Name: opsono-adherence assay to evaluate functional antibodies in vaccine development against bacillus anthracis and other encapsulated pathogens
Uploaded: 2020-05-19T13:30:00
Description: Watch this Scientific Journal Video about Opsono-Adherence Assay to Evaluate Functional Antibodies in Vaccine Development Against Bacillus anthracis and Other Encapsulated Pathogens at JoVE.com

J. Chua, D. Chabot og A. Friedlander designet prosedyrene som er beskrevet i manuskriptet. J. Chua og T. Putmon-Taylor utførte eksperimentene. D. Chabot utførte dataanalyse. J. Chua skrev manuskriptet.
Forfatterne takker Kyle J. Fitts for utmerket teknisk assistanse.
Arbeidet ble støttet av Defense Threat Reduction Agency grant CBM. VAXBT.03.10.RD.015, plannummer 921175.
Meninger, tolkninger, konklusjoner og anbefalinger er forfatternes og er ikke nødvendigvis godkjent av den amerikanske hæren. Innholdet i denne publikasjonen gjenspeiler ikke nødvendigvis synspunktene eller retningslinjene til Forsvarsdepartementet, og nevner heller ikke varenavn, kommersielle produkter eller organisasjoner som innebærer støtte fra den amerikanske regjeringen.

I samsvar med dyrevelferdsloven, folkehelsetjenestepolitikken og andre føderale vedtekter og forskrifter knyttet til dyr og eksperimenter som involverer dyr, ble forskningen beskrevet her utført under en godkjent protokoll for institusjonell dyreomsorgs- og brukskomité. Anlegget der denne forskningen ble utført er akkreditert av Association for Assessment and Accreditation of Laboratory Animal Care, International og overholder prinsippene som er angitt i Veiledningen for omsorg og bruk av laboratoriedyr, National Res...

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T., 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=Use+of+highly+encapsulated+Streptococcus+pneumoniae+strains+in+a+flow-cytometric+assay+for+assessment+of+the+phagocytic+capacity+of+serotype-specific+antibodies.">Use of highly encapsulated Streptococcus pneumoniae strains in a flow-cytometric assay for assessment of the phagocytic capacity of serotype-specific antibodies.</a> Clinical and Diagnostic Laboratory Immunology. 5 (5), 703-710 (1998).</li><li>Wang, T. T., Fellows, P. F., Leighton, T. J., Lucas, A. H. <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+opsonic+antibodies+to+the+gamma-D-glutamic+acid+capsule+of+Bacillus+anthracis+by+immunization+with+a+synthetic+peptide-carrier+protein+conjugate.">Induction of opsonic antibodies to the gamma-D-glutamic acid capsule of Bacillus anthracis by immunization with a synthetic peptide-carrier protein conjugate.</a> FEMS Immunology and Medical Microbiology. 40 (3), 231-237 (2004).</li><li>Chabot, D. 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=Protection+of+rhesus+macaques+against+inhalational+anthrax+with+a+Bacillus+anthracis+capsule+conjugate+vaccine.">Protection of rhesus macaques against inhalational anthrax with a Bacillus anthracis capsule conjugate vaccine.</a> Vaccine. 34 (34), 4012-4016 (2016).</li><li>Chabot, D. 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=Efficacy+of+a+capsule+conjugate+vaccine+against+inhalational+anthrax+in+rabbits+and+monkeys.">Efficacy of a capsule conjugate vaccine against inhalational anthrax in rabbits and monkeys.</a> Vaccine. 30 (5), 846-852 (2012).</li><li>Chua, 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=Formaldehyde+and+Glutaraldehyde+Inactivation+of+Bacterial+Tier+1+Select+Agents+in+Tissues.">Formaldehyde and Glutaraldehyde Inactivation of Bacterial Tier 1 Select Agents in Tissues.</a> Emerging Infectious Diseases. 25 (5), 919-926 (2019).</li><li>. Guide for the Care and Use of Laboratory Animals (Eighth Edition) Available from: <a target="_blank" href="https://grants.nih.gov/grants/olaw/guide-for-the-care-and-use-of-laboratory-animals.pdf">https://grants.nih.gov/grants/olaw/guide-for-the-care-and-use-of-laboratory-animals.pdf</a> (2011)</li><li>Durando, P., 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=Experience+with+pneumococcal+polysaccharide+conjugate+vaccine+(conjugated+to+CRM197+carrier+protein)+in+children+and+adults.">Experience with pneumococcal polysaccharide conjugate vaccine (conjugated to CRM197 carrier protein) in children and adults.</a> Clinical Microbiology and Infection. 19, 1-9 (2013).</li><li>Adams, W. 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V., Middleton, D. R., Avci, F. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Opsonophagocytic+Killing+Assay+to+Assess+Immunological+Responses+Against+Bacterial+Pathogens.">Opsonophagocytic Killing Assay to Assess Immunological Responses Against Bacterial Pathogens.</a> Journal of Visualized Experiments. (146), e59400 (2019).</li><li>Ezzell, J. W., Welkos, S. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+capsule+of+Bacillus+anthracis,+a+review.">The capsule of Bacillus anthracis, a review.</a> Journal of Applied Microbiology. 87 (2), 250 (1999).</li><li>Hanna, P. C., Acosta, D., Collier, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=On+the+role+of+macrophages+in+anthrax.">On the role of macrophages in anthrax.</a> Proceedings of the National Academies of Sciences of the United States of America. 90 (21), 10198-10201 (1993).</li><li>Fleck, R. A., Romero-Steiner, S., Nahm, M. 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J., Radu, C. G., Ghetie, V., Ward, E. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differences+in+promiscuity+for+antibody-FcRn+interactions+across+species:+implications+for+therapeutic+antibodies.">Differences in promiscuity for antibody-FcRn interactions across species: implications for therapeutic antibodies.</a> International Immunology. 13 (12), 1551-1559 (2001).</li><li>Sorman, A., Zhang, L., Ding, Z., Heyman, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=How+antibodies+use+complement+to+regulate+antibody+responses.">How antibodies use complement to regulate antibody responses.</a> Molecular Immunology. 61 (2), 79-88 (2014).</li><li>Henckaerts, I., Durant, N., De Grave, D., Schuerman, L., Poolman, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Validation+of+a+routine+opsonophagocytosis+assay+to+predict+invasive+pneumococcal+disease+efficacy+of+conjugate+vaccine+in+children.">Validation of a routine opsonophagocytosis assay to predict invasive pneumococcal disease efficacy of conjugate vaccine in children.</a> Vaccine. 25 (13), 2518-2527 (2007).</li><li>Wolf, J. 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Kapselbaserte vaksiner har vist seg å være effektive mot mange bakterielle patogener, og mange er lisensiert for bruk hos mennesker25,26,27. Disse vaksinene virker ved å generere antistoffer rettet motkapselen,og mange av disse studiene bruker OPKA til å vise opsono-fagocytiske funksjoner av antistoffene13,14,16,<sup clas...

Anerkjennelse, etterlevelse, internalisering og nedbrytning av et patogen er integrert i fagocytose1, en fremtredende vei i verten medfødt immunrespons først beskrevet av Ilya Metchnikoff i 18832,3. Fagocytiske leukocytter, så vel som andre celler i immunsystemet, er svært diskriminerende i deres valg av mål; de er i stand til å skille mellom "smittsomme ikke-selv" og "ikke-smittsomme selv" gjennom patogene assosiert molekylære mønstre ved deres repertoar av mønstergjenkjenning reseptorer (PRRs)4,5. Vertsgjenkjenning av et patogen kan også oppstå med binding av vertsopsoniner, for eksempel komplement ogantistoffer 6. Denne prosessen, kalt opsonisering, strøk patogenet med disse molekylene, og forbedrer internaliseringen ved binding til opsonisk reseptorer (f.eks. komplement og Fc reseptorer) på fagocytiske celler6. For at et patogen skal følge en fagocyte, er det nødvendig med kollektiv binding av flere reseptorer med sine kognate ligande. Først da kan etterlevelse utløse og opprettholde signalkaskader inne i vertscellen for å starte internalisering6.
På grunn av viktigheten av fagocytose i clearance av patogener og forebygging av infeksjon, har ekstracellulære patogener utviklet mange måter å undergrave denne prosessen for å forlenge overlevelsen. En strategi av betydning er produksjonen av en anionisk polymer (f.eks polysakkarid eller polyaminosyre) kapsel som er anti-fagocytisk i kraft av sin ladning, er dårlig immunogen, og beskytter molekyler på bakteriell konvolutt fra PRRs6,7. Patogener som Cryptococcus neoformans og Streptococcus pneumoniae har kapsler bestående av sakkaridpolymerer, mens Staphylococcus epidermidis og noen Bacillus arter produserer poly-ɣ-glutaminsyre (PGGA)7,8. Likevel produserer andre patogener kapsler som ligner det ikke-smittsomme selvet. For eksempel har Streptococcus pyogenes og en patogen stamme av B. cereus en hyaluronsyrekapsel som ikke bare er anti-phagocytic, men som heller ikke kan gjenkjennes som fremmed avimmunsystemet 9,10.
Bøyning av kapsel til bærerproteiner omdannes dem fra fattige, T-uavhengige antigener til svært immunogene T-avhengige antigener som kan indusere høy serum anti-kapsel antistoff titers11,12. Denne strategien er ansatt for lisensierte vaksiner mot S. pneumoniae, Haemophilus influenzae,og Neisseria meningitides11. De opsonisk aktivitetene til antikapselantistoffer har ofte blitt evaluert av opsono-fagocytiske drapsanalyser (OPKA)13,14,15,16. Disse analysene tester om funksjonelle antistoffer kan utløse fagocytose og drepe14. Imidlertid er bruk av OPKA med smittsomme patogener, som Tier 1 Biological Select Agents og Toxins (BSAT), inkludert B. anthracis17,farlig og utgjør sikkerhetsrisiko; disse analysene nødvendiggjør omfattende håndtering av en utvalgt agent. Utvalgte agenthåndtering kan bare gjøres i begrensede biosikkerhetsnivå 3 (BSL-3) laboratorier; arbeid på disse områdene krever langvarige driftsprosedyrer på grunn av de mange sikkerhets- og sikkerhetsforanstaltningene som må følges. BSL-3 laboratorier er heller ikke utstyrt med spesialutstyr som brukes til OPKA arbeid, for eksempel mikroskoper og cytometere. Dermed utviklet vi en alternativ analyse basert på bruk av inaktiverte bakterier18,19. Vi refererer til dette som en opsono-overholdelse analyse (OAA) som ikke er avhengig av internalisering og drap som analyse utganger; I stedet brukes overholdelse av opsoniserte inaktiverte patogener som en indeks av fagocytose. Mekanistisk er OAA en passende erstatning fordi etterlevelse skjer en priori og er nært sammenvevd med internalisering og intracellulært drap. Fra et biosikkerhetsperspektiv foretrekkes OAA fordi det krever minimal håndtering av et smittsomt middel, er eksperimentelt kortere varighet enn OPKA, og kan utføres i BSL-2-laboratorier etter at en bestand av det inaktiverte patogenet er produsert og overført.
Vi demonstrerer utnyttelsen av OAA for å undersøke opsonisk funksjon av antikapsel antistoffer som finnes i sera av ikke-menneskelige primater (NHPs) vaksinert med en kapsel konjugat [det vil si PGGA fra B. anthracis konjugert til ytre membran protein kompleks (OMPC) av Neisseria meningitides]20. Serum opsonisert bacilli ble inkubert med en tilhengermus makrofag cellelinje, RAW 264.7. Etter fiksering ble cellemonolaget og tilhengerbakilli avbildet av fluorescensmikroskopi. Bakteriell etterlevelse økte når bacilli ble inkubert med serum fra NHPs vaksinert med kapselkonjugatet sammenlignet med kontrollserum20. Overholdelse korrelert med overlevelse av miltbrann utfordring20,21. Dermed karakteriserte bruken av OAA funksjonen til antikapselantistoffer og sterkt tilrettelagt testing av vår vaksinekandidat.

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			<td height="20" style="height:20px;width:380px;">0.20 &#181;m syringe filter (25mm, regenerated cellulose)</td>
			<td style="width:483px;">Corning, Corning, NY</td>
			<td style="width:267px;">431222</td>
			<td style="width:391px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">10 mL syringe (Luer-Lok tip)</td>
			<td>BD, Franklin Lakes, NJ</td>
			<td>302995</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">15&#181; 96 well black plates (plate #1 for imaging)</td>
			<td>In Vitro Scientific, Sunnyvale, CA</td>
			<td>P96-1-N</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">16% paraformaldehyde</td>
			<td>Electron Microscopy Science, Hatfield, PA</td>
			<td>15710</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">75 cm sq. tissue culture treated flask</td>
			<td>Corning, Corning, NY</td>
			<td>430641</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Agar (powder)</td>
			<td>Sigma-Aldrich, St. Louis, MO</td>
			<td>A1296</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Baby Rabbit Complement</td>
			<td>Cedarlane Labs, Burlington, NC</td>
			<td>CL3441</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Bacto Yeast Extract</td>
			<td>BD, Sparks, MD</td>
			<td>288620</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">BBL Brain Heart Infusion (BHI)</td>
			<td>BD, Sparks, MD</td>
			<td>211059</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Blood Agar (TSA with Sheep Blood) plates</td>
			<td>Remel, Lenexa, KS</td>
			<td>R01198</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Cell scraper</td>
			<td>Sarstedt, Newton, NC</td>
			<td>83.183</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Costar 96 well cell culture plates (plates #2 &#38; 3 for dilutions)</td>
			<td>Corning, Corning, NY</td>
			<td>3596</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Cover glass</td>
			<td>Electron Microscopy Science, Hatfield, PA</td>
			<td>72200-10</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Difco Nutrient Broth</td>
			<td>BD, Sparks, MD</td>
			<td>234000</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Dulbecco&#39;s Modified Eagle Medium (DMEM), high glucose</td>
			<td>Gibco, Thermo Fisher Scientific, Waltham, MA</td>
			<td>11965-092</td>
			<td>contains 4500 mg/L glucose, 4 mM L-glutamine, Phenol Red</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">EVOS FL Auto Cell Imaging System (fluorescence microscope)</td>
			<td>Life Technologies, Thermo Fisher Scientific, Waltham, MA</td>
			<td>AMAFD1000</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Fetal Bovine Serum</td>
			<td>Hyclone, GE Healthcare Life Sciences, South Logan, UT</td>
			<td>SH30071.03</td>
			<td>not gamma irradiated, not heat inactivated</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Fluorescein isothiocyanate</td>
			<td>Invitrogen, Thermo Fisher Scientific, Waltham, MA</td>
			<td>F143</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">HCS Cell Mask Orange Cell Stain</td>
			<td>Invitrogen, Thermo Fisher Scientific, Waltham, MA</td>
			<td>H32713</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">hemocytometer (Improved Neubauer)</td>
			<td>Hausser Scientific, Horsham, PA</td>
			<td>3900</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">India Ink solution</td>
			<td>BD, Sparks, MD</td>
			<td>261194</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">L- glutamine (200 mM)</td>
			<td>Gibco, Thermo Fisher Scientific, Waltham MA</td>
			<td>25030081</td>
			<td>supplement medium with additional 2mM L-glutamine</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Nikon Eclipse TE2000-U (inverted compound microscope)</td>
			<td>Nikon Instruments, Melville, NY</td>
			<td>TE2000</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">PBS without Calcium or Magnesium</td>
			<td>Lonza, Walkersville, MD</td>
			<td>17-516F</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Penicillin-Streptomycin Solution, 100x</td>
			<td>Hyclone, GE Healthcare Life Sciences, South Logan, UT</td>
			<td>SV30010</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">petri dishes (100 x 15 mm)</td>
			<td>Falcon, Corning, Durham, NC</td>
			<td>351029</td>
			<td>for agar plates</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">RAW 264.7 macrophage cell line (Tib47)</td>
			<td>ATCC, Manassas, VA</td>
			<td>ATCC TIB-71</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Slides</td>
			<td>VWR, Radnor, PA</td>
			<td>16004-422</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Sodium Bicarbonate</td>
			<td>Sigma-Aldrich, St. Louis, MO</td>
			<td>S5761</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Trypan Blue Solution (0.4%)</td>
			<td>Sigma-Aldrich, St. Louis, MO</td>
			<td>T8154</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Zeiss 700 Laser Scanning Microscopy (confocal microscope)</td>
			<td>Carl Zeiss Microimaging, Thornwood, NY</td>
			<td>4109001865956000</td>
			<td></td>
		</tr>
	</tbody>
</table>

opsono-adherence assay to evaluate functional antibodies in vaccine development against bacillus anthracis and other encapsulated pathogens

Opsono-etterlevelsesanalysen er en funksjonell analyse som nummererer vedlegget av bakterielle patogener til profesjonelle fagocytter. Fordi overholdelse er nødvendig for fagocytose og drap, er analysen en alternativ metode for opsono-phagocytic drapsanalyser. En fordel med opsono-overholdelsesanalysen er muligheten til å bruke inaktiverte patogener og pattedyrcellelinjer, noe som tillater standardisering på tvers av flere eksperimenter. Bruken av et inaktivert patogen i analysen letter også arbeid med biosikkerhetsnivå 3 smittsomme stoffer og andre virulent patogener. I vårt arbeid ble opsono-adherence analysen brukt til å vurdere antistoffenes funksjonelle evne, fra sera av dyr vaksinert med en miltbrannkapselbasert vaksine, for å indusere overholdelse av fast Bacillus-anthracis til en musemakrofagcellelinje, RAW 264.7. Automatisert fluorescensmikroskopi ble brukt til å ta bilder av bacilli som følge makrofager. Økt etterlevelse var korrelert med tilstedeværelsen av antistoffer mot kapsel i serumet. Ikke-menneskelige primater som viste høye serum anti-kapsel antistoffkonsentrasjoner ble beskyttet mot miltbrann utfordring. Dermed kan opsono-overholdelsesanalysen brukes til å belyse de biologiske funksjonene til antigenspesifikke antistoffer i sera, for å evaluere effekten av vaksinekandidater og andre terapeutiske midler, og for å tjene som en mulig korrelering av immunitet.

Denne delen viser representative mikrografer samlet under et OAA-eksperiment sammen med resultater som viser at OAA kan brukes til å undersøke antistoffenes biologiske funksjon. Her ble analysen vellykket brukt til å evaluere effekten av en miltbrannvaksinekandidat. Det er viktig å verifisere tilstanden av innkapsling på bacilli som lite eller ingen innkapsling fører dem til å følge vertsceller, produsere en høy bakgrunn. Figur 1 er et bilde av B. anthracis Ames negativt fa...

Watch this Scientific Journal Video about Opsono-Adherence Assay to Evaluate Functional Antibodies in Vaccine Development Against Bacillus anthracis and Other Encapsulated Pathogens at JoVE.com

Opsono-Adherence Assay to Evaluate Functional Antibodies in Vaccine Development Against Bacillus anthracis and Other Encapsulated Pathogens

Opsono-adherence analysen er en alternativ metode for opsono-fagocytisk drapsanalyse for å evaluere opsonisk funksjoner av antistoffer i vaksineutvikling.

Opsono-overholdelse analyse for å evaluere funksjonelle antistoffer i vaksineutvikling mot Bacillus anthracis og andre innkapslede patogener

The opsono-adherence assay is a functional assay that enumerates the attachment of bacterial pathogens to professional phagocytes. Because adherence is ...

Research

JoVE Journal

Immunology and Infection

Opsono-Adherence Assay to Evaluate Functional Antibodies in Vaccine Development Against Bacillus anthracis and Other Encapsulated Pathogens

Passive Administration of Monoclonal Antibodies Against H. capsulatum and Others Fungal Pathogens

Passive Administration of Monoclonal Antibodies Against H. capsulatum and Others Fungal Pathogens

The purpose of the use of this methodology is 1) to advance our capacity to protect individuals with antibody or vaccine for preventing or treating ...

Generation of Recombinant Arenavirus for Vaccine Development in FDA-Approved Vero Cells

Generasjon av rekombinant arenavirus for Vaccine Development i FDA-godkjente Vero Cells

The development and implementation of arenavirus reverse genetics represents a significant breakthrough in the arenavirus field  4. The use of cell-based ...

Using Click Chemistry to Measure the Effect of Viral Infection on Host-Cell RNA Synthesis

Ved hjelp Klikk kjemi å måle effekten av virusinfeksjon på Host-Cell RNA syntese

Many RNA viruses have evolved the ability to inhibit host cell transcription as a means to circumvent cellular defenses. For the study of these viruses, ...

Cell-based Flow Cytometry Assay to Measure Cytotoxic Activity

Cytolytic activity of CD8+ T cells is rarely evaluated. We describe here a new cell-based assay to measure the capacity of antigen-specific CD8+ T cells ...

A High-throughput Compatible Assay to Evaluate Drug Efficacy against Macrophage Passaged Mycobacterium tuberculosis

A High-throughput Compatible Assay to Evaluate Drug Efficacy against Macrophage Passaged Mycobacterium tuberculosis

En High-throughput Kompatibel Assay å evaluere Drug Effekt mot macrophage passert<em&gt; Mycobacterium tuberculosis</em

The early drug development process for anti-tuberculosis drugs is hindered by the inefficient translation of compounds with in vitro activity to ...

Generation of Monoclonal Antibodies Against Natural Products

Generasjon av monoklonale antistoffer mot naturlige produkter

The analysis of the bioactive components present in foods and natural products has become a popular area of study in many fields, including traditional ...

Opsonophagocytic Killing Assay to Assess Immunological Responses Against Bacterial Pathogens

Opsonophagocytic drepe analysen å vurdere immunologiske svar mot bakteriell patogener

A key aspect of the immune response to bacterial colonization of the host is phagocytosis. An opsonophagocytic killing assay (OPKA) is an experimental ...

Using Human Induced Pluripotent Stem Cell-derived Intestinal Organoids to Study and Modify Epithelial Cell Protection Against Salmonella and Other Pathogens

Using Human Induced Pluripotent Stem Cell-derived Intestinal Organoids to Study and Modify Epithelial Cell Protection Against Salmonella and Other Pathogens

Benytter Human indusert pluripotent stilk cellen-avledet intestinal Organoids å studere og endre epitel cellen sikringen imot Salmonella og annet patogener

The intestinal &#8216;organoid&#8217; (iHO) system, wherein 3-D structures representative of the epithelial lining of the human gut can be produced from ...

In Vitro ELISA Test to Evaluate Rabies Vaccine Potency

In Vitro ELISA Test for å evaluere Rabies vaksine styrke

The growing global concern for the animal welfare is encouraging manufacturers and the National Control Laboratories (OMCLs) to follow the 3Rs strategy ...

Use of an Influenza Antigen Microarray to Measure the Breadth of Serum Antibodies Across Virus Subtypes

Bruk av en influensa antigen Microarray å måle bredden av serum antistoffer over virus under typer

The influenza virus remains a significant cause of mortality worldwide due to the limited effectiveness of currently available vaccines. A key challenge ...