Non-canonical NFKB Signaling Endows Suppressive Function Through FOXP3-dependent Regulatory T Cell Program

Overview

Journal Heliyon

Specialty Social Sciences

Date 2023 Dec 21

PMID 38125410

Authors

Yohei Sato

Erika Osada

Yoshinobu Manome

Affiliations

Soon will be listed here.

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.

Citing Articles

A dataset on NFKB1/NFKB2 knockout Jurkat cells and primary CD4+ T cells generated by CRISPR/Cas9 mediated gene disruption.

Osada E, Manome Y, Sato Y Data Brief. 2025; 59:111359.

PMID: 40027254 PMC: 11870169. DOI: 10.1016/j.dib.2025.111359.

Defective kinase activity of IKKα leads to combined immunodeficiency and disruption of immune tolerance in humans.

Cildir G, Aba U, Pehlivan D, Tvorogov D, Warnock N, Ipsir C Nat Commun. 2024; 15(1):9944.

PMID: 39550372 PMC: 11569180. DOI: 10.1038/s41467-024-54345-4.

MALT1 substrate cleavage: what is it good for?.

Moud B, Ober F, ONeill T, Krappmann D Front Immunol. 2024; 15:1412347.

PMID: 38863711 PMC: 11165066. DOI: 10.3389/fimmu.2024.1412347.

References

Sato Y, Liu J, Lee E, Perriman R, Roncarolo M, Bacchetta R . Co-Expression of FOXP3FL and FOXP3Δ2 Isoforms Is Required for Optimal Treg-Like Cell Phenotypes and Suppressive Function. Front Immunol. 2021; 12:752394. PMC: 8560788. DOI: 10.3389/fimmu.2021.752394. View

Ziegler L, Gerner M, Schmidt R, Trapin D, Steinberger P, Pickl W . Attenuation of canonical NF-κB signaling maintains function and stability of human Treg. FEBS J. 2020; 288(2):640-662. PMC: 7891634. DOI: 10.1111/febs.15361. View

Sharfe N, Merico D, Karanxha A, Macdonald C, Dadi H, Ngan B . The effects of RelB deficiency on lymphocyte development and function. J Autoimmun. 2015; 65:90-100. DOI: 10.1016/j.jaut.2015.09.001. View

Maccari M, Scarselli A, Cesare S, Floris M, Angius A, Deodati A . Severe Toxoplasma gondii infection in a member of a NFKB2-deficient family with T and B cell dysfunction. Clin Immunol. 2017; 183:273-277. DOI: 10.1016/j.clim.2017.09.011. View

Dever D, Bak R, Reinisch A, Camarena J, Washington G, Nicolas C . CRISPR/Cas9 β-globin gene targeting in human haematopoietic stem cells. Nature. 2016; 539(7629):384-389. PMC: 5898607. DOI: 10.1038/nature20134. View

Lam A, Lin D, Gillies J, Uday P, Pesenacker A, Kobor M . Optimized CRISPR-mediated gene knockin reveals FOXP3-independent maintenance of human Treg identity. Cell Rep. 2021; 36(5):109494. DOI: 10.1016/j.celrep.2021.109494. View

Cui Y, Benamar M, Schmitz-Abe K, Poondi-Krishnan V, Chen Q, Jugder B . A Stk4-Foxp3-NF-κB p65 transcriptional complex promotes T cell activation and homeostasis. Sci Immunol. 2022; 7(75):eabl8357. DOI: 10.1126/sciimmunol.abl8357. View

Ferreira L, Muller Y, Bluestone J, Tang Q . Next-generation regulatory T cell therapy. Nat Rev Drug Discov. 2019; 18(10):749-769. PMC: 7773144. DOI: 10.1038/s41573-019-0041-4. View

Sun S . The non-canonical NF-κB pathway in immunity and inflammation. Nat Rev Immunol. 2017; 17(9):545-558. PMC: 5753586. DOI: 10.1038/nri.2017.52. View

10.

Hori S, Nomura T, Sakaguchi S . Control of regulatory T cell development by the transcription factor Foxp3. Science. 2003; 299(5609):1057-61. DOI: 10.1126/science.1079490. View

11.

Bennett C, Christie J, Ramsdell F, Brunkow M, Ferguson P, Whitesell L . The immune dysregulation, polyendocrinopathy, enteropathy, X-linked syndrome (IPEX) is caused by mutations of FOXP3. Nat Genet. 2001; 27(1):20-1. DOI: 10.1038/83713. View

12.

Notarangelo L, Bacchetta R, Casanova J, Su H . Human inborn errors of immunity: An expanding universe. Sci Immunol. 2020; 5(49). PMC: 7647049. DOI: 10.1126/sciimmunol.abb1662. View

13.

Oh H, Ghosh S . NF-κB: roles and regulation in different CD4(+) T-cell subsets. Immunol Rev. 2013; 252(1):41-51. PMC: 3576882. DOI: 10.1111/imr.12033. View

14.

Chen K, Coonrod E, Kumanovics A, Franks Z, Durtschi J, Margraf R . Germline mutations in NFKB2 implicate the noncanonical NF-κB pathway in the pathogenesis of common variable immunodeficiency. Am J Hum Genet. 2013; 93(5):812-24. PMC: 3824125. DOI: 10.1016/j.ajhg.2013.09.009. View

15.

Mailer R . Alternative Splicing of FOXP3-Virtue and Vice. Front Immunol. 2018; 9:530. PMC: 5859138. DOI: 10.3389/fimmu.2018.00530. View

16.

Sakaguchi S, Miyara M, Costantino C, Hafler D . FOXP3+ regulatory T cells in the human immune system. Nat Rev Immunol. 2010; 10(7):490-500. DOI: 10.1038/nri2785. View

17.

Cepika A, Sato Y, Liu J, Uyeda M, Bacchetta R, Roncarolo M . Tregopathies: Monogenic diseases resulting in regulatory T-cell deficiency. J Allergy Clin Immunol. 2018; 142(6):1679-1695. DOI: 10.1016/j.jaci.2018.10.026. View

18.

Fontenot J, Gavin M, Rudensky A . Foxp3 programs the development and function of CD4+CD25+ regulatory T cells. Nat Immunol. 2003; 4(4):330-6. DOI: 10.1038/ni904. View

19.

Hamano R, Wu X, Wang Y, Oppenheim J, Chen X . Characterization of MT-2 cells as a human regulatory T cell-like cell line. Cell Mol Immunol. 2014; 12(6):780-2. PMC: 4711675. DOI: 10.1038/cmi.2014.123. View

20.

Badran Y, Dedeoglu F, Leyva Castillo J, Bainter W, Ohsumi T, Bousvaros A . Human haploinsufficiency results in autosomal-dominant chronic mucocutaneous ulceration. J Exp Med. 2017; 214(7):1937-1947. PMC: 5502421. DOI: 10.1084/jem.20160724. View