Non-canonical NFKB signaling endows suppressive function through FOXP3-dependent regulatory T cell program

Yohei Sato; Erika Osada; Yoshinobu Manome

doi:10.1016/j.heliyon.2023.e22911

Non-canonical NFKB signaling endows suppressive function through FOXP3-dependent regulatory T cell program

Heliyon. 2023 Nov 26;9(12):e22911. doi: 10.1016/j.heliyon.2023.e22911. eCollection 2023 Dec.

Authors

Yohei Sato¹, Erika Osada¹, Yoshinobu Manome¹

Affiliation

¹ Core Research Facilities, Research Center for Medical Sciences, The Jikei University School of Medicine, Tokyo, Japan.

Abstract

Regulatory T cells (Tregs) play a central role in modulating adaptive immune responses in humans and mice. The precise biological role of non-canonical nuclear factor 'κ-light-chain-enhancer' of activated B cells (NFKB) signaling in human Tregs has yet to be fully elucidated. To gain insight into this process, a Treg-like cell line (MT-2) was genetically modified using CRISPR/Cas9. Interestingly, NFKB2 knockout MT-2 cells exhibited downregulation of FOXP3, while NFKB1 knockout did not. Additionally, mRNA expression of FOXP3-dependent molecules was significantly reduced in NFKB2 knockout MT-2 cells. To better understand the functional role of the NFKB signaling, the NFKB1/NFKB2 loci of human primary Tregs were genetically edited using CRISPR/Cas9. Similar to MT-2 cells, NFKB2 knockout human Tregs displayed significantly reduced FOXP3 expression. Furthermore, NFKB2 knockout human Tregs showed downregulation of FOXP3-dependent molecules and a diminished suppressive function compared to wild-type and NFKB1 knockout Tregs. These findings indicate that non-canonical NFKB signaling maintains a Treg-like phenotype and suppressive function in human Tregs through the FOXP3-dependent regulatory T cell program.

Keywords: CRISPR/Cas9; FOXP3; NFKB signaling; Regulatory T cells; Suppressive function.