Name: in vivo imaging systems (ivis) detection of a neuro-invasive encephalitic virus
Uploaded: 2012-12-02T14:00:00
Description: Watch this Scientific Journal Video about In Vivo Imaging Systems IVIS Detection of a Neuro-Invasive Encephalitic Virus at JoVE.com

Institut for Translationel Sciences UTMB-NIH tilskud 1UL1RR029876-01 og Alisha Prather for hendes hjælp med videoredigering for dette håndskrift.

1. Animal Forberedelse <ol><li> Animal ankomst: Ved ankomsten til dyret biosikkerhed niveau 2 (ABSL2) faciliteter, dyr tillade minimum 2 dage til at akklimatisere sig til deres nye omgivelser. Efter denne hvileperiode, dyrene inspicere at vurdere deres helbred og generelle fysiske udseende. <ol><li> Når der anvendes fluorescerende reportere, er det vigtigt at placere alle animalske emner på en lucerne fri kost til at begrænse mængden af ​​GI autofluorescens og baggrund signal. </li></ol></li><li> Shave dyr: At...

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Selv om denne protokol dækker de billeddannende aspekter til in vivo analyse, er det vigtigt at erkende bioluminescerende vektor som en nøglefaktor for fremtidige undersøgelser. Vores anvendelse af TC83, en svækket vaccinestamme af VEEV, som en vektor til ekspression af luciferase sikrer, at store mængder af enzymet bliver produceret grund af den høje replikationshastighed af virusset i CNS som tidligere beskrevet 1-4. Mens tilsætning af en anden subgenomisk promotor og luciferasegenet resulte...

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in vivo imaging systems (ivis) detection of a neuro-invasive encephalitic virus

Moderne fremskridt i imaging-teknologi tilskynde til yderligere udvikling og finjustering i den måde viral forskning opnås. Oprindelig blev foreslået af Russel og Burch i Humes 3R (erstatning, begrænsning, forfining), udnyttelse af dyremodeller inden for videnskabelig forskning er under konstant pres for at finde nye metoder til at mindske dyrs forbrug og samtidig forbedre den videnskabelige nøjagtighed og hastighed. En stor udfordring for Humes skoleledere er imidlertid, hvordan man kan sikre undersøgelserne er statistisk præcis samtidig reducere dyresygdom sygelighed og samlede tal. Vaccineeffektivitet undersøgelser øjeblikket kræver et stort antal dyr for at blive betragtet som statistisk signifikant, og resulterer ofte i høj sygelighed og dødelighed endpoints til identifikation af immunbeskyttelse. Vi udnyttet in vivo imaging system (IVIS) i forbindelse med en ildflue bioluminescerende enzym til gradvis spore invasionen af centralnervesystemet (CNS) af en ningphalitic virus i en murin model. Typisk, som sygdommen skrider relativt langsomt, men virusreplikation er hurtig, især i CNS, og kan føre til en ofte dødelig udgang. Efter intranasal infektion af mus med TC83-Luc, modificeret en svækket venezuelansk hesteencefalitis-virus-stamme til udtrykker et luciferasegenet, er vi i stand til at visualisere virusreplikation i hjernen mindst tre dage før udviklingen af ​​kliniske sygdomssymptomer. Udnytte CNS invasion som en central encephalitiske udvikling af sygdom endpoint er vi i stand til hurtigt at identificere terapeutiske og vaccine beskyttelse mod TC83-Luc infektion, før de kliniske symptomer. Med IVIS teknologi er vi i stand til at demonstrere en hurtig og præcis test af narkotika lægemidler og vacciner og samtidig reducere antallet af dyr og sygelighed.

Med et genetisk modificeret virus,, TC83-luciferase så vi en stigning i bioluminescerende signalstyrke som virusreplikation bevæger sig fra den nasale region i de centrale CNS (figur 1). På grund af den høje virusreplikation hastighed, forventer vi at se høje niveauer af bioluminescerende signal (figur 2A) er afhængige af vektoren og dyret immunrespons på vektoren. Vi forventer dette signal stigning til fortsat at en top, mellem dagene 5-7 efter infektion, i kombination med virus...

Watch this Scientific Journal Video about In Vivo Imaging Systems IVIS Detection of a Neuro-Invasive Encephalitic Virus at JoVE.com

In Vivo Imaging Systems IVIS Detection of a Neuro-Invasive Encephalitic Virus

Anvendelse af luciferase og in vivo billeddannelse systemer (IVIS) som et hidtil ukendt middel til at identificere sygdomme endpoints før kliniske udvikling forekommer. IVIS har tilladt os at visualisere i realtid invasionen af ​​encephalitiske vira over flere dage, hvilket giver en mere præcis sygdom model for fremtidige undersøgelser. Det har også givet os mulighed for at identificere de potentielle beskyttende egenskaber af antivirale lægemidler og vacciner hurtigere end aktuelt anvendte dyremodeller. Evnen til at udnytte de enkelte dyr over flere tidspunkter sikrer reducerede dyrenes behov, omkostninger og samlet sygelighed til dyrene udnyttede sikre en mere human og mere videnskabelig hjælp af sygdom undersøgelse.

In vivo Imaging Systems (IVIS) Påvisning af en neuro-Invasive encephalitiske Virus

Modern advancements in imaging technology encourage further development and refinement in the way viral research is accomplished. Initially proposed by ...

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JoVE Journal

Immunology and Infection

In Vivo Imaging Systems (IVIS) Detection of a Neuro-Invasive Encephalitic Virus

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