Figure 5. AAV2H22-CFTRΔR transduces pig airway epithelia in vivo.

(A–G) Gut-corrected CFTR-null pigs (1 week old) were transduced with 1.96 × 10¹² VG of AAV2H22-CFTRΔR. Two weeks later, pigs were studied and compared with age-matched WT and nontreated CFTR-null pigs. (A–D) Confocal images of CFTR expression. (A) WT. (B) CFTR-null control. (C) CFTR-null transduced with AAV2H22-CFTRΔR. (D) Confocal image from C superimposed on differential interference contrast (DIC) image to show cilia. Red, CFTR; green, ZO-1 in A and cadherin in B–D; blue, nuclei (DAPI). (E) Correction of cAMP-stimulated Isc in the trachea of CFTR-null pigs transduced with AAV2H22-CFTRΔR relative to untreated controls. (F) Short-circuit currents of nasal epithelia in a CFTR-null pig transduced with AAV2H22-CFTRΔR in the right nostril. Left nostril was used as control. Inhibition of Na⁺ current with amiloride (Amil.) (1 × 10⁻⁴ M) hyperpolarized apical membrane voltage and increased the driving force for Cl⁻ secretion. Inhibition of non-CFTR Cl⁻ channels with 4,4’-diisothiocyano-2,2’-stilbenedisulfonic acid (DIDS) (1 × 10⁻⁴ M). CFTR activity stimulation (ΔIsc_cAMP) by cAMP levels elevated via forskolin (1 × 10⁻⁵ M) and IBMX (1 × 10⁻⁴ M). (G) Airway surface liquid (ASL) pH was measured in vivo using pH-sensitive planar optical probe placed on tracheal surface. (H) S. aureus–coated grids were placed on tracheal surface of WT and CFTR-null pigs transduced with AAV2H22-CFTRΔR or control. Data are percentage dead bacteria. For each pig, 2–3 grids were used for 1 minute, 5–16 fields were counted per grid, each field contained ~100–1,000 bacteria, and data from each field were averaged. For A–H, n = 3 CFTR-null pigs transduced with AAV2H22; n = 6 untreated CFTR-null pigs; n = 6 non-CF pigs. *P < 0.05 vs. untreated CFTR-null *P < 0.01, Mann-Whitney nonparametric t test. For F, n = 1 nostril with contralateral nostril as a control. The non-CF control pigs (E, G, and H) were shared with a companion manuscript by Cooney et al. (58).