Name: evaluation of t follicular helper cells and germinal center response during influenza a virus infection in mice
Uploaded: 2020-06-27T13:00:00
Description: Watch this Scientific Journal Video about Evaluation of T Follicular Helper Cells and Germinal Center Response During Influenza A Virus Infection in Mice at JoVE.com

Vi takker stabe flow cytometry facilitet, ABSL2 facilitet og SPF dyrefacilitet institut Pasteur i Shanghai for deres tekniske hjælp og rådgivning. Dette arbejde blev støttet af følgende tilskud: Strategic Priority Research Program of the Chinese Academy of Sciences (XDB29030103), National Key R&D Program of China (2016YFA0502202), National Natural Science Foundation of China (31570886).

Dyreforsøg blev godkendt af Institutional Animal Care and Use Committee of Institut Pasteur i Shanghai, Kina. Alle forsøgene blev udført på grundlag af de institutionelle dyrepleje- og brugskomitégodkendte dyreprotokoller.
BEMÆRK: Virusinfektion hos mus og isolering af organer skal udføres under ABSL2-tilstand.
1. Podning af PR8-influenzavirus og registrering af muss vægt
<ol><li>Forbered 8 uger gamle mandlige C57BL/6 mus til infektion på...

<ol>
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I., 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=Bcl6+mediates+the+development+of+T+follicular+helper+cells.">Bcl6 mediates the development of T follicular helper cells.</a> Science. 325 (5943), 1001-1005 (2009).</li><li>Yu, D., 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=The+transcriptional+repressor+Bcl-6+directs+T+follicular+helper+cell+lineage+commitment.">The transcriptional repressor Bcl-6 directs T follicular helper cell lineage commitment.</a> Immunity. 31 (3), 457-468 (2009).</li><li>Pedros, 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=A+TRAF-like+motif+of+the+inducible+costimulator+ICOS+controls+development+of+germinal+center+TFH+cells+via+the+kinase+TBK1.">A TRAF-like motif of the inducible costimulator ICOS controls development of germinal center TFH cells via the kinase TBK1.</a> Nature Immunology. 17 (7), 825-833 (2016).</li><li>Xu, 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=Follicular+T-helper+cell+recruitment+governed+by+bystander+B+cells+and+ICOS-driven+motility.">Follicular T-helper cell recruitment governed by bystander B cells and ICOS-driven motility.</a> Nature. 496 (7446), 523-527 (2013).</li><li>Lee, S. 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=B+cell+priming+for+extrafollicular+antibody+responses+requires+Bcl-6+expression+by+T+cells.">B cell priming for extrafollicular antibody responses requires Bcl-6 expression by T cells.</a> The Journal of Experimental Medicine. 208 (7), 1377-1388 (2011).</li><li>Zotos, D., 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=IL-21+regulates+germinal+center+B+cell+differentiation+and+proliferation+through+a+B+cell-intrinsic+mechanism.">IL-21 regulates germinal center B cell differentiation and proliferation through a B cell-intrinsic mechanism.</a> The Journal of Experimental Medicine. 207 (2), 365-378 (2010).</li><li>Vogelzang, 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=A+fundamental+role+for+interleukin-21+in+the+generation+of+T+follicular+helper+cells.">A fundamental role for interleukin-21 in the generation of T follicular helper cells.</a> Immunity. 29 (1), 127-137 (2008).</li><li>Ansel, K. 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=A+chemokine-driven+positive+feedback+loop+organizes+lymphoid+follicles.">A chemokine-driven positive feedback loop organizes lymphoid follicles.</a> Nature. 406 (6793), 309-314 (2000).</li><li>Shi, J., 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=PD-1+Controls+Follicular+T+Helper+Cell+Positioning+and+Function.">PD-1 Controls Follicular T Helper Cell Positioning and Function.</a> Immunity. 49 (2), 264-274 (2018).</li><li>Methot, S. P., Di Noia, J. 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D., Ballesteros-Tato, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=FoxP3++regulatory+T+cells+promote+influenza-specific+Tfh+responses+by+controlling+IL-2+availability.">FoxP3+ regulatory T cells promote influenza-specific Tfh responses by controlling IL-2 availability.</a> Nature Communications. 5, 3495 (2014).</li><li>He, 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=Extracellular+matrix+protein+1+promotes+follicular+helper+T+cell+differentiation+and+antibody+production.">Extracellular matrix protein 1 promotes follicular helper T cell differentiation and antibody production.</a> Proceedings of the National Academy of Sciences of the United States of America. 115 (34), 8621-8626 (2018).</li><li>Le&#243;n, B., 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=Regulation+of+TH2+development+by+CXCR5++dendritic+cells+and+lymphotoxin-expressing+B+cells.">Regulation of TH2 development by CXCR5+ dendritic cells and lymphotoxin-expressing B cells.</a> Nature Immunology. 13 (7), 681-690 (2012).</li><li>Wang, 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=The+transcription+factor+Foxp1+is+a+critical+negative+regulator+of+the+differentiation+of+follicular+helper+T+cells.">The transcription factor Foxp1 is a critical negative regulator of the differentiation of follicular helper T cells.</a> Nature Immunology. 15 (7), 667-675 (2014).</li><li>Bouvier, N. M., Lowen, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+Models+for+Influenza+Virus+Pathogenesis+and+Transmission.">Animal Models for Influenza Virus Pathogenesis and Transmission.</a> Viruses. 2 (8), 1530-1563 (2010).</li><li>Miyauchi, 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=Protective+neutralizing+influenza+antibody+response+in+the+absence+of+T+follicular+helper+cells.">Protective neutralizing influenza antibody response in the absence of T follicular helper cells.</a> Nature Immunology. 17 (12), 1447-1458 (2016).</li><li>Rodriguez, L., Nogales, A., Martinez-Sobrido, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influenza+A+Virus+Studies+in+a+Mouse+Model+of+Infection.">Influenza A Virus Studies in a Mouse Model of Infection.</a> Journal of Visualized Experiments. (127), (2017).</li><li>Kitano, 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=Bcl6+protein+expression+shapes+pre-germinal+center+B+cell+dynamics+and+follicular+helper+T+cell+heterogeneity.">Bcl6 protein expression shapes pre-germinal center B cell dynamics and follicular helper T cell heterogeneity.</a> Immunity. 34 (6), 961-972 (2011).</li><li>Yusuf, I., 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=Germinal+center+T+follicular+helper+cell+IL-4+production+is+dependent+on+signaling+lymphocytic+activation+molecule+receptor+(CD150).">Germinal center T follicular helper cell IL-4 production is dependent on signaling lymphocytic activation molecule receptor (CD150).</a> The Journal of Immunology. 185 (1), 190-202 (2010).</li><li>Sun, J., Dodd, H., Moser, E. K., Sharma, R., Braciale, T. 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På grund af specialiserede roller i at give B-celle hjælp til at generere høj-affinitet antistoffer, Tfh celler er blevet grundigt undersøgt i mekanismerne for differentiering og manipulation til at give nye strategier for vaccine design. Influenzavirusinfektion fremkalder kraftige Tfh- og GC B-celler, hvilket er en passende model for dette forskningsområde. I dette papir beskrev vi protokoller for influenzavirusinfektion ved intranasal inokulering, evaluering af Tfh-associeret respons ved flowcytometry, immunfluore...

I det seneste årti har adskillige undersøgelser været fokuseret på den nyligt identificerede CD4+ T celle delmængde, Tfh celle delmængde, for sine væsentlige roller i germinal center (GC) B udvikling. B celle lymfom 6 (Bcl6), som hovedsagelig betragtes som et gen repressor, er afstamning-definerende faktor Tfh celler for beviser for, at ektopisk udtryk for Bcl6 er tilstrækkelig til at drive Tfh differentiering, mens mangel på Bcl6 resulterer i forsvundet Tfh differentiering1,2,3. I modsætning til andre CD4+ T hjælper delmængder udfører deres effector funktion ved migration til de steder af inflammation, Tfh celler giver B-celle hjælp primært i B celle follikulær zone milt og lymfeknude. Co-stimulerende molekyler ICOS og CD40L, spiller en væsentlig rolle i samspillet mellem Tfh og GC B celler. Under Tfh differentiering, ICOS sender nødvendige signaler fra cognate B-celler og fungerer også som en receptor modtager migration signaler fra tilskuer B-celler til B cellezone lokalisering4,5. CD40L er en mediator af signaler fra Tfh celler til B-celler spredning og overlevelse6. En anden faktor, der spiller den lignende rolle som CD40L, er cytokinEN IL21, som hovedsageligt udskilles af Tfh-celler. IL21 regulerer direkte GC B-cellers udvikling og produktion af antistoffer med høj affinitet, men dets rolle i Tfh-differentiering er stadig kontroversiel7,8. PD-1 og CXCR5, som nu hyppigst anvendes til at identificere Tfh-celler i flowcytometrianalyse, spiller også en væsentlig rolle i differentieringen og funktionen af denne delmængde. CXCR5 er receptoren af B-celle follikulært kemokin og formidler lokaliseringen af Tfh-celler i B-cellesække9. PD-1 er nu identificeret til ikke kun at have follikulær vejledning funktion, men også sende kritiske signaler i processen med GC B celler affinitet modning10. Baseret på disse fund kunne evaluering af ekspressionen af disse molekyler dybest set afspejle modningen og funktionen af Tfh-celler.
GC er en induceret forbigående mikroanatomisk struktur i sekundære lymfoide organer og meget afhængig af Tfh-celler, hvilket er en perfekt udlæsning til at evaluere Tfh-respons. I GC, efter at have modtaget signaler medieret af cytokiner og co-stimulerende molekyler, er B-celler underlagt klasseskift og somatisk hypermutation for at generere antistoffer med høj affinitet11. Differential antistof klasseskift forekommer i differentialcytokin niche, hvor IL4 og IL21 fremkaldeR IgG1 klasseskift, mens IFNγ inducerer IgG2 klasseskift12. Plasmaceller er producenter af udskillede antistoffer og er uhelbredeligt differentierede celler. Ligesom Tfh celler, udvikling af B-celler i GC er forbundet med dynamisk udtryk for mange væsentlige molekyler. Baseret på den aktuelle undersøgelse kan GC B-celler identificeres som B220+PNA+Fas+ eller B220+GL7+Fas+ celler og plasmaceller sammenlignet med deres prækursorer, nedregulerer ekspressionen af B220 og opregulerer CD138-udtryk13. Desuden kan begge disse egenskaber påvises i flowcytometry- og immunfluorescensanalyse, hvilket er en passende evaluering af GC-respons.
Robust cellulære og humoristiske reaktioner er induceret i influenzavirus infektion, med Tfh og Th1 celler dominerer CD4+ T cellerespons14, hvilket gør det til en perfekt model for Tfh celler differentiering undersøgelse. Influenza A/Puerto Rico/8/34 H1N1(PR8), som er en almindeligt anvendt musetilpasset stamme, anvendes ofte i denne undersøgelse14,15,16. Her beskriver vi nogle grundlæggende protokoller for Tfh-undersøgelsesrelevant analyse i influenzavirusinfektion: 1) intranasal podning af PR8-virus; 2) antigenspecifikke Tfh-celler, GC B- og plasmaceller og IL21-detektion med flowcytometri 3) histologisk visualisering af GC; 4) påvisning af antigenspecifik antistoftitler i serum med ELISA. Disse protokoller giver de nødvendige teknikker til nye forskere i Tfh-associeret undersøgelse.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Immunostaining of Tfh cells, NP-specific Tfh cells and Bcl-6</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>37% formaldehyde</td>
			<td>Sigma</td>
			<td>F1635</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-CD16/32 mouse</td>
			<td>Thermo Fisher Scientific</td>
			<td>14-0161-86</td>
			<td></td>
		</tr>
		<tr>
			<td>APC-conjugated-IAbNP311-325 MHC class II tetramer</td>
			<td>NIH</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Bcl-6 PE</td>
			<td>Biolegend</td>
			<td>358504</td>
			<td>clone:7D1</td>
		</tr>
		<tr>
			<td>Biotin-CXCR5</td>
			<td>Thermo Fisher Scientific</td>
			<td>13-7185-82</td>
			<td>clone: SPRCL5</td>
		</tr>
		<tr>
			<td>CD4 Percp-eFluor 710</td>
			<td>Thermo Fisher Scientific</td>
			<td>46-0041-82</td>
			<td>clone:GK1.5</td>
		</tr>
		<tr>
			<td>CD44 eVolve 605</td>
			<td>Thermo Fisher Scientifi</td>
			<td>83-0441-42</td>
			<td>clone:IM7</td>
		</tr>
		<tr>
			<td>CD44 FITC</td>
			<td>Thermo Fisher Scientifi</td>
			<td>11-0441-82</td>
			<td>clone:IM7</td>
		</tr>
		<tr>
			<td>CD62L FITC</td>
			<td>BD Pharmingen</td>
			<td>553150</td>
			<td>clone:MEL-14</td>
		</tr>
		<tr>
			<td>ICOS BV421</td>
			<td>Biolegend</td>
			<td>564070</td>
			<td>clone:7E.17G9</td>
		</tr>
		<tr>
			<td>PD1 PE/Cy7</td>
			<td>Biolegend</td>
			<td>135216</td>
			<td>clone:29F.1A12</td>
		</tr>
		<tr>
			<td>Streptavidin BV421</td>
			<td>BD Pharmingen</td>
			<td>563259</td>
			<td></td>
		</tr>
		<tr>
			<td>Streptavidin PE</td>
			<td>BD Pharmingen</td>
			<td>554081</td>
			<td></td>
		</tr>
		<tr>
			<td>Intracelluar staining of IL21</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>37% formaldehyde</td>
			<td>Sigma</td>
			<td>F1635</td>
			<td></td>
		</tr>
		<tr>
			<td>anti-human IgG</td>
			<td>Jackson ImmunoResearch Laboratories</td>
			<td>109-605-098</td>
			<td></td>
		</tr>
		<tr>
			<td>Brefeldin A</td>
			<td>Sigma</td>
			<td>B6542</td>
			<td></td>
		</tr>
		<tr>
			<td>human FCc IL-21 receptor</td>
			<td>R&#38;D System</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>ionomycin</td>
			<td>Sigma</td>
			<td>I0634</td>
			<td></td>
		</tr>
		<tr>
			<td>Live/Dead Fixable Aqua Dead Cell staining kit</td>
			<td>Thermo Fisher Scientific</td>
			<td>L34966</td>
			<td></td>
		</tr>
		<tr>
			<td>PMA</td>
			<td>Sigma</td>
			<td>P1585</td>
			<td></td>
		</tr>
		<tr>
			<td>Saponin</td>
			<td>MP</td>
			<td>102855</td>
			<td></td>
		</tr>
		<tr>
			<td>GC B and plasma cells staining</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>B220 APC</td>
			<td>Thermo Fisher Scientific</td>
			<td>17-0452-81</td>
			<td>clone:RA3-6B2</td>
		</tr>
		<tr>
			<td>CD138 PE</td>
			<td>BD Pharmingen</td>
			<td>561070</td>
			<td>clone:281-2</td>
		</tr>
		<tr>
			<td>CD95 (FAS) PE/Cy7</td>
			<td>BD Pharmingen</td>
			<td>557653</td>
			<td>clone:Jo2</td>
		</tr>
		<tr>
			<td>IgD eFluor 450</td>
			<td>Thermo Fisher Scientific</td>
			<td>48-5993-82</td>
			<td>clone:11-26c</td>
		</tr>
		<tr>
			<td>PNA FITC</td>
			<td>Sigma</td>
			<td>L7381</td>
			<td></td>
		</tr>
		<tr>
			<td>Assay of HA-specific antibody titer with ELISA</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>PR8-HA</td>
			<td>Sino Biological</td>
			<td>11684-V08H</td>
			<td></td>
		</tr>
		<tr>
			<td>BSA</td>
			<td>SSBC</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Goat anti mouse Ig (SBA Clonotyping System-HRP)</td>
			<td>SouthernBiotech</td>
			<td>5300-05</td>
			<td></td>
		</tr>
		<tr>
			<td>Goat anti mouse IgM (SBA Clonotyping System-HRP)</td>
			<td>SouthernBiotech</td>
			<td>5300-05</td>
			<td></td>
		</tr>
		<tr>
			<td>Goat anti mouse IgG1 (SBA Clonotyping System-HRP)</td>
			<td>SouthernBiotech</td>
			<td>5300-05</td>
			<td></td>
		</tr>
		<tr>
			<td>Goat anti mouse IgG2b (SBA Clonotyping System-HRP)</td>
			<td>SouthernBiotech</td>
			<td>5300-05</td>
			<td></td>
		</tr>
		<tr>
			<td>Goat anti mouse IgG2c (SBA Clonotyping System-HRP)</td>
			<td>SouthernBiotech</td>
			<td>5300-05</td>
			<td></td>
		</tr>
		<tr>
			<td>TMB Substrate Reagent Set</td>
			<td>BD Pharmingen</td>
			<td>555214</td>
			<td></td>
		</tr>
		<tr>
			<td>Histology</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Alexa Fluor 555-Goat-anti rat IgG</td>
			<td>Life Technology</td>
			<td>A21434</td>
			<td></td>
		</tr>
		<tr>
			<td>anti-mouse IgD</td>
			<td>Biolegend</td>
			<td>405702</td>
			<td></td>
		</tr>
		<tr>
			<td>biotinylated PNA</td>
			<td>Vector laboratories</td>
			<td>B-1075</td>
			<td></td>
		</tr>
		<tr>
			<td>dilute Alexa Fluor 488-streptavidin</td>
			<td>Life Technology</td>
			<td>S11223</td>
			<td></td>
		</tr>
		<tr>
			<td>normal goat serum</td>
			<td>SouthernBiotech</td>
			<td>0060-01</td>
			<td></td>
		</tr>
		<tr>
			<td>Pro-long gold antifade reagent</td>
			<td>Thermo Fisher Scientific</td>
			<td>P3630</td>
			<td></td>
		</tr>
		<tr>
			<td>STREPTAVIDIN/BIOTIN blocking kit</td>
			<td>Vector laboratories</td>
			<td>SP-2002</td>
			<td></td>
		</tr>
	</tbody>
</table>

evaluation of t follicular helper cells and germinal center response during influenza a virus infection in mice

T Follicular Helper (Tfh) celler er en uafhængig CD4+ T celle delmængde specialiseret i at yde hjælp til germinal center (GC) udvikling og produktion af høj-affinitet antistoffer. I influenzavirusinfektion induceres robuste Tfh- og GC B-celleresponser for at lette effektiv virusudryddelse, hvilket giver en kvalificeret musemodel til Tfh-associeret undersøgelse. I dette papir beskrev vi protokoller til påvisning af grundlæggende Tfh-associeret immunrespons under influenzavirusinfektion hos mus. Disse protokoller omfatter: intranasal podning af influenzavirus; cytometribejdning og analyse af polyklonale og antigenspecifikke Tfh-celler, GC B-celler og plasmaceller påvisning af immunfluorescens i GC'er enzymrelateret immunosorbentanalyse (ELISA) af influenzavirusspecifikt antistof i serum. Disse assays dybest set kvantificere differentiering og funktion af Tfh celler i influenzavirus infektion, hvilket giver hjælp til undersøgelser i belyse differentiering mekanisme og manipulation strategi.

Karakterisering af mus sygelighed i influenzavirus infektion Efter influenzavirusinfektion er mus mindre aktive og anorektiske på grund af sygdom, hvilket afspejles af alvorligt vægttab, et almindeligt anvendt symptom til at overvåge musens sygelighed19. Som vist i figur 1abegyndte PR8-virusinficerede mus at tabe sig på dag 6, nåede det højeste tabsniveau på dag 8 og vendte tilbage til det oprindelige niveau på dag 10. Som...

Watch this Scientific Journal Video about Evaluation of T Follicular Helper Cells and Germinal Center Response During Influenza A Virus Infection in Mice at JoVE.com

Evaluation of T Follicular Helper Cells and Germinal Center Response During Influenza A Virus Infection in Mice

Dette papir beskriver protokoller for evaluering af Tfh og GC B respons i mus model af influenzavirus infektion.

Evaluering af T Follikulære Hjælpeceller og Germinal Center Response Under Influenza A Virus Infektion i Mus

T Follicular Helper (Tfh) cells are an independent CD4+&#160;T cell subset specialized in providing help for germinal center (GC) development and ...

Research

JoVE Journal

Immunology and Infection

Using Bioluminescent Imaging to Investigate Synergism Between Streptococcus pneumoniae and Influenza A Virus in Infant Mice

Using Bioluminescent Imaging to Investigate Synergism Between Streptococcus pneumoniae and Influenza A Virus in Infant Mice

During the 1918 influenza virus pandemic, which killed approximately 50 million people worldwide, the majority of fatalities were not the result of ...

In Vitro Analysis of Myd88-mediated Cellular Immune Response to West Nile Virus Mutant Strain Infection

In Vitro Analysis of Myd88-mediated Cellular Immune Response to West Nile Virus Mutant Strain Infection

In vitro analyse af MyD88-medieret Cellular immunrespons til West Nile Virus mutantstamme Infektion

An attenuated West Nile virus (WNV), a nonstructural (NS) 4B-P38G mutant, induced higher innate cytokine and T cell responses than the wild-type WNV in ...

Influenza Virus Propagation in Embryonated Chicken Eggs

Influenza virusformering i befrugtede hønseæg

Influenza infection is associated with about 36,000 deaths and more than 200,000 hospitalizations every year in the United States. The continuous ...

Co-immunoprecipitation of the Mouse Mx1 Protein with the Influenza A Virus Nucleoprotein

Co-immunoudfældning af musen Mx1 Protein med influenza A-virus nukleoprotein

Studying the interaction between proteins is key in understanding their function(s). A very powerful method that is frequently used to study interactions ...

Measuring Attachment and Internalization of Influenza A Virus in A549 Cells by Flow Cytometry

Måling Attachment og internalisering af influenza A-virus i A549-celler ved flowcytometri

Attachment to target cells followed by internalization are the very first steps of the life cycle of influenza A virus (IAV). We provide here a detailed ...

Neutrophil Isolation and Analysis to Determine their Role in Lymphoma Cell Sensitivity to Therapeutic Agents

Neutrofil Isolering og analyse for at fastslå deres rolle i lymfom Cell Følsomhed til terapeutiske midler

Neutrophils are the most abundant (40% to 75%) type of white blood cells and among the first inflammatory cells to migrate towards the site of ...

In Vitro Disassembly of Influenza A Virus Capsids by Gradient Centrifugation

In Vitro Disassembly of Influenza A Virus Capsids by Gradient Centrifugation

In vitro Demontering af influenza A-virus capsider ved gradientcentrifugering

Acid-triggered molecular processes closely control cell entry of many viruses that enter through the endocytic system. In the case of influenza A virus ...

Influenza A Virus Studies in a Mouse Model of Infection

Influenza A Virus undersøgelser i en musemodel af infektion

Influenza viruses cause over 500,000 deaths worldwide1 and are associated with an annual cost of 12 - 14 billion USD in the United States alone ...

Induction of Paralysis and Visual System Injury in Mice by T Cells Specific for Neuromyelitis Optica Autoantigen Aquaporin-4

Induktion af lammelse og visuelle System skade i mus af T celler specifik for Neuromyelitis Optica Autoantigen Aquaporin-4

While it is recognized that aquaporin-4 (AQP4)-specific T cells and antibodies participate in the pathogenesis of neuromyelitis optica (NMO), a human ...

Evaluation of Extracellular Vesicle Function During Malaria Infection

Evaluering af ekstracellulære vesikel funktion under Malaria infektion

Malaria is a life-threatening disease caused by Plasmodium parasites, with P. falciparum being the most prevalent on the African continent and responsible ...