Abstract
Methods Using a combination of nucleotide sequence optimization and protein domain swapping, we have generated a panel of novel gene inserts for VSV vectors that encode chimeric HIV-1 and VSV glycoprotein immunogens (EnvG). A stable VERO cell line engineered to express human CD4 and CCR5 was used to rescue rVSV vectors in which the G gene was replaced with coding sequence for several different EnvG proteins.
Highlights
Our objective is to develop replicating recombinant vesicular stomatitis virus vectored HIV vaccine candidates that deliver membrane-bound trimeric HIV Env in a functional conformation
Analysis of cells transfected with plasmid DNA expressing encode chimeric HIV-1 and VSV glycoprotein immunogens (EnvG) constructs revealed abundant cell surface expression of chimeric glycoproteins
The chimeric EnvG in which the signal peptide (SP), transmembrane (TM) and cytoplasmic tail (CT) domains of HIV Env were replaced with functionally related domains of VSV G were expressed efficiently and supported vector propagation to high titer in CD4+CCR5+ cells
Summary
Our objective is to develop replicating recombinant vesicular stomatitis virus (rVSV) vectored HIV vaccine candidates that deliver membrane-bound trimeric HIV Env in a functional conformation. G TM and CT were incorporated efficiently into VSV particles. An EnvG in which the Env MPER domain was replaced with membrane-proximal sequence from G was more effectively processed and incorporated into virus particles
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