Thus, absence of ICAM-1 and ICAM-2 reduces the ability of MOGaa35−55-pulsed DCs in establishing long-lasting interactions with naïve 2D2 T cells and enhanced their migration speed in the LNs. (2001) 108:557–65. Although our experimental set up does not allow to identify those Th17 cells that were adoptively transferred for inducing EAE, based on our observations it is plausible to conclude that in ICAM-1/-2−/− C57BL/6J mice a lower number of transferred Th17 cells were able to cross the BBB, resulting in a significantly lower number of CD4+ IL-17+ T cells detected in the CNS of ICAM-1/-2−/− vs. WT C57BL/6J mice suffering from EAE. The cell specific involvement of ICAM-1 and ICAM-2 in EAE pathogenesis observed in the present study supports the notion that further studies may allow for therapeutic targeting adhesion molecules to block CNS entry of pathogenic T cells while leaving CNS immune surveillance intact. Interestingly, lack of ICAM-1 and ICAM-2 rather affects T-cell trafficking into the brain than into the spinal cord, thus differentially affecting atypical and typical signs of EAE. Klicke hier, um einen neuen Artikel im DocCheck Flexikon anzulegen. Tietz SM, Zwahlen M, Haghayegh Jahromi N, Baden P, Lazarevic I, Enzmann G, et al. Grifka-Walk HM, Giles DA, Segal BM. Different concentrations of MOGaa35−55 peptide were added to the co-cultures and incubated for 72 h before pulsing with [3H]-thymidine. The exact ratio between Th1 and Th17 cells was checked by flow cytometry before and after each experiment and the mean between the two measurements was used for quantitative analysis of the videos when correction for the mixing factor was necessary. Cell Rep. (2018) 22:849–59. Primary mouse brain microvascular endothelial cells (pMBMECs) were isolated from 7 to 9 weeks old WT or ICAM-1/-2−/− C57BL/6J mice and cultured exactly as described before (16, 33). J Immunol. Spleens and pLNs were homogenized and filtered through a sterile 100 μm nylon mesh to prepare a single cell suspension. Statistical analysis was performed using GraphPad Prism 6.0 software (Graphpad software, La Jolla, CA, USA). Epifluorescence-IVM was performed as described before (34, 36). (A–C) TCs from 2D2 C57BL/6J mice co-cultured with irradiated DCs isolated from WT, ICAM-1−/− (A), ICAM-2−/− (B), or ICAM-1/-2−/− (C) C57BL/6J mice. Figure 5. Blood vessels were labeled with fluorescently labeled (Alexa Fluor 633/Alexa Fluor 594) rat-anti-mouse endoglin antibody (CD105: 2 μg/mouse). Effector T cell interactions with meningeal vascular structures in nascent autoimmune CNS lesions. Jager A, Dardalhon V, Sobel RA, Bettelli E, Kuchroo VK. The naïve cells were cultured at a concentration of 1 × 105/ml in restimulation medium containing RPMI-1640 supplemented with 10% FBS (Thermo Fisher Scientific), 10 U/ml penicillin-streptomycin, 2 mM L-glutamine, 1% (v/v) non-essential amino acids, 1 mM sodium pyruvate, and 0.05 mM β-mercaptoethanol (Grogg Chemie AG). Immunol., 14 January 2020 Accumulation of Th1 and Th17 cells on WT or ICAM-1/-2−/− pMBMECs on the flow chamber was allowed for 5 min at a low shear (0.1 dyn/cm2), followed by physiological shear (1.5 dyn/cm2) for additional 25 min, for a total recording time of 30 min. (B) Number of adhered Th1 and Th17 cells in inflamed cervical spinal cord microvessels of WT and ICAM-1/-2−/− C57BL/6J mice with EAE counted at 10, 30, and 60 min after infusion of Th1 and Th17 cells. Becher B, Segal BM. PLoS ONE. Eighteen hours after DCs injection, naïve e670-labeled 2D2-GFP CD4+ T cells were injected and allowed to home to the dLNs and interact with WT and ICAM-1/2−/− DCs. We observed no significant difference in disease incidence of ICAM-1/-2−/− and WT C57BL/6J recipients (data not shown). ICAM-1 is a transmembrane protein possessing an amino-terminus extracellular domain, a single transmembrane domain, and a carboxy-terminus cytoplasmic domain. Interestingly, reduced T-cell proliferation induced by ICAM-1/-2−/− DCs was already observed from the second generation of proliferated CD4+ T cells (Supplementary Figure 1B). Domingues HS, Mues M, Lassmann H, Wekerle H, Krishnamoorthy G. Functional and pathogenic differences of Th1 and Th17 cells in experimental autoimmune encephalomyelitis. doi: 10.1006/cimm.2001.1895, 23. For in vitro live cell imaging experiments, naïve T cells were isolated from 8 to 10 weeks old 2D2 Ubi-GFP or 2D2 tdTomato mice and cultured as above. Each data point represents one individual CD4+ Th1 or Th17 cells track. J Immunol. ICAM-1 is the primary entry receptor for Coxsackievirus A21, an oncolytic virus (brand name Cavatak, being developed by Viralytics). We next asked how lack of ICAMs on DCs will influence CD4+ 2D2 T cell activation in vivo. Immunity. doi: 10.1038/nm1715, 45. Eur J Immunol. While we did not observe a difference in disease incidence (data not shown), ICAM-1/-2−/− C57BL/6J mice showed delayed onset (Figure 6I) and significant amelioration of atypical EAE (Figures 6G,H) compared to WT recipients. We thank Claudia Blatti, Therese Périnat, and Yannick Yersin for their expert technical assistance. T cells were then fixed with Cytofix buffer (BD Biosciences) for 15 min at 20°C, washed with PBS, permeabilized with perm/wash buffer (BD Biosciences) for 15 min at 20°C and stained with fluorophore-conjugated mAbs and the respective isotype control mAbs. Side-by-side comparison of the dynamic interactions of 2D2 T cells with WT or ICAM-1/-2−/− DCs using two-photon intravital microscopy (2P-IVM) of popliteal LNs (42, 43), showed that 2D2-GFP CD4+ T cells preferentially interacted with WT rather than ICAM-1/-2−/− DCs (Figures 1A,B; Supplementary Video 1). The results suggested that four key pathways (metabolic pathways, TNF signaling pathway, rRNA processing, and cytoplasm), the key gene ICAM1, three miRNAs (hsa-miR7-5p, hsa-miR-433-3p, and hsa-miR-133b), and six TFs (ZBTB7A, OVOL1, GATA3, TFDP1, SMAD1, and QSOX) might play important roles in PD development. Diese Seite wurde zuletzt am 22. 'Integrated Computer-Aided Manufacturing' is one option -- get in to view more @ The Web's largest and most authoritative acronyms and abbreviations resource. Englisch: intercellular adhesion molecule 1. Kroenke MA, Chensue SW, Segal BM. doi: 10.1038/s41593-019-0497-x, 57. While a role for ICAM-1 in IS formation and subsequent activation of naïve T cells is shown (7–9), the role of ICAM-2 on APCs in activation of autoaggressive T cells has not been addressed. T(H)17 cytokines in autoimmune neuro-inflammation. Fluorescent labeling of 2D2 T cells and 2P-IVM imaging of T-cell interactions with the cervical spinal cord microvessels during EAE was done as described previously (37). As lack of ICAM-1 and ICAM-2 equally affected the migration of Th1 and Th17 cells across the BBB, we next explored the impact of absence of ICAM-1 and ICAM-2 on the development of Th1 vs. Th17-cell mediated typical and atypical EAE. We did not observe any differences in the role of endothelial ICAM-1 and ICAM-2 in the post-arrest behavior of Th1 and Th17 cells suggesting that both T cell subsets rely on similar interactions to polarize, probe, crawl on and transmigrate across the BBB. These two concentrations, 2 and 100 μg/ml MOGaa35−55 peptide, were selected as low and high concentrations of peptide based on our in vitro results of T-cell proliferation in the presence of various concentrations of MOGaa35−55 peptide. Bitte logge Dich ein, um diesen Artikel zu bearbeiten. Madisen L, Zwingman TA, Sunkin SM, Oh SW, Zariwala HA, Gu H, et al. ICAM-2−/− (Icam2tm1Jcgr) mice were described previously (19). Ann Neurol. (D–I) Cell surface expression of CD69 and CD25 within all (D,G), divided (E,H), and undivided (F,I) 2D2 GFP CD4+ T cell-subsets shown as percentage after in vivo interaction with WT or ICAM-1/-2−/− DCs pulsed with 2, 100 μg/ml, or no (BASAL) MOGaa35−55 peptide. Program Cell Migration and Microscopy Imaging Center of the University of Bern. After washing, fluorescently labeled Th1 and Th17 cells were mixed 1:1 (5 × 106 from each cell subset) and were systemically injected via the carotid artery catheter into a surgically prepared mouse with a clinical EAE score of 0.5 (limp tail) to 2 (hind leg paraplegia). doi: 10.1182/blood-2008-05-078154, 11. Beyond its classically described functions as an adhesion and viral entry molecule, ICAM-1 has now been characterized convincingly as possessing a role in signal transduction. Eur J Immunol. Researchers reasoned that, should ICAM-1 signal transduction prove to occur, it would be necessary to identify the mechanism of that signaling, the conditions and environment in which the signaling would occur, and the biological endpoints of any signaling cascades involved. Briefly, LPS-activated DCs from WT and ICAM-1/-2−/− C57BL/6J mice were pulsed with 100 μg/ml of MOGaa35−55 peptide and fluorescently labeled with 2.5 μM CellTracker Red CMTMR (5-(and-6)-(((4-chloromethyl)benzoyl) amino)tetramethylrhodamine) or 20 μM CellTracker Blue CMAC (7-amino-4-chloromethylcoumarin) at 37°C for 20 min, with dyes swapped between experiments. After visualization of the vascular lumen via injection of 1% tetramethyl rhodamineisothiocyanate (TRITC)-conjugated Dextran into the right carotid artery catheter, 5 × 106 2D2 GFP Th1 or Th17 cells were infused via the right carotid artery of the anesthetized mouse with a clinical EAE score of 0.5 (limp tail) to 2 (hind leg paraplegia). J Immunol. Data were compared by Mann-Whitney test, Student's t-test, ANOVA-one way test followed by Bonferroni's post-test or repeated measure ANOVA with Tukey post-test. (A–I) Proliferation of e670-labeled 2D2-GFP CD4+ T cells 48 and 72 h after transfer into recipient mice prior injected with MOGaa35−55-pulsed WT or ICAM-1/-2−/− DCs. Tags: Diva software or CellQuest were used for data acquisition, FlowJo software (Version 10) was used for data analysis. Lack of ICAM-1 and ICAM-2 impairs firm adhesion and crawling of in vitro polarized Th1 and Th17 CD4+ cells with the inflamed BBB in vivo. Finally, diapedesis of both Th1 and Th17 cells across IL-1β-stimulated ICAM-1/-2−/− pMBMECs was significantly reduced when compared to WT pMBMECs (Figure 4C). Fluids Barriers CNS. In experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS), myelin-specific T cells are activated in the periphery and differentiate in T helper (Th) 1 and Th17 effector cells, which cross the blood-brain barrier (BBB) to reach the central nervous system (CNS), where they induce neuroinflammation. In contrast, lack of ICAM-1 and ICAM-2 significantly reduced crawling of Th1 and Th17 cells that still were able to arrest within ICAM-1/-2−/− spinal cord microvessels (Figure 5D, Supplementary Video 6). (2011) 12:560–7. Immune cell entry into the central nervous system: involvement of adhesion molecules and chemokines. Th17 cells induce ectopic lymphoid follicles in central nervous system tissue inflammation.
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