title: Immunogenicity of Brugia malayi Abundant Larval Transcript-2, a potential filarial vaccine candidate expressed in tobacco.
abstract: 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. 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.
authors.affiliation: Centre for Biotechnology, Anna University, Guindy, Chennai, 600025, Tamil Nadu, India, mathangig@gmail.com.
authors.forename: Mathangi
authors.initials: M
authors.lastname: Ganapathy
authors.forename: Adhiseshan
authors.initials: A
authors.lastname: Perumal
authors.forename: Chakravarthi
authors.initials: C
authors.lastname: Mohan
authors.forename: Harunipriya
authors.initials: H
authors.lastname: Palaniswamy
authors.forename: Kaliraj
authors.initials: K
authors.lastname: Perumal
mesh_terms: D000818:Animals; D000918:Antibody Specificity / Q000276:immunology; D000947:Antigens, Helminth / Q000276:immunology*; D017178:Brugia malayi / Q000276:immunology*; D049109:Cell Proliferation; D004591:Electrophoresis, Polyacrylamide Gel; D004605:Elephantiasis, Filarial / Q000276:immunology / Q000517:prevention & control; D005260:Female; D015801:Helminth Proteins / Q000302:isolation & purification / Q000378:metabolism; D007114:Immunization; D007814:Larva / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D030821:Plants, Genetically Modified; D010957:Plasmids / Q000378:metabolism; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D013154:Spleen / Q000166:cytology; D014026:Nicotiana / Q000235:genetics*; D014170:Transformation, Genetic; D014612:Vaccines / Q000276:immunology*
chemical_list: D000947:Antigens, Helminth; D015801:Helminth Proteins; D014612:Vaccines