In Vivo Visualization of (Auto)Immune Processes in the Central Nervous System of Rodents

Schläger, Christian; Litke, Tanja; Flügel, Alexander; Odoardi, Francesca

doi:10.1007/7651_2014_150

Cited by 10 publications

(9 citation statements)

References 14 publications

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“…Thus also in the absence of JAM-B the molecular mechanisms, e.g. interaction of T cell α4β1-and αLβ2-integrins with endothelial VCAM-1 and ICAM-1/ ICAM-2 respectively, mediating the initial T-cell migration across leptomeningeal vessels as visualized by intravital microscopy (Bartholomaus et al, 2009;Schlager et al, 2016;Vajkoczy et al, 2001) and subsequently across the BBB in parenchymal microvessels are intact.…”

Section: Discussionmentioning

confidence: 99%

Lack of junctional adhesion molecule (JAM)-B ameliorates experimental autoimmune encephalomyelitis

Tietz

Perinat

Greene

et al. 2018

Brain, Behavior, and Immunity

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In multiple sclerosis (MS) and its animal model experimental autoimmune encephalomyelitis (EAE) autoaggressive CD4 T cells cross the blood-brain barrier (BBB) and cause neuroinflammation. Therapeutic targeting of CD4 T-cell trafficking into the CNS by blocking α4-integrins has proven beneficial for the treatment of MS but comes with associated risks, probably due to blocking CD8 T cell mediated CNS immune surveillance. Our recent observations show that CD8 T cells also rely on α4β1-integrins to cross the BBB. Besides vascular cell adhesion molecule-1 (VCAM-1), we identified junctional adhesion molecule-B (JAM-B) as a novel vascular α4β1-integrin ligand involved in CD8 T-cell migration across the BBB. This prompted us to investigate, if JAM-B also mediates CD4 T-cell migration across the BBB. We first ensured that encephalitogenic T cells can bind to JAM-B in vitro and next compared EAE pathogenesis in JAM-B C57BL/6J mice and their wild-type littermates. Following immunization with MOG peptide, JAM-B mice developed ameliorated EAE compared to their wild-type littermates. At the same time, we isolated higher numbers of CD45 infiltrating immune cells from the CNS of JAM-B C57BL/6J mice suffering from EAE. Immunofluorescence staining revealed that the majority of CD45 inflammatory cells accumulated in the leptomeningeal and perivascular spaces of the CNS behind the BBB but do not gain access to the CNS parenchyma. Trapping of CNS inflammatory cells was not due to increased inflammatory cell proliferation. Neither a loss of BBB integrity or BBB polarity potentially affecting local chemokine gradients nor a lack of focal gelatinase activation required for CNS parenchymal immune cell entry across the glia limitans could be detected in JAM-B mice. Lack of a role for JAM-B in the effector phase of EAE was supported by the observation that we did not detect any role for JAM-B in EAE pathogenesis, when EAE was elicited by in vitro activated MOG specific CD4 effector T cells. On the other hand, we also failed to demonstrate any role of JAM-B in in vivo priming, proliferation or polarization of MOG-specific CD4 T cells in peripheral immune organs. Finally, our study excludes expression of and thus a role for JAM-B on peripheral and CNS infiltrating myeloid cells. Taken together, although endothelial JAM-B is not required for immune cell trafficking across the BBB in EAE, in its absence accumulation of inflammatory cells mainly in CNS leptomeningeal spaces leads to amelioration of EAE.

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Section: Discussionmentioning

confidence: 99%

Lack of junctional adhesion molecule (JAM)-B ameliorates experimental autoimmune encephalomyelitis

Tietz

Perinat

Greene

et al. 2018

Brain, Behavior, and Immunity

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“…From those studies using 2P-IVM to image cellular interactions in the spinal cord (14, 18), only a limited number have focused on employing 2P-IVM to study immune cell infiltration into the spinal cord during neuroinflammation (12, 19, 20). As pointed out above, all of these studies have solely focused on imaging the lower spinal cord based on the rationale that immune cell infiltration in EAE starts at the level of the lumbar spinal cord.…”

Section: Introductionmentioning

confidence: 99%

“…2P-IVM is characterized by low phototoxicity, a reported tissue penetration depth of 100 and 150 µm in the CNS (15, 19) combined with good resolution (23). It thus allows for continuous imaging of slow immune cell movements such as post-arrest crawling (velocity ≈ 12 μm/min) on and diapedesis across the endothelial wall of spinal cord blood vessels (2, 12).…”

Section: Introductionmentioning

confidence: 99%

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A Novel Cervical Spinal Cord Window Preparation Allows for Two-Photon Imaging of T-Cell Interactions with the Cervical Spinal Cord Microvasculature during Experimental Autoimmune Encephalomyelitis

et al. 2017

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T-cell migration across the blood–brain barrier (BBB) is a crucial step in the pathogenesis of experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS). Two-photon intravital microscopy (2P-IVM) has been established as a powerful tool to study cell–cell interactions in inflammatory EAE lesions in living animals. In EAE, central nervous system inflammation is strongly pronounced in the spinal cord, an organ in which 2P-IVM imaging is technically very challenging and has been limited to the lumbar spinal cord. Here, we describe a novel spinal cord window preparation allowing to use 2P-IVM to image immune cell interactions with the cervical spinal cord microvascular endothelium during EAE. We describe differences in the angioarchitecture of the cervical spinal cord versus the lumbar spinal cord, which will entail different hemodynamic parameters in these different vascular beds. Using T cells as an example, we demonstrate the suitability of this novel methodology in imaging the post-arrest multistep T-cell extravasation across the cervical spinal cord microvessels. The novel methodology includes an outlook to the analysis of the cellular pathway of T-cell diapedesis across the BBB by establishing visualization of endothelial junctions in this vascular bed.

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“…A common hurdle is to get optical access to the region of interest, for example, when aiming to image immune cell migration in the CNS, which is shielded by bone, the dura mater, and the leptomeningeal layers [11]. Nevertheless, different surgical preparations have been developed allowing for imaging of the rodent brain and spinal cord, such as cranial windows, thinned skull preparations, and lumbar and thoracic spinal cord windows [12][13][14][15][16][17]. Recently, we have developed a novel window preparation that allows visualization of immune cell migration across microvessels in the cervical spinal cord of mice using 2P-IVM [18].…”

Section: Introductionmentioning

confidence: 99%

VivoFollow 2: Distortion-Free Multiphoton Intravital Imaging

et al. 2020

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Intravital multiphoton microscopy has become one of the central tools used in the investigation of dynamic cellular activity and function in living animals under nearly physiological conditions and is particularly important for studying the dynamic immune system. During intravital imaging in mice, periodic motion of tissue caused by respiration, induces significant shifts of the imaged region. In slow laser scanning imaging modalities, such as multiphoton microscopy, this movement can lead to considerable distortion and discontinuity in three dimensions of the acquired images. Here, we introduce VivoFollow 2, a toolkit that concurrent with image acquisition performs a precise measurement of the respective image distortion, enabling subsequent automated correction of the imaging data. Recovery of one three-dimensional image stack, corresponding to the tomographic tissue sectioning by the optical plane from each single raw image stack, preserves the time continuity within each image stack. Implementation of VivoFollow 2 thus enables a minimization of motion artifacts in tissues exposed to periodic movements and allows for long-term time-lapse imaging and subsequent precise image analysis of the dynamics of cellular and humoral factors in vivo.

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In Vivo Visualization of (Auto)Immune Processes in the Central Nervous System of Rodents

Cited by 10 publications

References 14 publications

Lack of junctional adhesion molecule (JAM)-B ameliorates experimental autoimmune encephalomyelitis

Lack of junctional adhesion molecule (JAM)-B ameliorates experimental autoimmune encephalomyelitis

A Novel Cervical Spinal Cord Window Preparation Allows for Two-Photon Imaging of T-Cell Interactions with the Cervical Spinal Cord Microvasculature during Experimental Autoimmune Encephalomyelitis

VivoFollow 2: Distortion-Free Multiphoton Intravital Imaging

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