Biochemical and immunological characterization of the plant-derived candidate human immunodeficiency virus type 1 mucosal vaccine CTB-MPR 649-684

Nobuyuki Matoba, Hiroyuki Kajiura, Irene Cherni, Jeffrey D. Doran, Morgane Bomsel, Kazuhito Fujiyama, Tsafrir Leket-Mor

Research output: Contribution to journalArticlepeer-review

44 Scopus citations

Abstract

Plants are potentially the most economical platforms for the large-scale production of recombinant proteins. Thus, plant-based expression of subunit human immunodeficiency virus type 1 (HIV-1) vaccines provides an opportunity for their global use against the acquired immunodeficiency syndrome pandemic. CTB-MPR649-684 [CTB, cholera toxin B subunit; MPR, membrane proximal (ectodomain) region of gp41] is an HIV-1 vaccine candidate that has been shown previously to induce antibodies that block a pathway of HIV-1 mucosal transmission. In this article, the molecular characterization of CTB-MPR 649-684 expressed in transgenic Nicotiana benthamiana plants is reported. Virtually all of the CTB-MPR649-684 proteins expressed in the selected line were shown to have assembled into pentameric, GM1 ganglioside-binding complexes. Detailed biochemical analyses on the purified protein revealed that it was N-glycosylated, predominantly with high-mannose-type glycans (more than 75%), as predicted from a consensus asparagine-X-serine/threonine (Asn-X-Ser/Thr) N-glycosylation sequon on the CTB domain and an endoplasmic reticulum retention signal attached at the C-terminus of the fusion protein. Despite this modification, the plant-expressed protein retained the nanomolar affinity to GM1 ganglioside and the critical antigenicity of the MPR649-684 moiety. Furthermore, the protein induced mucosal and serum anti-MPR649-684 antibodies in mice after mucosal prime-systemic boost immunization. Our data indicate that plant-based expression can be a viable alternative for the production of this subunit HIV-1 vaccine candidate.

Original languageEnglish (US)
Pages (from-to)129-145
Number of pages17
JournalPlant Biotechnology Journal
Volume7
Issue number2
DOIs
StatePublished - Feb 2009

Keywords

  • HIV vaccine
  • N-glycosylation
  • Post-translational modification
  • Transgenic plants
  • gp41 membrane proximal region

ASJC Scopus subject areas

  • Biotechnology
  • Agronomy and Crop Science
  • Plant Science

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