CD3 × CD28 cross-interacting bispecific antibodies improve tumor cell dependent T-cell activation
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Bispecific antibodies (Bs-Abs) containing an anti-CD3 and an anti-TAA specificity can recruit T cells to the tumor for cancer immunotherapy. To be effective, efficient activation at the tumor site is a prerequisite. This can be achieved by triggering both the T-cell receptor and the co-stimulatory molecule CD28. We engineered two recombinant cross-interacting Bs-Abs (CriBs-Abs) by incorporating a peptide tag and its cognate single-chain variable fragment (scFv), respectively, into a pair of (tumor × CD3) and (tumor × CD28) binding Bs-Abs. A 30-fold lower concentration of the activating CriBs-Ab as compared to non interacting Bs-Ab was sufficient for strong T-cell activation in the presence of tumor cells. One thousand-fold higher concentrations of both CriBs-Abs were required for marginal T-cell activation (70-fold below maximal response) in the absence of tumor cells. An optimized stoichiometry (1 : 1000) of activating versus co-stimulating CriBs-Ab thus allowed low doses of activating CriBs-Ab to induce tumor-cell dependent T-cell activation when used in combination with high concentrations of the pre-targeted co-stimulating CriBs-Ab in vitro. This indicates a large window of operation in which only tumor cell dependent T-cell activation is induced and systemic tumor cell independent T-cell activation is avoided, while ensuring optimal activation with a low concentration of the activating CriBs-Ab, which has the highest potential to induce toxic effects in vivo.
KeywordsT lymphocytes Co-stimulation Bispecific antibodies Tumor immunotherapy Recombinant antibodies CD28
Cross-interacting bispecific Ab
Human placental alkaline phosphatase
Single-chain variable fragment
Tumor associated antigen
An Willems is a research associate with the Fonds Wetenschappelijk Onderzoek (FWO)—Vlaanderen. We thank T. Van Belle and D. Ginneberghe (Ghent University, Belgium) for the use of T-HA cells. Dr. M. Hall (University of Birmingham, U.K.) and Dr. M. De Broe (University of Antwerp, Belgium) are acknowledged for donating HEK293T cells and MO4I4 cells, respectively.
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