Highly sensitive in vitro cytokine release assay incorporating high-density preculture
Immunostimulatory effects of monoclonal antibodies (mAb) through binding to Fcγ receptors (FcγR) on immune cells are a likely cause of cytokine release syndrome. However, it is difficult to detect the potential risk of FcγR-dependent cytokine release associated with mAb in the current standard cytok...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2021-01-01
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| Series: | Journal of Immunotoxicology |
| Subjects: | |
| Online Access: | http://dx.doi.org/10.1080/1547691X.2021.1984617 |
| _version_ | 1818953441758674944 |
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| author | Shiho Ito Kyoko Miwa Chiharu Hattori Tetsuo Aida Yoshimi Tsuchiya Kazuhiko Mori |
| author_facet | Shiho Ito Kyoko Miwa Chiharu Hattori Tetsuo Aida Yoshimi Tsuchiya Kazuhiko Mori |
| author_sort | Shiho Ito |
| collection | DOAJ |
| description | Immunostimulatory effects of monoclonal antibodies (mAb) through binding to Fcγ receptors (FcγR) on immune cells are a likely cause of cytokine release syndrome. However, it is difficult to detect the potential risk of FcγR-dependent cytokine release associated with mAb in the current standard cytokine release assays (CRA), including the air-drying solid-phase method using human peripheral blood mononuclear cells (PBMC). To increase the sensitivity to detect FcγR-dependent cytokine release due to mAb, a high-density preculture (HDC) method was incorporated into the air-drying solid-phase CRA. Here, PBMC were exposed to panitumumab, trastuzumab, rituximab, or alemtuzumab at 0.1, 0.3, 1, and 3 μg/well for 24 or 48 hr under both non-HDC and HDC conditions. T-cell agonists (anti-CD3 mAb, anti-CD28 super-agonist [SA] mAb) were used as reference mAb. Panitumumab, trastuzumab, rituximab, or alemtuzumab induced cytokine release under both non-HDC and HDC conditions, and cytokine release caused by alemtuzumab was more pronounced under HDC conditions. To investigate FcγR involvement in cytokine release associated with panitumumab, trastuzumab, rituximab, and alemtuzumab, CRA of these four mAb were conducted with anti-FcγRI, -FcγRII, or -FcγRIII F(ab’)2 fragments. The results showed cytokine release caused by trastuzumab, rituximab, and alemtuzumab was significantly suppressed by anti-FcγRIII F(ab’)2 pretreatment, and slightly reduced by anti-FcγRI or anti-FcγRII pretreatment, indicating these mAb induced FcγR (especially FcγRIII)-dependent cytokine release from PBMC. Cytokine release caused by panitumumab was slightly suppressed by anti-FcγRIII F(ab’)2 pretreatment. Anti-CD3 mAb and anti-CD28 SA mAb also induced significant release of cytokines under HDC conditions compared with that under non-HDC conditions. In conclusion, CRA incorporating HDC into the air-drying solid-phase method using human PBMC could sensitively capture the FcγR-dependent cytokine release potential of mAb. |
| first_indexed | 2024-12-20T10:06:20Z |
| format | Article |
| id | doaj.art-4fa8d7b3d9334334a65c40ee72fd091d |
| institution | Directory Open Access Journal |
| issn | 1547-691X 1547-6901 |
| language | English |
| last_indexed | 2024-12-20T10:06:20Z |
| publishDate | 2021-01-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Journal of Immunotoxicology |
| spelling | doaj.art-4fa8d7b3d9334334a65c40ee72fd091d2022-12-21T19:44:13ZengTaylor & Francis GroupJournal of Immunotoxicology1547-691X1547-69012021-01-0118113614310.1080/1547691X.2021.19846171984617Highly sensitive in vitro cytokine release assay incorporating high-density precultureShiho Ito0Kyoko Miwa1Chiharu Hattori2Tetsuo Aida3Yoshimi Tsuchiya4Kazuhiko Mori5Medicinal Safety Research Laboratories, Daiichi Sankyo Co., LtdMedicinal Safety Research Laboratories, Daiichi Sankyo Co., LtdOncology Research Laboratories I, Daiichi Sankyo Co., LtdMedicinal Safety Research Laboratories, Daiichi Sankyo Co., LtdMedicinal Safety Research Laboratories, Daiichi Sankyo Co., LtdTransrational Research, Daiichi Sankyo RD Novare Co, LtdImmunostimulatory effects of monoclonal antibodies (mAb) through binding to Fcγ receptors (FcγR) on immune cells are a likely cause of cytokine release syndrome. However, it is difficult to detect the potential risk of FcγR-dependent cytokine release associated with mAb in the current standard cytokine release assays (CRA), including the air-drying solid-phase method using human peripheral blood mononuclear cells (PBMC). To increase the sensitivity to detect FcγR-dependent cytokine release due to mAb, a high-density preculture (HDC) method was incorporated into the air-drying solid-phase CRA. Here, PBMC were exposed to panitumumab, trastuzumab, rituximab, or alemtuzumab at 0.1, 0.3, 1, and 3 μg/well for 24 or 48 hr under both non-HDC and HDC conditions. T-cell agonists (anti-CD3 mAb, anti-CD28 super-agonist [SA] mAb) were used as reference mAb. Panitumumab, trastuzumab, rituximab, or alemtuzumab induced cytokine release under both non-HDC and HDC conditions, and cytokine release caused by alemtuzumab was more pronounced under HDC conditions. To investigate FcγR involvement in cytokine release associated with panitumumab, trastuzumab, rituximab, and alemtuzumab, CRA of these four mAb were conducted with anti-FcγRI, -FcγRII, or -FcγRIII F(ab’)2 fragments. The results showed cytokine release caused by trastuzumab, rituximab, and alemtuzumab was significantly suppressed by anti-FcγRIII F(ab’)2 pretreatment, and slightly reduced by anti-FcγRI or anti-FcγRII pretreatment, indicating these mAb induced FcγR (especially FcγRIII)-dependent cytokine release from PBMC. Cytokine release caused by panitumumab was slightly suppressed by anti-FcγRIII F(ab’)2 pretreatment. Anti-CD3 mAb and anti-CD28 SA mAb also induced significant release of cytokines under HDC conditions compared with that under non-HDC conditions. In conclusion, CRA incorporating HDC into the air-drying solid-phase method using human PBMC could sensitively capture the FcγR-dependent cytokine release potential of mAb.http://dx.doi.org/10.1080/1547691X.2021.1984617cytokine release syndromehigh-density preculturefcγrcytokine release assaytherapeutic monoclonal antibodyperipheral blood mononuclear cell |
| spellingShingle | Shiho Ito Kyoko Miwa Chiharu Hattori Tetsuo Aida Yoshimi Tsuchiya Kazuhiko Mori Highly sensitive in vitro cytokine release assay incorporating high-density preculture Journal of Immunotoxicology cytokine release syndrome high-density preculture fcγr cytokine release assay therapeutic monoclonal antibody peripheral blood mononuclear cell |
| title | Highly sensitive in vitro cytokine release assay incorporating high-density preculture |
| title_full | Highly sensitive in vitro cytokine release assay incorporating high-density preculture |
| title_fullStr | Highly sensitive in vitro cytokine release assay incorporating high-density preculture |
| title_full_unstemmed | Highly sensitive in vitro cytokine release assay incorporating high-density preculture |
| title_short | Highly sensitive in vitro cytokine release assay incorporating high-density preculture |
| title_sort | highly sensitive in vitro cytokine release assay incorporating high density preculture |
| topic | cytokine release syndrome high-density preculture fcγr cytokine release assay therapeutic monoclonal antibody peripheral blood mononuclear cell |
| url | http://dx.doi.org/10.1080/1547691X.2021.1984617 |
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