Stability and characterization of infectious monkeypox virus extracellular virions in vitro2and vivo; implications for transmission, pathogenesis and treatment

Spik, K.; Davis, C. I.; Bixler, S.; Garrison, A. R.; Golden, J. W.; Piper, A. E.; Koehler, J. W.; Williams, J. A.; RICKS, K. M.; Mucker, E. M.

doi:10.1101/2025.03.12.642804

bioRxiv · 10.1101/2025.03.12.642804

Stability and characterization of infectious monkeypox virus extracellular virions in vitro2and vivo; implications for transmission, pathogenesis and treatment

Spik, K.·Davis, C. I.·Bixler, S.·Garrison, A. R.·Golden, J. W.·Piper, A. E.·Koehler, J. W.·Williams, J. A.·RICKS, K. M.·Mucker, E. M.

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Abstract

Abstract/SummaryPoxviruses exist as multiple infectious morphogenic forms commonly simplified as mature virions (MV) and extracellular virions (EV). The roles of morphogenic subtypes as related to disease and transmission are enigmatic as EVs can exist both as cell associated (CEV) or released particles (rCEV) each with potentially unique biochemical properties impacting stability and infectivity. In vitro analysis of prototypical poxviruses is commonly utilized to infer larger conclusions about the in vivo function of all EV-like particles. Here we show that infectious EV of MPXV and VACV strains are stable for [≥] 14 weeks and are more sensitive to human or non-human primate complement compared to rabbit-derived complement. We also characterize the levels of EV produced during MPXV infection in NHPs and found temporal differences in production that may influence spread. Also, we present data characterizing and contrasting the EV from monkeypox (MPXV) and vaccinia virus (VACV) strains. Specifically, we quantified infectious EV quantities produced by different cultured cell lines and characterized the infectious properties and composition of the released (extracellular) virions. We conclude that A33 neutralizable cell associated-like virions (CEV-like), a form of EV, can significantly increase, depending on the strain of VACV or MPXV and the cell lines from which they were released. Based on the outcomes of our studies, the importance of understanding specific orthopoxvirus EV roles in a host-specific manner, as it relates to pathogenesis, stability, and transmission, is warranted and requires further study.

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Spik, K., Davis, C. I., Bixler, S., Garrison, A. R., Golden, J. W., Piper, A. E., Koehler, J. W., Williams, J. A., RICKS, K. M., Mucker, E. M.. 2025-03-12. Stability and characterization of infectious monkeypox virus extracellular virions in vitro2and vivo; implications for transmission, pathogenesis and treatment. https://doi.org/10.1101/2025.03.12.642804

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