Production of a cellular product consisting of monocytes stimulated with Sylatron® (Peginterferon alfa-2b) and Actimmune® (Interferon gamma-1b) for human use
Abstract Background Monocytes are myeloid cells that reside in the blood and bone marrow and respond to inflammation. At the site of inflammation, monocytes express cytokines and chemokines. Monocytes have been shown to be cytotoxic to tumor cells in the presence of pro-inflammatory cytokines such a...
| Main Authors: | , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2019-03-01
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| Series: | Journal of Translational Medicine |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12967-019-1822-6 |
| _version_ | 1831638402007564288 |
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| author | Daniel S. Green Ana T. Nunes Kevin W. Tosh Virginia David-Ocampo Vicki S. Fellowes Jiaqiang Ren Jianjian Jin Sue-Ellen Frodigh Chauha Pham Jolynn Procter Celina Tran Irene Ekwede Hanh Khuu David F. Stroncek Steven L. Highfill Kathryn C. Zoon Christina M. Annunziata |
| author_facet | Daniel S. Green Ana T. Nunes Kevin W. Tosh Virginia David-Ocampo Vicki S. Fellowes Jiaqiang Ren Jianjian Jin Sue-Ellen Frodigh Chauha Pham Jolynn Procter Celina Tran Irene Ekwede Hanh Khuu David F. Stroncek Steven L. Highfill Kathryn C. Zoon Christina M. Annunziata |
| author_sort | Daniel S. Green |
| collection | DOAJ |
| description | Abstract Background Monocytes are myeloid cells that reside in the blood and bone marrow and respond to inflammation. At the site of inflammation, monocytes express cytokines and chemokines. Monocytes have been shown to be cytotoxic to tumor cells in the presence of pro-inflammatory cytokines such as Interferon Alpha, Interferon Gamma, and IL-6. We have previously shown that monocytes stimulated with both interferons (IFNs) results in synergistic killing of ovarian cancer cells. We translated these observations to an ongoing clinical trial using adoptive cell transfer of autologous monocytes stimulated ex vivo with IFNs and infused into the peritoneal cavity of patients with advanced, chemotherapy resistant, ovarian cancer. Here we describe the optimization of the monocyte elutriation protocol and a cryopreservation protocol of the monocytes isolated from peripheral blood. Methods Counter flow elutriation was performed on healthy donors or women with ovarian cancer. The monocyte-containing, RO-fraction was assessed for total monocyte number, purity, viability, and cytotoxicity with and without a cryopreservation step. All five fractions obtained from the elutriation procedure were also assessed by flow cytometry to measure the percent of immune cell subsets in each fraction. Results Both iterative monocyte isolation using counter flow elutriation or cryopreservation following counter flow elutriation can yield over 2 billion monocytes for each donor with high purity. We also show that the monocytes are stable, viable, and retain cytotoxic functions when cultured with IFNs. Conclusion Large scale isolation of monocytes from both healthy donors and patients with advanced, chemotherapy resistant ovarian cancer, can be achieved with high total number of monocytes. These monocytes can be cryopreserved and maintain viability and cytotoxic function. All of the elutriated cell fractions contain ample immune cells which could be used for other cell therapy-based applications. |
| first_indexed | 2024-12-19T07:11:15Z |
| format | Article |
| id | doaj.art-895d8b2b169c4356b08a7ca746b3f353 |
| institution | Directory Open Access Journal |
| issn | 1479-5876 |
| language | English |
| last_indexed | 2024-12-19T07:11:15Z |
| publishDate | 2019-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Translational Medicine |
| spelling | doaj.art-895d8b2b169c4356b08a7ca746b3f3532022-12-21T20:31:12ZengBMCJournal of Translational Medicine1479-58762019-03-0117111310.1186/s12967-019-1822-6Production of a cellular product consisting of monocytes stimulated with Sylatron® (Peginterferon alfa-2b) and Actimmune® (Interferon gamma-1b) for human useDaniel S. Green0Ana T. Nunes1Kevin W. Tosh2Virginia David-Ocampo3Vicki S. Fellowes4Jiaqiang Ren5Jianjian Jin6Sue-Ellen Frodigh7Chauha Pham8Jolynn Procter9Celina Tran10Irene Ekwede11Hanh Khuu12David F. Stroncek13Steven L. Highfill14Kathryn C. Zoon15Christina M. Annunziata16Women’s Malignancy Branch, Center for Cancer Research, National Cancer Institute, National Institutes of HealthWomen’s Malignancy Branch, Center for Cancer Research, National Cancer Institute, National Institutes of HealthLaboratory of Parasitic Diseases, National Institutes of Allergy and Infectious Diseases, National Institutes of HealthCell Processing Section, Department of Transfusion Medicine, Clinical Center, National Institutes of HealthCell Processing Section, Department of Transfusion Medicine, Clinical Center, National Institutes of HealthCell Processing Section, Department of Transfusion Medicine, Clinical Center, National Institutes of HealthCell Processing Section, Department of Transfusion Medicine, Clinical Center, National Institutes of HealthCell Processing Section, Department of Transfusion Medicine, Clinical Center, National Institutes of HealthCell Processing Section, Department of Transfusion Medicine, Clinical Center, National Institutes of HealthCell Processing Section, Department of Transfusion Medicine, Clinical Center, National Institutes of HealthCell Processing Section, Department of Transfusion Medicine, Clinical Center, National Institutes of HealthWomen’s Malignancy Branch, Center for Cancer Research, National Cancer Institute, National Institutes of HealthCell Processing Section, Department of Transfusion Medicine, Clinical Center, National Institutes of HealthCell Processing Section, Department of Transfusion Medicine, Clinical Center, National Institutes of HealthCell Processing Section, Department of Transfusion Medicine, Clinical Center, National Institutes of HealthLaboratory of Infectious Diseases, National Institutes of Allergy and Infectious Diseases, National Institutes of HealthWomen’s Malignancy Branch, Center for Cancer Research, National Cancer Institute, National Institutes of HealthAbstract Background Monocytes are myeloid cells that reside in the blood and bone marrow and respond to inflammation. At the site of inflammation, monocytes express cytokines and chemokines. Monocytes have been shown to be cytotoxic to tumor cells in the presence of pro-inflammatory cytokines such as Interferon Alpha, Interferon Gamma, and IL-6. We have previously shown that monocytes stimulated with both interferons (IFNs) results in synergistic killing of ovarian cancer cells. We translated these observations to an ongoing clinical trial using adoptive cell transfer of autologous monocytes stimulated ex vivo with IFNs and infused into the peritoneal cavity of patients with advanced, chemotherapy resistant, ovarian cancer. Here we describe the optimization of the monocyte elutriation protocol and a cryopreservation protocol of the monocytes isolated from peripheral blood. Methods Counter flow elutriation was performed on healthy donors or women with ovarian cancer. The monocyte-containing, RO-fraction was assessed for total monocyte number, purity, viability, and cytotoxicity with and without a cryopreservation step. All five fractions obtained from the elutriation procedure were also assessed by flow cytometry to measure the percent of immune cell subsets in each fraction. Results Both iterative monocyte isolation using counter flow elutriation or cryopreservation following counter flow elutriation can yield over 2 billion monocytes for each donor with high purity. We also show that the monocytes are stable, viable, and retain cytotoxic functions when cultured with IFNs. Conclusion Large scale isolation of monocytes from both healthy donors and patients with advanced, chemotherapy resistant ovarian cancer, can be achieved with high total number of monocytes. These monocytes can be cryopreserved and maintain viability and cytotoxic function. All of the elutriated cell fractions contain ample immune cells which could be used for other cell therapy-based applications.http://link.springer.com/article/10.1186/s12967-019-1822-6Cell therapyCellular immunotherapyMonocytesInterferonsInnate immunity |
| spellingShingle | Daniel S. Green Ana T. Nunes Kevin W. Tosh Virginia David-Ocampo Vicki S. Fellowes Jiaqiang Ren Jianjian Jin Sue-Ellen Frodigh Chauha Pham Jolynn Procter Celina Tran Irene Ekwede Hanh Khuu David F. Stroncek Steven L. Highfill Kathryn C. Zoon Christina M. Annunziata Production of a cellular product consisting of monocytes stimulated with Sylatron® (Peginterferon alfa-2b) and Actimmune® (Interferon gamma-1b) for human use Journal of Translational Medicine Cell therapy Cellular immunotherapy Monocytes Interferons Innate immunity |
| title | Production of a cellular product consisting of monocytes stimulated with Sylatron® (Peginterferon alfa-2b) and Actimmune® (Interferon gamma-1b) for human use |
| title_full | Production of a cellular product consisting of monocytes stimulated with Sylatron® (Peginterferon alfa-2b) and Actimmune® (Interferon gamma-1b) for human use |
| title_fullStr | Production of a cellular product consisting of monocytes stimulated with Sylatron® (Peginterferon alfa-2b) and Actimmune® (Interferon gamma-1b) for human use |
| title_full_unstemmed | Production of a cellular product consisting of monocytes stimulated with Sylatron® (Peginterferon alfa-2b) and Actimmune® (Interferon gamma-1b) for human use |
| title_short | Production of a cellular product consisting of monocytes stimulated with Sylatron® (Peginterferon alfa-2b) and Actimmune® (Interferon gamma-1b) for human use |
| title_sort | production of a cellular product consisting of monocytes stimulated with sylatron r peginterferon alfa 2b and actimmune r interferon gamma 1b for human use |
| topic | Cell therapy Cellular immunotherapy Monocytes Interferons Innate immunity |
| url | http://link.springer.com/article/10.1186/s12967-019-1822-6 |
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