Glucocorticoid-Resistant B-Lymphoblast Cell Line Derived from the Bolivian Squirrel Monkey (Saimiri boliviensis boliviensis)

摘要

Thegoalofthestudyreportedherewastodevelopacontinuouscelllinefromthesquirrelmonkeythatexpressesthespecies-specificphenotypeofimpairedsensitivitytoglucocorticoids.ThirtymillilitersofbloodfromamaleBoliviansquirrelmonkey(Saimiriboliviensisboliviensis)wasfractionated,andthebuffycoatwasobtainedandincubatedinthepresenceofB95-8cell-conditionedmedium,anabundantsourceofEpstein-Barrvirus(EBV),and2gofcyclosporinA/ml.Cellgrowthwasdetectedwithin8weeks,afterwhichthecellswereclonedbyuseofthelimitingdilutionmethod.Oneclone(4D8)wascharacterizedindetail.ThechromosomalcountandG-bandingpatternconfirmedthatthecellswereofBoliviansquirrelmonkeyorigin.TheB-celloriginofthesecellswasindicatedbyelectronmicroscopicanalysisandwasconfirmedbyexpressionofCD20.ThecellsstainedstronglyforLMP1,amarkeroflatentEBVinfection,andoccasionallyforthelyticinfectionmarkerZEBRA(BZLF1).Theresponsivenessofclone4D8cellstoglucocorticoidswasdeterminedbycomparingtheeffectsofdexamethasoneoncellgrowthandtheinductionofaglucocorticoid-induciblemRNAin4D8cellswiththeeffectsonahumanEBV-transformedB-lymphoblastcellline(HL).DexamethasoneinhibitedthegrowthofHLcells,withICsubxmlns="http://pub2web.metastore.ingenta.coms/"50/subofapproximately9nM,buthadnoeffectonthegrowthof4D8cells.TheinductionofFK506-bindingproteinFKBP51mRNAbydexamethasonewasalsosignificantlybluntedin4D8cells.Thus,wehavedevelopedandcharacterizedasquirrelmonkeylympho-blasticcelllinederivedbytransformationofB-lymphocyteswithEBV;thecelllinehasdiminishedgrowthandtranscriptionalresponsestoglucocorticoids.