Epithelial organoid supports resident memory CD8 T cell differentiation
Max R. Ulibarri,
Ying Lin,
Julian C. Ramprashad,
Geongoo Han,
Mohammad H. Hasan,
Farha J. Mithila,
Chaoyu Ma,
Smita Gopinath,
Nu Zhang,
J. Justin Milner,
Lalit K. Beura
Affiliations
Max R. Ulibarri
Department of Molecular Microbiology and Immunology, Brown University, Providence, RI 02912, USA
Ying Lin
Department of Molecular Microbiology and Immunology, Brown University, Providence, RI 02912, USA; Pathobiology Graduate Program, Brown University, Providence, RI 02912, USA
Julian C. Ramprashad
Department of Molecular Microbiology and Immunology, Brown University, Providence, RI 02912, USA
Geongoo Han
Department of Molecular Microbiology and Immunology, Brown University, Providence, RI 02912, USA
Mohammad H. Hasan
Department of Molecular Microbiology and Immunology, Brown University, Providence, RI 02912, USA
Farha J. Mithila
Department of Molecular Microbiology and Immunology, Brown University, Providence, RI 02912, USA; Molecular Biology, Cell Biology and Biochemistry Graduate Program, Brown University, Providence, RI 02912, USA
Chaoyu Ma
Department of Microbiology, Immunology and Molecular Genetics, University of Texas Health Science Center, San Antonio, TX 78229, USA
Smita Gopinath
Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health, Cambridge, MA 02115, USA
Nu Zhang
Department of Microbiology, Immunology and Molecular Genetics, University of Texas Health Science Center, San Antonio, TX 78229, USA; South Texas Veterans Health Care System, San Antonio, TX 78229, USA
J. Justin Milner
Department of Microbiology and Immunology, University of North Carolina, Chapel Hill, NC 27599, USA
Lalit K. Beura
Department of Molecular Microbiology and Immunology, Brown University, Providence, RI 02912, USA; Corresponding author
Summary: Resident memory T cells (TRMs) play a vital role in regional immune defense. Although laboratory rodents have been extensively used to study fundamental TRM biology, poor isolation efficiency and low cell survival rates have limited the implementation of TRM-focused high-throughput assays. Here, we engineer a murine vaginal epithelial organoid (VEO)-CD8 T cell co-culture system that supports CD8 TRM differentiation. These in-vitro-generated TRMs are phenotypically and transcriptionally similar to in vivo TRMs. Pharmacological and genetic approaches showed that transforming growth factor β (TGF-β) signaling plays a crucial role in their differentiation. The VEOs in our model are susceptible to viral infections and the CD8 T cells are amenable to genetic manipulation, both of which will allow a detailed interrogation of antiviral CD8 T cell biology. Altogether we have established a robust in vitro TRM differentiation system that is scalable and can be subjected to high-throughput assays that will rapidly add to our understanding of TRMs.
