Influence of cell isolation method on the optimization of CD4+ T cell expansion using anti-CD3/CD28 coated beads
Backgrounds: Activation of CD4+ T lymphocytes with anti-CD3/CD28 coated magnetic beads promotes intrinsic resistance to HIV as well as cell expansion. The propose of this study is to define the optimal cell isolation protocol for expansion of CD4+ T lymphocytes by using anti-CD3/CD28 coated bead sti...
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th-mahidol.318852018-10-19T12:18:38Z Influence of cell isolation method on the optimization of CD4+ T cell expansion using anti-CD3/CD28 coated beads Nattawat Onlamoon Michittra Boonchan Petai Unpol Narakorn Khunweeraphong Kasama Sukapirom Palanee Ammaranond Kovit Pattanapanyasat Mahidol University Chulalongkorn University Immunology and Microbiology Medicine Backgrounds: Activation of CD4+ T lymphocytes with anti-CD3/CD28 coated magnetic beads promotes intrinsic resistance to HIV as well as cell expansion. The propose of this study is to define the optimal cell isolation protocol for expansion of CD4+ T lymphocytes by using anti-CD3/CD28 coated bead stimulation with an ultimate goal of using these cells for adoptive immunotherapy. Methods: CD4+ T cells were isolated from healthy donor blood samples using three different methods including immunorosette formation, negative selection and CD8 depletion using immunomagnetic beads. These cells were activated with anti-CD3/CD28 coated beads at a bead to cell ratio of 1:1 and cell expansion was carried for 3 weeks. Cell numbers, cell viability and phenotypic characterization were determined by trypan blue exclusion and flow cytometry. Results: Purified CD4+ T lymphocytes which were isolated via immunorosette formation can be expanded up to 1000-fold within 3 weeks with high viability (90%) and high purity of CD4+ T lymphocytes (>95%). However, cell expansion from purified CD4+ T lymphocytes which were isolated by negative selection and CD8-depletion provided approximately 300-fold expansion. Conclusions: The results demonstrate that purified CD4+ T lymphocytes from immunorosette formation provided the highest CD4+ T lymphocyte expansion when stimulated with anti-CD3/CD28 coated beads. This method can be used to obtain a large number of expanded CD4+ T cells for adoptive immunotherapy. 2018-10-19T05:02:08Z 2018-10-19T05:02:08Z 2013-08-20 Article Asian Pacific Journal of Allergy and Immunology. Vol.31, No.2 (2013), 99-105 10.12932/AP0222.31.2.2013 22288694 0125877X 2-s2.0-84881524631 https://repository.li.mahidol.ac.th/handle/123456789/31885 Mahidol University SCOPUS https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84881524631&origin=inward |
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Immunology and Microbiology Medicine Nattawat Onlamoon Michittra Boonchan Petai Unpol Narakorn Khunweeraphong Kasama Sukapirom Palanee Ammaranond Kovit Pattanapanyasat Influence of cell isolation method on the optimization of CD4+ T cell expansion using anti-CD3/CD28 coated beads |
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Backgrounds: Activation of CD4+ T lymphocytes with anti-CD3/CD28 coated magnetic beads promotes intrinsic resistance to HIV as well as cell expansion. The propose of this study is to define the optimal cell isolation protocol for expansion of CD4+ T lymphocytes by using anti-CD3/CD28 coated bead stimulation with an ultimate goal of using these cells for adoptive immunotherapy. Methods: CD4+ T cells were isolated from healthy donor blood samples using three different methods including immunorosette formation, negative selection and CD8 depletion using immunomagnetic beads. These cells were activated with anti-CD3/CD28 coated beads at a bead to cell ratio of 1:1 and cell expansion was carried for 3 weeks. Cell numbers, cell viability and phenotypic characterization were determined by trypan blue exclusion and flow cytometry. Results: Purified CD4+ T lymphocytes which were isolated via immunorosette formation can be expanded up to 1000-fold within 3 weeks with high viability (90%) and high purity of CD4+ T lymphocytes (>95%). However, cell expansion from purified CD4+ T lymphocytes which were isolated by negative selection and CD8-depletion provided approximately 300-fold expansion. Conclusions: The results demonstrate that purified CD4+ T lymphocytes from immunorosette formation provided the highest CD4+ T lymphocyte expansion when stimulated with anti-CD3/CD28 coated beads. This method can be used to obtain a large number of expanded CD4+ T cells for adoptive immunotherapy. |
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Mahidol University |
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Mahidol University Nattawat Onlamoon Michittra Boonchan Petai Unpol Narakorn Khunweeraphong Kasama Sukapirom Palanee Ammaranond Kovit Pattanapanyasat |
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Article |
author |
Nattawat Onlamoon Michittra Boonchan Petai Unpol Narakorn Khunweeraphong Kasama Sukapirom Palanee Ammaranond Kovit Pattanapanyasat |
author_sort |
Nattawat Onlamoon |
title |
Influence of cell isolation method on the optimization of CD4+ T cell expansion using anti-CD3/CD28 coated beads |
title_short |
Influence of cell isolation method on the optimization of CD4+ T cell expansion using anti-CD3/CD28 coated beads |
title_full |
Influence of cell isolation method on the optimization of CD4+ T cell expansion using anti-CD3/CD28 coated beads |
title_fullStr |
Influence of cell isolation method on the optimization of CD4+ T cell expansion using anti-CD3/CD28 coated beads |
title_full_unstemmed |
Influence of cell isolation method on the optimization of CD4+ T cell expansion using anti-CD3/CD28 coated beads |
title_sort |
influence of cell isolation method on the optimization of cd4+ t cell expansion using anti-cd3/cd28 coated beads |
publishDate |
2018 |
url |
https://repository.li.mahidol.ac.th/handle/123456789/31885 |
_version_ |
1763496595478806528 |