人鼻粘膜类器官EB病毒感染模型的建立

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Abstract：ObjectiveTodevelopancost-effectiveandconvenient methodforculturing humannasalorganoids toestablishan in vitroEpstein-Barrvirus （EBV）infection model.Methods Nasal polyptisueobtained fromsurgerywasroutinely washed， cut，digestedandfiedtoainellustersecellstesretexpaeditoietedsaanis throughdynamicsuspensioncultureundermatrix-freeconditions，followedbya14-daydiferentiationinductiontreatmento obtaindiferentiatednasalorganoids.Themajorcellularcomponentsoftheorganoidswereidentifiedbyimmunofluorescence stainingandimmunohistochemistry.ThenasalorganoidswereinfectedbyEBVinvitroandviralreplicationwasverifiedby detecting the expresions of the virus-specific genes EBNA1 and BALF5 using RT-qPCR and immunofluorescence staining. ResultsNasal organoidsconsisting ofbasalcells，mucouscells，andciliatedcellsinamartigel-free systemwereobtained successfully bydynamicsuspensionculture.Tediferentiatednasal organoids expressedhigh levelsofEBV-associated receptorsEphA2，NRP1，andNMHI-A.Boththeundierentiatednderentiatednasalorganoidsouldbifetedby EBV.Viralreplicationinterganoidsicreasedwiththeviralexposureload，andthediferentiatedrganoidsappeardore permissive toviralreplication.Conclusion The matrigel-free dynamic suspension culture methd is economicaland simple forconstructingnasalorganoids，whichcanbeusedasamodelofEBVinfectionforstudiesofepithelialEBVinfection. Kevwords： organoids; Epstein-Barr virus; nasal mucosa

EB病毒（EBV）也称为人类疱疹病毒4（HHV-4），是γ⋅ -疱疹病毒家族的成员，于1964年在非洲儿童的伯基特淋巴瘤中被首次发现。（剩余12945字）