Surface display of IgG Fc on baculovirus vectors enhances binding to antigen-presenting cells and cell lines expressing Fc receptors
Recombinant baculoviruses (recBV) were constructed with dual cassettes for constitutive expression of human IgG Fc following infection of insect cells and the structural proteins of hepatitis C virus (core, E1 and E2) following transduction of mammalian cells. The IgG Fc was expressed in insect cells as a fusion protein with the signal sequence and transmembrane region of either the native baculovirus envelope protein gp64 or the human transferrin receptor as a type I or type II integral membrane protein, respectively. The IgG Fc fusion proteins formed functional homodimers on the surface of recBV-infected insect cells and were incorporated into the envelope of recBV particles during egress from the infected cell. Both pseudotyped recBV bound specifically to recombinant soluble FcγRIIα receptor and to cell lines and antigen-presenting cells expressing Fc receptors (FcRs). These novel baculoviral vectors, which target cells of the immune system that express FcRs, have potential applications for vaccination or gene therapy.
KeywordsInsect Cell Huh7 Cell Mean Fluorescence Intensity Recombinant Baculoviruses Gp64 Envelope Protein
We thank Dr. Patrick Condreay for the kind gift of plasmid pFastBacMam (pFBM) and Professor Stan Lemon for plasmid pHCV-N. This study was supported by a grant from the National Health and Medical Research Council of Australia. The HCV Laboratory at the Burnet Institute is a member of the Australian Centre for Hepatitis Virology and the Australian Centre for HIV and Hepatitis Virology.
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