Preclinical efficacy and immunological safety of FR104, an antagonist anti-CD28 monovalent Fab' antibody

Am J Transplant. 2012 Oct;12(10):2630-40. doi: 10.1111/j.1600-6143.2012.04164.x. Epub 2012 Jul 3.


Antagonist anti-CD28 antibodies prevent T cell costimulation and differentiate from CTLA4Ig since they cannot block CTLA-4 and PDL-1 coinhibitory signals. They demonstrated efficacy in suppressing effector T cells while enhancing regulatory T cells function and immune tolerance. However, anti-CD28 antibodies devoid of immunotoxicity and with a good pharmacokinetic profile have not yet been developed. Here, we describe FR104, a novel humanized pegylated anti-CD28 Fab' antibody fragment presenting a long elimination half-life in monkeys. In vitro, FR104 failed to induce human T cell proliferation and cytokines secretion, even in the presence of anti-CD3 antibodies or when cross-linked with secondary antibodies. Furthermore, in humanized NOD/SCID mice adoptively transferred with human PBMC, whereas superagonist and divalent antibodies elicited rapid cytokines secretion and human T cell activation, FR104 did not. These humanized mice developed a florid graft-versus-host disease, which was prevented by administration of FR104 in a CTLA4-dependent manner. Interestingly, administration of high doses of CTLA4-Ig was ineffective to prevent GVHD, whereas administration of low doses was partially effective. In conclusion, we demonstrated that FR104 is devoid of agonist activity on human T cells and thus compatible with a clinical development that might lead to higher therapeutic indexes, by sparing CTLA-4, as compared to CD80/CD86 antagonists.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CD28 Antigens / immunology*
  • CTLA-4 Antigen / immunology
  • Drug Evaluation, Preclinical
  • Flow Cytometry
  • Immunoglobulin Fab Fragments / adverse effects
  • Immunoglobulin Fab Fragments / immunology
  • Immunoglobulin Fab Fragments / therapeutic use*
  • Immunohistochemistry
  • Lymphocyte Culture Test, Mixed
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID


  • CD28 Antigens
  • CTLA-4 Antigen
  • Immunoglobulin Fab Fragments