Figure 1.

Characterization of targeted lentiviral vectors. (a) Schematic drawing of plasmids used for production of targeted vectors. Targeted vectors were generated by six-plasmid cotransfection using the (i) pαThy1.1/pαp75^NTR/pαCAR plasmids, (ii) Igαβ plasmids required for surface expression of antibody molecules (iii) SINmu(SGN) envelope plasmid, the (iv) pRRLsincppt_CMV_eGFP-WPRE genome plasmid and the pMD2-LgpRRE and pRSV-Rev packaging plasmids. (b) Schematic representation of the virus staining method for visualizing individual particles. (c) αp75^NTR, αΤhy1.1, αCAR, 5pl, and VSVG particles were overlaid upon coverslips and stained for the presence of SGN fusogen (anti-HA-FITC), αp75^NTR/αThy1.1/αCAR (Alexa 594 anti-human IgG) and HIV p24 (anti-p24). The insert shows zoomed in view of the indicated region. Colocalizing pixels are indicated by a white signal. Bars = 10 μm. (d) Quantification of triple positive particles. For α75^NTR 49.8% of particles codisplay antibody and fusogen molecule, while for αThy1.1 and for αCAR 31 and 53% of total particles are respectively triple positive. CAR, coxsackievirus and adenovirus receptor; CMV, cytomegalovirus; EGFP, enhanced GFP.