A comprehensive battery of flow cytometric immunoassays for the in vitro testing of chemical effects in human blood cells
BackgroundThere is a growing need for immunological assays to test toxic and modulatory effects of chemicals. The assays should be easy to use, reproducible and superior to cell line-based assays. We have therefore developed a comprehensive portfolio of assays based on primary human blood cells that...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2024-01-01
|
| Series: | Frontiers in Immunology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1327960/full |
| _version_ | 1797368856811929600 |
|---|---|
| author | Arkadiusz Pierzchalski Ana C. Zenclussen Ana C. Zenclussen Gunda Herberth |
| author_facet | Arkadiusz Pierzchalski Ana C. Zenclussen Ana C. Zenclussen Gunda Herberth |
| author_sort | Arkadiusz Pierzchalski |
| collection | DOAJ |
| description | BackgroundThere is a growing need for immunological assays to test toxic and modulatory effects of chemicals. The assays should be easy to use, reproducible and superior to cell line-based assays. We have therefore developed a comprehensive portfolio of assays based on primary human blood cells that are suitable for testing chemical effects.MethodsThe flow cytometry-based assays were designed to target a wide range of human peripheral blood mononuclear cells and whole blood, including T cells, NK cells, B cells, basophils and innate-like T cells such as γδT, MAIT and NKT cells. We have selected a set of activation markers for each immune cell, e.g: CD154 (T cells), CD137, CD107a (NK cells), CD63 (basophils), CD69, CD83 (B cells), CD69, IFN-γ (MAIT cells) and we selected cell specific stimuli: aCD3 antibodies (T cells); E. coli and cytokines IL-12/15/18 (MAIT cells); CpG ODN2006, R848 or aCD40 antibodies (B cells), fMLP or aFcϵR1 (basophils) or K562 cells (NK cells).ResultsBy selecting immune cell-specific markers and cell-specific stimuli, we were able to induce particular immune responses from the targeted immune cells. For example, the response to stimulation with anti-CD3 antibodies was in 36.8% of CD107a+CD8+ cells. Cytokine stimulation induced the production of IFN-γ in 30% of MAIT cells. After stimulation with E. coli, around 50% of MAIT cells produced TNF. About 40% of basophils responded to aFcƐR1 stimulation. Similar activation ranges were achieved in K562-stimulated NK cells.ConclusionOur test portfolio covers the most relevant immune cells present in human blood, providing a solid basis for in vitro toxicity and immunomodulatory testing of chemicals. By using human blood, the natural composition of cells found in the blood can be determined and the effects of chemicals can be detected at the cellular level. |
| first_indexed | 2024-03-08T17:38:43Z |
| format | Article |
| id | doaj.art-2dcbf435b6aa4f3a87207f27b2444367 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-03-08T17:38:43Z |
| publishDate | 2024-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-2dcbf435b6aa4f3a87207f27b24443672024-01-02T11:18:43ZengFrontiers Media S.A.Frontiers in Immunology1664-32242024-01-011410.3389/fimmu.2023.13279601327960A comprehensive battery of flow cytometric immunoassays for the in vitro testing of chemical effects in human blood cellsArkadiusz Pierzchalski0Ana C. Zenclussen1Ana C. Zenclussen2Gunda Herberth3Helmholtz Centre for Environmental Research - UFZ, Department of Environmental Immunology, Leipzig, GermanyHelmholtz Centre for Environmental Research - UFZ, Department of Environmental Immunology, Leipzig, GermanyPerinatal Immunology Research Group, Medical Faculty, Saxonian Incubator for Clinical Translation (SIKT), University of Leipzig, Leipzig, GermanyHelmholtz Centre for Environmental Research - UFZ, Department of Environmental Immunology, Leipzig, GermanyBackgroundThere is a growing need for immunological assays to test toxic and modulatory effects of chemicals. The assays should be easy to use, reproducible and superior to cell line-based assays. We have therefore developed a comprehensive portfolio of assays based on primary human blood cells that are suitable for testing chemical effects.MethodsThe flow cytometry-based assays were designed to target a wide range of human peripheral blood mononuclear cells and whole blood, including T cells, NK cells, B cells, basophils and innate-like T cells such as γδT, MAIT and NKT cells. We have selected a set of activation markers for each immune cell, e.g: CD154 (T cells), CD137, CD107a (NK cells), CD63 (basophils), CD69, CD83 (B cells), CD69, IFN-γ (MAIT cells) and we selected cell specific stimuli: aCD3 antibodies (T cells); E. coli and cytokines IL-12/15/18 (MAIT cells); CpG ODN2006, R848 or aCD40 antibodies (B cells), fMLP or aFcϵR1 (basophils) or K562 cells (NK cells).ResultsBy selecting immune cell-specific markers and cell-specific stimuli, we were able to induce particular immune responses from the targeted immune cells. For example, the response to stimulation with anti-CD3 antibodies was in 36.8% of CD107a+CD8+ cells. Cytokine stimulation induced the production of IFN-γ in 30% of MAIT cells. After stimulation with E. coli, around 50% of MAIT cells produced TNF. About 40% of basophils responded to aFcƐR1 stimulation. Similar activation ranges were achieved in K562-stimulated NK cells.ConclusionOur test portfolio covers the most relevant immune cells present in human blood, providing a solid basis for in vitro toxicity and immunomodulatory testing of chemicals. By using human blood, the natural composition of cells found in the blood can be determined and the effects of chemicals can be detected at the cellular level.https://www.frontiersin.org/articles/10.3389/fimmu.2023.1327960/fullimmune assayshuman bloodflow cytometrychemical testingactivation markerimmune cells |
| spellingShingle | Arkadiusz Pierzchalski Ana C. Zenclussen Ana C. Zenclussen Gunda Herberth A comprehensive battery of flow cytometric immunoassays for the in vitro testing of chemical effects in human blood cells Frontiers in Immunology immune assays human blood flow cytometry chemical testing activation marker immune cells |
| title | A comprehensive battery of flow cytometric immunoassays for the in vitro testing of chemical effects in human blood cells |
| title_full | A comprehensive battery of flow cytometric immunoassays for the in vitro testing of chemical effects in human blood cells |
| title_fullStr | A comprehensive battery of flow cytometric immunoassays for the in vitro testing of chemical effects in human blood cells |
| title_full_unstemmed | A comprehensive battery of flow cytometric immunoassays for the in vitro testing of chemical effects in human blood cells |
| title_short | A comprehensive battery of flow cytometric immunoassays for the in vitro testing of chemical effects in human blood cells |
| title_sort | comprehensive battery of flow cytometric immunoassays for the in vitro testing of chemical effects in human blood cells |
| topic | immune assays human blood flow cytometry chemical testing activation marker immune cells |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1327960/full |
| work_keys_str_mv | AT arkadiuszpierzchalski acomprehensivebatteryofflowcytometricimmunoassaysfortheinvitrotestingofchemicaleffectsinhumanbloodcells AT anaczenclussen acomprehensivebatteryofflowcytometricimmunoassaysfortheinvitrotestingofchemicaleffectsinhumanbloodcells AT anaczenclussen acomprehensivebatteryofflowcytometricimmunoassaysfortheinvitrotestingofchemicaleffectsinhumanbloodcells AT gundaherberth acomprehensivebatteryofflowcytometricimmunoassaysfortheinvitrotestingofchemicaleffectsinhumanbloodcells AT arkadiuszpierzchalski comprehensivebatteryofflowcytometricimmunoassaysfortheinvitrotestingofchemicaleffectsinhumanbloodcells AT anaczenclussen comprehensivebatteryofflowcytometricimmunoassaysfortheinvitrotestingofchemicaleffectsinhumanbloodcells AT anaczenclussen comprehensivebatteryofflowcytometricimmunoassaysfortheinvitrotestingofchemicaleffectsinhumanbloodcells AT gundaherberth comprehensivebatteryofflowcytometricimmunoassaysfortheinvitrotestingofchemicaleffectsinhumanbloodcells |