Adenovirus-mediated expresssion of the murine ecotropic receptor facilitates transduction of human hematopoietic cells with an ecotropic retroviral vector

Abstract

One factor limiting the ability to modify human repopulating to 48%) by replacing the RSV promoter with the CMV E1a hematopoietic cells genetically with retroviral vectors is the promoter, resulting in a parallel increase in retroviral transrelatively low expression of the cognate viral receptor. We duction efficiency. Replacing the head portion of the fiber have tested sequential transduction of human hematopoprotein in conventional adenoviral vectors (serotype 5) with ietic cells with an adenoviral vector encoding the ecotropic the corresponding portion from an adenoviral 3 serotype retroviral receptor followed by transduction with an ecoresulted in ecotropic receptor expression in 60% of CD34 + tropic retroviral vector. Adenoviral transduction of K562 cells at an MOI of 10 4 and a retroviral transduction of 60% erythroleukemia cells was highly efficiently with Ͼ95% of of hematopoietic clonogenic progenitors. The sequential cells expressing the ecotropic receptor at a multiplicity of transduction strategy also resulted in efficient transduction infection (MOI) of 10 3 with a correspondingly high transof the primitive CD34 + CD38 − subset suggesting that it may duction with a retroviral vector. Ecotropic receptor hold promise for genetic modification of human hematopoexpression in CD34 + cells following transduction with adenietic stem cells. oviral vectors was increased by at least two-fold (from 20

Enhancing transduction of the liver by adeno-associated viral vectors

“…Next the influence of AAV promoter was assessed using AV5GFP, which is identical to AV5LacZ except that the β-galactosidase gene has been replaced with the green fluorescent protein cDNA. (25) 4×10 9 vg/kg of AV5GFP was administered into the tail vein of mice at 20 weeks after gene transfer with rAAV2-LPS-FIX, a vector in which the CAGG proviral promoter has been replaced with the α1-microglobulin / bikunin enhancer and thyroid hormone-binding globulin promoter (also referred to as liver specific promoter, LSP). (24;26) A three-fold increase in steady state hFIX from 0.02±0.007μg/ml to 0.07±0.006μg/ml (p=0.005 by student T test) was observed following delayed administration of recombinant adenovirus.…”

“…(38),(23) Adenoviral vectors, AV5LacZ and AV, vector stocks were prepared as described previously. (25)…”

Adaptation of Chimeric Retroviruses In Vitro and In Vivo: Isolation of Avian Retroviral Vectors with Extended Host Range

“…Ecotropic Moloney MLV use the murine cationic amino acid transporter MCAT-1 as a cellular receptor. Most nonrodent cells are not permissive for ecotropic MLV infection; however, these cells become permissive if they express MCAT-1 (5,8,28,51,58,67,68). Since avian cells lack a functional receptor for ecotropic murine retroviruses, they are not efficiently infected by ecotropic murine retroviruses.…”

Acute intermittent porphyria: vector optimization for gene therapy

“…While these results provided the rationale for gene therapy in AIP, the firstgeneration adenoviral vectors used in this study are not practical for therapy as they trigger a cytotoxic immune response, resulting in vector shut-down [12]. In contrast, adeno-associated virus (AAV) vectors provide an alternative vector for efficient liver transduction with reduced risk of the immune consequences [13,14].…”