Expression of the recombinant tE2 antigen of bovine viral diarrhea virus (BVDV) in Nicotiana tabacum plants and biosafety studies in mice

RUBIO CECILIA; LAGUIA MELINA; SANJUAN PAULA; LANGLE YANINA; BENAVIDES PATRICIA; SANJUAN NORBERTO; MARCONI PATRICIA; GALLO MARINA; MARTINEZ SONIA; ALVAREZ, MARÍA ALEJANDRA

Biotecnología Aplicada

Editorial Elfos Scientiae

Lugar: La Habana; Año: 2016 vol. 33 p. 2401 - 2405

Antigenic glycoprotein tE2 from bovine viral diarrhea virus (BVDV) and the fluorescence marker EGFP were transiently expressed in leaves of Nicotiana tabacum using Agrobacterium tumefaciens as vector. Analyzing the Agrobacterium and EGFP RNA/protein persistence allowed us to estimate an environmental exposure to recombinant material during 6 days after agroinfiltration. Time course analysis for the expression of EGFP shows silencing after 6 days post-infiltration with recombinant Agrobacterium strain in treated leaves. The leaf extracts expressing tE2 and EGFP were clarified at 6 days after agroinfiltration. The extracts were mixed with oil or aqueous adjuvants and injected to Balb/c mice in order to establish if the plant extracts are innocuous. Four doses were applied, followed by necropsy. No significant differences between non-recombinant extracts (control treatment) or recombinant leaf extracts in clinical and biochemical analysis were observed. In all mice injected, the adjuvant caused the classical pathological lesions observed in typical inflammatory processes at the site of inoculation. Histological sections of these lesions and major organs showed no further injury.