Antioxidant enzyme gene delivery to protect from HIV-1 gp120-induced neuronal apoptosis

Intracisternal rSV40 administration provides effective pan-CNS transgene expression

“…12,13 Combination NT+antibody staining was performed using primary and secondary antibodies staining first (see above), followed by staining with the NT fluorescent stain. All experiments were repeated three times and test and control slides were stained the same day.…”

“…12 Intracerebral injection of rSV40s carrying antioxidant enzymes, Cu/Zn superoxide dismutase (SOD1) or glutathione peroxidase (GPx1), SV(SOD1) or SV(GPx1), into the rat caudate putamen (CP), significantly protected neurons from apoptosis caused by injection of recombinant HIV-1 envelope glycoprotein, gp120 or Tat at the same location. [12][13][14] Moreover, intra-CP SV40-mediated gene delivery of antioxidant enzymes largely protected against several deleterious consequences elicited by injecting gp120 into the CP. [15][16][17] Vector administration into the lateral ventricle, particularly if preceded by mannitol intraperitoneal, protected from intra-CP gp120-induced neurotoxicity comparably to intra-CP vector administration.…”

Gene transfer to the rhesus monkey brain using SV40-derived vectors is durable and safe

“…21 Intracerebral or intraventricular injection of recombinant SV40 vectors carrying the antioxidant enzymes, Cu/Zn superoxide dismutase (SOD1) or glutathione peroxidase (GPx1), SV(SOD1) or SV(GPx1), significantly protected neurons from oxidative injury and apoptosis elicited by HIV-1 envelope glycoprotein, gp120 or Tat. [22][23][24][25] Intracerebroventricular administration of SV(SOD1) or SV(GPx1), particularly if preceded by systemic mannitol, is protected from local gp120-induced neurotoxicity as well as did intraparenchymal administration of antioxidant enzymes to the site of subsequent challenge (490% protection). 24 Previous personal reports from our group have shown the transduction efficiency and distribution of rSV40 viral vectors in the rodent brain.…”

“…21,22,24,57 After rehydration in 0.1 M PBS, pH 7.2, sections were treated with PBS plus 0.1% Triton X-100 10 min, washed twice for 5 min in PBS and then stained by NT (Molecular Probes Inc., Eugene, OR, USA) (1:100) for 20 min at room temperature. Sections were washed in PBS plus 0.1% Triton X-100, then 2Â with PBS and finally let to stand for 2 h at room temperature in PBS before being counterstained with 4¢,6-diamidino-2-phenylindole.…”

“…Lysates were prepared as reported previously. 22 Total protein was estimated using BCA protein assay kit (Pierce, Rockford, IL, USA). Total cell proteins (100 mg) were loaded in each well, and then transferred to Immobilon-P poly vinylidine difluoride membrane (Millipore Inc., Billerica, MA, USA).…”