title: In vitro activation of CMV-specific human CD8(+) T cells by adenylate cyclase toxoids delivering pp65 epitopes.
abstract: Human CMV infects between 50-85% of healthy individuals and can cause live-threatening infections in immunocompromised patients. Therefore, peptide vaccination is being developed as a promising immunotherapeutic approach for treatment of patients at risk of CMV disease. The enzymatically inactive toxoid of Bordetella adenylate cyclase (CyaA-AC(-)) was shown to be an efficient tool for delivery of peptide epitopes and stimulation of Ag-specific T-cell immune responses. We investigated here the capacity of two CyaA-AC(-) constructs to deliver epitopes derived from the CMV phosphoprotein pp65 for activation of human T cells in vitro. Expansion of γ-IFN-secreting CMV-specific CD8(+) T cells, as well as increase of total IFN-γ and TNF-α production by PBMCs from CMV-seropositive donors were observed after in vitro stimulation with CyaA-AC(-) constructs carrying CMV epitopes, whereas limited activation of immune response occurred with free peptides. The activation of immune response was confirmed by expansion of CMV-specific T-cell clones and anti-CMV cytotoxic effect of stimulated PBMCs. These data open the way to clinical evaluation of CyaA-AC(-) constructs as tools for detection and expansion of CMV-specific T-cell immune responses for diagnostic and immunotherapeutic applications against CMV-associated diseases.
authors: University Cell Immunotherapy Center, Babak Research Institute, Faculty of Medicine, Masaryk University, Brno, Czech Republic. jjelinekucbi@email.cz
authors: J
authors: J
authors: Jelinek
authors: I
authors: I
authors: Adkins
authors: Z
authors: Z
authors: Mikulkova
authors: J
authors: J
authors: Jagosova
authors: R
authors: R
authors: Pacasova
authors: S
authors: S
authors: Michlickova
authors: P
authors: P
authors: Sebo
authors: J
authors: J
authors: Michalek
mesh_terms: D000262:Adenylyl Cyclases / Q000235:genetics / Q000276:immunology*; D000595:Amino Acid Sequence; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D003587:Cytomegalovirus / Q000276:immunology*; D022482:Cytomegalovirus Vaccines / Q000235:genetics / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D006801:Humans; D008213:Lymphocyte Activation; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology; D010750:Phosphoproteins / Q000276:immunology*; D014763:Viral Matrix Proteins / Q000276:immunology*
chemical_list: D022482:Cytomegalovirus Vaccines; D018984:Epitopes, T-Lymphocyte; D010446:Peptide Fragments; D010750:Phosphoproteins; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa; D000262:Adenylyl Cyclases