Optimisation of herpes simplex virus-based vectors for delivery to human peripheral blood mononuclear cells.
Peripheral blood mononuclear cells (PBMCs) represent a significant target for gene delivery both for therapeutic and experimental purposes. Thus far however, it has proved difficult to develop vectors capable of high efficient gene delivery to unstimulated PBMCs. We have tested a range of different vectors derived from herpes simplex virus (HSV) which differ in their degree of disablement in terms of their gene delivery efficiency to unstimulated human PBMCs and ability to deliver a reporter gene. None of the viruses had any significant toxic effect in PBMCs. However, optimal gene delivery to unstimulated PBMCs was obtained with a semidisabled virus lacking functional genes encoding ICP34.5 and Vmw65 which was more efficient than either nondisabled or more extremely disabled viruses. Expression of green fluorescent protein (GFP) with this virus was observed in up to 50% of PBMCs 1 day after infection, and reporter gene expression was detectable by Western blotting and immunofluorescence at undiminished levels at the longest time points tested, up to 5 days after infection. This optimised HSV vector may thus represent an effective tool for gene delivery to unstimulated PBMCs in culture.
Papageorgiou
Konstantina
K
Institute of Child Health, University College London, 30 Guilford Street, London WC1N 1EH, UK.
Isenberg
David A
DA
Latchman
David S
DS
D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D006224:Cricetinae; D017353:Gene Deletion; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics / Q000502:physiology*; D049452:Green Fluorescent Proteins; D017405:Herpes Simplex Virus Protein Vmw65 / Q000235:genetics / Q000502:physiology*; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000502:physiology*; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics / Q000502:physiology*; D007963:Leukocytes, Mononuclear / Q000166:cytology; D008164:Luminescent Proteins / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics / Q000502:physiology*
D017405:Herpes Simplex Virus Protein Vmw65; C057120:ICP27 protein, human herpesvirus 1; D017874:Immediate-Early Proteins; D008164:Luminescent Proteins; D014764:Viral Proteins; C066469:gamma 34.5 protein, Human herpesvirus 1; C048925:herpes simplex virus, type 1 protein ICP4; D049452:Green Fluorescent Proteins