Streptamer versus tetramer-based selection of functional cytomegalovirus-specific T cells
Cytomegalovirus (CMV) disease constitutes a serious complication after stem cell transplantation and has been treated by adoptive transfer of donor-derived CMV-specific CD8+ T cells. CMV-specific CD8+ T cells were selected by multimers, and the technologies may alter the function of these T cells. T...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2013-06-01
|
| Series: | Journal of the Formosan Medical Association |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0929664612001866 |
| _version_ | 1818980262324731904 |
|---|---|
| author | Xin-Chao Wang Hua Pang Xun Xu Anita Schmitt Mathias Freund Michael Schmitt Bao-An Chen |
| author_facet | Xin-Chao Wang Hua Pang Xun Xu Anita Schmitt Mathias Freund Michael Schmitt Bao-An Chen |
| author_sort | Xin-Chao Wang |
| collection | DOAJ |
| description | Cytomegalovirus (CMV) disease constitutes a serious complication after stem cell transplantation and has been treated by adoptive transfer of donor-derived CMV-specific CD8+ T cells. CMV-specific CD8+ T cells were selected by multimers, and the technologies may alter the function of these T cells. Therefore, here we evaluated the impact of multimer reagents on the function of CD8+ T lymphocytes.
Methods: CMV-specific CD8+ T cells were purified from the peripheral blood of donors using tetra- and streptamer technologies. The functional status of purified CMV-specific CD8+ T cells was assessed by multiparametric immunophenotyping and carboxyfluorescein succinimidyl ester proliferation assays as well as by enzyme-linked immunospot assays.
Results: A similar percentage of CMV-specific CD8+ T cells could be purified by both tetra- (90%) and streptamer (92%) technologies. That constitutes a 30- to 50-fold concentration of CMV-specific CD8+CD45RA+CCR7−effector T cells. Selected cells secreted interferon-gamma and granzyme B upon stimulation with CMVpp65 peptide, thus demonstrating their functionality.
Conclusion: Our study demonstrated that both tetra- and streptamer technologies can be used to purify CMV-specific cytotoxic CD8+ T cells for adoptive T-cell transfer. Both multimer technologies did not have any negative influence on the proliferation of selected T cells. Importantly, streptamer technology is available at good manufacturing practice level. |
| first_indexed | 2024-12-20T17:12:38Z |
| format | Article |
| id | doaj.art-86b45fbc4066495d82b7d2d4b80680ac |
| institution | Directory Open Access Journal |
| issn | 0929-6646 |
| language | English |
| last_indexed | 2024-12-20T17:12:38Z |
| publishDate | 2013-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of the Formosan Medical Association |
| spelling | doaj.art-86b45fbc4066495d82b7d2d4b80680ac2022-12-21T19:32:06ZengElsevierJournal of the Formosan Medical Association0929-66462013-06-01112633834510.1016/j.jfma.2012.02.020Streptamer versus tetramer-based selection of functional cytomegalovirus-specific T cellsXin-Chao Wang0Hua Pang1Xun Xu2Anita Schmitt3Mathias Freund4Michael Schmitt5Bao-An Chen6Department of Hematology, The Affiliated Zhongda Hospital of Southeast University, Nanjing, ChinaDepartment of Hematology, The Fourth Center Clinical College of Tianjin Medical University, Tianjin, ChinaDepartment of Internal Medicine III, University of Rostock, Rostock, GermanyDepartment of Internal Medicine V, Hematology, Oncology and Rheumatology, University of Heidelberg, Heidelberg, GermanyDepartment of Internal Medicine III, University of Rostock, Rostock, GermanyDepartment of Internal Medicine V, Hematology, Oncology and Rheumatology, University of Heidelberg, Heidelberg, GermanyDepartment of Hematology, The Affiliated Zhongda Hospital of Southeast University, Nanjing, ChinaCytomegalovirus (CMV) disease constitutes a serious complication after stem cell transplantation and has been treated by adoptive transfer of donor-derived CMV-specific CD8+ T cells. CMV-specific CD8+ T cells were selected by multimers, and the technologies may alter the function of these T cells. Therefore, here we evaluated the impact of multimer reagents on the function of CD8+ T lymphocytes. Methods: CMV-specific CD8+ T cells were purified from the peripheral blood of donors using tetra- and streptamer technologies. The functional status of purified CMV-specific CD8+ T cells was assessed by multiparametric immunophenotyping and carboxyfluorescein succinimidyl ester proliferation assays as well as by enzyme-linked immunospot assays. Results: A similar percentage of CMV-specific CD8+ T cells could be purified by both tetra- (90%) and streptamer (92%) technologies. That constitutes a 30- to 50-fold concentration of CMV-specific CD8+CD45RA+CCR7−effector T cells. Selected cells secreted interferon-gamma and granzyme B upon stimulation with CMVpp65 peptide, thus demonstrating their functionality. Conclusion: Our study demonstrated that both tetra- and streptamer technologies can be used to purify CMV-specific cytotoxic CD8+ T cells for adoptive T-cell transfer. Both multimer technologies did not have any negative influence on the proliferation of selected T cells. Importantly, streptamer technology is available at good manufacturing practice level.http://www.sciencedirect.com/science/article/pii/S0929664612001866adoptive immunotherapycytomegaloviruscytomegalovirus-specific T cellsstem cell transplantationstreptamerstetramer |
| spellingShingle | Xin-Chao Wang Hua Pang Xun Xu Anita Schmitt Mathias Freund Michael Schmitt Bao-An Chen Streptamer versus tetramer-based selection of functional cytomegalovirus-specific T cells Journal of the Formosan Medical Association adoptive immunotherapy cytomegalovirus cytomegalovirus-specific T cells stem cell transplantation streptamers tetramer |
| title | Streptamer versus tetramer-based selection of functional cytomegalovirus-specific T cells |
| title_full | Streptamer versus tetramer-based selection of functional cytomegalovirus-specific T cells |
| title_fullStr | Streptamer versus tetramer-based selection of functional cytomegalovirus-specific T cells |
| title_full_unstemmed | Streptamer versus tetramer-based selection of functional cytomegalovirus-specific T cells |
| title_short | Streptamer versus tetramer-based selection of functional cytomegalovirus-specific T cells |
| title_sort | streptamer versus tetramer based selection of functional cytomegalovirus specific t cells |
| topic | adoptive immunotherapy cytomegalovirus cytomegalovirus-specific T cells stem cell transplantation streptamers tetramer |
| url | http://www.sciencedirect.com/science/article/pii/S0929664612001866 |
| work_keys_str_mv | AT xinchaowang streptamerversustetramerbasedselectionoffunctionalcytomegalovirusspecifictcells AT huapang streptamerversustetramerbasedselectionoffunctionalcytomegalovirusspecifictcells AT xunxu streptamerversustetramerbasedselectionoffunctionalcytomegalovirusspecifictcells AT anitaschmitt streptamerversustetramerbasedselectionoffunctionalcytomegalovirusspecifictcells AT mathiasfreund streptamerversustetramerbasedselectionoffunctionalcytomegalovirusspecifictcells AT michaelschmitt streptamerversustetramerbasedselectionoffunctionalcytomegalovirusspecifictcells AT baoanchen streptamerversustetramerbasedselectionoffunctionalcytomegalovirusspecifictcells |