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Immunogenicity of Brugia malayi Abundant Larval Transcript-2, a potential filarial vaccine candidate expressed in tobacco

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Transgenic tobacco plants with Bm ALT-2, a filarial vaccine candidate, were developed. The plant-produced antigen showed immunogenicity on par with the E.coli product.

Abstract

Transgenic tobacco plants were developed using Brugia malayi Abundant Larval Transcript-2 (Bm ALT-2), a major antigen produced from recombinant E.coli found to be experimentally successful as potential vaccine candidate against lymphatic filariasis. Results of experiments on the transformation and expression of the Bm ALT-2 in tobacco plant to produce plant-based vaccine are presented here. We have successfully transformed the tobacco plant with Bm ALT-2 and confirmed that the plants expressed the filarial protein by PCR analysis and Western blotting. The level of expression varied from 50 to 90 ng/μg of total soluble protein for ALT-2. Immunization of mice with plant-extracted protein indicated that the plant-produced protein had immunological characteristics similar to the E.coli-produced protein. Antibody titres produced by plant-produced recombinant ALT 2-immunized mice were on par with those immunized with recombinant protein produced by E.coli. Antibody isotype assay showed that plant-produced recombinant ALT-2 induced significant IgG1, whereas E.coli-produced recombinant ALT-2 induced IgG3. This result is a step forward towards the development of a model eukaryotic system for the production of recombinant filarial proteins, which can be utilized to produce therapeutic and diagnostic molecules against lymphatic filariasis, a neglected tropical infectious disease which has a negative impact on socioeconomic development. In addition, this is the first report of the immunogenicity of a plant-derived filarial antigen.

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Acknowledgments

This research was funded by a grant (LS-30/2010) from WOS-A section of Department of Science &Technology, Government of India. The authors would like to thank The Director, Centre for Biotechnology, Anna University, Chennai for allowing access to the lab and the infrastructure. Thanks are also due to Dr. N. Subramoniam, Sugarcane Breeding Institute, Coimbatore, India for his expertise and to Mrs. Mahalakshmi Natarajan for her help in statistical analysis.

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Authors and Affiliations

  1. Centre for Biotechnology, Anna University, Guindy, Chennai, 600025, Tamil Nadu, India

    Mathangi Ganapathy, Adhiseshan Perumal & Kaliraj Perumal

  2. Sugarcane Breeding Institute, Coimbatore, 641007, India

    Chakravarthi Mohan & Harunipriya Palaniswamy

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  1. Mathangi Ganapathy
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  2. Adhiseshan Perumal
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  4. Harunipriya Palaniswamy
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Correspondence to Mathangi Ganapathy.

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Communicated by B. Li.

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Ganapathy, M., Perumal, A., Mohan, C. et al. Immunogenicity of Brugia malayi Abundant Larval Transcript-2, a potential filarial vaccine candidate expressed in tobacco. Plant Cell Rep 33, 179–188 (2014). https://doi.org/10.1007/s00299-013-1521-3

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