title: An in vitro immune response model to determine tetanus toxoid antigen (vaccine) specific immunogenicity: Selection of sensitive assay criteria.
abstract: Many vaccines employed in childhood vaccination programmes are produced by conventional techniques, resulting in complex biological mixtures for which batch-related quality control requires in vivo potency testing. Monitoring consistency via in vitro tests during the vaccine production has the capacity to replace certain of the in vivo methods. In this respect, determining vaccine antigen immunogenicity through functional immunological tests has high potential. Advances in immunology have made it possible to analyse this biological activity by in vitro means. The present study established such an in vitro test system for tetanus toxoid (TT). This measured vaccine immunogenicity through an antigen-specific secondary (recall) response in vitro, using a porcine model growing in value for its closeness to human immune response characteristics. Discrimination between the specific recall TT antigen and diphtheria toxoid (DT) was possible using both peripheral blood mononuclear cell cultures and monocyte-derived dendritic cells in co-culture with autologous specific lymphocytes. TT-specific activation was detected with highest discrimination capacity using proliferation assays, as well as IFN-gamma and TT-specific antibody ELISPOTS (measuring secreting T and B lymphocytes, respectively). These in vitro systems show a high potential for replacing animal experimentation to evaluate the immunogenicity of complex vaccines.
authors.affiliation: Institute of Virology and Immunoprophylaxis (IVI), Sensemattstrasse 293, CH-3147 Mittelhäusern, Switzerland.
authors.forename: Sytse J
authors.initials: SJ
authors.lastname: Piersma
authors.forename: Marlies P P A M
authors.initials: MP
authors.lastname: Leenaars
authors.forename: Laurence
authors.initials: L
authors.lastname: Guzylack-Piriou
authors.forename: Artur
authors.initials: A
authors.lastname: Summerfield
authors.forename: Coenraad F M
authors.initials: CF
authors.lastname: Hendriksen
authors.forename: Ken C
authors.initials: KC
authors.lastname: McCullough
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis; D049109:Cell Proliferation; D018920:Coculture Techniques; D003713:Dendritic Cells / Q000276:immunology; D004168:Diphtheria Toxoid / Q000276:immunology; D007118:Immunoassay / Q000379:methods*; D007371:Interferon-gamma / Q000096:biosynthesis; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008213:Lymphocyte Activation; D023421:Models, Animal; D015375:Receptors, Interleukin-2 / Q000032:analysis; D012680:Sensitivity and Specificity; D013552:Swine; D013601:T-Lymphocytes / Q000276:immunology; D013745:Tetanus Toxoid / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D004168:Diphtheria Toxoid; D015375:Receptors, Interleukin-2; D013745:Tetanus Toxoid; D007371:Interferon-gamma