A surface-modified baculovirus vector with improved gene delivery to B-lymphocytic cells.
A short peptide motif from gp350/220 of Epstein-Barr virus, EDPGFFNVEI, which was known to bind to CD21, a surface protein on B-lymphocyte, was inserted into the baculovirus surface protein gp64. The recombinant virus carrying the hybrid gp64/gp350 gene, vAc-gp350EGFP, was obtained, and the expression of gp64/gp350 protein was confirmed with immunoblot using anti-gp350 antibody. When compared with a control virus with wild type gp64, vAc-gp350EGFP showed increased transduction efficiency in B cell lines Raji, HR1, B95-8, BJAB, and DG75, regardless of their being EBV-positive or EBV-negative. No such increase was seen in non-B cell lines HEK293 and HeLa. When Raji cells were transduced with increased amount of vAc-gp350EGFP, transduction became saturated when the multiplicity of infection was higher than 20pfu/cell. The transduction of Raji cells by vAc-gp350EGFP was dose-dependently inhibited by pre-treatment of cells with anti-CD21 antibody. These results showed that vAc-gp350EGFP entered B cells by interacting with CD21.
Ge
Jing
J
Department Of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University, Shanghai 200433, People's Republic of China.
Huang
Yishu
Y
Hu
Xiaotong
X
Zhong
Jiang
J
D001402:B-Lymphocytes / Q000378:metabolism*; D016367:Baculoviridae / Q000235:genetics* / Q000378:metabolism; D001709:Biotechnology / Q000379:methods*; D015153:Blotting, Western; D002460:Cell Line; D017931:DNA Primers; D005434:Flow Cytometry; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D004854:Herpesvirus 4, Human / Q000235:genetics; D006801:Humans; D009842:Oligopeptides / Q000235:genetics; D014161:Transduction, Genetic / Q000379:methods*; D014764:Viral Proteins / Q000235:genetics
D017931:DNA Primers; D009842:Oligopeptides; D014764:Viral Proteins