Experiment Overview
| Repository ID: | FR-FCM-ZY3S | Experiment name: | ENUMERATION OF WT1-SPECIFIC CD8+ T CELLS | MIFlowCyt score: | 46.94% |
| Primary researcher: | Sarah Matko | PI/manager: | Sarah Matko | Uploaded by: | Sarah Matko |
| Experiment dates: | 2012-02-28 - 2016-09-15 | Dataset uploaded: | Mar 2017 | Last updated: | Jun 2017 |
| Keywords: | [Wilms’ tumor antigen (WT1)] [antigen-specific CD8+ T cells] [MHC Streptamer] [singleplatform] | Manuscripts: | [28544366] |
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| Organizations: | None | ||||
| Purpose: | The advent of novel strategies to generate leukemia-associated-antigen (LAA)-specific T cells for adoptive immunotherapies creates a demand for standardized good laboratory practice (GLP)-compliant enumeration assays to provide a secure clinical environment - whether it is to identify potential donors, define therapeutic doses for transplantation or monitor clinical success. Here, we introduce a no-wash assay based on single-platform cell enumeration and Streptamer staining to determine the Wilms? tumor antigen 1 (WT1)-specific T cell immunity in clinical samples. | ||||
| Conclusion: | The accuracy of the assay was high for all performed experiments. Differences were apparent regarding the limit of detection/quantification (LOD/LOQ). Results obtained for WT1 yielded a notably higher LOD/LOQ than for virus-specific multimers. However, subsequent screening of 22 healthy individuals revealed significantly higher values staining for WT1 than for HIV pol-specific T cells. These differences could not be observed applying a classical wash-protocol. Therefore, we strongly suggest the use of no-wash single-platform assays in combination with MHC Streptamer staining for the detection of low affinity T cells. | ||||
| Comments: | None | ||||
| Funding: | This work was supported by a grant from the Bundesministerium fuer Bildung und Forschung (BMBF) - (joint project: Streptamer-based selection of leukemia-specific T cells for adoptive cellular therapy after stem cell transplantation [Stratos], grant no.: 01GU1108A). | ||||
| Quality control: | Triplicates, Limit of detection, limit of quantification, goodness of fit, recovery, linearity, intra-assay-deviation, inter-assay-deviation | ||||
