Rapid generation of combined CMV-specific CD4+ and CD8+ T-cell lines for adoptive transfer into recipients of allogeneic stem cell transplants
| Autor: | Christian Sinzger, Gabriele Kuntz, Mario Assenmacher, Hermann Einsele, Max S. Topp, Dorothee Wernet, John D.M. Campbell, Georg Rauser |
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| Rok vydání: | 2003 |
| Předmět: |
CD4-Positive T-Lymphocytes
Adoptive cell transfer Immunology Cell Culture Techniques Cytomegalovirus Blood Donors CD8-Positive T-Lymphocytes Major histocompatibility complex Lymphocyte Activation Biochemistry Interleukin 21 Antigen MHC class I Cytotoxic T cell Humans Transplantation Homologous Antigens Viral biology Hematopoietic Stem Cell Transplantation virus diseases Cell Biology Hematology T lymphocyte Virology Adoptive Transfer biology.protein Peptides CD8 |
| Zdroj: | Blood. 103(9) |
| ISSN: | 0006-4971 |
| Popis: | Adoptive transfer of cytomegalovirus (CMV)-specific T cells can restore long-lasting, virus-specific immunity and clear CMV viremia in recipients of allogeneic stem cell transplants if CD4+ and CD8+ CMV-specific T cells are detected in the recipient after transfer. Current protocols for generating virus-specific T cells use live virus, require leukapheresis of the donor, and are time consuming. To circumvent these limitations, a clinical-scale protocol was developed to generate CMV-specific T cells by using autologous cellular and serum components derived from a single 500-mL blood draw. CMV-specific T cells were stimulated simultaneously with CMV-specific major histocompatibility complex class I (MHC I)- restricted peptides and CMV antigen. Activated T cells were isolated with the interferon-γ (IFN-γ) secretion assay and expanded for 10 days. In 8 randomly selected, CMV-seropositive donors, 1.34 × 108 combined CD4+ and CD8+ CMV-specific T cells, on average, were generated, as determined by antigen-triggered IFN-γ production. CMV-infected fibroblasts were efficiently lysed by the generated T cells, and CMV-specific CD4+ and CD8+ T cells expanded if they were stimulated with natural processed antigen. On the other hand, CD4+ and CD8+ T cell-mediated alloreactivity of generated CMV-specific T-cell lines was reduced compared with that of the starting population. In conclusion, the culture system developed allowed the rapid generation of allodepleted, highly enriched, combined CD4+ and CD8+ CMV-specific T cells under conditions mimicking good manufacturing practice. (Blood. 2004; 103:3565-3572) |
| Databáze: | OpenAIRE |
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