Efficient presentation of known HLA class II-restricted MAGE-A3 epitopes by dendritic cells electroporated with messenger RNA encoding an invariant chain with genetic exchange of class II-associated invariant chain peptide.
For the induction of an optimal immune response against cancer or infections not only CD8(+) CTLs but also CD4(+) T helper cells must be induced, in particular IFN-gamma-secreting type 1 T helper cells. Several strategies have been explored to target tumor-associated antigens to the HLA class II processing compartments. We engineered a genetic construct encoding an invariant chain (Ii) protein where the CLIP region has been replaced by sequences encoding HLA class II-restricted MAGE-A3 epitopes. Monocyte-derived dendritic cells (DCs) were electroporated with in vitro transcribed mRNA encoding a modified Ii protein containing the HLA-DP4-restricted MAGE-A3 epitope. The capacity of these electroporated DCs to stimulate a MAGE-A3-specific T-cell clone was compared at different stages of DC maturation with the T-cell stimulatory capacity of DCs pulsed with the synthetic peptide. After electroporation, the T-cell stimulatory capacity was shown to be high and long lasting, whereas the stimulatory capacity of peptide-pulsed DCs decreased rapidly. Upon coculture with epitope-specific T cells, electroporated immature DCs expressed enhanced levels of costimulatory molecules, HLA class II molecules, and CD83, suggesting the induction of maturation. The electroporated DCs can be frozen and thawed without losing their capability to stimulate the specific T-cell clone in vitro, and they are able to stimulate unprimed CD4(+) T cells specific to the HLA-DP4-restricted MAGE-A3 epitope in vitro. Similar results were obtained with a recombinant Ii containing the MAGE-A3 epitope presented in the context of HLA-DR13.
D000595:Amino Acid Sequence; D017951:Antigen Presentation / Q000235:genetics / Q000276:immunology*; D000944:Antigens, Differentiation, B-Lymphocyte / Q000235:genetics / Q000276:immunology*; D000951:Antigens, Neoplasm / Q000235:genetics / Q000276:immunology*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D003713:Dendritic Cells / Q000276:immunology*; D018274:Electroporation; D000939:Epitopes / Q000235:genetics / Q000276:immunology; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D006682:HLA-DP Antigens / Q000235:genetics / Q000276:immunology*; D059868:HLA-DP beta-Chains; D006684:HLA-DR Antigens / Q000235:genetics / Q000276:immunology*; D059810:HLA-DR Serological Subtypes; D000949:Histocompatibility Antigens Class II / Q000235:genetics / Q000276:immunology*; D006801:Humans; D016130:Immunophenotyping; D008213:Lymphocyte Activation / Q000276:immunology; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins; D012333:RNA, Messenger / Q000235:genetics / Q000276:immunology
D000944:Antigens, Differentiation, B-Lymphocyte; D000951:Antigens, Neoplasm; D000939:Epitopes; D018984:Epitopes, T-Lymphocyte; D006682:HLA-DP Antigens; D059868:HLA-DP beta-Chains; C056284:HLA-DPw4 antigen; D006684:HLA-DR Antigens; D059810:HLA-DR Serological Subtypes; C097337:HLA-DR13 antigen; D000949:Histocompatibility Antigens Class II; C072013:MAGEA3 protein, human; D009363:Neoplasm Proteins; D012333:RNA, Messenger; C045084:invariant chain