Development and Characterization of Recombinant Human Fc:OX40L Fusion Protein Linked Via a Coiled-coil Trimerization Domain

Overview

Journal Mol Immunol

Specialties Allergy & Immunology
Molecular Biology

Date 2007 Mar 22

PMID 17374396

Citations 34

Authors

Nicholas P Morris

Carmen Peters

Ryan Montler

Hong-Ming Hu

Brendan D Curti

Walter J Urba

Andrew D Weinberg

Affiliations

Soon will be listed here.

Abstract

OX40 (CD134) is a potent costimulatory molecule found on the surface of activated CD4(+) and CD8(+) T cells. Immunotherapy with OX40 agonists administered in vivo has demonstrated efficacy in several murine tumor models. A phase I clinical trial is currently underway in patients with advanced cancer using a mouse anti-CD134 monoclonal antibody. Therapy with this antibody will likely be limited to one cycle because patients develop neutralizing human anti-mouse antibody (HAMA). Therefore, we developed a humanized OX40 agonist that links the extracellular domain of human OX40L to the Fc domain of human IgG(1) via a trimerizing isoleucine zipper domain (ILZ). Physical characterization by velocity sedimentation revealed that this novel construct, hFcILZOX40L, was assembled into hexamers in which the Fc domains formed three disulfide-bonded dimers and the ILZ-OX40L domains formed two trimers. Trimerization of the ILZ domain was necessary to achieve appropriate assembly. In vitro biologic activity of the hFcILZOX40L hexamer was equivalent to the activity of agonist antibodies in plate-bound assays and was superior when the agonists were tested as soluble agents. Our ultimate goal is to use this recombinant molecule in a future clinical trial, and we feel that the OX40L hexamer will have equivalent or superior agonist activity in vivo when compared to an anti-OX40 antibody.

Citing Articles

Generation and characterization of OX40-ligand fusion protein that agonizes OX40 on T-Lymphocytes.

Sato A, Nagai H, Suzuki A, Ito A, Matsuyama S, Shibui N Front Immunol. 2025; 15:1473815.

PMID: 39867912 PMC: 11757143. DOI: 10.3389/fimmu.2024.1473815.

OX40 agonist stimulation increases and sustains humoral and cell-mediated responses to SARS-CoV-2 protein and saRNA vaccines.

Duhen R, Beymer M, Jensen S, Abbina S, Abraham S, Jain N Front Immunol. 2022; 13:896310.

PMID: 36238275 PMC: 9551348. DOI: 10.3389/fimmu.2022.896310.

Development of OX40 agonists for canine cancer immunotherapy.

Ruiz D, Haynes C, Marable J, Pundkar C, Nance R, Bedi D iScience. 2022; 25(10):105158.

PMID: 36217551 PMC: 9547195. DOI: 10.1016/j.isci.2022.105158.

Trimeric receptor-binding domain of SARS-CoV-2 acts as a potent inhibitor of ACE2 receptor-mediated viral entry.

Basavarajappa S, Liu A, Bruchez A, Li Z, Suzart V, Liu Z iScience. 2022; 25(8):104716.

PMID: 35813876 PMC: 9251894. DOI: 10.1016/j.isci.2022.104716.

Activation of OX40 and CD27 Costimulatory Signalling in Sheep through Recombinant Ovine Ligands.

Rojas J, Alejo A, Avia J, Rodriguez-Martin D, Sanchez C, Alcami A Vaccines (Basel). 2020; 8(2).

PMID: 32580486 PMC: 7350415. DOI: 10.3390/vaccines8020333.

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