title: Ex vivo induction of viral antigen-specific CD8 T cell responses using mRNA-electroporated CD40-activated B cells.
abstract: Cell-based immunotherapy, in which antigen-loaded antigen-presenting cells (APC) are used to elicit T cell responses, has become part of the search for alternative cancer and infectious disease treatments. Here, we report on the feasibility of using mRNA-electroporated CD40-activated B cells (CD40-B cells) as alternative APC for the ex vivo induction of antigen-specific CD8(+) T cell responses. The potential of CD40-B cells as APC is reflected in their phenotypic analysis, showing a polyclonal, strongly activated B cell population with high expression of MHC and co-stimulatory molecules. Flow cytometric analysis of EGFP expression 24 h after EGFP mRNA-electroporation showed that CD40-B cells can be RNA transfected with high gene transfer efficiency. No difference in transfection efficiency or postelectroporation viability was observed between CD40-B cells and monocyte-derived dendritic cells (DC). Our first series of experiments show clearly that peptide-pulsed CD40-B cells are able to (re)activate both CD8+ and CD4(+) T cells against influenza and cytomegalovirus (CMV) antigens. To demonstrate the ability of viral antigen mRNA-electroporated CD40-B cells to induce virus-specific CD8+ T cell responses, these antigen-loaded cells were co-cultured in vitro with autologous peripheral blood mononuclear cells (PBMC) for 7 days followed by analysis of T cell antigen-specificity. These experiments show that CD40-B cells electroporated with influenza M1 mRNA or with CMV pp65 mRNA are able to activate antigen-specific interferon (IFN)-gamma-producing CD8(+) T cells. These findings demonstrate that mRNA-electroporated CD40-B cells can be used as alternative APC for the induction of antigen-specific (memory) CD8(+) T cell responses, which might overcome some of the drawbacks inherent to DC immunotherapy protocols.
authors.affiliation: Laboratory of Experimental Hematology, Antwerp University Hospital (UZA), University of Antwerp (UA), Belgium.
authors.forename: G A
authors.initials: GA
authors.lastname: Van den Bosch
authors.forename: P
authors.initials: P
authors.lastname: Ponsaerts
authors.forename: G
authors.initials: G
authors.lastname: Nijs
authors.forename: M
authors.initials: M
authors.lastname: Lenjou
authors.forename: G
authors.initials: G
authors.lastname: Vanham
authors.forename: D R
authors.initials: DR
authors.lastname: Van Bockstaele
authors.forename: Z N
authors.initials: ZN
authors.lastname: Berneman
authors.forename: V F I
authors.initials: VF
authors.lastname: Van Tendeloo
mesh_terms: D000938:Antigen-Presenting Cells / Q000276:immunology*; D000956:Antigens, Viral / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology*; D019013:CD40 Antigens / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D018929:Cell Culture Techniques; D002460:Cell Line; D003587:Cytomegalovirus / Q000276:immunology; D018274:Electroporation; D049452:Green Fluorescent Proteins / Q000235:genetics; D006650:Histocompatibility Testing; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D008213:Lymphocyte Activation; D012333:RNA, Messenger / Q000032:analysis
chemical_list: D000956:Antigens, Viral; D019013:CD40 Antigens; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins
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