dr Rebecca Boston, University of North Carolina, USA, for the generous gift of anti-calnexin antibody.

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Authors and Affiliations

  1. National Botanical Research Institute, Rana Pratap Marg, Lucknow, 226001, India

    Dinesh K. Yadav, Shadma Ashraf, Pradhyumna K. Singh & Rakesh Tuli

  2. Amity Education Valley, Amity University, Haryana, Gurgaon, 122413, Haryana, India

    Dinesh K. Yadav

  3. National Agri-Food Biotechnology Institute, C-127, Industrial Area, Phase VIII, SAS Nagar, Mohali, 160071, Punjab, India

    Rakesh Tuli

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  1. Dinesh K. Yadav
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  2. Shadma Ashraf
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  3. Pradhyumna K. Singh
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Correspondence to Dinesh K. Yadav.

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Yadav, D.K., Ashraf, S., Singh, P.K. et al. Localization of Rabies Virus Glycoprotein into the Endoplasmic Reticulum Produces Immunoprotective Antigen. Protein J 31, 447–456 (2012). https://doi.org/10.1007/s10930-012-9420-y

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