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Rapid, Seamless Generation of Recombinant Poxviruses using Host Range and Visual Selection

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Published on: May 24, 2020

Recombinant myxoma virus lacking all poxvirus ankyrin-repeat proteins stimulates multiple cellular anti-viral

Stephanie A Lamb¹, Masmudur M Rahman¹, Grant McFadden¹

¹Department of Molecular Genetics and Microbiology, College of Medicine, University of Florida, 1600 SW Archer Road, PO Box 100266, Gainesville FL 32610, USA.

|July 29, 2014

Summary

Myxoma virus (MYXV) ankyrin-repeat (ANK-R) proteins collectively regulate host immune responses. Disrupting these MYXV ANK-R proteins attenuated the virus and activated immune pathways.

Keywords:

Ankyrin-repeat Interleukin 6 M-T5 M148 M149 M150 Myxoma virus Myxomatosis Nuclear factor kappa B Poxvirus

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Dissecting Innate Immune Signaling in Viral Evasion of Cytokine Production

Dissecting Innate Immune Signaling in Viral Evasion of Cytokine Production

Published on: March 2, 2014

Area of Science:

Virology
Immunology
Molecular Biology

Background:

Studying viral gene functions is challenging due to gene redundancy.
Myxoma virus (MYXV) has a complex genome with potential for overlapping gene functions.

Purpose of the Study:

To investigate the collective functions of the poxviral ankyrin-repeat (ANK-R) protein superfamily in MYXV.
To characterize the in vitro and in vivo effects of disrupting multiple ANK-R genes.

Main Methods:

Genetic engineering of MYXV to create a complete ANK-R knockout (vMyx-ANKsKO).
In vitro studies using A549 cells to assess immune pathway activation (NF-κB) and cytokine release (IL-6).
In vivo studies in rabbits to evaluate viral attenuation and lesion development.

Main Results:

Infection with vMyx-ANKsKO robustly activated the NF-κB pathway in A549 cells.
Increased IL-6 release was observed specifically upon vMyx-ANKsKO infection.
vMyx-ANKsKO exhibited the most significant attenuation and smallest lesion size in rabbits.

Conclusions:

The poxviral ANK-R protein superfamily plays a crucial, shared role in modulating host immune responses.
Disruption of ANK-R proteins leads to viral attenuation and altered immune signaling.
This study provides novel insights into the collective functions of MYXV ANK-R proteins.