Sendai virus‐based expression of HIV‐1 gp120: reinforcement by the V(−) version

Abstract

Background: We have established a system for recovering Sendai virus (SeV), a nonsegmented negative strand RNA virus, entirely from cDNA at an extremely high rate, and have succeeded in creating a V(-) SeV whose gene expression was greatly enhanced by the deletion of the nonessential V gene. Because of its extreme medical importance, there has been a strong need for the establishment of a better system to express the gp120 envelope glycoprotein of the human immunodeficiency virus type 1 (HIV-1) in sufficient quantity and purity. It also remains to be established to produce gp120 in in vitro natural host cells for HIV-1 such as human primary blood mononuclear cells, macrophages or established T cell lines.

Creation of an infectious recombinant Sendai virus expressing the firefly luciferase gene from the 3' proximal first locus.

“…Insertion as a NotI-tagged cassette into the first locus should be the principal method for construction of a recombinant SeV expressing other foreign genes. Indeed, we have also recovered SeV recombinants with the gene encoding the gp120 envelope glycoprotein of human immunodeficiency virus type 1 inserted into the same site (Yu et al, 1997). In this case, the length of inserted gene was 1608 nucleotides, only slightly shorter than the luciferase gene, and the SeV\gp120 recombinant displayed a very similar attenuated phenotype to that of SeV\luc.…”

Antibody-Dependent Cellular Cytotoxicity via Humoral Immune Epitope of Nef Protein Expressed on Cell Surface

“…Nef proteins were expressed in these cells by using a recombinant Sendai virus system, which has been shown to express large amounts of heterologous recombinant proteins in 24 h after infection in suspension cells (42). CEM-NK R cells were infected with SeVNef to express HIV-1 (NL43 strain) Nef proteins or wild SeV at a multiplicity of infection of 10 for 1 h at 37 o C, as previously described (43), and cultured for 24 h in RPMI 10.…”

A Cytoplasmic RNA Vector Derived from Nontransmissible Sendai Virus with Efficient Gene Transfer and Expression

“…Such reverse genetics technology has enabled the construction of genetically engineered viruses which carry additional foreign genes and opened the way for the development of gene transfer vectors from RNA viruses of this type (24). The vectors prepared by this method have shown a high efficiency of gene transfer and expression of foreign proteins in vitro (3,12,18,21,28,32,36). However, the recombinant paramyxoviruses constructed to date have contained all the viral structural genes and thus are replication competent, giving rise to fully infectious progeny capable of spreading in the body.…”