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Volume 31, Number 1—January 2025
Research Letter

Replication Restriction of Influenza A(H5N1) Clade 2.3.4.4b Viruses by Human Immune Factor, 2023–2024

Jakob Ankerhold1, Susanne Kessler1, Martin Beer, Martin Schwemmle, and Kevin CiminskiComments to Author 
Author affiliation: University of Freiburg Spemann Graduate School of Biology and Medicine, Freiburg, Germany (J. Ankerhold); University Medical Centre and Faculty of Medicine Freiburg, Freiburg (J. Ankerhold, S. Kessler, M. Schwemmle, K. Ciminski); Friedrich-Loeffler-Institut, Greifswald–Insel Riems, Germany (M. Beer)

Main Article

Figure 2

In vitro testing of bovine influenza A(H5N1) restriction through replacement of individual viral polymerase complex components from a human-adapted MxA-resistant strain. A) HEK293T cells were transfected with expression plasmids encoding the indicated pH1N1 or bovine H5N1 polymerase subunits PB2, PB1, PA, and NP together with expression plasmids encoding antivirally active MxA or the inactive MxAT103A variant. After 24 hours, we determined the relative polymerase activity as the ratio of MxA to MxAT103A. Data are mean ± SD of n = 4 independent experiments. B) HEK293T cells were transfected with expression plasmids encoding the pH1N1, KAN-1 H5N1, or bovine H5N1 polymerase subunits PB2, PB1, and PA together with the respective NP. After 24 hours, we determined the polymerase activity in presence of the indicated MxA/Mx1 variant, normalized to a transfection control, and calculated relative to the empty vector control. Data are mean ± SD of n = 4 independent experiments. We used 2-tailed t-tests for statistical analysis. MxA, human myxovirus resistance protein 1; NP, nucleoprotein; PA, polymerase; PB, polymerase basic; pH1N1, pandemic H1N1.

Figure 2. In vitro testing of bovine influenza A(H5N1) restriction through replacement of individual viral polymerase complex components from a human-adapted MxA-resistant strain. A) HEK293T cells were transfected with expression plasmids encoding the indicated pH1N1 or bovine H5N1 polymerase subunits PB2, PB1, PA, and NP together with expression plasmids encoding antivirally active MxA or the inactive MxAT103A variant. After 24 hours, we determined the relative polymerase activity as the ratio of MxA to MxAT103A. Data are mean ± SD of n = 4 independent experiments. B) HEK293T cells were transfected with expression plasmids encoding the pH1N1, KAN-1 H5N1, or bovine H5N1 polymerase subunits PB2, PB1, and PA together with the respective NP. After 24 hours, we determined the polymerase activity in presence of the indicated MxA/Mx1 variant, normalized to a transfection control, and calculated relative to the empty vector control. Data are mean ± SD of n = 4 independent experiments. We used 2-tailed t-tests for statistical analysis. MxA, human myxovirus resistance protein 1; NP, nucleoprotein; PA, polymerase; PB, polymerase basic; pH1N1, pandemic H1N1.

Main Article

1These first authors contributed equally to this article.

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