title: High thioredoxin-1 levels in rheumatoid arthritis patients diminish binding and signalling of the monoclonal antibody Tregalizumab.
abstract: The humanized non-depleting anti-CD4 monoclonal antibody Tregalizumab (BT-061) is able to selectively activate the suppressive function of regulatory T cells and has been investigated up to phase IIb in clinical trials in patients suffering from rheumatoid arthritis (RA). A pharmacokinetic-pharmacodynamic model based on clinical data from RA and healthy volunteers, which used the cell surface CD4 downmodulation as marker of activity, confirmed a stronger effect in healthy volunteers compared with RA patients. We tried to understand this phenomenon and evaluated the influence of the small oxidoreductase thioredoxin-1 (Trx1). To counteract oxidative stress that is strongly associated with RA pathophysiology, the organism employs Trx1. Therefore, increased expression and secretion of Trx1 is found in the synovial fluid and plasma of RA patients. Moreover, the binding site of Tregalizumab is in close proximity to a disulphide bond in domain 2 (D2) of CD4, which is a known target for a reduction by oxidoreductase Trx1. With the experiments reported herein, we demonstrated that specific reduction of the D2 disulphide bond by Trx1 led to diminished binding of Tregalizumab to recombinant human soluble CD4 and membrane-bound CD4 on T cells. Moreover, we showed that this caused changes in the Tregalizumab-induced CD4 signalling pathway via the lymphocyte-specific protein tyrosine kinase p56Lckand CD4 downmodulation. In summary, we provide evidence that high Trx1 levels in RA patients compared with healthy subjects are a potential reason for diminished binding of Tregalizumab to CD4-positive T cells and offer an explanation for the observed decreased CD4 downmodulation in RA patients in comparison to healthy subjects.
authors.affiliation: Biotest AG , Dreieich, Germany.
authors.forename: Katharina
authors.initials: K
authors.lastname: Heim
authors.affiliation: Biotest AG , Dreieich, Germany.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Dälken
authors.affiliation: Biotest AG , Dreieich, Germany.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Faust
authors.affiliation: Biotest AG , Dreieich, Germany.
authors.forename: Faiza
authors.initials: F
authors.lastname: Rharbaoui
authors.affiliation: Biotest AG , Dreieich, Germany.
authors.forename: Andre
authors.initials: A
authors.lastname: Engling
authors.affiliation: Condor Scientific Computing and Consulting , Sulzbach, Germany.
authors.forename: Holger
authors.initials: H
authors.lastname: Wallmeier
authors.affiliation: Institute of Pharmaceutical Biology, Goethe University Frankfurt , Frankfurt, Germany.
authors.forename: Theodor
authors.initials: T
authors.lastname: Dingermann
authors.affiliation: Institute of Pharmacology and Toxicology/ZAFES, Clinic of the Goethe University , Frankfurt, Germany.
authors.forename: Heinfried H
authors.initials: HH
authors.lastname: Radeke
authors.affiliation: Biotest AG , Dreieich, Germany.
authors.forename: Jörg
authors.initials: J
authors.lastname: Schüttrumpf
authors.affiliation: Biotest AG , Dreieich, Germany.
authors.forename: Marcus
authors.initials: M
authors.lastname: Gutscher