Bone marrow derived dendritic cells (BM-derived DCs) culture was described before (28). (2011) 12:255–63. (2004) 200:847–56. 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. ICAM-1 besteht aus 505 Aminosäuren und ist ein Typ-1-Transmembranprotein der Plasmamembran. Bettelli E, Pagany M, Weiner HL, Linington C, Sobel RA, Kuchroo VK. Mouse syngenic in vitro blood-brain barrier model: a new tool to examine inflammatory events in cerebral endothelium. (D–F) Representative clinical course (D), sum of the AUC (E), and disease onset (F) of Th17 cell mediated typical and atypical EAE (triangles) in WT (blue) and ICAM-1/-2−/− (red) mice. JS, FS, UD, and CP provided the methodology. IL-12-polarized Th1 cells produce GM-CSF and induce EAE independent of IL-23. Kandula S, Abraham C. LFA-1 on CD4+ T cells is required for optimal antigen-dependent activation in vivo. 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. Steiner O, Coisne C, Cecchelli R, Boscacci R, Deutsch U, Engelhardt B, et al. Clinical disease course is shown as mean ± SEM of 11 WT and 10 ICAM-1/-2−/− mice. Krishnamoorthy G, Saxena A, Mars LT, Domingues HS, Mentele R, Ben-Nun A, et al. ICAM-1 and soluble ICAM-1 have antagonistic effects on the tight junctions forming the blood-testis barrier, thus playing a major role in spermatogenesis.. (C) Graphs show speed of individual WT and ICAM-1/-2−/− DCs within the PLN and relative frequency of the speeds of the DCs as monitored by 2P-IVM imaging. To prevent fluorescence artifacts on T-cells' behavior, Th1 and Th17 cells were alternatively isolated from 2D2 Ubi-GFP or 2D2 tdTomato in different replicates of the experiment. Imaging was performed in the T cell area identified by the presence of HEVs labeled with Alexa Fluor 633–conjugated MECA-79 (15 μg/mouse). Diese Seite wurde zuletzt am 22. Hereby the T cells engaged in longer interactions (≥20 min) with WT DCs than with ICAM-1/-2−/− DCs (Figure 1D and Supplementary Video 2). Vajkoczy P, Laschinger M, Engelhardt B. Alpha4-integrin-VCAM-1 binding mediates G protein-independent capture of encephalitogenic T cell blasts to CNS white matter microvessels. 50. J Immunol. 5 × 105 APCs with a ratio of 5:1 APC/T cell and 1 × 104 DCs with a ratio of 1:10 DC/T cell were seeded per well in restimulation medium before MOGaa35−55 peptide was added. Results are shown as mean ± SEM and are pooled from 4 to 6 experiments comparing a total of 26 WT mice and 28 ICAM-1/-2−/− mice. Zhu J, Paul WE. Absence of ICAM-1 and ICAM-2 ameliorates Th1 and Th17 mediated EAE and reduces brain infiltration of cytokine-producing T cells. tdTomato mice were generated by breeding Ai14 mice (23) with ZP3-Cre mice (24) [Gt(ROSA)26Sortm14(CAG−tdTomato)Hze; Tg(Zp3-cre)93Knw] (MGI ID: 3809524; 2176187). 'Integrated Computer-Aided Manufacturing' is one option -- get in to view more @ The Web's largest and most authoritative acronyms and abbreviations resource. doi: 10.1002/eji.201040489, 63. Received: 04 November 2019; Accepted: 16 December 2019; Published: 14 January 2020. (2007) 179:8470–9. Ge AZ, Butcher EC. To this end, we pulsed LPS-matured ICAM-1/-2−/− or WT DCs with low (2 μg/ml) or high (100 μg/ml) concentrations of MOGaa35−55 peptide or no peptide, and allowed them after subcutaneous (s.c.) injection into WT C57BL/6J mice to reach the dLNs. Lack of endothelial ICAM-1 and ICAM-2 significantly altered Th1 and Th17 cells post-arrest behavior, again in a comparable manner. Time shown in minutes and seconds. Figure 4. doi: 10.1073/pnas.101130498, 56. (B) Arrow points to a 2D2 GFP CD4+ T cell positioned between one WT DC (blue) and one ICAM-1/-2−/− DC (red) after migration across the HEV and subsequently interacting preferentially with the WT DC. The interaction between the integrin lymphocyte function associated antigen-1 (LFA-1) on the T cells and its ligand intercellular adhesion molecule-1 (ICAM-1) on the APCs is suggested to be involved in modulating the IS (7). The vast majority of Th1 and Th17 cells able to arrest on IL-1β stimulated ICAM-1/-2−/− pMBMECs were unable to polarize and resist shear forces (Supplementary Video 4), resulting in their detachment (Figure 4C). 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). Immunity. Cloning and expression of a cDNA encoding mouse endoglin, an endothelial cell TGF-beta ligand. T-cell priming by dendritic cells in lymph nodes occurs in three distinct phases. Absence of ICAM-1 and ICAM-2 affects the interaction of in vitro polarized Th1 and Th17 CD4+ cells with an in vitro model of the BBB. Genesis. doi: 10.1002/(SICI)1526-968X(200002)26:2<110::AID-GENE2>3.0.CO;2-8, 25. Epifluorescence-IVM was performed as described before (34, 36). (1998) 102:2096–105. LM wrote and edited the second draft of the manuscript. Briefly, BM cell suspensions from WT, ICAM-1−/−, ICAM-2−/−, and ICAM-1/-2−/− C57BL/6J mice were obtained by centrifugation (4,000 rpm, 4 min) of femurs and tibiae. Furthermore, adoptive transfer of Th1 cells into ICAM-1/-2−/− C57BL/6J mice results in ameliorated EAE compared to WT recipients. Here we induced Th1 and Th17 cell-mediated tEAE in immune competent WT or ICAM-1/-2−/− C57BL/6J mice and observed the development of typical signs of EAE upon transfer of Th1 cells, while transfer of Th17 cells caused both typical and atypical signs of EAE. In the spinal cord lack of ICAM-1 and ICAM-2 moderately reduced the number of CD4+ T cells in mice with typical but not with atypical EAE (Figure 6K). J Immunol. We thank Claudia Blatti, Therese Périnat, and Yannick Yersin for their expert technical assistance. (2008) 274:23–6. Imaging the single cell dynamics of CD4+ T cell activation by dendritic cells in lymph nodes. I have found it very effective for many incident investigations and was a significant improvement on older methodologies that we all used some 20 years ago. (2016) 46:2481–3. Eur J Immunol. Researchers began to question the role of ICAM-1 as a simple adhesion molecule upon discovering that ICAM-1 serves as the binding site for entry of the major group of human rhinovirus (HRV) into various cell types. 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. Remarkably, we found significantly lower numbers and percentage of CD4+ IL-17+ cells in the brain of ICAM-1/-2−/− vs. WT C57BL/6J mice irrespective of typical or atypical signs of (Figures 6N,O), underscoring that absence of ICAM-1 and ICAM-2 preferentially affects brain infiltration by Th17 cells. We previously reported that endothelial ICAM-1 and ICAM-2 are essential for Th1 cell polarization and crawling on the BBB in vitro (14, 16). Arthritis Res. (B,C) Post-arrest dynamic behavior during 30 min of recording time of in vitro polarized CD4+ Th1 and Th17 cells on TNFα (B) or IL-1β (C) stimulated pMBMECs. Data are pooled from 2 to 3 experiments (26 WT mice and 22 in ICAM-1/-2−/− mice). (2014) 122:173–89. We next asked how lack of ICAMs on DCs will influence CD4+ 2D2 T cell activation in vivo. Each dot represents one mouse. doi: 10.1084/jem.20110434, 18. Copyright ©2020 DocCheck Medical Services GmbH |, ICAM-1 molecular mechanism and genome wide SNP's association studies. The editor and reviewers' affiliations are the latest provided on their Loop research profiles and may not reflect their situation at the time of review. Each dot in the plots represents pLN T cells pooled from one mouse. To further characterize the CNS infiltrating CD4+ T cells, we measured in a non-exclusive manner the expression of Th1 and Th17 signature cytokines IFN-γ and IL-17, respectively, in addition to GM-CSF, which has been proposed as a pathogenic mediator in MS and EAE (46–49). Glatigny S, Duhen R, Oukka M, Bettelli E. Cutting edge: loss of alpha4 integrin expression differentially affects the homing of Th1 and Th17 cells. into WT and ICAM-1/-2−/− recipient C57BL/6J mice. To investigate how lack of ICAM-1 and ICAM-2 affect T-cell infiltration into the brain vs. the spinal cord, we isolated CD45+ infiltrating cells from the brains and spinal cords of mice suffering from atypical or typical Th17 cell-mediated EAE.