The novel CD16A/anti-CD3 bifunctional protein, eCD16A/anti-CD3-BFP, redirects T cell cytotoxicity toward antibody-bound target cells
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 antib...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2025-12-01
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| Series: | Human Vaccines & Immunotherapeutics |
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| Online Access: | https://www.tandfonline.com/doi/10.1080/21645515.2024.2447141 |
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| author | Chien-Ting Lin Shang-Ju Wu Cheng Hao Liao Reyhung Roc Weng Chun-Ming Lin |
| author_facet | Chien-Ting Lin Shang-Ju Wu Cheng Hao Liao Reyhung Roc Weng Chun-Ming Lin |
| author_sort | Chien-Ting Lin |
| collection | DOAJ |
| description | 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 in vitro, ex vivo, and in vivo 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 ex vivo 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. |
| format | Article |
| id | doaj-art-f43305e7624f431d91a55d5143ba724d |
| institution | Kabale University |
| issn | 2164-5515 2164-554X |
| language | English |
| publishDate | 2025-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Human Vaccines & Immunotherapeutics |
| spelling | doaj-art-f43305e7624f431d91a55d5143ba724d2025-01-07T08:12:16ZengTaylor & Francis GroupHuman Vaccines & Immunotherapeutics2164-55152164-554X2025-12-0121110.1080/21645515.2024.2447141The novel CD16A/anti-CD3 bifunctional protein, eCD16A/anti-CD3-BFP, redirects T cell cytotoxicity toward antibody-bound target cellsChien-Ting Lin0Shang-Ju Wu1Cheng Hao Liao2Reyhung Roc Weng3Chun-Ming Lin4Department of Internal Medicine, National Taiwan University Hospital, Taipei, TaiwanDepartment of Internal Medicine, National Taiwan University Hospital, Taipei, TaiwanDepartment of Research and Development, ManySmart Therapeutics, Taipei, TaiwanDepartment of Internal Medicine, National Taiwan University Hospital, Taipei, TaiwanDepartment of Research and Development, ManySmart Therapeutics, Taipei, TaiwanMonoclonal 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 in vitro, ex vivo, and in vivo 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 ex vivo 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.https://www.tandfonline.com/doi/10.1080/21645515.2024.2447141Bifunctional proteinbispecific antibodyCD16anti-CD3lymphoma |
| spellingShingle | Chien-Ting Lin Shang-Ju Wu Cheng Hao Liao Reyhung Roc Weng Chun-Ming Lin The novel CD16A/anti-CD3 bifunctional protein, eCD16A/anti-CD3-BFP, redirects T cell cytotoxicity toward antibody-bound target cells Human Vaccines & Immunotherapeutics Bifunctional protein bispecific antibody CD16 anti-CD3 lymphoma |
| title | The novel CD16A/anti-CD3 bifunctional protein, eCD16A/anti-CD3-BFP, redirects T cell cytotoxicity toward antibody-bound target cells |
| title_full | The novel CD16A/anti-CD3 bifunctional protein, eCD16A/anti-CD3-BFP, redirects T cell cytotoxicity toward antibody-bound target cells |
| title_fullStr | The novel CD16A/anti-CD3 bifunctional protein, eCD16A/anti-CD3-BFP, redirects T cell cytotoxicity toward antibody-bound target cells |
| title_full_unstemmed | The novel CD16A/anti-CD3 bifunctional protein, eCD16A/anti-CD3-BFP, redirects T cell cytotoxicity toward antibody-bound target cells |
| title_short | The novel CD16A/anti-CD3 bifunctional protein, eCD16A/anti-CD3-BFP, redirects T cell cytotoxicity toward antibody-bound target cells |
| title_sort | novel cd16a anti cd3 bifunctional protein ecd16a anti cd3 bfp redirects t cell cytotoxicity toward antibody bound target cells |
| topic | Bifunctional protein bispecific antibody CD16 anti-CD3 lymphoma |
| url | https://www.tandfonline.com/doi/10.1080/21645515.2024.2447141 |
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