Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Jan;3(1):15-23.
doi: 10.47936/encephalitis.2022.00052. Epub 2023 Jan 3.

Regulatory T cell profiles in patients with N-methyl-ᴅ-aspartate receptor-antibody encephalitis

Mildred A Iro  1   2 , Christine S Rollier  3   4 , Sarosh R Irani  5   6 , Manish Sadarangani  7   8 , Adam Al-Diwani  5   9 , Andrew J Pollard  3 , Elizabeth A Clutterbuck  3
Affiliations

Regulatory T cell profiles in patients with N-methyl-ᴅ-aspartate receptor-antibody encephalitis

Mildred A Iro et al. Encephalitis. 2023 Jan.

Abstract

Purpose: Purpose Regulatory T cells (Tregs) have been implicated in the pathogenesis of several autoimmune disorders and used in adoptive cell transfer therapies. Neither have been explored in patients with autoimmune encephalitis where treated patient outcomes remain suboptimal with frequent relapses. Here, to identify new treatment strategies for autoimmune encephalitis, we sought to evaluate the proportion of circulating Tregs and Treg subpopulations in peripheral blood of patients with N-methyl-ᴅ-aspartate receptor-antibody encephalitis (NMDAR-Ab-E) and compared this with healthy controls.

Methods: We compared the phenotype of peripheral blood Tregs in four adult NMDAR-Ab-E patients and four age- and sex-matched healthy controls using an 11-color flow cytometry assay panel for characterization of Tregs (CD4+ CD25+ FoxP3+) cells into naïve (chemokine receptor [CCR] 7+ CD45RA+), central memory (CCR7+ CD45RA-), and effector memory (CCR7- CD45RA-) cells. We also examined and compared the expression of the CCR6 by circulating Tregs and the respective Treg subpopulations between the study groups.

Results: The proportion of circulating Tregs was similar between patients with NMDAR-Ab-E and healthy controls but the proportion of naïve Tregs was lower in NMDAR-Ab-E patients (p = 0.0026). Additionally, the frequency of circulating effector memory Tregs was higher, and the proportion of circulating effector memory Tregs expressing CCR6 was lower, in NMDAR-Ab-E patients compared with healthy controls (p = 0.0026).

Conclusion: Altered Treg homeostasis may be a feature of patients with NMDAR-Ab-E. Future studies with larger samples are warranted to validate these findings.

Keywords: Brain inflammation; CCR6 receptors; Regulatory T cells; Suppressor T cells.

PubMed Disclaimer

Conflict of interest statement

Conflicts of Interest Irani SR is an inventor of ‘Diagnostic Strategy to improve specificity of CASPR2 antibody detection (PCT/G82019/051257)' and receives royalties on a licensed patent application for LGI1/CASPR2 testing as co-applicant (PCT/GB2009/051441) entitled “Neurological Autoimmune Disorders.” Irani SR has received honoraria and/or research support from UCB, Immunovant, MedImmun, Roche, Janssen, Cerebral therapeutics, CSL Behring, and ONO Pharma. Iro MA is funded by Health Education England (HEE) / NIHR for this research project. No other potential conflict of interest relevant to this article was reported.

Figures

Figure 1.
Figure 1.. Gating strategy
(A) Gating strategy for fluorescence minus one control for intracellular markers. (B) Gating strategy used for identification of regulatory T cells (Tregs) and the different subpopulations. Subsequent gating is done on the population highlighted either in the circle of rectangle in the preceding figure. CTLA, cytotoxic T-lymphocyte associated protein 4; FMO, fluorescence minus one control; SSC, side scatter; FSC, forward scatter; PECF, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(carboxyfluorescein); APC, allophycocyanin; CCR, chemokine receptor. Tregs, CD3+ CD4+ CD25+ FoxP3+; memory Tregs, CD3+ CD4+ CD25+ FoxpP3+ CD45RA–; effector memory (EM) Tregs, CD3+ CD4+ CD25+ FoxP3+ CD45RA– CCR7–; central memory (CM) Tregs, CD3+ CD4+ CD25+ FoxP3+ CD45RA– CCR7+.
Figure 2.
Figure 2.. Comparison of different cell populations between NMDAR-Ab-E patients and healthy controls
(A) Lymphocyte comparison between NMDAR-Ab-E patients and healthy controls. (B) Comparison of proportion of CD3+ CD4+ T cells between NMDAR-Ab-E patients and healthy controls. (C) Comparison of proportion of Tregs between NMDAR-Ab-E patients and healthy controls. (D) Comparison of proportion of naïve Tregs between NMDAR-Ab-E patients and healthy controls. (E) Comparison of proportion of Tregs expressing CCR6 between NMDAR-Ab-E patients and healthy controls. (F) Comparison of proportion of effector memory Tregs between NMDAR-Ab-E patients and healthy controls. (G) Comparison of proportion of effector memory Tregs expressing CCR6 between NMDAR-Ab-E patients and healthy controls. (H) Comparison of proportion of central memory Tregs between NMDAR-Ab-E patients and healthy controls. NMDAR-Ab-E, N-methyl-ᴅ-aspartate receptor-antibody encephalitis; PBMC, peripheral blood mononuclear cell; CCR, chemokine receptor; Tregs, regulatory T cells.
See this image and copyright information in PMC

References

    1. Sun B, Ramberger M, O’Connor KC, Bashford-Rogers RJ, Irani SR. The B cell immunobiology that underlies CNS autoantibody-mediated diseases. Nat Rev Neurol. 2020;16:481–492. - PMC - PubMed
    1. Al-Diwani A, Theorell J, Damato V, et al. Cervical lymph nodes and ovarian teratomas as germinal centres in NMDA receptor-antibody encephalitis. Brain. 2022;145:2742–2754. - PMC - PubMed
    1. Zeng C, Chen L, Chen B, et al. Th17 cells were recruited and accumulated in the cerebrospinal fluid and correlated with the poor prognosis of anti-NMDAR encephalitis. Acta Biochim Biophys Sin (Shanghai) 2018;50:1266–1273. - PubMed
    1. Byun JI, Lee ST, Moon J, et al. Distinct intrathecal interleukin-17/interleukin-6 activation in anti-N-methyl-d-aspartate receptor encephalitis. J Neuroimmunol. 2016;297:141–147. - PubMed
    1. Miyara M, Gorochov G, Ehrenstein M, Musset L, Sakaguchi S, Amoura Z. Human FoxP3+ regulatory T cells in systemic autoimmune diseases. Autoimmun Rev. 2011;10:744–755. - PubMed