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. 2011 Apr 12;96(7):1024–1032. doi: 10.3324/haematol.2010.037481

Figure 2.

Figure 2.

HLA class I allele-specific allorecognition of CD8 T-cell subsets. MLR cultures were tested for allo-HLA reactivity in an IFN-γ ELISpot assay on d12 (i.e. 5 days after first allo-HLA restimulation on d7). (A) Reactivity to original K562-HLA mismatch stimulator cells, as well as to parental K562 cells and K562 transfectant cells carrying an irrelevant HLA allele. To demonstrate HLA-restriction of detected reactivity, monoclonal antibodies blocking T-cell receptor-HLA interactions (and IgG isotype controls) were used. Representative results with naïve-enriched CD8 T-cell subsets of donor 898 for allo-HLA-A*02:01 are shown. (B) Numbers of spot-forming cells from d12 cultures initiated with sorted subsets (CD45RA, CD45RO, CD62L, CCR7) or entire CD8 T cells from six healthy donors. Allo-HLA mismatch alleles used for in vitro stimulation were HLA-A*02:01 (upper panels), HLA-B*35:03 and HLA-C*03:03 (lower panels). (C) Box plots and P values of data presented in (B). Median (line), 25th to 75th percentile (box), minimum and maximum values (error bars) are indicated. If PBMC numbers were limited, MLR stimulations were restricted to fewer allo-HLA alleles or T-cell subsets.