Recombinant antigen tE2 from bovine viral diarrhea virus (BVDV) expressed in Nicotiana tabacum plants: Biosafety studies

LAGUIA BECHER, MELINA; GALLO CALDERON, MARINA; RUBIO, CECILIA; YANINA LANGLE; MARTINEZ, S; SAN JUAN, P; SAN JUAN, N; ALVAREZ, ALEJANDRA; BENAVIDEZ, MP; MARCONI, PATRICIA

Biotecnologia Aplicada

Elfos Scientia

Año: 2016

1027-2852

Antigenicglycoprotein tE2 from bovine viral diarrhea virus (BVDV) and thefluorescence marker EGFP were transiently expressed in leaves of Nicotiana tabacum using Agrobacterium tumefaciens as vector. Analyzing the Agrobacterium and EGFP RNA/protein persistenceallowed us to estimate an environmental exposure to recombinant material during6 days after agroinfiltration. Time course analysis for the expression of EGFPshows silencing after 6 days post-infiltration with recombinant Agrobacteriumstrain in treated leaves.The leaf extracts expressing tE2 and EGFP wereclarified at 6 days after agroinfiltration. The extracts were mixed with oil oraqueous adjuvants and injected to Balb/c mice in order to establish if theplant extracts are innocuous. Four doses were apply followed by a necropsy. Nosignificant differences between non-recombinant extracts (control treatment) orrecombinant leaf extracts in clinical and biochemical analysis were observed.In all mice injected the adjuvant caused the classical pathological lesionsobserved in typical inflammatory processes at the site of inoculation.Histological sections of these lesions and major organs showed no furtherinjury.