Table 1.
Summary of multiplex MHC-based strategies for detection of antigen-responsive T cells
| Strategy for “high-throughput” detection of specific CD8 T cell responses | References | Reported sensitivity (frequency of specific T cells) | Reported order of complexity | Combined with functional and/or phenotypic readout | Offer recovery of AG specific T cells |
|---|---|---|---|---|---|
|
DNA barcode labeling of pMHC multimers Detection by sequencing |
Bentzen et al. [28] | 0.005% of CD8+ T cells | 1031 different specificities per sample (0.20 × 106–10 × 106 PBMCs) | Yes | Yes |
|
Three-dimensional combinatorial encoding of metal labeled pMHC multimers Detection by cytometry by time-of-flight |
Newell et al. [25] | 0.001% of CD8+ T cells (after MHC tetramer-based enrichment) | 109 different specificities per sample (16 × 106–302 × 106 PBMCs) | Yes | No |
|
Two-dimensional combinatorial encoding of fluorescent labeled pMHC multimers Detection by flow cytometry |
Andersen et al. [31], Hadrup et al. [20] | 0.002% of CD8+ T cells | 36 different specificities per sample (2 × 106 PBMCs) | Yes | Yes |
|
Combinatorial pooling of fluorescent labeled pMHC multimers and splitting of sample material into each pool Detection by flow cytometry |
Klinger et al. [27] | Not reported | 16 different specificities per analysisa | Noa | Yes |
|
Multi-dimensional combinatorial encoding of fluorescent labeled pMHC multimers Detection by flow cytometry |
Newell et al. [21] | 0.01% of CD8+ T cells | 15 different specificities per sample (2 × 106 PBMCs) | No | Yes |
| MHC microarray: pMHC tetramers spotted onto polyacrylamide slides combined with an aggressive washing scheme | Brooks et al. [19] | 0.02% of CD8+ T cells (after CD8+ T cell enrichment) | 40 different specificities per assay (0.8–1.2 × 106 T cells) | No | No |
| MHC microarray: DNA-probed pMHC tetramers spotted onto complementary DNA probe printed glass slide | Kwong et al. [18] | 0.1% of CD8+ T cells | 3 different specificities per assay (106 T cells) | No | Yes |
| MHC microarray: Dimeric pMHCs spotted onto antibody-coated polyacrylamide microscopy slide | Deviren et al. [17] | 0.01% of CD8+ T cells | 2 different specificities per assay (106 cells) | No | No |
| MHC microarray: Dimeric pMHCs co-spotted with cytokine capture antibodies onto polyacrylamide microscopy slide | Chen et al. [16] | 0.01% of CD8+ T cells | 7 different specificities per assay (CD8+ T cells isolated from 5 × 107 PBMCs) | Yes | No |
| MHC microarray: pMHCs co-spotted onto polystyrene microscopy slide with cytokine capture antibodies and co-stimulatory molecules | Stone et al. [15] | 0.1% of CD8+ T cells | 30 different specificities per assay (106 cells) | Yes | No |
| MHC microarray: pMHCs tetramers spotted onto polyacrylamide microscopy slide | Soen et al. [14] | 0.1% of CD8+ T cells | 7 different specificities per assay (106 cells) | Yes (Ca2+ levels) | No |
aA non-MHC variation of this approach based on activation of T cells has been applied to screen for 30 specificities in parallel