ThymoSpheres culture: A model to study human polyclonal unconventional T cells
In vitro cultures remain crucial for studying the fundamental mechanisms of human T-cell development. Here, we introduce a novel in vitro cultivation system based on ThymoSpheres (TS): dense spheroids consisting of DLL4-expressing stromal cells and human hematopoietic precursor cells, in the absence of thymic epithelial cells. These spheroids are subsequently cultured at the air-liquid interphase. TS generate large numbers of mature T cells, are easy to manipulate, scalable, and can be repeatably sampled to monitor T-cell differentiation. The mature T cells generated from primary human hematopoietic precursor cells were extensively characterized using single-cell RNA and combined T-cell receptor (TCR) sequencing. These predominantly CD8α T cells exhibit transcriptional and TCR CDR3 characteristics similar to the recently described human polyclonal αβ unconventional T cell (UTC) lineage. This includes the expression of hallmark genes associated with agonist selection, such as IKZF2 (Helios), and the expression of various natural killer receptors. The TCR repertoire of these UTCs is polyclonal and enriched for CDR3-associated autoreactive features and early rearrangements of the TCR-α chain. In conclusion, TS cultures offer an intriguing platform to study the development of this human polyclonal UTC lineage and its inducing selection mechanisms.
Authors
- Tom Taghon (ORCID: https://orcid.org/0000-0002-5781-0288)
- Bart Vandekerckhove (ORCID: https://orcid.org/0000-0003-3828-4195)
- Niels Vandamme (ORCID: https://orcid.org/0000-0001-6582-0040)
- Laurenz De Cock (ORCID: https://orcid.org/0000-0002-4935-6337)
- Lucas Deseins (ORCID: https://orcid.org/0000-0002-1623-9521)
- Stijn De Munter (ORCID: https://orcid.org/0000-0003-3821-0620)
- Melissa Pille (ORCID: https://orcid.org/0000-0002-2757-6486)
- David Vermijlen (ORCID: https://orcid.org/0000-0002-7605-8845)
- Lena Boehme (ORCID: https://orcid.org/0000-0001-7593-7533)
- Thomas Michiels (ORCID: https://orcid.org/0000-0001-9615-8053)
- Joline Ingels (ORCID: https://orcid.org/0000-0001-7030-0373)
- Glenn Goetgeluk (ORCID: https://orcid.org/0000-0002-9123-5707)
- Tessa Kerre (ORCID: https://orcid.org/0000-0003-2114-2844)
- Jana Roels (ORCID: https://orcid.org/0000-0001-6037-8767)
- Hanne Jansen (ORCID: https://orcid.org/0000-0002-6636-6916)
- Ruslan I. Dmitriev (ORCID: https://orcid.org/0000-0002-0347-8718)
- Guillem Sanchez Sanchez (ORCID: https://orcid.org/0000-0001-9919-1748)
- Eva Pascal (ORCID: https://orcid.org/0000-0002-7582-1490)
- Lore Billiet (ORCID: https://orcid.org/0000-0001-9372-1920)
- Amin Zolfaghari
- Robrecht De Vos
Institutions
- Université Libre de Bruxelles (BE)
- Ghent University Hospital (BE)
- Ghent University (BE)
- Cancer Research Institute Ghent (BE)
- VIB-VUB Center for Structural Biology (BE)
- Walloon Excellence in Lifesciences and Biotechnology (BE)
- VIB Single Cell Core
Publication Details
- Journal
- European Journal of Immunology
- Published
- 2024-09-09
- DOI
- https://doi.org/10.1002/eji.202451265
- Citations
- 1
- Primary Topic
- Immune Cell Function and Interaction
- Type
- article
- Field-Weighted Citation Impact
- 0.24
Funders
- Fonds Wetenschappelijk Onderzoek
- Universiteit Gent
- Stichting Tegen Kanker
- Vlaamse regering