Fig. 2
Human PCLS are productively infected by pandemic and avian IAV strains. A PCLS were infected one to seven days after generation with 106 PFU of IAV H1N1 A/Hamburg/4/2009 or H5N1 A/turkey/Turkey/2005. After 2 h PCLS were washed twice with PBS. Culture medium was changed daily. 4, 24, 48, and 72 h p.i. supernatants and PCLS were harvested for further analysis. B Representative confocal microscopy evaluation of Mock- and H1N1-infected PCLS, 4, 24, 48, and 72 h p.i. a, DAPI; b, IAV nucleoprotein (NP); c, F-actin; d, merge: DAPI, blue; IAV NP, green; F-actin, red. Scale bars, 100 µm. C Supernatants of infected PCLS were analyzed with a PFU assay. Each line represents an individual donor. D Tissue-associated viral RNA loads per 18S rRNA in PCLS over time following IAV infection. Dotted line represents the detection limit. E Correlation of tissue-associated viral RNA and donor age. F Correlation of tissue-associated viral RNA and deltaCt CDKN2A (p16) (deltaCt = Ct CDKN2A-Ct B2M). Association was tested using the Spearman rank correlation test. Each symbol represents an individual donor at 24 h (circle), 48 h (square), or 72 h p.i. (triangle). IAV H1N1, n = 3 and H5N1, n = 4