A simple method for measuring immune complex-mediated, Fc gamma receptor dependent antigen-specific activation of primary human T cells

J Immunol Methods. 2018 Mar;454:32-39. doi: 10.1016/j.jim.2017.12.002. Epub 2017 Dec 16.


Immune complex (IC) deposition of IgG containing autologous antigens has been observed in autoimmunity. This can lead to IC-mediated antigen uptake and presentation by antigen presenting cells (APC) driving T cell dependent inflammation. IgG receptors (FcγRs) have been suggested to be involved in this process. Since ICs have been linked to autoimmune diseases, interfering with IC mediated effects on APCs and subsequent autoimmune T cell activation via FcγR blockade may be therapeutically beneficial. However, this is currently challenging due to a lack of translatable animal models and specific human in vitro assays to study IC-driven T cell responses. Here, we developed a simple cellular assay to study IC-mediated T cell activation in vitro using human peripheral blood mononuclear cells and tetanus toxoid as a model antigen. We observed that tetanus ICs led to a strong induction of T cell proliferation and release of pro-inflammatory cytokines, which are hallmarks of chronic inflammation. This process was exacerbated when compared to tetanus toxoid challenge alone. IC-mediated T cell effects were FcγR dependent and inhibited by high-dose intravenous IgG (IVIg), a drug often used for the clinical treatments of autoimmune diseases. Similar effects were also seen using a hepatitis antigen. Consequently, we propose our assay as a rapid yet robust alternative to more labour-intense and time-consuming protocols, for example involving separate maturation of dendritic cells followed by T cell co-culture to study antigen specific primary T cell activation.

Keywords: Antigen-specific T cell memory recall; Autoimmunity; FcγR; Hepatitis; Immune complexes; Tetanus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigen Presentation
  • Antigen-Antibody Complex / metabolism
  • Autoantigens / metabolism
  • Autoimmune Diseases / diagnosis*
  • Autoimmunity
  • Cell Proliferation
  • Cells, Cultured
  • Cytokines / metabolism
  • Disease Models, Animal
  • Humans
  • Immunoassay / methods*
  • Inflammation Mediators / metabolism
  • Lymphocyte Activation
  • Primary Cell Culture
  • Receptors, IgG / metabolism
  • T-Cell Antigen Receptor Specificity
  • T-Lymphocytes / immunology*


  • Antigen-Antibody Complex
  • Autoantigens
  • Cytokines
  • Inflammation Mediators
  • Receptors, IgG