title: Dynamics of APC recruitment at the site of injection following injection of vaccine adjuvants.
abstract: Vaccines often contain adjuvants to strengthen the response to the vaccine antigen. However, their modes of action at the site of injection (SOI) are poorly understood. Therefore, we assessed the local effects of adjuvant on the innate immune system in mice. We investigated the safe, widely used adjuvants MF59 and aluminum hydroxide (alum), as well as trehalose-6,6'-dibehenate (TDB), Complete Freund's Adjuvant (CFA) and the Toll-Like-Receptor-ligands lipopolysaccharide (LPS) and Pam3CysSerLys4 (Pam3CSK4). We assessed muscle immune cell infiltration after adjuvant injection and observed 16h post immunization (hpi) an increased influx with CFA, MF59 and TDB, but not with alum, LPS or Pam3CSK4. An elevated influx with the latter three became visible only 72hpi. Contribution of granulocytes, macrophages and dendritic cells to the influx differed per adjuvant and in time. Adjuvants generally induced a local pro-inflammatory micro-milieu that was transient except for CFA and TDB. The gene expression of CXCL-1, CCL-2 and CCL-5, involved in recruitment of immune cells, varied per adjuvant and corresponded grossly with the observed influx of granulocytes and monocytes/macrophages. Muscles injected with CFA or MF59 (when co-injected with peptide) resulted in APC ex vivo capable to induce proliferation of peptide-specific T-cells. By adding in vitro an excess of peptide to the APC/T cell co-cultures, we observed an adjuvant-enhanced co-stimulation or antigen presentation by APC after CFA- but not MF59-injection. After TDB-injection this effect was observed only at 72hpi, but not 24hpi. Thus the cellular influx profile and the local cytokine and chemokine micro-milieu in the muscle were strongly influenced by the type of adjuvant. Additionally, the capacity of muscle APC to load and present antigen was affected by the adjuvant. These findings may assist the development of novel adjuvanted vaccines in a more rational manner.
authors.affiliation: Department of Infectious Diseases and Immunology, Utrecht University, Yalelaan 1, 3584 CL Utrecht, The Netherlands. Electronic address: s.vanaalst@uu.nl.
authors.forename: Susan
authors.initials: S
authors.lastname: van Aalst
authors.affiliation: Department of Infectious Diseases and Immunology, Utrecht University, Yalelaan 1, 3584 CL Utrecht, The Netherlands. Electronic address: i.s.ludwig@uu.nl.
authors.forename: Irene Stephanie
authors.initials: IS
authors.lastname: Ludwig
authors.affiliation: Department of Infectious Diseases and Immunology, Utrecht University, Yalelaan 1, 3584 CL Utrecht, The Netherlands. Electronic address: p.j.s.vankooten@uu.nl.
authors.forename: Peter Johannes Sylvester
authors.initials: PJS
authors.lastname: van Kooten
authors.affiliation: Department of Infectious Diseases and Immunology, Utrecht University, Yalelaan 1, 3584 CL Utrecht, The Netherlands. Electronic address: r.vanderzee@uu.nl.
authors.forename: Ruurd
authors.initials: R
authors.lastname: van der Zee
authors.affiliation: Department of Infectious Diseases and Immunology, Utrecht University, Yalelaan 1, 3584 CL Utrecht, The Netherlands. Electronic address: w.vaneden@uu.nl.
authors.forename: Willem
authors.initials: W
authors.lastname: van Eden
authors.affiliation: Department of Infectious Diseases and Immunology, Utrecht University, Yalelaan 1, 3584 CL Utrecht, The Netherlands. Electronic address: f.broere@uu.nl.
authors.forename: Femke
authors.initials: F
authors.lastname: Broere
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage*; D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology*; D005260:Female; D007113:Immunity, Innate; D007273:Injections, Intramuscular; D008807:Mice, Inbred BALB C; D009132:Muscles / Q000276:immunology
chemical_list: D000276:Adjuvants, Immunologic
keywords: APC; Innate immunity; Mechanism of action; Muscle immunity; Site of injection; Vaccine adjuvant