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Example Pro5® MHC Class I Pentamer Staining:
Webb, et al. Profound elevation of CD8+ T cells expressing the intraepithelial lymphocyte marker CD103 (αE/β7 Integrin) in high-grade serous ovarian cancer Gynecological Oncology (2010) doi:10.1016/j.ygyno.2010.05.016
Figure 2. CD8+ T cells specific for the tumor antigen NY-ESO-1 are predominantly CD103+. Flow cytometry analysis of T cells derived from the malignant ascites of an HLA-A2+ EOC patient (IROC013) who demonstrated reactivity to an HLA-A2-restricted epitope of NY-ESO-1. The analysis shows the frequency of CD8+ lymphocytes staining positive with an A*02:01/SLLMWITQV (NY-ESO-1157–165) Pro5® Pentamer. Pentamer-positive CD8+ T cells were further characterized for CD103 expression (right panel). All events were first gated on total lymphocyte populations by forward and side scatter.
Example ProT2® MHC Class II Tetramer Staining:
Kramer C. et al. (2020), "Generation and reactivity analysis of human recombinant monoclonal antibodies directed against epitopes on HLA-DR"
AJT DOI: 10.1111/ajt.15950
Figure 1. HLA-DR-specific memory B cell clones isolated from peripheral blood using ProT2® MHC Class II Tetramers. A, Representative example of 3 independent experiments depicting the flow cytometry gating strategy to live single cell sort CD3-CD27+IgD-HLA-DR tetramer double positive B cells from PBMC. B, IgG antibody production by the clones was determined by ELISA. C, IgG positive clones were screened with HLA class II Lifecodes Lifescreen Deluxe kit to detect HLA antibody. The kit contains 5 groups of HLA class II beads and each data point represents a single bead group. D, HLA-specific B cell clones were tested with SAB assays to confirm tetramer specificity used for cell sorting. Each dot presents 1 clone and only MFI of bead with tetramer specificity is depicted. E, Percentage of sorted HLA-specific B cells from total memory B cells. F, Percentage of HLA antibody-producing B cell clones from sorted B cells. On the x-axis are the specificity of the tetramers used depicted.
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