Regulatory T cells modulate inflammation and reduce infarct volume in experimental brain ischaemia

David Brea; Jesús Agulla; Manuel Rodríguez‐Yáñez; David Barral; Pedro Ramos‐Cabrer; Francisco Campos; Angeles Almeida; Antoni Dávalos; José Castillo

doi:10.1111/jcmm.12304

Regulatory T cells participate in the recovery of ischemic stroke patients

10.21203/rs.2.16371/v4 ◽

2020 ◽

Author(s):

María Santamaría-Cadavid ◽

Emilio Rodríguez-Castro ◽

Manuel Rodríguez-Yáñez ◽

Susana Arias-Rivas ◽

Iria López-Dequidt ◽

...

Keyword(s):

T Cells ◽

Ischemic Stroke ◽

Regulatory T Cells ◽

Functional Outcome ◽

Infarct Volume ◽

Control Subjects ◽

Good Functional Outcome ◽

Early Neurological Deterioration ◽

The Relationship

Abstract Background: Recent preclinical studies have shown that regulatory T cells (Treg) play a key role in the immune response after ischemic stroke (IS). However, the role of Treg in human acute IS has been poorly investigated. Our aim was to study the relationship between circulating Treg and outcome in human IS patients. Methods: A total of 204 IS patients and 22 control subjects were recruited. The main study variable was good functional outcome at 3 months (modified Rankin scale ≤2) considering infarct volume, Early Neurological Deterioration (END) and risk of infections as secondary variables. The percentage of circulating Treg was measured at admission, 48, 72h and at day 7 after stroke onset. Results: Circulating Treg levels were higher in IS patients compared to control subjects. Treg at 48h were independently associated with good functional outcome (OR, 3.5; CI: 1.9-7.8) after adjusting by confounding factors. Patients with lower Treg at 48h showed higher frequency of END and risk of infections. In addition, a negative correlation was found between circulating Treg at 48h (r=-0.414) and 72h (r=-0.418) and infarct volume. Conclusions: These findings suggest that Treg may participate in the recovery of IS patients. Therefore, Treg may be considered a potential therapeutic target in acute ischemic stroke.

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Abstract 113: Regulatory T Cells Contribute to Sexual Dimorphic Outcomes After Acute Ischemic Brain Injury

Stroke ◽

10.1161/str.51.suppl_1.113 ◽

2020 ◽

Vol 51 (Suppl_1) ◽

Author(s):

Zeyu Sun ◽

Wei Su ◽

Ligen Shi ◽

Jie Chen ◽

Xiaoming Hu

Keyword(s):

T Cells ◽

Regulatory T Cells ◽

Immune Cell ◽

Infarct Volume ◽

Young Male ◽

Young Female ◽

Male Mice ◽

Stroke Outcomes ◽

Female Mice ◽

Ifn Γ

Introduction: The contribution of CD4 + Foxp3 + regulatory T cells (Treg) to acute stroke outcomes has been controversially reported in young male murine models of stroke, their effects in female stroke mice, however, are not characterized. This study explored the sexual dimorphisms in Treg and its contribution to acute stroke outcomes. Methods: Cerebral ischemia was induced by 60 min tMCAO in male or female (Young: 10-week, aged: 15-month) wild type (WT) mice and DTR mice expressing the diphtheria toxin (DT) receptor under the control of Foxp3 promoter. Tregs depletion was achieved by DT injection in DTR mice for 3 days prior to tMCAO. Infarct volume, sensorimotor functions and peripheral immune cell populations were assessed up to 5d after stroke. For RNA sequencing analysis, Tregs were sorted from blood of male or female DTR mice at 5d after sham or tMCAO surgery. Results: Young Treg competent (WT mice or DTR mice without DT) female mice exhibited significantly reduced infarct volume, as assessed by MRI T2 scanning and MAP2 staining, and greatly improved sensorimotor functions (rotarod test and adhesive removal test) compared to age- and genotype-matched male mice (n=8/group) 5d after tMCAO. Treg depletion deprived the neuroprotection in young female, while showed no significant effect on young male or aged female mice (n=8/group). RNA-seq analysis showed that IFN-γ signaling was downregulated in female Treg while upregulated in male Treg, suggesting a sexual difference in Treg-mediated immune response after stroke. Flow cytometry revealed ameliorated immune cell activation in blood and brain in female vs male mice 5d after stroke. Furthermore, Treg were isolated from young female, young male, aged female or female mice subjected to ovariectomy, and adoptively transferred (1 million cell/animal, iv) to young male mice 1 hour after tMCAO. Only young female Treg significantly reduced the infarct volume and improved sensorimotor functions compared to other treatment groups (n=7-8/group). Conclusion: Treg contribute to the neuroprotection in young female vs male in an age- and hormone-dependent manner. Transcriptomic analysis uncovered sexual differences in an IFN-γ centered regulatory pathways in Tregs, which keep post-stroke immune responses in check.

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The effect of fingolimod on regulatory T cells in a mouse model of brain ischaemia

Journal of Neuroinflammation ◽

10.1186/s12974-021-02083-5 ◽

2021 ◽

Vol 18 (1) ◽

Author(s):

Kyle Malone ◽

Andrea C. Diaz Diaz ◽

Jennifer A. Shearer ◽

Anne C. Moore ◽

Christian Waeber

Keyword(s):

T Cells ◽

Regulatory T Cells ◽

Mouse Model ◽

Cell Population ◽

Neuroprotective Activity ◽

Left Middle Cerebral Artery ◽

Brain Ischaemia ◽

Aged Mice ◽

Treg Frequency ◽

Ischaemic Brain

Abstract Background The role of the immune system in stroke is well-recognised. Fingolimod, an immunomodulatory agent licensed for the management of relapsing-remitting multiple sclerosis, has been shown to provide benefit in rodent models of stroke. Its mechanism of action, however, remains unclear. We hypothesised fingolimod increases the number and/or function of regulatory T cells (Treg), a lymphocyte population which promotes stroke recovery. The primary aim of this study was to rigorously investigate the effect of fingolimod on Tregs in a mouse model of brain ischaemia. The effect of fingolimod in mice with common stroke-related comorbidities (ageing and hypercholesteremia) was also investigated. Methods Young (15–17 weeks), aged C57BL/6 mice (72–73 weeks), and ApoE−/− mice fed a high-fat diet (20–21 weeks) underwent permanent electrocoagulation of the left middle cerebral artery. Mice received either saline or fingolimod (0.5 mg/kg or 1 mg/kg) at 2, 24, and 48 h post-ischaemia via intraperitoneal injection. Another cohort of young mice (8–9, 17–19 weeks) received short-term (5 days) or long-term (10 days) fingolimod (0.5 mg/kg) treatment. Flow cytometry was used to quantify Tregs in blood, spleen, and lymph nodes. Immunohistochemistry was used to quantify FoxP3+ cell infiltration into the ischaemic brain. Results Fingolimod significantly increased the frequency of Tregs within the CD4+ T cell population in blood and spleen post-ischaemia in all three mouse cohorts compared to untreated ischemic mice. The highest splenic Treg frequency in fingolimod-treated mice was observed in ApoE−/− mice (9.32 ± 1.73% vs. 7.8 ± 3.01% in young, 6.09 ± 1.64% in aged mice). The highest circulating Treg frequency was also noted in ApoE−/− mice (8.39 ± 3.26% vs. 5.43 ± 2.74% in young, 4.56 ± 1.60% in aged mice). Fingolimod significantly increased the number of FoxP3+ cells in the infarct core of all mice. The most pronounced effects were seen when mice were treated for 10 days post-ischaemia. Conclusions Fingolimod increases Treg frequency in spleen and blood post-ischaemia and enhances the number of FoxP3+ cells in the ischaemic brain. The effect of fingolimod on this regulatory cell population may underlie its neuroprotective activity and could be exploited as part of future stroke therapy.

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Regulatory T cells participate in the recovery of ischemic stroke patients

10.21203/rs.2.16371/v3 ◽

2019 ◽

Author(s):

María Santamaría-Cadavid ◽

Emilio Rodríguez-Castro ◽

Manuel Rodríguez-Yáñez ◽

Susana Arias-Rivas ◽

Iria López-Dequidt ◽

...

Keyword(s):

T Cells ◽

Ischemic Stroke ◽

Regulatory T Cells ◽

Functional Outcome ◽

Infarct Volume ◽

Control Subjects ◽

Good Functional Outcome ◽

Early Neurological Deterioration ◽

The Relationship

Abstract Background: Recent preclinical studies have shown that regulatory T cells (Treg) play a key role in the immune response after ischemic stroke (IS). However, the role of Treg in human acute IS has been poorly investigated. Our aim was to study the relationship between circulating Treg and outcome in human IS patients. Methods: A total of 204 IS patients and 22 control subjects were recruited. The main study variable was good functional outcome at 3 months (modified Rankin scale ≤2) considering infarct volume, Early Neurological Deterioration (END) and risk of infections as secondary variables. The percentage of circulating Treg was measured at admission, 48, 72h and at day 7 after stroke onset. Results: Circulating Treg levels were higher in IS patients compared to control subjects. Treg at 48h were independently associated with good functional outcome (OR, 3.5; CI: 1.9-7.8) after adjusting by confounding factors. Patients with lower Treg at 48h showed higher frequency of END and risk of infections. In addition, a negative correlation was found between circulating Treg at 48h (r=-0.414) and 72h (r=-0.418) and infarct volume. Conclusions: These findings suggest that Treg may participate in the recovery of IS patients. Therefore, Treg may be considered a potential therapeutic target in acute ischemic stroke.

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Abstract 3856: Adoptive Transfer Of Regulatory T Cells Confers Long Term Neuroprotection Against Cerebral Ischemic Stroke

Stroke ◽

10.1161/str.43.suppl_1.a3856 ◽

2012 ◽

Vol 43 (suppl_1) ◽

Author(s):

Peiying Li ◽

Xiaoming Hu ◽

Yu Gan ◽

Feng Zhang ◽

Yanqin Gao ◽

...

Keyword(s):

T Cells ◽

Ischemic Stroke ◽

Regulatory T Cells ◽

Infarct Volume ◽

The Body ◽

Neurological Deficits ◽

Adoptive Therapy ◽

Ischemic Brain ◽

Cell Based Therapy

Stroke is the leading cause of serious long-term disability in adults. Activation and mobilization of CD4+CD25+ regulatory T cells (Tregs) is an intrinsic mechanism the body uses to restrict pro-inflammatory response, one of the well-established contributing factors for secondary neuronal injury and long-term neurological deficits after stroke. The current study explores the protective effect of Tregs adoptive therapy against post-ischemic brain damage and investigated the mechanisms underlying the action of Tregs. Using a mouse model of focal transient ischemia, we found that intravenous injections of Tregs (2 x 10 6 /animal) within 24 hours (2, 6, and 24 hours) after ischemia resulted in marked reduction of brain infarct. The maximal protection occurred upon earlier Tregs transfer with 2-hour delay after MCAO, which resulted in approximately 30% reduction of infarct volume. Post-ischemic sensorimotor dysfunction significantly improved during both the acute and late recovery after MCAO in Treg-treated mice as assessed by corner test, forelimb placing and cylinder test up to 28 days after ischemic stroke. Furthermore, Tregs treatment inhibited the up-regulation of IL-6, IL-1β, IL-17 and TNF-α in the ischemic brain and mitigated the cerebral infiltration of peripheral immune cells, including neutrophil, macrophage and T cells early after MCAO. Taken together, our study demonstrates that adoptive therapy with Tregs is a novel and potent cell-based therapy targeting post-stroke inflammatory dysregulation.

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