The Hemagglutinin of Bat-Associated Influenza Viruses Is Activated by TMPRSS2 for pH-Dependent Entry into Bat but Not Human Cells

Hoffmann, Markus; Krüger, Nadine; Zmora, Pawel; Wrensch, Florian; Herrler, Georg; Pöhlmann, Stefan

The NA proteins of human FLUAV facilitate release of progeny particles from infected cells by removing sialic acids from the cell surface. To study the impact of the batFLUAV-NAL on transduction efficiency, we produced pseudotypes bearing batFLUAV-HAL, -NAL or both proteins. For comparison, we generated pseudotypes harboring WSN-HA, WSN-NA or both proteins. These pseudotypes were then used for inoculation of MDCK (inoculated with pseudotypes bearing WSN proteins) and EpoNi/22.1 (inoculated with pseudotypes bearing WSN or batFLUAV proteins) cells. Pseudotypes harboring only WSN-NA were not able to transduce target cells (Fig 3) while pseudotypes bearing either WSN-HA alone or in combination with WSN-NA transduced both MDCK and EpoNi/22.1, as expected. Transduction efficiency was 500-1,500-fold higher when WSN-NA was expressed in cells used for pseudotype production, in keeping with the findings that the presence of NA is required for efficient release of HA-bearing vectors and infectious FLUAV [32, 45, 46] . Pseudotypes harboring NAL were not infectious while pseudotypes bearing HAL robustly transduced EpoNi/22.1 cells, indicating that batFLUAV-HAL, like WSN-HA, is sufficient to mediate host cell entry. However, unlike WSN-NA, the expression of NAL in pseudotype producer cells did not increase transduction efficiency of HAL-harboring pseudotypes (Fig 3) , suggesting that NAL is not required for release and/or infectivity of HAL containing particles, at least in the experimental system chosen.