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  4. The novel CD16A/anti-CD3 bifunctional protein, eCD16A/anti-CD3-BFP, redirects T cell cytotoxicity toward antibody-bound target cells.
 
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The novel CD16A/anti-CD3 bifunctional protein, eCD16A/anti-CD3-BFP, redirects T cell cytotoxicity toward antibody-bound target cells.

Journal
Human vaccines & immunotherapeutics
Journal Volume
21
Journal Issue
1
Pages
2447141
ISSN
2164-554X
Date Issued
2025-12
Author(s)
Lin, Chien-Ting
SHANG-JU WU  
Liao, Cheng Hao
Weng, Reyhung Roc
Lin, Chun-Ming
DOI
10.1080/21645515.2024.2447141
URI
https://scholars.lib.ntu.edu.tw/handle/123456789/729338
Abstract
Monoclonal antibodies enhance innate immunity, while bispecific T cell engager antibodies redirect adaptive T cell immunity. To stimulate both innate and adaptive mechanisms, we created a bifunctional eCD16A/anti-CD3-BFP adapter protein for combined use with clinically approved monoclonal IgG1 antibodies. The adaptor protein contains the extracellular domain of the human CD16A high-affinity variant, which binds the Fc domain of IgG1 antibodies, and an anti-human CD3 single-chain variable fragment that redirects T cell cytotoxicity. Functional characterization of eCD16A/anti-CD3-BFP was performed using , , and animal assays. Combination treatments of eCD16A/anti-CD3-BFP with rituximab (anti-CD20), cetuximab (anti-EGFR), trastuzumab (anti-HER2) or anti-PD-L1 IgG1 led to specific killing of antigen-expressing tumor cells. In an assay, eCD16A/anti-CD3-BFP combined with rituximab effectively eliminated B cells within peripheral blood samples from healthy donors and cancer patients; the effectiveness of this treatment was further enhanced by addition of either autologous or allogeneic γ9δ2 T cells. In mouse xenograft studies, the combination of eCD16A/anti-CD3-BFP and rituximab led to rapid depletion of tumor cells. The presence of non-targeting competing serum immunoglobulins interfered with efficacy, but this interference could be overcome by eCD16A/anti-CD3-BFP dose escalation or by using a Fc-glycoengineered version of rituximab. Moreover, combining eCD16A/anti-CD3-BFP with anti-Epstein Barr virus (EBV) antibodies directed T cell cytotoxicity toward EBV-infected cells. These findings support further development of eCD16A/anti-CD3-BFP as a novel anti-cancer therapeutic to stimulate both innate and adaptive immunity.
Subjects
Bifunctional protein
CD16
anti-CD3
bispecific antibody
lymphoma
Type
journal article

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