Application of the Human Hepatitis B Virus Core Antigen from Transgenic Tobacco Plants for Serological Diagnosis

Abstract

Background and objectives: The aim was to produce HBcAg from plants more cheaply than can be done by other currently available means, and to apply such antigen to immunoassay procedures for pretransfusion testing of donor blood. Materials and methods: Transgenic Nicotiana tabacum cv. SR-1 plants expressing the human hepatitis B virus (HBV) core antigen (HBcAg) gene were generated by Agrobacterium-mediated transformation. The recombinant product, called tHBcAg, can assemble itself into a spherical particle with a diameter of 25 to 30 nm, and can maintain two antigenic determinants of HBcAg, namely HBc/α and HBc/β. Partly purified tHBcAg was used in the hemagglutination-inhibition (HI) test, as routinely used by the Japanese Blood Center, to test a panel of 524 blood units taken from HBV-positive donors. Results: In the HI test, tHBcAg showed serologic properties comparable to that from Escherichia coli, the standard antigen used in the Japanese Blood Center. Conclusions: Transgenic plants can produce reagents for serologic testing and perhaps even such medical materials as oral vaccines.

HBV Core Particles as a Carrier for B Cell/T Cell Epitopes

“…Correct folding of the HBc protein and formation of authentic HBc particles have been documented in various mammalian cell cultures [13][14][15][16][17][18], retrovirus [19], vaccinia virus [20,21] and adenovirus [22] expression systems, frog Xenopus oocytes [23], insect Spodoptera cells [24][25][26][27], yeast Saccharomyces cerevisiae [28][29][30][31], in plants Nicotiana tabacum [32], and in bacteria such as Escherichia coli [33][34][35][36][37][38][39][40][41][42], Bacillus subtilis [43], Salmonella [44] and Acetobacter [45]. Electron microscopy revealed the ultrastructural identity of the HBc particles derived from either HBV virions and infected hepatocytes, or from E. coli [46] or yeast [47].…”