title: T cell response to human papillomavirus 16 E7 in mice: comparison of Cr release assay, intracellular IFN-gamma production, ELISPOT and tetramer staining.
abstract: Successful vaccination against infections by high-risk papillomaviruses aiming at the prevention of cervical cancer most likely requires the induction of neutralizing antibodies and human papillomavirus (HPV)-specific T cells directed against early viral proteins such as E7. Whereas the technology for detection of antibodies is well established, measurement of T cells is more cumbersome and standardization of assays is difficult. By using chromium release assay, ELISPOT, tetramer staining and intracellular IFN-gamma assay, we compared the levels of HPV 16 E7-specific T cells obtained after immunization of C57BL/6 mice with different DNA expression vectors. We found that all four assays gave highly comparable results. ELISPOT can be recommended for future studies as it indicates the presence of activated (i.e. IFN-gamma-secreting) T cells in a quantitative manner and combines high sensitivity with relatively low T cell demand.
authors.affiliation: Deutsches Krebsforschungszentrum, Heidelberg, Germany.
authors.forename: Nico
authors.initials: N
authors.lastname: Michel
authors.forename: Peter
authors.initials: P
authors.lastname: Ohlschläger
authors.forename: Wolfram
authors.initials: W
authors.lastname: Osen
authors.forename: Eva-Jasmin
authors.initials: EJ
authors.lastname: Freyschmidt
authors.forename: Heidrun
authors.initials: H
authors.lastname: Guthöhrlein
authors.forename: Andreas M
authors.initials: AM
authors.lastname: Kaufmann
authors.forename: Martin
authors.initials: M
authors.lastname: Müller
authors.forename: Lutz
authors.initials: L
authors.lastname: Gissmann
mesh_terms: D000818:Animals; D003601:Cytotoxicity Tests, Immunologic; D007371:Interferon-gamma / Q000096:biosynthesis*; D051379:Mice; D008810:Mice, Inbred C57BL; D009856:Oncogene Proteins, Viral / Q000276:immunology*; D027383:Papillomaviridae / Q000276:immunology*; D050725:Papillomavirus E7 Proteins; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D019444:Vaccines, DNA / Q000276:immunology*; D014765:Viral Vaccines / Q000276:immunology*
chemical_list: D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D019444:Vaccines, DNA; D014765:Viral Vaccines; C059731:oncogene protein E7, Human papillomavirus type 16; D007371:Interferon-gamma