Highlights
- •HAV undergoes limited replication in B6 mice, inducing an antiviral transcriptional response that restricts infection.
- •Genetically engineered Mavsvs/vs mice express murine MAVS protein (mMAVS-VS) that is cleaved by HAV 3ABC protease.
- •3ABC cleavage of mMAVS-VS enhances virus replication and reduces liver inflammation in HAV-infected Ifnar1-/- mice.
- •By itself, 3ABC cleavage of MAVS is not sufficient to overcome the host species barrier to HAV infection in B6 mice.
Background & Aims
Consistent with its relatively narrow host species range, hepatitis A virus (HAV)
cannot infect C57BL/6 mice. However, in Mavs-/- mice with genetic deficiency of the innate immune signaling adaptor MAVS, HAV replicates
robustly in the absence of disease. The HAV 3ABC protease cleaves MAVS in human cells,
thereby disrupting virus-induced IFN responses, but it cannot cleave murine MAVS (mMAVS)
due to sequence differences at the site of scission. Here, we sought to elucidate
the role of 3ABC MAVS cleavage in determining HAV pathogenesis and host species range.
Methods
Using CRISPR/Cas9 gene editing, we established two independent lineages of C57BL/6
mice with knock-in mutations altering two amino acids in mMAVS (‘mMAVS-VS’), rendering
it susceptible to 3ABC cleavage without loss of signaling function. We challenged
homozygous Mavsvs/vs mice with HAV, and compared infection outcomes with C57BL/6 and genetically deficient
Mavs-/- mice.
Results
The humanized murine mMAVS-VS protein was cleaved as efficiently as human MAVS when
co-expressed with 3ABC in Huh-7 cells. In embyronic fibroblasts from Mavsvs/vs mice, mMAVS-VS was cleaved by ectopically expressed 3ABC, significantly disrupting
Sendai virus-induced IFN responses. However, in contrast to Mavs-/- mice with genetic MAVS deficiency, HAV failed to establish infection in Mavsvs/vs mice, even with additional genetic knockout of Trif or Irf1. Nonetheless, when crossed with permissive Ifnar1-/- mice lacking type I IFN receptors, Mavsvs/vsIfnar1-/- mice demonstrated enhanced viral replication coupled with significant reductions
in serum alanine aminotransferase, hepatocellular apoptosis, and intrahepatic inflammatory
cell infiltrates compared with Ifnar1-/- mice.
Conclusions
MAVS cleavage by 3ABC boosts viral replication and disrupts disease pathogenesis,
but it is not by itself sufficient to break the host-species barrier to HAV infection
in mice.
Impact and implications
The limited host range of human hepatitis viruses could be explained by species-specific
viral strategies that disrupt innate immune responses. Both hepatitis A virus (HAV)
and hepatitis C virus express viral proteases that cleave the innate immune adaptor
protein MAVS, in human but not mouse cells. However, the impact of this immune evasion
strategy has never been assessed in vivo. Here we show that HAV 3ABC protease cleavage of MAVS enhances viral replication
and lessens liver inflammation in mice lacking interferon receptors, but that it is
insufficient by itself to overcome the cross-species barrier to infection in mice.
These results enhance our understanding of how hepatitis viruses interact with the
host and their impact on innate immune responses.
Graphical abstract
Graphical Abstract
Keywords
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Article info
Publication history
Published online: September 21, 2022
Accepted:
September 14,
2022
Received in revised form:
August 17,
2022
Received:
May 20,
2022
Footnotes
Author names in bold designate shared co-first authorship
Identification
Copyright
© 2022 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.
