High precision-cut liver slice model to study cell-autonomous antiviral defense of hepatocytes within their microenvironment.
IP3, inositol-3-phosphate
LCMV, lymphocytic choriomeningitis virus
PCLS, precision-cut liver slices
PLCg, phospholipase C gamma
ROS, reactive oxygen species
TNF, tumor necrosis factor
TNF-induced apoptosis
anti-viral immunity
precision-cut liver slices
Journal
JHEP reports : innovation in hepatology
ISSN: 2589-5559
Titre abrégé: JHEP Rep
Pays: Netherlands
ID NLM: 101761237
Informations de publication
Date de publication:
May 2022
May 2022
Historique:
received:
21
10
2021
revised:
07
02
2022
accepted:
14
02
2022
entrez:
25
4
2022
pubmed:
26
4
2022
medline:
26
4
2022
Statut:
epublish
Résumé
Increased sensitivity towards tumor necrosis factor (TNF)-induced cell death in virus-infected hepatocytes has revealed a so far unrecognized hepatocyte-intrinsic antiviral immune surveillance mechanism, for which no Preparation of PCLS from mouse and human liver tissue was optimized for minimal procedure-associated apoptosis. Functionality of liver cells in PCLS was characterized using extracellular flux analysis to determine mitochondrial respiration, and viral infection with recombinant adenovirus and lymphocytic choriomeningitis virus (LCMV) was used to probe for hepatocyte-intrinsic sensitivity towards apoptosis in PCLS. Apoptosis was detected by immunohistochemical staining for cleaved-caspase 3 and quantified by detection of effector caspase activity in PCLS. We established an optimized protocol for preparation of PCLS from human and mouse models using agarose-embedding of liver tissue to improve precision cutting and using organ-protective buffer solutions to minimize procedure-associated cell death. PCLS prepared from virus-infected livers showed preserved functional metabolic properties. Importantly, in PCLS from adenovirus- and LCMV-infected livers we detected increased induction of apoptosis after TNF challenge We conclude that PCLS can be used as model system to Virus-infected hepatocytes
Sections du résumé
Background & Aims
UNASSIGNED
Increased sensitivity towards tumor necrosis factor (TNF)-induced cell death in virus-infected hepatocytes has revealed a so far unrecognized hepatocyte-intrinsic antiviral immune surveillance mechanism, for which no
Methods
UNASSIGNED
Preparation of PCLS from mouse and human liver tissue was optimized for minimal procedure-associated apoptosis. Functionality of liver cells in PCLS was characterized using extracellular flux analysis to determine mitochondrial respiration, and viral infection with recombinant adenovirus and lymphocytic choriomeningitis virus (LCMV) was used to probe for hepatocyte-intrinsic sensitivity towards apoptosis in PCLS. Apoptosis was detected by immunohistochemical staining for cleaved-caspase 3 and quantified by detection of effector caspase activity in PCLS.
Results
UNASSIGNED
We established an optimized protocol for preparation of PCLS from human and mouse models using agarose-embedding of liver tissue to improve precision cutting and using organ-protective buffer solutions to minimize procedure-associated cell death. PCLS prepared from virus-infected livers showed preserved functional metabolic properties. Importantly, in PCLS from adenovirus- and LCMV-infected livers we detected increased induction of apoptosis after TNF challenge
Conclusion
UNASSIGNED
We conclude that PCLS can be used as model system to
Lay summary
UNASSIGNED
Virus-infected hepatocytes
Identifiants
pubmed: 35462860
doi: 10.1016/j.jhepr.2022.100465
pii: S2589-5559(22)00037-4
pmc: PMC9019249
doi:
Types de publication
Journal Article
Langues
eng
Pagination
100465Informations de copyright
© 2022 The Authors.
Déclaration de conflit d'intérêts
The authors declare no conflict of interest. Please refer to the accompanying ICMJE disclosure forms for further details.
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