HIV-1 p24-immunoglobulin fusion molecule: a new strategy for plant-based protein production.
We describe the engineering of a human immunodeficiency virus-1 (HIV-1) p24-immunoglobulin A (IgA) antigen-antibody fusion molecule for therapeutic purposes and its enhancing effect on fused antigen expression in tobacco plants. Although many recombinant proteins have been expressed in transgenic plants as vaccine candidates, low levels of expression are a recurring problem. In this paper, using the HIV p24 core antigen as a model vaccine target, we describe a strategy for increasing the yield of a recombinant protein in plants. HIV p24 antigen was expressed as a genetic fusion with the alpha2 and alpha3 constant region sequences from human Ig alpha-chain and targeted to the endomembrane system. The expression of this fusion protein was detected at levels approximately 13-fold higher than HIV p24 expressed alone, and a difference in the behaviour of the two recombinant proteins during trafficking in the plant secretory pathway has been identified. Expressing the antigen within the context of alpha-chain Ig sequences resulted in the formation of homodimers and the antigen was correctly recognized by specific antibodies. Furthermore, the HIV p24 elicited T-cell and antibody responses in immunized mice. The use of Ig fusion partners is proposed as a generic platform technology for up-regulating the expression of antigens in plants, and may represent the first step in a strategy to design new vaccines with enhanced immunological properties.
Obregon
Patricia
P
Department of Cellular and Molecular Medicine, Infectious Diseases, Immunology Unit, St. George's Hospital Medical School, Cranmer Terrace, London SW17 0RE, UK. pobregon@sgul.ac.uk
Chargelegue
Daniel
D
Drake
Pascal M W
PM
Prada
Alessandra
A
Nuttall
James
J
Frigerio
Lorenzo
L
Ma
Julian K-C
JK
D000818:Animals; D003001:Cloning, Molecular; D019281:Dimerization; D005260:Female; D016655:HIV Core Protein p24 / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D015497:HIV-1 / Q000235:genetics / Q000276:immunology*; D007070:Immunoglobulin A / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D030821:Plants, Genetically Modified / Q000378:metabolism* / Q000502:physiology; D017510:Protein Folding; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D012038:Regeneration; D013601:T-Lymphocytes / Q000276:immunology; D014026:Nicotiana / Q000235:genetics*
D016655:HIV Core Protein p24; D007070:Immunoglobulin A; D011993:Recombinant Fusion Proteins