Single-cell analysis of Kaposi's sarcoma-associated herpesvirus infection in three-dimensional air-liquid interface culture model

The oral cavity is the major site for transmission of Kaposi's sarcoma-associated herpesvirus (KSHV), but how KSHV establishes infection and replication in the oral epithelia remains unclear. Here, we report a KSHV spontaneous lytic replication model using fully differentiated, three-dimensiona...

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Bibliographic Details
Published in:PLoS pathogens 2022-08, Vol.18 (8), p.e1010775-e1010775
Main Authors: Jung, Kyle L, Choi, Un Yung, Park, Angela, Foo, Suan-Sin, Kim, Stephanie, Lee, Shin-Ae, Jung, Jae U
Format: Article
Language:English
Subjects:
Acquired Immunodeficiency Syndrome
Biology and Life Sciences
Cell culture
Cell death
Cell differentiation
Development and progression
Differentiation
Distribution
Epithelial cells
Epithelium
Gene expression
Gene Expression Regulation, Viral
Gene sequencing
Herpesviridae Infections
Herpesvirus 8, Human - physiology
Humans
Infections
Kaposi's sarcoma
Kaposis sarcoma
Keratin
Medicine and Health Sciences
Oral cavity
Organoids
Quorum sensing
Replication
Sarcoma
Single-Cell Analysis
Virus Latency
Virus Replication
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Summary:The oral cavity is the major site for transmission of Kaposi's sarcoma-associated herpesvirus (KSHV), but how KSHV establishes infection and replication in the oral epithelia remains unclear. Here, we report a KSHV spontaneous lytic replication model using fully differentiated, three-dimensional (3D) oral epithelial organoids at an air-liquid interface (ALI). This model revealed that KSHV infected the oral epithelia when the basal epithelial cells were exposed by damage. Unlike two-dimensional (2D) cell culture, 3D oral epithelial organoid ALI culture allowed high levels of spontaneous KSHV lytic replication, where lytically replicating cells were enriched at the superficial layer of epithelial organoid. Single cell RNA sequencing (scRNAseq) showed that KSHV infection induced drastic changes of host gene expression in infected as well as uninfected cells at the different epithelial layers, resulting in altered keratinocyte differentiation and cell death. Moreover, we identified a unique population of infected cells containing lytic gene expression at the KSHV K2-K5 gene locus and distinct host gene expression compared to latent or lytic infected cells. This study demonstrates an in vitro 3D epithelial organoid ALI culture model that recapitulates KSHV infection in the oral cavity, where KSHV undergoes the epithelial differentiation-dependent spontaneous lytic replication with a unique cell population carrying distinct viral gene expression.
ISSN:1553-7374
1553-7366
1553-7374
DOI:10.1371/journal.ppat.1010775