Abnormal effector and regulatory T cell subsets in paediatric-onset multiple sclerosis

Mexhitaj, Ina; Nyirenda, Mukanthu; Li, Rui; O’Mahony, Julia; Rezk, Ayman; Rozenberg, Ayal; Moore, Craig S.; Johnson, Trina; Sadovnick, Dessa; Collins, D. Louis; Arnold, Douglas L.; Gran, Bruno; Yeh, E. Ann; Marrie, Ruth Ann; Banwell, Brenda; Bar‐Or, Amit

doi:10.1093/brain/awz017

Cited by 69 publications

(50 citation statements)

References 51 publications

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“…Future biomarker discovery studies could include more extensive marker panels covering multiple immune cell types and their functional molecules in similar analyses by using mass cytometry, which allows for many more markers to be measured simultaneously, or by utilising alternative cell phenotyping approaches such as single-cell sequencing. This would allow assessment of treatment effects and potential predictive utility of the balance between subsets of T cells, B cells and myeloid cells considered increasingly relevant to defining the state of MS disease activity 35,36 . Our study involved multiple comparisons, which carries a risk of false positive discoveries.…”

Section: Discussionmentioning

confidence: 99%

“…PBMC were isolated from whole blood (Ficoll density centrifugation; GE Healthcare) and cryopreserved using rigorous standardized operating procedures (SOPs) developed and validated by our Experimental Therapeutics Program for all steps of sample procurement, processing, cryopreservation, storage and subsequent thawing. Following our strict SOPs for PBMC isolation and cryopreservation 36 , the majority of thawed samples had viabilities of over 90%. Any sample with cell viability of less than 70% was excluded from analysis.…”

Section: Mri Acquisitionmentioning

confidence: 99%

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Pre-treatment T-cell subsets associate with fingolimod treatment responsiveness in multiple sclerosis

Ghadiri

Rezk

et al. 2020

Sci Rep

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Bar-or 1,3* Biomarkers predicting fingolimod (FTY) treatment response in relapsing-remitting multiple sclerosis (RRMS) are lacking. Here, we performed extensive functional immunophenotyping using multiparametric flow cytometry to examine peripheral immune changes under FTY treatment and explore biomarkers of FTY treatment response. From among 135 RRMS patients who initiated FTY in a 2-year multicentre observational study, 36 were classified as 'Active' or 'Stable' based on clinical and/ or radiological activity on-treatment. Flow cytometric analysis of immune cell subsets was performed on pre-and on-treatment peripheral blood mononuclear cells (PBMC) samples. Decreased absolute counts of B cells and most T-cell subsets were seen on-treatment. Senescent CD8 + T cells, CD56 + t cells, CD56 dim natural killer cells, monocytes and dendritic cells were not reduced in number and hence relatively increased in frequency on-treatment. An unbiased multiparametric and traditional manual analysis of T-cell subsets suggested a higher pre-treatment frequency of CD4 + central memory T cells (TCM) in patients who were subsequently Active versus Stable on-treatment. Lower pre-treatment terminally differentiated effector memory (TEMRA) cell frequencies were also seen in the subsequently Active cohort. Together, our data highlight differential effects of FTY on peripheral immune cell subsets and suggest that pre-treatment T-cell subset frequencies may have value in predicting FTY treatment response.

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Section: Discussionmentioning

confidence: 99%

Section: Mri Acquisitionmentioning

confidence: 99%

Pre-treatment T-cell subsets associate with fingolimod treatment responsiveness in multiple sclerosis

Ghadiri

Rezk

et al. 2020

Sci Rep

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“…The study was approved by and carried out in accordance with the guidelines and regulations of the Institutional Review Board of the Montreal Neurological Institute (McGill University) and University of Pennsylvania and in accordance with the Declaration of Helsinki. Peripheral blood mononuclear cells (PBMC) were isolated from whole blood using Ficoll-Paque density gradient centrifugation (GE Healthcare) and a strict standardized protocol 53 . B cells were selected from PBMC using CD19 microbeads (Miltenyi Biotec) according to manufacturer’s recommendations.…”

Section: Methodsmentioning

confidence: 99%

Multiplexed detection and isolation of viable low-frequency cytokine-secreting human B cells using cytokine secretion assay and flow cytometry (CSA-Flow)

Rezk

Bar‐Or

2020

Sci Rep

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The ability to functionally characterize cytokine-secreting immune cells has broad implications in both health and a range of immune-mediated and auto-immune diseases. Low-frequency cytokine-defined immune-cell subsets can play key immune-regulatory roles, yet their detailed study is often hampered by limited clinical sample availability. Commonly used techniques including intracellular cytokine staining require cell fixation, precluding subsequent functional interrogation. The cytokine-secretion assay (CSA) can overcome this limitation, though has mostly been used for detection of relatively high-frequency, single-cytokine secreting cells. We examined how adaptation of the CSA in combination with multiparametric flow-cytometry (CSA-Flow) may enable simultaneous isolation of multiple, low-frequency, cytokine-secreting cells. Focusing on human B cells (traditionally recognized as harder to assay than T cells), we show that single-capture CSA-Flow allows for isolation of highly-purified populations of both low-frequency (IL-10+; GM-CSF+) and high-frequency (TNF+) cytokine-defined B cells. Simultaneous detection and isolation of up to three viable and highly-purified cytokine-secreting B-cell subpopulations is feasible, albeit with some signal loss, with fractions subsequently amenable to gene expression analysis and in vitro cell culture. This multiplexing CSA-Flow approach will be of interest in many human cellular immunology contexts aiming to functionally characterize cytokine-secreting immune cells, especially when sample volumes and cell numbers are limited.

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“…As well as conventional αβT cells, the thymus also produces γδT cells, natural killer T (NKT) cells and mucosal-associated invariant T cells. These are not discussed in depth here, but all are associated with CNS autoimmune disease and require intact thymopoiesis for their development [18][19][20]. Positive selection of conventional T cells occurs in the cortex and is mediated exclusively by cortical TECs (cTECs).…”

Section: Thymic Development and Functionmentioning

confidence: 99%

The contribution of thymic tolerance to central nervous system autoimmunity

Alberti

Handel

2020

Semin Immunopathol

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Autoimmune diseases of the central nervous system (CNS) are associated with high levels of morbidity and economic cost. Research efforts have previously focused on the contribution of the peripheral adaptive and innate immune systems to CNS autoimmunity. However, a failure of thymic negative selection is a necessary step in CNS-reactive T cells escaping into the periphery. Even with defective thymic or peripheral tolerance, the development of CNS inflammation is rare. The reasons underlying this are currently poorly understood. In this review, we examine evidence implicating thymic selection in the pathogenesis of CNS autoimmunity. Animal models suggest that thymic negative selection is an important factor in determining susceptibility to and severity of CNS inflammation. There are indirect clinical data that suggest thymic function is also important in human CNS autoimmune diseases. Specifically, the association between thymoma and paraneoplastic encephalitis and changes in T cell receptor excision circles in multiple sclerosis implicate thymic tolerance in these diseases. We identify potential associations between CNS autoimmunity susceptibility factors and thymic tolerance. The therapeutic manipulation of thymopoiesis has the potential to open up new treatment modalities, but a better understanding of thymic tolerance in CNS autoimmunity is required before this can be realised.

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Abnormal effector and regulatory T cell subsets in paediatric-onset multiple sclerosis

Cited by 69 publications

References 51 publications

Pre-treatment T-cell subsets associate with fingolimod treatment responsiveness in multiple sclerosis

Pre-treatment T-cell subsets associate with fingolimod treatment responsiveness in multiple sclerosis

Multiplexed detection and isolation of viable low-frequency cytokine-secreting human B cells using cytokine secretion assay and flow cytometry (CSA-Flow)

The contribution of thymic tolerance to central nervous system autoimmunity

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