Alopecia areata patients show deficiency of FOXP3+CD39+ T regulatory cells and clonotypic restriction of Treg TCRβ-chain, which highlights the immunopathological aspect of the disease

Fatma N Hamed; Annika Åstrand; Marta Bertolini; Alfredo Rossi; Afsaneh Maleki-Dizaji; Andrew G Messenger; Andrew J G McDonagh; Rachid Tazi-Ahnini

doi:10.1371/journal.pone.0210308

Alopecia areata patients show deficiency of FOXP3+CD39+ T regulatory cells and clonotypic restriction of Treg TCRβ-chain, which highlights the immunopathological aspect of the disease

PLoS One. 2019 Jul 5;14(7):e0210308. doi: 10.1371/journal.pone.0210308. eCollection 2019.

Authors

Fatma N Hamed¹, Annika Åstrand², Marta Bertolini³, Alfredo Rossi⁴, Afsaneh Maleki-Dizaji⁵, Andrew G Messenger⁶, Andrew J G McDonagh⁵, Rachid Tazi-Ahnini¹

Affiliations

¹ Department of Infection, Immunity and Cardiovascular disease, The Medical School, University of Sheffield, Sheffield, United Kingdom.
² Respiratory, Inflammation and Autoimmunity IMED Biotech Unit, AstraZeneca, Gothenburg, Sweden.
³ Laboratory for Hair Research and Regenerative Medicine, Department of Dermatology, University of Münster, Germany.
⁴ Dermatology Centre, University of Manchester, Salford Royal NHS Foundation Trust, Salford, United Kingdom.
⁵ Dermatology department, Sheffield Teaching Hospitals, Sheffield, United Kingdom.
⁶ Department of Dermatology, Royal Hallamshire Hospital, Sheffield, United Kingdom.

Abstract

Alopecia areata (AA) is a hair loss disorder resulting from an autoimmune reaction against hair follicles. T-helper 1 cells are a major contributor to this disorder, but little is known about the role of T-regulatory cells (Tregs) in AA. Here, we analysed the distribution of circulating Treg subsets in twenty AA patients with active hair loss and fifteen healthy subjects by flow cytometry. The Treg suppressor HLA-DR+ subpopulation was significantly reduced in the patients (P<0.001) and there were significantly fewer cells expressing CD39 among the CD4+CD25+Foxp3+ Treg subpopulation in patients (P = 0.001). FOXP3 CD39 Treg cells were also reduced in hair follicles; by 75% in non-lesional skin and 90% in lesional skin, when compared to control healthy skin. To further characterise Treg cells in AA; Tregs (CD4+CD25+FOXP3+) were investigated for their TCRβ sequence. PCR products analysed by Next Generation Sequencing techniques, showed that all frequent public clonotypes in AA Tregs were also present in controls at relatively similar frequencies, excepting two public clonotypes: CATSRDEGGLDEKLFF (V15 D1 J1-4) and CASRDGTGPSNYGYTF (V2 D1 J1-2), which were exclusively present in controls. This suggests that these Treg clonotypes may have a protective effect and that they may be an exciting subject for future therapeutic applications.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adult
Alopecia Areata / immunology*
Alopecia Areata / metabolism
Antigens, CD / metabolism*
Female
Forkhead Transcription Factors / metabolism*
Humans
Male
T-Lymphocytes, Regulatory / metabolism*

Substances

Antigens, CD
Forkhead Transcription Factors

Grants and funding

Financial support was provided under the Contract Research Organization agreement between AstraZeneca and University of Sheffield (no. X/009300 to RT-A). FNH was awarded a studentship (no. 313821 MED) from the Libyan Ministry of Education. Discussion between author AÅ, AstraZeneca, and author RT-A from University of Sheffield lead to the hypothesis that Tregs, and especially CD39 & CD73, were altered in AA patients. AstraZeneca was thus involved in the design of the study and sponsored with a oneoff payment to cover approximately 50% of consumables associated with the experiments. The funders had no additional role in data collection and analysis, decision to publish, or preparation of the manuscript. Author AÅ receives salary from AstraZeneca. Authors AM-D receives salary from AF ChemPharm Ltd, author RT-A receives salary from University of Sheffield and authors AGM and AJGM from National Health Services (NHS)/Sheffield Teaching Hospital (STH). The specific roles of these authors are articulated in the ‘author contributions.