Deciphering the heterogeneity of the Lyve1+ perivascular macrophages in the mouse brain

Perivascular macrophages (pvMs) are associated with cerebral vasculature and mediate brain drainage and immune regulation. Here, using reporter mouse models, whole brain and section immunofluorescence, flow cytometry, and single cell RNA sequencing, besides the Lyve1+F4/80+CD206+CX3CR1+ pvMs, we identify a CX3CR1– pvM population that shares phagocytic functions and location. Furthermore, the brain parenchyma vasculature mostly hosts Lyve1+MHCII– pvMs with low to intermediate CD45 expression. Using the double Cx3cr1GFP x Cx3cr1-Cre;RosatdT reporter mice for finer mapping of the lineages, we establish that CD45lowCX3CR1– pvMs are derived from CX3CR1+ precursors and require PU.1 during their ontogeny. In parallel, results from the Cxcr4-CreErt2;Rosa26tdT lineage tracing model support a bone marrow-independent replenishment of all Lyve1+ pvMs in the adult mouse brain. Lastly, flow cytometry and 3D immunofluorescence analysis uncover increased percentage of pvMs following photothrombotic induced stroke. Our results thus show that the parenchymal pvM population is more heterogenous than previously described, and includes a CD45low and CX3CR1– pvM population

The unique immune ecosystems in pediatric brain tumors: integrating single-cell and bulk RNA-sequencing

BackgroundThe significant progress of immune therapy in non-central nervous system tumors has sparked interest in employing the same strategy for adult brain tumors. However, the advancement of immunotherapy in pediatric central nervous system (CNS) tumors is not yet on par. Currently, there is a lack of comprehensive comparative studies investigating the immune ecosystem in pediatric and adult CNS tumors at a high-resolution single-cell level.MethodsIn this study, we comprehensively analyzed over 0.3 million cells from 171 samples, encompassing adult gliomas (IDH wild type and IDH mutation) as well as four major types of pediatric brain tumors (medulloblastoma (MB), ependymoma (EPN), H3K27M-mutation (DIPG), and pediatric IDH-mutation glioma (P-IDH-M)). Our approach involved integrating publicly available and newly generated single-cell datasets. We compared the immune landscapes in different brain tumors, as well as the detailed functional phenotypes of T-cell and myeloid subpopulations. Through single-cell analysis, we identified gene sets associated with major cell types in the tumor microenvironment (gene features from single-cell data, scFes) and compared them with existing gene sets such as GSEA and xCell. The CBTTC and external GEO cohort was used to analyze and validate the immune-stromal-tumor patterns in pediatric brain tumors which might potentially respond to the immunotherapy.ResultsFrom the perspective of single-cell analysis, it was observed that major pediatric brain tumors (MB, EPN, P-IDH-M, DIPG) exhibited lower immune contents compared with adult gliomas. Additionally, these pediatric brain tumors displayed diverse immunophenotypes, particularly in regard to myeloid cells. Notably, the presence of HLA-enriched myeloid cells in MB was found to be independently associated with prognosis. Moreover, the scFes, when compared with commonly used gene features, demonstrated superior performance in independent single-cell datasets across various tumor types. Furthermore, our study revealed the existence of heterogeneous immune ecosystems at the bulk-RNA sequencing level among different brain tumor types. In addition, we identified several immune-stromal-tumor patterns that could potentially exhibit significant responses to conventional immune checkpoint inhibitors.ConclusionThe single-cell technique provides a rational path to deeply understand the unique immune ecosystem of pediatric brain tumors. In spite of the traditional attitudes of “cold” tumor towards pediatric brain tumor, the immune-stroma-tumor patterns identified in this study suggest the feasibility of immune checkpoint inhibitors and pave the way for the upcoming tide of immunotherapy in pediatric brain tumors

Early-start antiplatelet therapy after operation in patients with spontaneous intracerebral hemorrhage and high risk of ischemic events (E-start):Protocol for a multi-centered, prospective, open-label, blinded endpoint randomized controlled trial

BACKGROUND: For severe spontaneous intracerebral hemorrhage (sSICH) patients with high risk of ischemic events, the incidence of postoperative major cardiovascular/cerebrovascular and peripheral vascular events (MACCPE) is notable. Although antiplatelet therapy is a potential way to benefit these patients, the severe hemorrhagic complications, e.g., intracranial re-hemorrhage, is a barrier for early starting antiplatelet therapy. OBJECTIVES: This randomized controlled trial aims to identify the benefit and safety of early starting antiplatelet therapy after operation for sSICH patients with high risk of ischemic events. METHODS: This study is a multicenter, prospective, randomized, open-label, blinded-endpoint trial. We will enroll 250 sSICH patients with a high risk of ischemic events (including cerebral infarcts, transient ischemic attack, myocardial infarction, pulmonary embolism, and deep venous thrombosis). The participants will be randomized in a 1:1 manner to early-start group (start antiplatelet therapy at 3 days after operation) and normal-start group (start antiplatelet therapy at 30 days after operation). The early-start group will receive aspirin 100 mg daily. The control group will not receive antithrombotic therapy until 30 days after operation. The efficacy endpoint is the incidence of MACCPE, and the safety endpoint is the incidence of intracranial re-hemorrhage. DISCUSSION: The Early-Start antiplatelet therapy after operation in patients with spontaneous intracerebral hemorrhage trial (E-start) is the first randomized trial about early start antiplatelet therapy for operated sSICH patients with a high risk of ischemic events. This study will provide a new strategy and evidence for postoperative management in the future. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov, identifier NCT04820972; Available at: https://clinicaltrials.gov/ct2/show/NCT04820972?term=NCT04820972&draw=2&rank=1. Chinese Clinical Trial Registry, identifier ChiCTR2100044560; Available at: http://www.chictr.org.cn/showproj.aspx?proj=123277

Hydrogen Sulfide Mitigates Kidney Injury in High Fat Diet-Induced Obese Mice

Obesity is prevalent worldwide and is a major risk factor for the development and progression of kidney disease. Hydrogen sulfide (H2S) plays an important role in renal physiological and pathophysiological processes. However, whether H2S is able to mitigate kidney injury induced by obesity in mice remains unclear. In this study, we demonstrated that H2S significantly reduced the accumulation of lipids in the kidneys of high fat diet- (HFD-) induced obese mice. The results of hematoxylin and eosin, periodic acid-Schiff, and Masson’s trichrome staining showed that H2S ameliorated the kidney structure, decreased the extent of interstitial injury, and reduced the degree of kidney fibrosis in HFD-induced obese mice. We found that H2S decreased the expression levels of tumor necrosis factor-α, interleukin- (IL-) 6, and monocyte chemoattractant protein-1 but increased the expression level of IL-10. Furthermore, H2S treatment decreased the protein expression of p50, p65, and p-p65 in the kidney of HFD-induced obese mice. In conclusion, H2S is able to mitigate renal injury in HFD-induced obese mice through the reduction of kidney inflammation by downregulating the expression of nuclear factor-kappa B. H2S or its releasing compounds may serve as a potential therapeutic molecule for obesity-induced kidney injury

Vertical distribution of suspended particulate matter and its response to river discharge and seawater intrusion: a case study in the Pearl River Estuary during the 2020 dry season

The vertical distribution of suspended particulate matter (SPM) in the Pearl River Estuary (PRE) during winter has not been widely reported. The aim of this paper is to describe the high-resolution vertical distribution of SPM along the transect based on the in-situ observations (including SPM, attenuation coefficient, and particle backscattering coefficient) from three transects of the winter cruise in the northern South China Sea in 2020. The empirical relationship between SPM and bio-optical parameters with correlation coefficients greater than 0.7 is also established and combined with model data to further discuss the mechanism of river discharge and seawater intrusion effects on the vertical distribution of SPM. In the horizontal distribution, the mass concentration of SPM was high in the nearshore region and was low in the offshore region. In the vertical direction, the mass concentration of SPM in the offshore region was more homogeneous, while the mass concentration of SPM in the nearshore region varied greatly, showing a pattern of high bottom and middle layer or high bottom and surface layer. The difference in the vertical distribution of SPM in the nearshore area is the combined effect of river discharge and seawater intrusion on the resuspension of sediment and the inhibition of the spread of high SPM

Dephosphorylated Polymerase I and Transcript Release Factor Prevents Allergic Asthma Exacerbations by Limiting IL-33 Release

BackgroundAsthma is a chronic inflammatory disease characterized by airway inflammation and airway hyperresponsiveness (AHR). IL-33 is considered as one of the most critical molecules in asthma pathogenesis. IL-33 is stored in nucleus and passively released during necrosis. But little is known about whether living cells can release IL-33 and how this process is regulated.ObjectiveWe sought to investigate the role of polymerase I and transcript release factor (PTRF) in IL-33 release and asthma pathogenesis.MethodsOvalbumin (OVA)-induced asthma model in PTRF+/− mice were employed to dissect the role of PTRF in vivo. Then, further in vitro experiments were carried out to unwind the potential mechanism involved.ResultsIn OVA asthma model with challenge phase, PTRF+/− mice showed a greater airway hyper-reaction, with an intense airway inflammation and more eosinophils in bronchoalveolar lavage fluid (BALF). Consistently, more acute type 2 immune response in lung and a higher IL-33 level in BALF were found in PTRF+/− mice. In OVA asthma model without challenge phase, airway inflammation and local type 2 immune responses were comparable between control mice and PTRF+/− mice. Knockdown of PTRF in 16HBE led to a significantly increased level of IL-33 in cell culture supernatants in response to LPS or HDM. Immunoprecipitation assay clarified Y158 as the major phosphorylation site of PTRF, which was also critical for the interaction of IL-33 and PTRF. Overexpression of dephosphorylated mutant Y158F of PTRF sequestered IL-33 in nucleus together with PTRF and limited IL-33 extracellular secretion.ConclusionPartial loss of PTRF led to a greater AHR and potent type 2 immune responses during challenge phase of asthma model, without influencing the sensitization phase. PTRF phosphorylation status determined subcellular location of PTRF and, therefore, regulated IL-33 release

Présence de cellules lymphoïdes innées dans le cerveau après un AVC ischémique

L'AVC ischémique déclenche l'activation de la microglie cérébrale et le recrutement continu de leucocytes, y compris les neutrophiles, les monocytes, les cellules dendritiques (DC) et les lymphocytes. Les cellules lymphoïdes innées (ILC) sont les contreparties innées des cellules T, exprimant des facteurs de transcription et des cytokines, qui sont essentiels pour le maintien de l'homéostasie et du remodelage tissulaire ainsi que pour la réparation tissulaire dans certaines circonstances. Cependant, leur rôle dans les attaques cérébrales ischémiques reste inconnue. Dans la thèse, nous avons induit un modèle d'accident vasculaire cérébral photothrombotique en activant par la lumière le Rose Bengal injecté de et nous avons découvert que les cellules NK, les ILC1, les ILC2 et les ILC3 étaient recrutées dans la lésion d'AVC comme observé par analyse d'imagerie confocale et 3D. Nous avons par la suite observé que les membres de la famille ILC s’accumulent progressivement dans le cerveau, la majorité de ces cellules sont des cellules NK et des ILC1. En outre, l'augmentation des cellules NK dans le cerveau atteint est causée par un afflux mais pas par une prolifération in situ.Grâce à l’utilisation du modèle murin Cdh5CreERT2 ; Cxcl12 fl et Ncr1iCre ; Cxcr4fl, nous avons observé que l’axe CXCL12-CXCR4 induit la migration des cellules NK dans le cerveau atteint d'accident vasculaire cérébral ischémique. Pour étudier les déficits neurologiques de ces souris atteintes d’AVC et le rôle des cellules NK, des expériences de “beam walk” ont été mises en place. Les souris NK déficientes apparaissent avoir le plus de déficits neurologiques.Ischemic stroke triggers the activation of brain-resident microglia and continuous recruitment of leukocytes, including neutrophils, monocytes, dendritic cells (DC) and lymphocytes. Innate lymphoid cells (ILCs) are the innate counterparts of T cells by expressing transcription factors and cytokines, which are critical for the maintenance of homeostasis and tissue remodeling as well as tissue repair under certain circumstances. However, their involvement in ischemic stroke brain insult is unknown.In the thesis, we induced photothrombotic stroke model by light-activating injected Rose Bengal and found that NK cells, ILC1s, ILC2s and ILC3s were recruited to the stroke lesion as observed by confocal and 3D imaging analysis. We subsequently observed that ILC family members progressively accumulated in the stroke brain, of which the majority were NK cells and ILC1s. Furthermore, the increase of NK cells in the stroke brain was caused by influx but not in situ proliferation.Using Cdh5CreERT2 ; Cxcl12fl and Ncr1iCre ; Cxcr4fl mouse models, we observed that CXCL12-CXCR4 axis mediated the migration of NK cells into the ischemic stroke brain. We further performed beam-walk to study the role of NK cells in the ischemic stroke brain and observed that the NK-deficient mice had more neurological deficit

Innate Lymphoid Cells in the Central Nervous System

International audienceImmune cells are present within the central nervous system and play important roles in neurological inflammation and disease. As relatively new described immune cell population, Innate Lymphoid Cells are now increasingly recognized within the central nervous system and associated diseases. Innate Lymphoid Cells are generally regarded as tissue resident and early responders, while conversely within the central nervous system at steady-state their presence is limited. This review describes the current understandings on Innate Lymphoid Cells in the central nervous system at steady-state and its borders plus their involvement in major neurological diseases like ischemic stroke, Alzheimer’s disease and Multiple Sclerosis