Background: Multimer technology is widely used to screen antigen-specific immune recovery after allogeneic hematopoietic stem cell transplantation (allo-HSCT) as it enables identification, enumeration, phenotypic characterization and isolation of virus-specific T-cells. Novel approaches of multimerization might improve on classical tetramer staining; however, their use as standard monitoring technique to quantify antigen-specific cells has not been validated yet. We have compared two of these available multimeric complexes: pentamer and streptamer to select the best strategy for the incorporation into clinical monitoring practice.
Methods: CMVpp65 -specific HLA-A*02:01 CD8 T lymphocytes (CTL * -CMVpp65 ) were examined with pentamer and streptamer in peripheral blood cells of 77 healthy volunteers. Quantitative and qualitative analyses were performed to compare the precision and repeatability, sensitivity and accuracy and specificity of both technologies by flow cytometry.
Results: Standard deviation for both techniques was less than 0.05 showing that they are repetitive and precise. Both techniques significantly correlated at high frequencies (r = 0.9422; P < 0.0001) but it was lost at lower levels (<1%) of CTL * -CMVpp65 (r = 0.3351; P = 0.1376). Streptamer is more accurate for the detection of CTL * -CMVpp65 providing significantly closer values to the theoretical ones (P < 0.0001) as pentamer binds unspecifically to a notable proportion of non-CMV-specific CD8 T-cells.
Conclusion: Our results suggest that streptamer multimer provides precise, accurate and specific results to detect CTL * -CMVpp65 by flow cytometry. Streptamer multimer can be used not only for the monitoring of early CTL * -CMVpp65 reconstitution in immunosuppressed patients following allo-HSCT but also, in conjunction with its reversibility role, for the isolation of CTL * -CMVpp65 for its future use in adoptive immunotherapy. © 2016 International Clinical Cytometry Society.
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Streptamer technology allows accurate and specific detection of CMV-specific HLA-A*02 CD8 T cells by flow cytometry.
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Oncohematology Research Group, Navarrabiomed-Miguel Servet Foundation, IDISNA (Navarra's Health Research Institute), Pamplona, Spain.
Background: Multimer technology is widely used to screen antigen-specific immune recovery after allogeneic hematopoietic stem cell transplantation (allo-HSCT) as it enables identification, enumeration, phenotypic characterization and isolation of virus-specific T-cells. Novel approaches of multimerization might improve on classical tetramer staining; however, their use as standard monitoring technique to quantify antigen-specific cells has not been validated yet. We have compared two of these available multimeric complexes: pentamer and streptamer to select the best strategy for the incorporation into clinical monitoring practice.
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Multimer monitoring has become a standard technique for detection of antigen-specific T cells. The term "multimer" refers to a group of reagents based on the multimerisation of molecules in order to raise avidity and thus stabilize binding to their ligand. Multimers for detection of antigen-specific T-cell responses are based on major histocompatibility complex class I peptide complexes.
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Laboratory of Experimental Tumor Immunology, Department of Medical Oncology, Erasmus University Medical Center-Daniel den Hoed Cancer Center, 3015 GE Rotterdam, The Netherlands.
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Department of Internal Medicine III, Clinical Stem Cell Transplantation and Immunotherapy, University Clinic Rostock, 18055 Rostock, Germany.
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