Name: an ex vivo model of an oligodendrocyte-directed t-cell attack in acute brain slices
Uploaded: 2015-02-05T13:00:00
Description: Watch this Scientific Journal Video about An Ex vivo Model of an Oligodendrocyte-directed T-Cell Attack in Acute Brain Slices at JoVE.com

This work was supported by the Interdisciplinary Center for Clinical Research (IZKF) M&#252;nster (SEED 03/12, SB), Deutsche Forschungsgemeinschaft (SFB TR128, TP B6 to S.G.M. ME3283/2-1 to S.G.M.) and by Innovative Medizinische Forschung, M&#252;nster (I-BI111316, SB and SGM).

Alle forsøg med mus bør udføres i overensstemmelse med retningslinjerne i den respektive institutionelle pasning af dyr og brug udvalg. 1. Generelle kommentarer Mouse Eksperimenter <ol><li> Hold musene under patogenfrie forhold og give dem adgang til foder og vand ad libitum. BEMÆRK: Det er vigtigt at bruge alders- og sex-matchede mus i eksperimentelle grupper, fordi immunologiske mønstre kan variere med alder og køn. </li></ol> 2. Fremstilling og aktiver...

<ol>
	<li>Moll, N. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cortical+demyelination+in+PML+and+MS:+Similarities+and+differences.">Cortical demyelination in PML and MS: Similarities and differences.</a> Neurology. 70 (5), 336-343 (2008).</li><li>Peterson, J. W., Bo, L., Mork, S., Chang, A., Trapp, B. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transected+neurites,+apoptotic+neurons,+and+reduced+inflammation+in+cortical+multiple+sclerosis+lesions.">Transected neurites, apoptotic neurons, and reduced inflammation in cortical multiple sclerosis lesions.</a> Ann Neurol. 50 (3), 389-400 (2001).</li><li>Bo, L., Vedeler, C. A., Nyland, H. I., Trapp, B. D., Mork, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Subpial+demyelination+in+the+cerebral+cortex+of+multiple+sclerosis+patients.">Subpial demyelination in the cerebral cortex of multiple sclerosis patients.</a> J Neuropathol Exp Neurol. 62 (7), 723-732 (2003).</li><li>Babbe, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clonal+expansions+of+CD8(+)+T+cells+dominate+the+T+cell+infiltrate+in+active+multiple+sclerosis+lesions+as+shown+by+micromanipulation+and+single+cell+polymerase+chain+reaction.">Clonal expansions of CD8(+) T cells dominate the T cell infiltrate in active multiple sclerosis lesions as shown by micromanipulation and single cell polymerase chain reaction.</a> J Exp Med. 192 (3), 393-404 (2000).</li><li>Junker, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiple+sclerosis:+T-cell+receptor+expression+in+distinct+brain+regions.">Multiple sclerosis: T-cell receptor expression in distinct brain regions.</a> Brain. 130 (11), 2789-2799 (2007).</li><li>Skulina, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiple+sclerosis:+brain-infiltrating+CD8++T+cells+persist+as+clonal+expansions+in+the+cerebrospinal+fluid+and+blood.">Multiple sclerosis: brain-infiltrating CD8+ T cells persist as clonal expansions in the cerebrospinal fluid and blood.</a> Proc Natl Acad Sci U S A. 101 (8), 2428-2433 (2004).</li><li>Friese, M. A., Fugger, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Autoreactive+CD8++T+cells+in+multiple+sclerosis:+a+new+target+for+therapy.">Autoreactive CD8+ T cells in multiple sclerosis: a new target for therapy.</a> Brain. 128 (8), 1747-1763 (2005).</li><li>Melzer, N., Meuth, S. G., Wiendl, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CD8++T+cells+and+neuronal+damage:+direct+and+collateral+mechanisms+of+cytotoxicity+and+impaired+electrical+excitability).">CD8+ T cells and neuronal damage: direct and collateral mechanisms of cytotoxicity and impaired electrical excitability).</a> FASEB J. 23 (11), 3659-3673 (2009).</li><li>Booss, J., Esiri, M. M., Tourtellotte, W. W., Mason, D. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunohistological+analysis+of+T+lymphocyte+subsets+in+the+central+nervous+system+in+chronic+progressive+multiple+sclerosis.">Immunohistological analysis of T lymphocyte subsets in the central nervous system in chronic progressive multiple sclerosis.</a> J Neurol Sci. 62 (1-3), 219-232 (1983).</li><li>Hauser, S. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunohistochemical+analysis+of+the+cellular+infiltrate+in+multiple+sclerosis+lesions.">Immunohistochemical analysis of the cellular infiltrate in multiple sclerosis lesions.</a> Ann Neurol. 19 (6), 578-587 (1986).</li><li>Hoftberger, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression+of+major+histocompatibility+complex+class+I+molecules+on+the+different+cell+types+in+multiple+sclerosis+lesions.">Expression of major histocompatibility complex class I molecules on the different cell types in multiple sclerosis lesions.</a> Brain Pathol. 14 (1), 43-50 (2004).</li><li>G&#246;bel, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Collateral+neuronal+apoptosis+in+CNS+gray+matter+during+an+oligodendrocyte-directed+CD8(+)+T+cell+attack.">Collateral neuronal apoptosis in CNS gray matter during an oligodendrocyte-directed CD8(+) T cell attack.</a> Glia. 58, 469-480 (2010).</li><li>Melzer, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Excitotoxic+neuronal+cell+death+during+an+oligodendrocyte-directed+CD8++T+cell+attack+in+the+CNS+gray+matter.">Excitotoxic neuronal cell death during an oligodendrocyte-directed CD8+ T cell attack in the CNS gray matter.</a> J Neuroinflammation. 10, 121 (2013).</li><li>Huseby, E. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+pathogenic+role+for+myelin-specific+CD8(+)+T+cells+in+a+model+for+multiple+sclerosis.">A pathogenic role for myelin-specific CD8(+) T cells in a model for multiple sclerosis.</a> J Exp Med. 194 (5), 669-676 (2001).</li><li>McPherson, S. W., Heuss, N. D., Roehrich, H., Gregerson, D. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bystander+killing+of+neurons+by+cytotoxic+T+cells+specific+for+a+glial+antigen.">Bystander killing of neurons by cytotoxic T cells specific for a glial antigen.</a> Glia. 53 (5), 457-466 (2006).</li><li>Hogquist, K. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=T+cell+receptor+antagonist+peptides+induce+positive+selection.">T cell receptor antagonist peptides induce positive selection.</a> Cell. 76 (1), 17-27 (1994).</li><li>Cao, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+experimental+autoimmune+encephalomyelitis+in+transgenic+mice+expressing+ovalbumin+in+oligodendrocytes.">Induction of experimental autoimmune encephalomyelitis in transgenic mice expressing ovalbumin in oligodendrocytes.</a> Eur J Immunol. 36 (1), 207-215 (2006).</li><li>G&#246;bel, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CD4(+)+CD25(+)+FoxP3(+)+regulatory+T+cells+suppress+cytotoxicity+of+CD8(+)+effector+T+cells:+implications+for+their+capacity+to+limit+inflammatory+central+nervous+system+damage+at+the+parenchymal+level.">CD4(+) CD25(+) FoxP3(+) regulatory T cells suppress cytotoxicity of CD8(+) effector T cells: implications for their capacity to limit inflammatory central nervous system damage at the parenchymal level.</a> J Neuroinflammation. 9, 41 (2012).</li><li>Edwards, F. A., Konnerth, A., Sakmann, B., Takahashi, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+thin+slice+preparation+for+patch+clamp+recordings+from+neurones+of+the+mammalian+central+nervous+system.">A thin slice preparation for patch clamp recordings from neurones of the mammalian central nervous system.</a> Pflugers Arch. 414 (5), 600-612 (1989).</li><li>Meuth, S. G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contribution+of+TWIK-related+acid-sensitive+K++channel+1+(TASK1)+and+TASK3+channels+to+the+control+of+activity+modes+in+thalamocortical+neurons.">Contribution of TWIK-related acid-sensitive K+ channel 1 (TASK1) and TASK3 channels to the control of activity modes in thalamocortical neurons.</a> J Neurosci. 23 (16), 6460-6469 (2003).</li><li>Misgeld, U., Frotscher, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dependence+of+the+viability+of+neurons+in+hippocampal+slices+on+oxygen+supply.">Dependence of the viability of neurons in hippocampal slices on oxygen supply.</a> Brain Res Bull. 8 (1), 95-100 (1982).</li><li>Ling, C., Verbny, Y. I., Banks, M. I., Sandor, M., Fabry, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+situ+activation+of+antigen-specific+CD8++T+cells+in+the+presence+of+antigen+in+organotypic+brain+slices.">In situ activation of antigen-specific CD8+ T cells in the presence of antigen in organotypic brain slices.</a> J Immunol. 180, 8393-8399 (2008).</li></ol>

Forskellige dyremodeller er blevet beskrevet i de seneste årtier for at løse de patologiske træk ved inflammatoriske demyeliniseringssygdomme som MS. In vivo mus og rotte-modeller er almindeligt anvendt til at efterligne patofysiologiske funktioner i sygdommen, nemlig analyse af konsekvenserne af demyeliniserende og remyelinering processer og blandet episoder af inflammation og neurodegeneration. Alligevel kun en ex vivo fremgangsmåde gør det muligt at dissekere de nøjagtige underliggende mekanis...

Death of oligodendrocytes and destruction of the myelin sheath accompanied by loss of neurons and axons are typical pathological findings in grey matter lesions in individuals suffering from multiple sclerosis (MS)1,2. Cortical lesions can be divided so far in three different subtypes2: subpial, intracortical and leukocortical lesions. In comparison to white matter plaques, infiltrates are characterized by a predominance of CD8+ T-cells, suggesting their possible decisive role in grey matter inflammation3. Furthermore, oligoclonal expansions in blood, cerebrospinal fluid (CSF) and within inflammatory lesions can be found for CD8+ T-cells themselves4-6. 
In line with this, it is assumed that CD8+ T-cells may be specific for different myelin proteins7,8. Indeed, CD8+ T-cells are found near oligodendrocytes and myelin sheaths9,10 that show MHC I expression11 and might therefore be responsible for the loss of the myelin sheath. This process is often seen together with extensive &#8216;&#8216;collateral&#8217;&#8217; neuronal and axonal damage within the central nervous system (CNS) grey matter1,2. In fact, direct and indirect death of oligodendrocytes and neurons is induced by CD8+ T-cells via two different mechanism: (i) cell membrane swelling and rupture due to the formation of cytotoxic granules following the release of perforins and granzymes and (ii) ligation to the Fas receptors or exposition of FasL on their surface8,12,13.
Different types of animal models are established to study the underlying mechanism of the mentioned processes. In this respect, primed CD8+ T-cells specific for autoantigens with induced expression in CNS glial cells, like oligodendrocytes or astrocytes, can be adoptively transferred to analyse &#8216;&#8216;collateral&#8217;&#8217; neuronal and axonal death in grey matter subsequently14,15. To perform such in vivo experiments is a big help to mimic some pathophysiological aspects of MS, however, this approach is not suited to resolve the underlying mechanism and kinetics of axonal damage and neuronal apoptosis.
To overcome these restrictions, an ex vivo approach was established to study the mechanisms and time course of neuronal cell death following a oligondendrocytes-directed CD8+ T-cell attack. Since only oligodendrocytes and therefore myelin sheath production should be targeted by immune cells, MHC class-I-restricted, ovalbumin (OVA)-reactive OT-I Tcells are used16. These cells are subsequently transferred into brain slices obtained from mice selectively expressing OVA in oligodendrocytes (ODC-OVA mice)17.

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			<td height="20" style="height:20px;width:216px;">70 &#181;m strainer</td>
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			<td height="26" style="height:26px;width:216px;">Isofluran</td>
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			<td style="width:119px;">05260-05</td>
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			<td height="26" style="height:26px;width:216px;">KCl</td>
			<td style="width:187px;">Merck</td>
			<td style="width:119px;">1.04933.0500</td>
			<td>potassium chloride</td>
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			<td height="26" style="height:26px;width:216px;">KHCO3</td>
			<td style="width:187px;">Sigma</td>
			<td style="width:119px;">P9144</td>
			<td>potassium hydrogen carbonate</td>
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			<td height="26" style="height:26px;width:216px;">L-Glutamine</td>
			<td style="width:187px;">Gibco</td>
			<td style="width:119px;">35050-038</td>
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			<td height="20" style="height:20px;width:216px;">MgSO4</td>
			<td style="width:187px;">Merck</td>
			<td style="width:119px;">1.05886.0500</td>
			<td>magnesium sulfate</td>
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			<td height="20" style="height:20px;width:216px;">NaCl</td>
			<td style="width:187px;">Sigma</td>
			<td style="width:119px;">31434</td>
			<td>sodium chloride</td>
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			<td style="width:187px;">Merck</td>
			<td style="width:119px;">1.06346.0500</td>
			<td>sodium hydrogen phosphate</td>
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			<td height="20" style="height:20px;width:216px;">NaHCO3</td>
			<td style="width:187px;">Merck</td>
			<td style="width:119px;">1.06329.0500</td>
			<td>sodium hydrogen carbonate</td>
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			<td height="20" style="height:20px;width:216px;">NaOH</td>
			<td style="width:187px;">Merck</td>
			<td style="width:119px;">1.09137.1000</td>
			<td>sodium hydroxide</td>
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			<td height="40" style="height:40px;width:216px;">NH4Cl</td>
			<td style="width:187px;">Sigma</td>
			<td style="width:119px;">213330</td>
			<td>ammonium chloride</td>
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			<td height="20" style="height:20px;width:216px;">Non Essential Amino Acid</td>
			<td style="width:187px;">Gibco</td>
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			<td height="20" style="height:20px;width:216px;">OVA (257-264)</td>
			<td style="width:187px;">Genscript</td>
			<td style="width:119px;">RP10611</td>
			<td>ovalbumin</td>
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			<td height="20" style="height:20px;width:216px;">PIPES</td>
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			<td height="26" style="height:26px;width:216px;">Sukrose</td>
			<td style="width:187px;">Merck</td>
			<td style="width:119px;">1.07687.1000</td>
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			<td height="26" style="height:26px;width:216px;">Tissue-Tek OCT</td>
			<td style="width:187px;">Sakura</td>
			<td style="width:119px;">4583</td>
			<td></td>
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an ex vivo model of an oligodendrocyte-directed t-cell attack in acute brain slices

Death of oligodendrocytes accompanied by destruction of neurons and axons are typical histopathological findings in cortical and subcortical grey matter lesions in inflammatory demyelinating disorders like multiple sclerosis (MS). In these disorders, mainly CD8+ T-cells of putative specificity for myelin- and oligodendrocyte-related antigens are found, so that neuronal apoptosis in grey matter lesions may be a collateral effect of these cells. Different types of animal models are established to study the underlying mechanisms of the mentioned pathophysiological processes. However, although they mimic some aspects of MS, it is impossible to dissect the exact mechanism and time course of &#8216;&#8216;collateral&#8217;&#8217; neuronal cell death. To address this course, here we show a protocol to study the mechanisms and time response of neuronal damage following an oligodendrocyte-directed CD8+ T cell attack. To target only the myelin sheath and the oligodendrocytes, in vitro activated oligodendrocyte-specific CD8+ T-cells are transferred into acutely isolated brain slices. After a defined incubation period, myelin and neuronal damage can be analysed in different regions of interest. Potential applications and limitations of this model will be discussed.

Efter inkubation af hjerneskiver med oligodendrocyt-directed CD8 + T-celler, oligodendrocytter samt neuroner undergå apoptose (figur 2A og 1C, henholdsvis). Histologiske tegn på apoptose (f.eks Caspase-3, Tunel) kan tidligst påvises efter 3 timers inkubation. Inkubationstid, ikke være længere end 8 timer for at sikre en god kvalitet af præparatet og reproducerbare resultater. Apoptotiske celler kan findes over hele skive med overvægt i myelinerede områder. Eksempler histologi...

Watch this Scientific Journal Video about An Ex vivo Model of an Oligodendrocyte-directed T-Cell Attack in Acute Brain Slices at JoVE.com

An Ex vivo Model of an Oligodendrocyte-directed T-Cell Attack in Acute Brain Slices

En<em&gt; Ex vivo</em&gt; Model af en oligodendrocyt rettet T-celle Angrib akut Brain Skiver

To address mechanisms of demyelination and neuronal apoptosis in cortical lesions of inflammatory demyelinating disorders, different animal models are used. We here describe an ex vivo approach by using oligodendrocyte-specific CD8+ T-cells on brain slices, resulting in oligodendroglial and neuronal death. Potential applications and limitations of the model are discussed.

Death of oligodendrocytes accompanied by destruction of neurons and axons are typical histopathological findings in cortical and subcortical grey matter ...

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An Ex vivo Model of an Oligodendrocyte-directed T-Cell Attack in Acute Brain Slices

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Following in Real Time the Impact of Pneumococcal Virulence Factors in an Acute Mouse Pneumonia Model Using Bioluminescent Bacteria

Efter i Real Time konsekvenserne af Pneumokok virulensfaktorer i en akut Mouse Lungebetændelse Model Brug Bioluminescent Bakterier

Pneumonia is one of the major health care problems in developing and industrialized countries and is associated with considerable morbidity and mortality. ...

Isolation and Ex Vivo Culture of V&#948;1+CD4+&#947;&#948; T Cells, an Extrathymic &#945;&#946;T-cell Progenitor

Isolation and Ex Vivo Culture of VÃƒÅ½Ã‚Â´1+CD4+ÃƒÅ½Ã‚Â³ÃƒÅ½Ã‚Â´ T Cells, an Extrathymic ÃƒÅ½Ã‚Â±ÃƒÅ½Ã‚Â²T-cell Progenitor

Isolering og<em&gt; Ex vivo</em&gt; Kultur Vδ1<sup&gt; +</sup&gt; CD4<sup&gt; +</sup&gt; γδ T-celler, en Extrathymic αβT-celleprogenitor

The thymus, the primary organ for the generation of &#945;&#946; T cells and backbone of the adaptive immune system in vertebrates, has long been ...

Establishment of a High-throughput Setup for Screening Small Molecules That Modulate c-di-GMP Signaling in Pseudomonas aeruginosa

Establishment of a High-throughput Setup for Screening Small Molecules That Modulate c-di-GMP Signaling in Pseudomonas aeruginosa

Etablering af en High-throughput Opsætning til screening små molekyler, der modulerer c-di-GMP Signalering i<em&gt; Pseudomonas aeruginosa</em

Bacterial resistance to traditional antibiotics has driven research attempts to identify new drug targets in recently discovered regulatory pathways. ...

An In Vitro Model of the Blood-brain Barrier Using Impedance Spectroscopy: A Focus on T Cell-endothelial Cell Interaction

An In Vitro Model of the Blood-brain Barrier Using Impedance Spectroscopy: A Focus on T Cell-endothelial Cell Interaction

en<em&gt; In vitro</em&gt; Model af blodhjernebarrieren Brug Impedans Spektroskopi: En Fokus på T-celle-endotel Cell Interaction

Breakdown of the blood-brain barrier (BBB) is a critical step in the development of autoimmune diseases such as multiple sclerosis (MS) and its animal ...

Isolation of the Side Population in Myc-induced T-cell Acute Lymphoblastic Leukemia in Zebrafish

Isolering af Side Population i Myc-induceret T-celle akut lymfoblastisk leukæmi i zebrafisk

Heterogeneous cell populations, from either healthy or malignant tissues, may contain a population of cells characterized by a differential ability to ...

An In Vivo Mouse Model to Measure Na&#239;ve CD4 T Cell Activation, Proliferation and Th1 Differentiation Induced by Bone Marrow-derived Dendritic Cells

An In Vivo Mouse Model to Measure NaÃƒÆ’Ã†'Ãƒâ€šÃ‚Â¯ve CD4 T Cell Activation, Proliferation and Th1 Differentiation Induced by Bone Marrow-derived Dendritic Cells

En In Vivo musemodel foranstaltning naive CD4 T-celle aktivering, spredning og Th1 differentiering induceret af knoglemarv-afledte dendritiske celler

Quantification of na&#239;ve CD4 T cell activation, proliferation, and differentiation to T helper 1 (Th1) cells is a useful way to assess the role played ...

An Ex Vivo Chicken Primary Bursal-cell Culture Model to Study Infectious Bursal Disease Virus Pathogenesis

An Ex Vivo Chicken Primary Bursal-cell Culture Model to Study Infectious Bursal Disease Virus Pathogenesis

En Ex Vivo kylling primære Bursal-cellekultur Model til at studere smitsomme Bursal Disease Virus patogenese

Infectious bursal disease virus (IBDV) is a birnavirus of economic importance to the poultry industry. The virus infects B cells, causing morbidity, ...

Multicolor Flow Cytometry-based Quantification of Mitochondria and Lysosomes in T Cells

Flerfarvet Flow flowcytometri-baserede kvantificering af mitokondrier og lysosomer i T celler

T cells utilize different metabolic programs to match their functional needs during differentiation and proliferation. Mitochondria are crucial cellular ...