Abstract
Brain-infiltrating lymphocytes (BILs) were isolated from resected brain tissue from 10 pediatric epilepsy patients who had undergone surgery for Hemimegalencephaly (HME) (n = 1), Tuberous sclerosis complex (TSC) (n = 2), Focal cortical dysplasia (FCD) (n = 4), and Rasmussen encephalitis (RE) (n = 3). Peripheral blood mononuclear cells (PBMCs) were also isolated from blood collected at the time of the surgery. Cells were immunostained with a panel of 20 antibody markers, and analyzed by mass cytometry. To identify and quantify the immune cell types in the samples, an unbiased clustering method was applied to the entire data set. More than 85 percent of the CD45+ cells isolated from resected RE brain tissue comprised T cells; by contrast NK cells and myeloid cells constituted 80–95 percent of the CD45+ cells isolated from the TSC and the FCD brain specimens. Three populations of myeloid cells made up >50 percent of all of the myeloid cells in all of the samples of which a population of HLA-DR+ CD11b+ CD4− cells comprised the vast majority of myeloid cells in the BIL fractions from the FCD and TSC cases. CD45RA+ HLA-DR− CD11b+ CD16+ NK cells constituted the major population of NK cells in the blood from all of the cases. This subset also comprised the majority of NK cells in BILs from the resected RE and HME brain tissue, whereas NK cells defined as CD45RA− HLA-DR+ CD11b− CD16− cells comprised 86–96 percent of the NK cells isolated from the FCD and TSC brain tissue. Thirteen different subsets of CD4 and CD8 αβ T cells and γδ T cells accounted for over 80% of the CD3+ T cells in all of the BIL and PBMC samples. At least 90 percent of the T cells in the RE BILs, 80 percent of the T cells in the HME BILs and 40–66 percent in the TSC and FCD BILs comprised activated antigen-experienced (CD45RO+ HLA-DR+ CD69+) T cells. We conclude that even in cases where there is no evidence for an infection or an immune disorder, activated peripheral immune cells may be present in epileptogenic areas of the brain, possibly in response to seizure-driven brain inflammation.
Highlights
It has been estimated that by 15 years of age, approximately one percent of children will have experienced at least one seizure [1]
In the present study we report on the characterization, by mass cytometry, of brain-infiltrating lymphocytes (BILs) isolated from surgical resections of epileptogenic tissue to treat Focal Cortical Dysplasia (FCD), Tuberous Sclerosis Complex (TSC), and HME, as well as Rasmussen encephalitis (RE), and of peripheral blood mononuclear cells (PBMCs) prepared from blood collected at the time of surgery from the same cases
PBMCs and BILs isolated from 10 pediatric epilepsy cases were analyzed by cytometry time of flight (CyTOF) using a panel of antibodies designed to identify populations of adaptive lymphoid cells, innate lymphoid cells, and myeloid cells
Summary
It has been estimated that by 15 years of age, approximately one percent of children will have experienced at least one seizure [1]. For children with drug-resistant epilepsy, surgery may be the only option to obtain seizure freedom, but will result in neurological deficits if the zone of resection involves eloquent cerebral cortex. Many of the children who are candidates for epilepsy surgery suffer from rare neurological disorders including Rasmussen encephalitis (RE), Tuberous Sclerosis Complex (TSC), Focal Cortical Dysplasia (FCD), and Hemimegalencephaly (HME). RE patients present with partial (focal) seizures; magnetic resonance images (MRI) may indicate inflammation and atrophy in the affected cerebral hemisphere [3]. Histopathological examination of resected brain tissue and brain biopsies show T cells in perivascular spaces, leptomeninges, and in small clusters scattered throughout the affected gray and white matter [5, 6]. Focused T cells have been found in resected RE brain tissue strongly implicating an antigen driven immune response in disease etiology [7,8,9,10]
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