Murinization and H Chain Isotype Matching of the Anti-GITR Antibody DTA-1 Reduces Immunogenicity-Mediated Anaphylaxis in C57BL/6 Mice

J Immunol. 2017 Jun 1;198(11):4502-4512. doi: 10.4049/jimmunol.1601512. Epub 2017 Apr 26.


Recent advances in immuno-oncology have shown that the immune system can be activated to induce long-term, durable antitumor responses. For immuno-oncology drug development, immune activation is often explored using rat Abs in immunocompetent mouse models. Although these models can be used to show efficacy, antidrug immune responses to experimental protein-based therapeutics can arise. Immunogenicity of surrogate Abs may therefore represent an important obstacle to the evaluation of the antitumor efficacy of immunomodulator Abs in syngeneic models. A recent publication has shown that anti-glucocorticoid-induced TNFR family-related protein agonistic Ab DTA-1 (rat or murinized IgG2a) can induce the development of anaphylaxis in C57BL/6 mice upon repeated i.p. dosing because of an anti-idiotypic anti-drug Ab immune response. This study was undertaken to address the impact of the immunogenicity derived from the Fc and variable domains. To this end, chimerized (rat V domains/mouse constant regions) and murinized (95% mouse sequence) DTA-1-based surrogate Abs with a murine IgG2c H chain isotype were created. Chimerization and murinization of DTA-1 did not affect receptor binding and glucocorticoid-induced TNFR family-related protein-induced T cell agonistic properties. Similar in vivo antitumor efficacy and intratumoral CD8+/regulatory T cells were also observed. Finally, treatment of C57BL/6 mice with the chimerized and murinized DTA-1 Abs on a C57BL/6-matched IgG2c isotype resulted in reduced development and severity of anaphylaxis as measured by decline of body temperature, behavioral effects, serum IL-4, IgE, and anti-drug Ab levels. These results suggest that careful murinization and selection of a strain-matched H chain isotype are critical to generate ideal surrogate Abs for testing immuno-oncology mechanisms in vivo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anaphylaxis / immunology*
  • Animals
  • CD8-Positive T-Lymphocytes / immunology
  • Cell Line, Tumor
  • Glucocorticoid-Induced TNFR-Related Protein / immunology*
  • Immunoglobulin Isotypes / immunology*
  • Interleukin-4 / immunology
  • Mice
  • Mice, Inbred C57BL
  • Rats
  • Receptors, IgG / immunology
  • T-Lymphocytes, Regulatory / immunology


  • Glucocorticoid-Induced TNFR-Related Protein
  • IgG2c receptor
  • Immunoglobulin Isotypes
  • Receptors, IgG
  • Tnfrsf18 protein, mouse
  • Interleukin-4