Streptamer-based selection of WT1-specific CD8+ T cells for specific donor lymphocyte infusions

Wang, Xinchao; Schmitt, Anita; Chen, Baoan; Xu, Xun; Mani, Jiju; Linnebacher, Michael; Freund, Mathias; Schmitt, Michael

doi:10.1016/j.exphem.2010.07.002

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Volume 38, Issue 11 , Pages 1066-1073, November 2010

Streptamer-based selection of WT1-specific CD8⁺ T cells for specific donor lymphocyte infusions

Xinchao Wang
- Department of Internal Medicine III, University of Rostock, Rostock, Germany
- Department of Hematology and Oncology, Zhongda Hospital, Southeast University Medical School, Nanjing, P.R. China
- Department of Oncology, The Fourth Center Clinical College of Tianjin Medical University, Tianjin, P.R. China
Anita Schmitt
- Department of Internal Medicine III, University of Rostock, Rostock, Germany
Baoan Chen
- Department of Hematology and Oncology, Zhongda Hospital, Southeast University Medical School, Nanjing, P.R. China
Xun Xu
- Department of Internal Medicine III, University of Rostock, Rostock, Germany
- Department of Immunology, Jiangsu University, Zhenjiang, P.R. China
Jiju Mani
- Department of Internal Medicine III, University of Rostock, Rostock, Germany
Michael Linnebacher
- Department of Surgery, University of Rostock, Rostock, Germany
Mathias Freund
- Department of Internal Medicine III, University of Rostock, Rostock, Germany
Michael Schmitt
- Department of Internal Medicine III, University of Rostock, Rostock, Germany
- Offprint requests to: Michael Schmitt, M.D., Ph.D., Clinical Stem Cell Transplantation and Immunotherapy, Department of Internal Medicine III, University of Rostock, Ernst-Heydemann-Str. 6, Rostock 18055, Germany

Received 26 December 2009; received in revised form 2 July 2010; accepted 6 July 2010. published online 16 July 2010.

Objective

Donor lymphocyte infusions may generate a desirable graft-versus-leukemia effect, but also elicit a noxious graft-versus-host disease. A positive selection of leukemia (antigen)-specific T cells would be highly desirable. In this study, we focused on the immunogenic leukemia antigen Wilms’ Tumor gene 1 (WT1).

Materials and Methods

We employed the technology of streptamers available at good manufacturing practice level to first determine the frequency of human leukocyte antigen−A2 restricted WT1-specific CD8⁺ T cells. Then, specific cells were labeled with streptamers and selected by magnetic cell separation. Purity and immunophenotype of selected cells were analyzed.

Results

Twenty-one of 40 healthy donors had naïve WT1-specific CD8⁺ T-cell frequencies of >0.5%, and 8 of 40 even >1.0% of all CD8⁺ T cells. In 7 of 10 acute myeloid leukemia patients, the frequencies were 0.5% to 3.65%. After positive selection by magnetic cell separation, a 60-fold increase with a purity of up to 17.79% in the lymphocyte gate and 86.18% in the CD8⁺ T-cell gate could be achieved for CD8⁺WT1streptamer⁺CD28^+/−CD45RA⁺CCR7⁻ effector T cells.

Conclusions

Streptamer technology allows selection of pure WT1-specific effector T cells. This is a prerequisite for clinical applications targeting tumor-specific antigens, such as adoptive T-cell transfer.