Ex-vivo Clonally Expanded B Lymphocytes Infiltrating Colorectal Carcinoma Are of Mature Immunophenotype and Produce Functional IgG 英文参考文献.docVIP

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Ex-vivo Clonally Expanded B Lymphocytes Infiltrating Colorectal Carcinoma Are of Mature Immunophenotype and Produce Functional IgG 英文参考文献.doc

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Ex-vivo Clonally Expanded B Lymphocytes Infiltrating Colorectal Carcinoma Are of Mature Immunophenotype and Produce Functional IgG 英文参考文献

Ex-vivoClonallyExpandedBLymphocytesInfiltrating ColorectalCarcinomaAreofMatureImmunophenotype andProduceFunctionalIgG ClaudiaMaletzki1.,AnnikaJahnke1.,ChristianeOstwald2,ErnstKlar1,FriedrichPrall2 ,Michael Linnebacher1* 1DivisionofMolecularOncologyandImmunotherapy,DepartmentofGeneralSurgery,UniversityofRostock,Rostock,Germany,2InstituteofPathology,Universityof Rostock,Rostock,Germany Abstract Background:TumorinfiltratingBcells(TiBc)havenotyetbeeninvestigatedindetail.Thismayatleastinpartbedueto technicaldifficulties.HerewedescribeastraightforwardandreproduciblemethodtoisolateandcultureTiBcfromprimary colorectalcarcinomas(CRC). Methods/Results:TiBcculturesweregeneratedbyEpstein-Barrvirus(EBV)immortalization.Withthismethod,monoclonal TiBc cultures were obtained for 14/19 CRCs. As assessed by flow cytometry and ELISA, TiBc showed an activated immunophenotype(CD23+,CD80 )andproducedimmunoglobulin(Ig;IgGsecretionin55%ofthecultures).Infunctionalin vitroanalysis,mostoftheIgGsspecificallyboundtoallogeneicCRCtargetcells.ThesedatasuggestthatTiBcareantigen- experiencedandthusmayexhibitfunctionalityinsitu.Additionally,mini-culturesgeneratedfrom12furtherCRCsrevealed TiBcoutgrowthexclusivelyinthepresenceofEBV. + Conclusion:Insummary,thissimplemethodprovidesacellulartoolandourdatasetthestageforanalysingthebivalent role of TiBc; being antigen-presenting cells on the one hand and tumor-specific antibody producers on the other. Additionally,thegenerationoflong-termTiBcculturesandtheirmonoclonalIgmayservetoidentifynoveltumor-specific antigens. Citation:MaletzkiC,JahnkeA,OstwaldC,KlarE,PrallF,etal.(2012)Ex-vivoClonallyExpandedBLymphocytesInfiltratingColorectalCarcinomaAreofMature ImmunophenotypeandProduceFunctionalIgG.PLoSONE7(2):e32639.doi:10.1371/journal.pone.0032639 Editor:AnnM.Moormann,UniversityofMassachusettsMedicalSchool,UnitedStatesofAmerica ReceivedNovember15,2011;AcceptedJanuary28,2012;PublishedFebruary29,2012 Copyright: ? 2012 Maletzki et al. This is an open-access article