Screening of Human Tumor Antigens for CD4+ T Cell Epitopes by Combination of HLA-Transgenic Mice, Recombinant Adenovirus and Antigen Peptide Libraries 英文参考文献.docVIP

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Screening of Human Tumor Antigens for CD4+ T Cell Epitopes by Combination of HLA-Transgenic Mice, Recombinant Adenovirus and Antigen Peptide Libraries 英文参考文献.doc

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ScreeningofHumanTumorAntigensforCD4TCellEpitopesbyCombinationofHLA-TransgenicMice,RecombinantAdenovirusandAntigenPeptideLibraries英文参考文献

ScreeningofHumanTumorAntigensforCD4+TCell EpitopesbyCombinationofHLA-TransgenicMice, RecombinantAdenovirusandAntigenPeptideLibraries WolframOsen1,SabineSoltek1.,MingxiaSong3.,BarbaraLeuchs2,JuliaSteitz4,ThomasTu¨ting5, StefanB.Eichmu¨ller1,Xuan-DucNguyen6,DirkSchadendorf3,AnnettePaschen3* 1Clinical Cooperation for Unit Dermato-Oncology, German Cancer Research Center, Heidelberg, Germany, 2Infection and Cancer Program, German Cancer Research Center, Heidelberg, Germany, 3Department of Dermatology, University Hospital Essen, Essen, Germany, 4Institute of Laboratory Animal Science and Experimental Surgery, RWTH-Aachen University, Aachen, Germany, 5Department of Dermatology, University of Bonn, Bonn, Germany, 6Institute for Immunology and Transfusion Medicine,Mannheim,Germany Abstract Background: As tumor antigen-specific CD4+ T cells can mediate strong therapeutic anti-tumor responses in melanoma patients we set out to establish a comprehensive screening strategy for the identification of tumor-specific CD4 T cell epitopessuitablefordetection,isolationandexpansionoftumor-reactiveTcellsfrompatients. + MethodsandFindings:ToscanthehumanmelanomadifferentiationantigensTRP-1andTRP-2forHLA-DRB1*0301-restricted CD4 Tcellepitopesweappliedthefollowingmethodology:SplenocytesofHLA-DRB1*0301-transgenicmiceimmunizedwith + recombinant adenovirus encoding TRP-1 (Ad5.TRP-1) or TRP-2 (Ad5.TRP-2) were tested for their T cell reactivity against combinatorialTRP-1-andTRP-2-specificpeptidelibraries.CD4 Tcellepitopesthusidentifiedwerevalidatedinthehuman systembystimulationofperipheralbloodmononuclearcells(PBMC)fromhealthydonorsandmelanomapatients.Usingthis + + strategyweobservedthatrecombinantAd5inducedstrongCD4 Tcellresponsesagainsttheheterologoustumorantigens.In Ad5.TRP-2-immunized mice CD4+ T cell reactivity was detected against the known HLA-DRB1*0301-restricted TRP-260–74 epitopeandagainstthenewepitopeTRP-2149–163.Importantly,humanTcellsspecificallyrecognizingtargetcellsloadedwi