TheMIT.HTMIDResearchProjectsaredependentonacontinuoussupplyofvirusesforexperimentalwork directedatdevelopingHumanTissueModelsforInfectiousDiseases.ThepurposeoftheVirologyCoreto providetheauthenticatedandquantifiedvirusesforthework.Authenticationreferstodefining:a)thesourceof thevirus,b)thenucleotidesequenceofthevirus,c)thecelltypeusedforpassageandamplifiction,d)the passagenumber,e)thestorageconditions.Quantifiedreferstodefiningthetitersofinfectiousvirusesby plaqueassayorflowcytometry,orbyquantifyingviralRNAgenomesbyqRTPCR.InresponsetoAI16022, weintendtotest2Dand3Dhumancerebralorganoidculturesusingseveralviruses?specifically,Dengue Virus,WestNileVirus,andZikavirus?moreover,weproposetousethe2Dand3Dculturestotestthe tropismsofanumberofotherneurotropicvirusesusingpseudotypedvesicularstomatitisviruses(VSV).The rigoroftheMIT.HTMIDCenterwillbeenhancedsignificantlybyperformingcarefulvalidationand quantificationoftheviruspreparations.Theflavivirusesaresignificantforthisprojectbecauseoftheir neurotropicproperties?moreover,theuseofpseudotypedVSVswillenabletheMIT.HTMIDCentertosafely evaluateselectagentvirusesusingthenearphysiologicalcerebralorganoids.Zikavirusstudiesaresignificant becauseZikahasbeenlinkedtothedevelopmentoffetalmicrocephaly.Dengue,Zika,andWestNileviruses areallmembersofthe?Flaviviridae?familyandareimportantpathogensthatcausesignificantmorbidityand mortality.Becausethethreeflaviviruseshavedifferentinfectionphenotypesandpathogenesisoutcomes,a comparisonofvirusreplicationinthe2Dand3Dcultureswillbeextremelyinformative,andwillprovide ongoingpositiveandnegativecontrolstoenhancetherigoroftheexperimentalwork.TheVirologyCorewill alsoberesponsibleforbiosafetyoversightsothatallpersonnelintheMIT.HTMIDCenterfollowguidelines requiredforusingBSL2andBSL3viruses.
