title: Highly aggregated antibody therapeutics can enhance the in vitro innate and late-stage T-cell immune responses.
abstract: Aggregation of biotherapeutics has the potential to induce an immunogenic response. Here, we show that aggregated therapeutic antibodies, previously generated and determined to contain a variety of attributes (Joubert, M. K., Luo, Q., Nashed-Samuel, Y., Wypych, J., and Narhi, L. O. (2011) J. Biol. Chem. 286, 25118-25133), can enhance the in vitro innate immune response of a population of naive human peripheral blood mononuclear cells. This response depended on the aggregate type, inherent immunogenicity of the monomer, and donor responsiveness, and required a high number of particles, well above that detected in marketed drug products, at least in this in vitro system. We propose a cytokine signature as a potential biomarker of the in vitro peripheral blood mononuclear cell response to aggregates. The cytokines include IL-1β, IL-6, IL-10, MCP-1, MIP-1α, MIP-1β, MMP-2, and TNF-α. IL-6 and IL-10 might have an immunosuppressive effect on the long term immune response. Aggregates made by stirring induced the highest response compared with aggregates made by other methods. Particle size in the 2-10 μm range and the retention of some folded structure were associated with an increased response. The mechanism of aggregate activation at the innate phase was found to occur through specific cell surface receptors (the toll-like receptors TLR-2 and TLR-4, FcγRs, and the complement system). The innate signal was shown to progress to an adaptive T-cell response characterized by T-cell proliferation and secretion of T-cell cytokines. Investigating the ability of aggregates to induce cytokine signatures as biomarkers of immune responses is essential for determining their risk of immunogenicity.
authors.affiliation: Department of Product Attribute Sciences, Amgen Inc., Thousand Oaks, California 91320, USA. mjoubert@amgen.com
authors.forename: Marisa K
authors.initials: MK
authors.lastname: Joubert
authors.forename: Martha
authors.initials: M
authors.lastname: Hokom
authors.forename: Catherine
authors.initials: C
authors.lastname: Eakin
authors.forename: Lei
authors.initials: L
authors.lastname: Zhou
authors.forename: Meghana
authors.initials: M
authors.lastname: Deshpande
authors.forename: Matthew P
authors.initials: MP
authors.lastname: Baker
authors.forename: Theresa J
authors.initials: TJ
authors.lastname: Goletz
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Kerwin
authors.forename: Naren
authors.initials: N
authors.lastname: Chirmule
authors.forename: Linda O
authors.initials: LO
authors.lastname: Narhi
authors.forename: Vibha
authors.initials: V
authors.lastname: Jawa
mesh_terms: D000906:Antibodies / Q000276:immunology / Q000494:pharmacology*; D002478:Cells, Cultured; D016207:Cytokines / Q000276:immunology; D006801:Humans; D007111:Immunity, Cellular / Q000187:drug effects*; D007113:Immunity, Innate / Q000187:drug effects*; D007167:Immunotherapy / Q000379:methods*; D017452:Receptors, IgG / Q000276:immunology; D015398:Signal Transduction / Q000187:drug effects / Q000276:immunology; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*; D051195:Toll-Like Receptor 2 / Q000276:immunology; D051197:Toll-Like Receptor 4 / Q000276:immunology
chemical_list: D000906:Antibodies; D016207:Cytokines; D017452:Receptors, IgG; C495342:TLR2 protein, human; C495345:TLR4 protein, human; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4