Gastroenterology

Gastroenterology

Volume 124, Issue 7, June 2003, Pages 1774-1785
Gastroenterology

Clinical-alimentary tract
Infliximab but not etanercept induces apoptosis in lamina propria T-lymphocytes from patients with Crohn’s disease

https://doi.org/10.1016/S0016-5085(03)00382-2Get rights and content

Abstract

Background & Aims:

Steroid-refractory Crohn’s disease responds to therapy with the chimeric anti-tumor necrosis factor (TNF)-α antibody infliximab. Etanercept, a recombinant TNF receptor/immunoglobulin G fusion protein, is highly effective in rheumatoid arthritis but not in Crohn’s disease. Because both infliximab and etanercept are TNF-α-neutralizing drugs, we investigated the differences in TNF-α-neutralizing capacity and human lymphocyte binding and apoptosis-inducing capacity of both molecules.

Methods:

We used a nuclear factor κB reporter assay and a cytotoxicity bioassay to study TNF-α neutralization by infliximab and etanercept. Lymphocyte binding and apoptosis-inducing capacity was investigated using fluorescence-activated cell sorter analysis, annexin V staining, and cleaved caspase-3 immunoblotting using mixed lymphocyte reaction-stimulated peripheral blood lymphocytes (PBL) from healthy volunteers and lamina propria T cells from patients with Crohn’s disease.

Results:

Both infliximab and etanercept neutralized TNF-α effectively. Infliximab bound to activated PBL and lamina propria T cells, whereas binding of etanercept was equal to a nonspecific control antibody. Infliximab but not etanercept induced peripheral and lamina propria lymphocyte apoptosis when compared with a control antibody. Infliximab activated caspase 3 in a time-dependent manner, whereas etanercept did not.

Conclusions:

Although both infliximab and etanercept showed powerful TNF-α neutralization, only infliximab was able to bind to PBL and lamina propria T cells and subsequently to induce apoptosis of activated lymphocytes. These data may provide a biological basis for the difference in efficacy of the 2 TNF-α-neutralizing drugs.

Section snippets

Cell culture

All cells were cultured at 37°C, 5% CO2, and 99% humidity. Three culture media were used. Complete Iscove’s modified Dulbecco’s medium (Life Technologies, Paisley, Scotland) contained gentamicin (86 μg/mL; Duchefa, Haarlem, The Netherlands), 1% heat inactivated fetal calf serum (HyClone, Logan, UT), and l-glutamine (29.2 mg/mL; Gibco, Rhode Island, NY) for monocyte-derived dendritic cells. Complete Dulbecco’s modified Eagle medium (BioWhittaker, Verviers, Belgium) was supplemented with 5% fetal

Infliximab and etanercept neutralize recombinant human TNF-α effectively

A reason for the difference between infliximab and etanercept in Crohn’s disease could be related to a difference in TNF-α-neutralizing potential between the 2 drugs. Hence, we assayed the TNF-α-neutralizing potential of infliximab and etanercept both with respect to TNF-α-dependent transactivation of a NF-κB-driven reporter construct as well as in the WEHI assay for TNF-α-induced cell death. Infliximab abolished TNF-α-induced GFP expression in NF-κB reporter construct transfected HeLa cells at

Discussion

The cytokine TNF-α is believed to play a key role in the pathogenesis of acute and chronic inflammatory diseases. TNF-α-neutralizing antibodies have proved to be effective in various animal models; 2 therapeutic agents, infliximab (a chimeric monoclonal anti-TNF-α antibody) and etanercept (a recombinant human TNF receptor fusion protein), have been approved for clinical use. Both TNF-α-neutralizing agents have shown high efficacy in rheumatoid arthritis,17, 33 but only infliximab is efficacious

Acknowledgements

The authors thank Dr. David Shealy for providing the C17-1a antibody as well as for helpful discussions and critical reading of the manuscript, Prof. Marc Feldmann for the WEHI cells, Frederik Slors for providing intestinal specimen, and Eric van Tol, Hilde Croes, Inge Pronk and Ludo Evers for their assistance with the experiments.

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    Supported by an unrestricted educational grant from Johnson & Johnson (to J.M.H.V.), an unrestricted educational grant from Numico Research b.v. (to H.B.), the Amsterdam Medical Center Research b.v. (to G.R.V. and H.H.V.), the Meelmeijer Foundation (to C.V.), the Dutch Heart Foundation (99.188 to I.H.), and the Dutch Cancer Foundation (99.30 to M.P.P.).

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