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. 2009 Nov 6;4(11):e7788. doi: 10.1371/journal.pone.0007788

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This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.

This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.

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Human bronchi tissues were collected from patients undergoing lung resection and immediately placed on air-liquid interface membranes for culturing. Within 12 hours the tissues were pretreated with PBS (“control”) or DAS181 (10 µg/cm², “2 h DAS181 pretreatment”) for 2 hours at 37°C. After washing, the tissues were infected with pandemic influenza A(H1N1) A/California/04/2009 (10⁶ TCID₅₀/tissue) for 1 hr and then washed. Tissues were then incubated at 37°C for 24 or 48 hrs before washing the apical surface and determining viral titer in either the apical wash by TCID₅₀ on MDCK cells (top panel), or within tissue homogenate by RT-PCR analysis of viral M-gene RNA level normalized to cellular β-actin RNA level (bottom panel). Dotted line represents limit of detection of the TCID₅₀ assay.