Name: genome-wide analysis of hdac inhibitor-mediated modulation of micrornas and mrnas in b cells induced to undergo class-switch dna recombination and plasma cell differentiation
Uploaded: 2017-09-20T12:30:00
Description: Watch this Scientific Journal Video about Genome-wide Analysis of HDAC Inhibitor-mediated Modulation of microRNAs and mRNAs in B Cells Induced to Undergo Class-switch DNA Recombination and Plasma Cell

Dit werk werd gesteund door de NIH grants AI 105813 en AI 079705 (op PC), toekennen het Bondgenootschap voor Lupus onderzoek Target identificatie in Lupus Grant ALR 295955 (op PC), en het onderzoek van de artritis National Research Foundation (HZ). TS werd gesteund door de kindergeneeskunde Medical Center, tweede Xiangya ziekenhuis, centrale Zuid-Universiteit, Changsha (China), in het kader van het Xiangya-UT School van geneeskunde San Antonio medisch student bezoekende programma.

het protocol volgt de richtlijnen van de verzorging van de dieren van de institutionele zorg van het dier en gebruik commissies van de University of Texas Health Science Center in San Antonio.
 1. stimulatie van de muis B cellen voor MVO, Plasma celdifferentiatie en HDI behandeling <ol><li>voorbereiding van milt cel schorsingen Opmerking: alle stappen in een laminaire flow-kap met uitzondering van de muis euthanasie en dissectie.
	<ol><li>De specifieke pathogenen vrij...

<ol>
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Dit protocol biedt uitgebreide benaderingen om te induceren van B-cel klasse schakelen en plasma celdifferentiatie; voor het analyseren van de gevolgen daarvan door epigenetische modulatoren, namelijk HDI; en het effect van HDI op mRNA en miRNA expressie in deze cellen worden opgespoord. De meeste van deze benaderingen kan ook worden gebruikt voor het analyseren van de impact van epigenetische factor op menselijke expressie van B-cel functie en mRNA/miRNA. De qRT-PCR en mRNA-seq/miRNA-seq benaderingen kunnen ook worden g...

Epigenetische merken of factoren, zoals methylation van DNA, histone posttranslationele modificaties en niet-coderende RNAs (inclusief microRNAs), moduleren functie cel door gene expression1te wijzigen. Epigenetische aanpassingen reguleren B-lymfocyt functie, zoals immunoglobulin klasse-switch DNA recombinatie (CSR), somatische hypermutation (SHM) en differentiatie geheugencellen B of plasmacellen, waardoor het moduleren van het antilichaam en autoantibody Reacties2,3. MVO en SHM vereisen kritisch activering-geïnduceerde cytosinetrifosfaat Dipyridamol (steun, gecodeerd als Aicda), die zeer wordt geïnduceerd in B cellen in reactie op T-afhankelijke en T-onafhankelijke antigenen4. Klasse-geschakeld/hypermutated B cellen verder differentiëren tot plasmacellen, die grote volumes van antilichamen in een mode kritisch afhankelijk van rijping B-lymfocyt-geïnduceerde proteïne afscheiden 1 (Blimp1, gecodeerd als Prdm1)5. Abnormale epigenetische veranderingen in de B-cellen kunnen leiden tot afwijkende antilichaam/autoantibody reacties, die tot allergische reactie of autoimmuniteit1,4 leiden kunnen. Begrijpen hoe epigenetische factoren, zoals miRNAs, moduleren B-cel-intrinsieke genexpressie is niet alleen belangrijk voor de ontwikkeling van een vaccin, maar is ook essentieel om te onthullen van de mechanismen van potentieel abnormale antilichaam/autoantibody reacties.
Histon acetylation en deacetylation zijn wijzigingen van de lysine residuen op Histon eiwitten meestal gekatalyseerd door Histon acetyltransferase (hoed) en histone deacetylase (HDAC). Deze wijzigingen leiden tot de toenemende of dalende toegankelijkheid van de chromatine en verdere toestaan of voorkomen van de binding van transcriptiefactoren of proteïnen aan DNA en de wijziging van gen expressie5,6, 7 , 8. HDAC-remmers (HDI) vormen een klasse van verbindingen die met de functie van HDACs interfereren. Hier, we gebruikten HDI (VPA) inspelen op de regulering van HDAC op de intrinsieke genexpressie profiel van B-cellen en zijn mechanisme.
miRNAs zijn kleine, niet-coderende RNAs ongeveer 18 tot en met 22 nucleotiden in lengte die worden gegenereerd door verschillende stadia. miRNA host genen worden getranscribeerd en vormen haarspeld primaire microRNAs (pri-miRNAs). Ze worden geëxporteerd naar het cytoplasma waar pri-miRNAs verder worden verwerkt tot voorloper miRNAs (pre-miRNAs). Ten slotte, volwassen miRNAs worden gevormd door de splitsing van de pre-miRNAs. miRNAs herkennen de aanvullende reeksen binnen de 3' niet-vertaalde regio van hun doel mRNAs6,7. Via post-transcriptional tot zwijgen brengen, regelen miRNAs cellulaire activiteit, zoals de proliferatie en differentiatie apoptosis10,11. Aangezien meerdere miRNAs zich op de dezelfde mRNA richten kunt en één enkele miRNA mogelijk meerdere mRNAs kunt richten, is het belangrijk dat een weergave van de in het kader van de miRNA expressie profiel te begrijpen de waarde van het individu en het collectief effect van miRNAs. miRNAs is aangetoond te worden betrokken bij de ontwikkeling van de B-cel en perifere differentiatie, evenals fase-specifieke B-celdifferentiatie, antistofrespons tegen en autoimmuniteit1,4,9. In de 3' UTR van Aicda en Prdm1zijn er verschillende gevalideerde of voorspelde evolutionair geconserveerde sites die kunnen worden gericht door miRNAs8.
Epigenetische modulatie, met inbegrip van Histon na transcriptie wijziging en miRNAs, een celtype en cel fase-specifieke reglementering patroon van gen expressie9weergeven We beschrijven hier, methoden om te definiëren van de HDI-gemedieerde modulatie van miRNA en mRNA uitdrukking, MVO en plasma celdifferentiatie. Deze omvatten protocollen voor inducerende B cellen ondergaan van MVO en plasma celdifferentiatie; voor de behandeling van de B-cellen met HDI; en voor het analyseren van miRNA en mRNA uitdrukking door qRT-PCR, miRNA-seq en mRNA-seq10,11,12,8,13.

<table border="0" cellpadding="0" cellspacing="0" width="854">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:528px;">C57BL/6 mice</td>
			<td style="width:152px;">Jackson Labs</td>
			<td align="right" style="width:111px;">664</td>
			<td style="width:64px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Corning cellgro RPMI 1640 Medium (Mod.) 1X with L-Glutamine (Size: 6 x 500mL; With L-Glutamine)</td>
			<td>Fisher Scientific</td>
			<td>MT 10-040-CV&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">FBS</td>
			<td>Hyclone</td>
			<td>SH300</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">HyClone Antibiotic Antimycotic Solution 100 mL</td>
			<td>Fisher Scientific - Hyclone</td>
			<td>SV3007901&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#946;-Mercaptoethanol</td>
			<td>Fisher Scientific</td>
			<td>44-420-3250ML</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Falcon Cell Strainers</td>
			<td>Fisher Scientific</td>
			<td>21008-952&#160;&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Trypan Blue Stain 0.04%</td>
			<td>GIBCO/Life Technologies/Inv</td>
			<td align="right">15250</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">ACK Lysis Buffer&#160;</td>
			<td>Fisher Scientific</td>
			<td>BW10-548E&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hausser Scientific Bright-Line Counting Chamber</td>
			<td>Fisher Scientific</td>
			<td>02-671-51B</td>
			<td></td>
		</tr>
		<tr height="35">
			<td height="35" style="height:35px;width:528px;">EasySep Magnet</td>
			<td>Stem Cell Technologies</td>
			<td align="right">18000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Falcon Round-Bottom Polystyrene Tubes with Cap</td>
			<td>Fisher Scientific</td>
			<td>14-959-1A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">EasySep Mouse B cell Isolation Kit</td>
			<td>Stem Cell Tech</td>
			<td align="right">19854</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">BD Needle Only 18 Gauge 1.5 inch SHORT BEVEL 100/box&#160;</td>
			<td>BD Biosciences</td>
			<td>&#160;305199</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PE/Cy7 anti-mouse CD138 (Syndecan-1) Antibody</td>
			<td>BioLegend&#160;</td>
			<td>142513 (25 ug)</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PE-Cy7 B220 antibody</td>
			<td>BioLegend</td>
			<td align="right">103222</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">7-AAD (1 mg)</td>
			<td>Sigma Aldrich</td>
			<td>A9400-1MG</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">APC anti-mouse/human CD45R/B220 antibody</td>
			<td>Biolegend</td>
			<td align="right">103212</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Mouse APC-IgG1 200 &#181;g</td>
			<td>Biolegend</td>
			<td align="right">406610</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">FITC anti-mouse IgM Antibody</td>
			<td>Biolegend</td>
			<td align="right">406506</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">FITC anti-mouse/human CD45R/B220 Antibody</td>
			<td>Biolegend</td>
			<td align="right">103206</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PE Anti-Human/Mouse CD45R (B220) (RA3-6B2)</td>
			<td>Biolegend</td>
			<td align="right">103208</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">HBSS 1X</td>
			<td>Fisher Scientific</td>
			<td>MT-21-022-CM</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Bovine Serum Albumin, Fraction V, Heat Shock Treated&#160;</td>
			<td>Fisher Scientific</td>
			<td>BP1600-100</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">LPS 25mg (Lipopolysaccharides from Escherichia coli 055:B5)</td>
			<td>Sigma Aldrich</td>
			<td>L2880-25MG</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Recombinant mouse IL-4 (carrier-free)</td>
			<td>BioLegend</td>
			<td>574302 (size: 10 ug)</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Valproic acid sodium salt</td>
			<td>Sigma Aldrich</td>
			<td>P4543</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">SterilGARD e3 Class II Type A2 Biosafety Cabinet</td>
			<td>The Baker Company</td>
			<td>SG404</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Large-Capacity Reach-In CO2 Incubator&#160;</td>
			<td>Thermo Scientific</td>
			<td align="right">3950</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Isotemp Digital-Control Water Baths: Model 205</td>
			<td>Fisher Scientific</td>
			<td>15-462-5Q</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">5mL Round Bottom Polystyrene Test Tube</td>
			<td>Fisher Scientific</td>
			<td>&#160;14-959-5</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Corning CentriStar 15ml Centrifuge Tubes&#160;</td>
			<td>Fisher Scientific&#160;</td>
			<td>05-538-59A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">1.7 mL Microtube, clear</td>
			<td>Genesee</td>
			<td>22-282</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Higher-Speed Easy Reader Plastic Centrifuge Tubes 50ml</td>
			<td>Fisher Scientific</td>
			<td>06-443-18</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">ELMI SkySpin CM-6MT</td>
			<td>ELMI</td>
			<td>CM-6MT</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Rotor 6M</td>
			<td>ELMI</td>
			<td>6M</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Rotor 6M.06</td>
			<td>ELMI</td>
			<td>6M.06</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Drummond Portable Pipet-Aid XP Pipet Controller</td>
			<td>Drummond Scientific</td>
			<td>4-000-101</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">25 mL serological pipette tips</td>
			<td>Fisher Scientific</td>
			<td>89130-900</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">10 mL serological pipette tips</td>
			<td>Fisher Scientific</td>
			<td>89130-898</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">5 mL serological pipette tips</td>
			<td>Fisher Scientific</td>
			<td>898130-896</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">48-well plates</td>
			<td>Fisher Scientific</td>
			<td>07-200-86</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Allegra 6 Benchtop Centrifuge, Non-Refrigerated</td>
			<td>Beckman Coulter</td>
			<td align="right">366802</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">GH-3.8A Rotor, Horizontal, ARIES Smart Balance</td>
			<td>Beckman Coulter</td>
			<td align="right">366650</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Allegra 25R Benchtop Centrifuge, Refrigerated</td>
			<td>Beckman Coulter</td>
			<td align="right">369434</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">TA-15-1.5 Rotor, Fixed Angle</td>
			<td>Beckman Coulter</td>
			<td align="right">368298</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fisher Scientific AccuSpin Micro 17</td>
			<td>Fisher Scientific</td>
			<td>13-100-675</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fisher Scientific Analog Vortex Mixer&#160;</td>
			<td>Fisher Scientific</td>
			<td>02-215-365</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">miRNeasy Mini Kit (50)</td>
			<td>Qiagen</td>
			<td align="right">217004</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Direct-zol RNA MiniPrep kit</td>
			<td>Zymo Research</td>
			<td>R2050</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Chloroform (Approx. 0.75% Ethanol as Preservative/Molecular Biology)</td>
			<td>Fisher Scientific</td>
			<td>BP1145-1</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Rnase-Free Dnase set (50)</td>
			<td>QIAGEN</td>
			<td align="right">79254</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">NanoDrop 2000 Spectrophotometers</td>
			<td>Thermo Scientific</td>
			<td>ND-2000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Superscript III First-strand Synthesis System RT-PCR</td>
			<td>Invitrogen</td>
			<td align="right">175013897</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">iTaq Universal SYBR Green Supermix</td>
			<td>Bio-rad&#160;</td>
			<td>172-5121</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fisherbrand 96-Well Semi-Skirted PCR Plates, case of 25</td>
			<td>Fisher</td>
			<td style="width:111px;">14-230-244</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Microseal &#39;B&#39; Adhesive Seals</td>
			<td>Bio-Rad</td>
			<td>MSB-1001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">MyiQ Optics Module</td>
			<td>Bio-Rad</td>
			<td>170-9744</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">iCycler Chassis</td>
			<td>Bio-Rad</td>
			<td>170-8701</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Optical Kit</td>
			<td>Bio-Rad</td>
			<td>170-9752</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">BD LSR II Flow Cytometry Analyzer</td>
			<td>BD Biosciences</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">FACSDiva software</td>
			<td>BD Biosciences</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">FlowJo 10</td>
			<td>BD Biosciences</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">2100 Bioanalyzer</td>
			<td>Agilent Technologies</td>
			<td>G2943CA</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">S200 Focused-ultrasonicator</td>
			<td>Covaris</td>
			<td>S200</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">SPRIworks Fragment Library System I for Illumina</td>
			<td>Beckman Coulter</td>
			<td>A288267</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">cBot Cluster Generation Station</td>
			<td>illumina</td>
			<td>SY-312-2001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">HiSeq 2000 Genome Sequencer</td>
			<td>Illumina</td>
			<td>SY-401-1001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">TruSeq RNA Library Prep Kit v2</td>
			<td>Illumina</td>
			<td>RS-122-2001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">TruSeq Small RNA Library Prep Kit</td>
			<td>Illumina</td>
			<td>RS-200-0012</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">NEXTflex Illumina Small RNA Sequencing Kit v3</td>
			<td>Bioo Scientific</td>
			<td>5132-05</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">2200 TapeStation</td>
			<td>Agilent</td>
			<td>G2964AA</td>
			<td></td>
		</tr>
	</tbody>
</table>

genome-wide analysis of hdac inhibitor-mediated modulation of micrornas and mrnas in b cells induced to undergo class-switch dna recombination and plasma cell differentiation

Antilichaam reacties zijn bereikt door verschillende kritische B-cel-intrinsieke processen, met inbegrip van somatische hypermutation (SHM), klasse-switch DNA recombinatie (CSR) en plasma celdifferentiatie. In de afgelopen jaren is epigenetische veranderingen of factoren, zoals Histon deacetylation en microRNAs (miRNAs), gebleken om te interageren met de B-cel genetische programma's vorm antilichaam reacties, terwijl de disfunctie van epigenetische factoren blijkt te leiden tot autoantibody reacties. Het analyseren van genoom-brede miRNA en mRNA uitdrukking in B cellen in reactie op de epigenetische modulatoren is belangrijk voor het begrijpen van de Epigenetische regulatie van de B-cel functie en antilichaam reactie. Hier tonen we een protocol voor inducerende B cellen ondergaan van MVO en plasma cel differentiatie, behandelen deze B-cellen met histone deacetylase (HDAC) remmers (Hotel) en analyseren van mRNA en microRNA expressie. In dit protocol, analyseren we rechtstreeks aanvullende opeenvolgingen van DNA (cDNA) met behulp van volgende-generatie mRNA sequencing (mRNA-seq) en miRNA-seq technologieën, in kaart brengen van de volgorde leest aan het genoom, en kwantitatieve omgekeerde transcriptie (qRT)-PCR. Met deze aanpak, die we hebben gedefinieerd dat in B cellen geïnduceerde ondergaan MVO en plasma celdifferentiatie, HDI, een epigenetische regulator, selectief miRNA en mRNA uitdrukking moduleert en verandert van MVO en plasma cel differentiatie.

Met behulp van onze protocol, gezuiverd B cellen geplaatst met LPS (3 µg/mL) en IL-4 (5 ng/mL) voor 96 h 30-40% van MVO te IgG1 en ~ 10% van plasma celdifferentiatie kan veroorzaken. Na behandeling met HDI (500µM VPA), de CSR aan IgG1 verlaagd naar 10-20%, terwijl plasma celdifferentiatie tot ~ 2 daalde % (Figuur 1). HDI-gemedieerde remming van MVO was verder bevestigd door verminderde aantallen na recombinatie Iμ-Cγ1 en volwassen VH-DJH-afschrif...

Watch this Scientific Journal Video about Genome-wide Analysis of HDAC Inhibitor-mediated Modulation of microRNAs and mRNAs in B Cells Induced to Undergo Class-switch DNA Recombination and Plasma Cell

Genome-wide Analysis of HDAC Inhibitor-mediated Modulation of microRNAs and mRNAs in B Cells Induced to Undergo Class-switch DNA Recombination and Plasma Cell Differentiation

Epigenetische factoren kunnen communiceren met genetische programma's voor het moduleren van genexpressie en de regulering van de B-cel functie. Door stimulatie van de in vitro B-cel, qRT-PCR, en high-throughput microRNA-sequence en mRNA-sequence benaderingen te combineren, kunnen we analyseren de epigenetische modulatie van miRNA en gen-expressie in B-cellen.

Genoom-brede analyse van HDAC-Inhibitor-gemedieerde modulatie van microRNAs en mRNAs in B cellen geïnduceerde Undergo klasse-switch DNA recombinatie en Plasma celdifferentiatie

Antibody responses are accomplished through several critical B cell-intrinsic processes, including somatic hypermutation (SHM), class-switch DNA ...

Research

JoVE Journal

Immunology and Infection

Induction and Assessment of Class Switch Recombination in Purified Murine B Cells

Inductie en beoordeling van klasse Switch recombinatie in gezuiverd Murine B-cellen

Humoral immunity is the branch of the immune system maintained by B cells and mediated through the secretion of antibodies. Upon B cell activation, the ...

Directed Differentiation of Induced Pluripotent Stem Cells towards T Lymphocytes

Geregisseerd Differentiatie van geïnduceerde pluripotente stamcellen naar T lymfocyten

Adoptive cell transfer (ACT) of antigen-specific CD8+  cytotoxic T lymphocytes (CTLs) is a promising treatment for a variety of  malignancies 1. CTLs can  ...

Establishment of an In vitro System to Study Intracellular Behavior of Candida glabrata in Human THP-1 Macrophages

Establishment of an In vitro System to Study Intracellular Behavior of Candida glabrata in Human THP-1 Macrophages

Oprichting van een<em&gt; In vitro</em&gt; Systeem om Intracellular Gedrag van Studie<em&gt; Candida glabrata</em&gt; In Human THP-1 macrofagen

A cell culture model system, if a close mimic of host environmental conditions, can serve as an inexpensive, reproducible and easily manipulatable ...

Rapid Molecular Detection and Differentiation of Influenza Viruses A and B

Snelle Moleculaire detectie en differentiatie influenzavirussen A en B

Influenza is a contagious respiratory illness caused by influenza viruses A and B in humans and causes a significant amount of morbidity and mortality ...

The Isolation, Differentiation, and Quantification of Human Antibody-secreting B Cells from Blood: ELISpot as a Functional Readout of Humoral Immunity

De isolatie, differentiatie en kwantificering van menselijk antilichaam-uitscheidende B-cellen uit bloed: ELISpot als functionele Uitlezing van humorale immuniteit

The hallmark of humoral immunity is to generate functional ASCs, which synthesize and secrete Abs specific to an antigen (Ag), such as a pathogen, and are ...

Detection and Enrichment of Rare Antigen-specific B Cells for Analysis of Phenotype and Function

Detectie en Verrijking van zeldzame antigeen-specifieke B-cellen voor analyse van fenotype en functie

B cells reactive with a specific antigen usually occur at a frequency of &lt;0.05% of lymphocytes. For decades researchers have sought methods to isolate ...

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

Een muismodel In Vivo maatregel naïef CD4 T-cel activatie, proliferatie en differentiatie van de Th1 geïnduceerd door het beenmerg-afgeleide dendritische cellen

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 ...

Analysis of Shear Flow-induced Migration of Murine Marginal Zone B Cells In Vitro

Analyse van Shear Flow-geïnduceerde migratie van RattenUitrustingen Marginal Zone B cellen In Vitro

Marginal zone B cells (MZBs) are a population of B cells that reside in the mouse splenic marginal zones that envelop follicles. To reach the follicles, ...

In Vitro Differentiation Model of Human Normal Memory B Cells to Long-lived Plasma Cells

In Vitro differentiatie Model van menselijke normale B geheugencellen te langlevende plasmacellen

Plasma cells (PCs) secrete large amounts of antibodies and develop from B cells that have been activated. PCs are rare cells located in the bone marrow or ...