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. 2014 Sep;88(17):10066–10077. doi: 10.1128/JVI.02300-13

FIG 4.

FIG 4

MLV-based retroviral vector suppression of IFN-α production by human pDCs is sensitive to enzyme treatment. (A) pDCs isolated from healthy individuals were stimulated with ODN2395 or Toca 511 at an MOI of 1, 10, and 100 or lentiviral vector pseudotyped with VSV-G at an MOI of 20. Production of IFN-α was measured 30 h after stimulation. (B) Western blot of Toca 511 and Toca 511 pseudotyped with a VSV-G (Toca 511-G) vector by using antibodies against the amphotropic envelope glycoprotein (gp70), VSV-G protein, and the capsid protein (CA). (C) IFN-α production of pDCs stimulated with Toca 511 or Toca 511-G at an MOI of 20. (D) IFN-α production of pDCs stimulated with Toca 511 or Toca 511-G at an MOI of 20 with or without heat inactivation. (E) Blockade of IFN-α production in pDCs stimulated and coincubated with Toca 511, Toca 511 HI (heat treated), or Toca 511 and LV-G. (F) Immunoblots of heat- and non-heat-treated Toca 511 and enzyme digestion with deglycodiases, phospholipase C, or trypsin. D, deglysidases; P, phospholipase C; T, trypsin. (G) Blockade of IFN-α production in pDCs is removed by coincubation of trypsin-treated and heat-treated Toca 511.