Dataset Information

Human regulatory T cells locally differentiate and are functionally heterogeneous within the inflamed arthritic joint.

ABSTRACT:

Objective

Tregs are crucial for immune regulation, and environment-driven adaptation of effector (e)Tregs is essential for local functioning. However, the extent of human Treg heterogeneity in inflammatory settings is unclear.

Methods

We combined single-cell RNA- and TCR-sequencing on Tregs derived from three to six patients with juvenile idiopathic arthritis (JIA) to investigate the functional heterogeneity of human synovial fluid (SF)-derived Tregs from inflamed joints. Confirmation and suppressive function of the identified Treg clusters was assessed by flow cytometry.

Results

Four Treg clusters were identified; incoming, activated eTregs with either a dominant suppressive or cytotoxic profile, and GPR56⁺CD161⁺CXCL13⁺ Tregs. Pseudotime analysis showed differentiation towards either classical eTreg profiles or GPR56⁺CD161⁺CXCL13⁺ Tregs supported by TCR data. Despite its most differentiated phenotype, GPR56⁺CD161⁺CXCL13⁺ Tregs were shown to be suppressive. Furthermore, BATF was identified as an overarching eTreg regulator, with the novel Treg-associated regulon BHLHE40 driving differentiation towards GPR56⁺CD161⁺CXCL13⁺ Tregs, and JAZF1 towards classical eTregs.

Conclusion

Our study reveals a heterogeneous population of Tregs at the site of inflammation in JIA. SF Treg differentiate to a classical eTreg profile with a more dominant suppressive or cytotoxic profile that share a similar TCR repertoire, or towards GPR56⁺CD161⁺CXCL13⁺ Tregs with a more distinct TCR repertoire. Genes characterising GPR56⁺CD161⁺CXCL13⁺ Tregs were also mirrored in other T-cell subsets in both the tumor and the autoimmune setting. Finally, the identified key regulators driving SF Treg adaptation may be interesting targets for autoimmunity or tumor interventions.

SUBMITTER: Lutter L

PROVIDER: S-EPMC9525321 | biostudies-literature | 2022

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Human regulatory T cells locally differentiate and are functionally heterogeneous within the inflamed arthritic joint.

Lutter Lisanne L van der Wal M Marlot MM Brand Eelco C EC Maschmeyer Patrick P Vastert Sebastiaan S Mashreghi Mir-Farzin MF van Loosdregt Jorg J van Wijk Femke F

Clinical & translational immunology 20220930 10

<h4>Objective</h4>Tregs are crucial for immune regulation, and environment-driven adaptation of effector (e)Tregs is essential for local functioning. However, the extent of human Treg heterogeneity in inflammatory settings is unclear.<h4>Methods</h4>We combined single-cell RNA- and TCR-sequencing on Tregs derived from three to six patients with juvenile idiopathic arthritis (JIA) to investigate the functional heterogeneity of human synovial fluid (SF)-derived Tregs from inflamed joints. Confirma ...[more]

PMID: 36204213

Similar Datasets

Project description:BACKGROUND:While multiple in vitro studies examined mesenchymal stromal cells (MSCs) derived from bone marrow or hyaline cartilage, there is little to no data about the presence of MSCs in the joint capsule or the ligamentum capitis femoris (LCF) of the hip joint. Therefore, this in vitro study examined the presence and differentiation potential of MSCs isolated from the bone marrow, arthritic hyaline cartilage, the LCF and full-thickness samples of the anterior joint capsule of the hip joint. METHODS:MSCs were isolated and multiplied in adherent monolayer cell cultures. Osteogenesis and adipogenesis were induced in monolayer cell cultures for 21?days using a differentiation medium containing specific growth factors, while chondrogenesis in the presence of TGF-ß1 was performed using pellet-culture for 27?days. Control cultures were maintained for comparison over the same duration of time. The differentiation process was analyzed using histological and immunohistochemical stainings as well as semiquantitative RT-PCR for measuring the mean expression levels of tissue-specific genes. RESULTS:This in vitro research showed that the isolated cells from all four donor tissues grew plastic-adherent and showed similar adipogenic and osteogenic differentiation capacity as proven by the histological detection of lipid droplets or deposits of extracellular calcium and collagen type I. After 27?days of chondrogenesis proteoglycans accumulated in the differentiated MSC-pellets from all donor tissues. Immunohistochemical staining revealed vast amounts of collagen type II in all differentiated MSC-pellets, except for those from the LCF. Interestingly, all differentiated MSCs still showed a clear increase in mean expression of adipogenic, osteogenic and chondrogenic marker genes. In addition, the examination of an exemplary selected donor sample revealed that cells from all four donor tissues were clearly positive for the surface markers CD44, CD73, CD90 and CD105 by flow cytometric analysis. CONCLUSIONS:This study proved the presence of MSC-like cells in all four examined donor tissues of the hip joint. No significant differences were observed during osteogenic or adipogenic differentiation depending on the source of MSCs used. Further research is necessary to fully determine the tripotent differentiation potential of cells isolated from the LCF and capsule tissue of the hip joint.

Dataset Information

Human regulatory T cells locally differentiate and are functionally heterogeneous within the inflamed arthritic joint.

Objective

Methods

Results

Conclusion

Publications

Human regulatory T cells locally differentiate and are functionally heterogeneous within the inflamed arthritic joint.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets