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. 2018 Jul 17;9:1664. doi: 10.3389/fimmu.2018.01664

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Copyright © 2018 Westenius, Mäkelä, Julkunen and Österlund.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Infectivity and the cleavage of the HA protein of A/Hong Kong/156/1997 (H5N1) virus. (A) Monocyte-derived macrophages (Mɸs) or (B) monocyte-derived DCs (moDCs) were infected with different doses of a human isolate of avian influenza A/Hong Kong/156/1997 (H5N1) virus for 6 or 24 h. Results are the mean values from four donors infected and analyzed separately. Infected cells were detected with antibodies against H5N1 vaccine antigens (A) using immunofluorescence microscopy or (B) with flow cytometry. From each donor, at least 200 cells (average ca. 420) were counted (A) or 10,000 events were analyzed (B). The titers for infective particles per ml (FFU/ml) and representative multiplicity of infection (MOI) values were calculated for Mɸs and moDCs. (C) moDCs were infected with H5N1/1997 virus at different MOI values for 48 h. The cells were stained with virus-specific antibodies and analyzed with flow cytometry. From each donor, 10,000 events were analyzed. Results are the mean values of four donors analyzed separately. (D) moDCs from four donors were infected with indicated MOI values of H5N1/1997 virus for 24 h with or without added TPCK-trypsin and for protein samples cellular lysates from different donors were pooled. Cleavage of HA protein was detected by Western blotting with antibodies against the glycoproteins of H5N1 virus to detect the HA0, HA1, and HA2 of the H5N1 virus.