Figure 6: Actin cytoskeletal disruption restrains initial viral dissemination.

(A) ORF diagrams of HIV Nef_WT and HIV Nef_F191A.

(B) 10⁴ IUs each of each clone were intravaginally co-inoculated into BLT NSG mice at 50:50 ratio (based on I.U.). Plasma viremia was measured at week two, followed the next day by tissue harvest or, in non-viremic animals, repeat intravaginal inoculation and tissue harvest at week three.

(C) Plasma viremia. Dashed line and grey-shaded area indicate mean and range of background signals in three uninfected control animals.

(D) Ratio of nef_WT (red) or nef_F191A (green) NGS reads (~2 × 10⁴ total reads/sample) from plasma vRNA obtained at the time of initial viremia. Numbers indicate individual animals. HIV nef_WT vRNA was undetectable at any time in plasma or any tissue analyzed in the four animals grouped on the left.

(E) ORF diagrams of HIV Nef_WT and HIV ΔNef.

(F) 10⁴ IUs each of HIV Nef_WT and HIV ΔNef were intravaginally inoculated into BLT NSG mice at 50:50 ratio (based on I.U.). Plasma viremia was measured weekly starting at week 2. Non-viremic animals received repeat intravaginal inoculations the next day.

(G) Plasma viremia. Dotted line and grey-shaded area indicate mean and range of background signals.

(H, I) Ratio of NGS reads (~2 × 10⁴ total reads/sample) for nef_WT (red) or Δnef (green) from plasma vRNA obtained at the time of initial viremia (H) or at the time of sacrifice (I). Numbers indicate individual animals. Either nef_WT or Δnef vRNA were undetectable in any samples in the animals grouped on the left (n=1) or on the right (n=2), respectively.