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. 2017 Apr 25;101(4):e86–e92. doi: 10.1097/TP.0000000000001646

FIGURE 3.

FIGURE 3.

Some sera contain antibodies which bind class I SLAs. Flow cytometry evaluated IgG binding from 119 sera to PBMC collected from swine lacking GGTA1/CMAH/B4GalNT2 genes before (intact y axis) and after (absorbed x axis) depletion of antibodies recognizing pig-specific carbohydrate xenoantigens. The preabsorption and postabsorption MFI of 73 sera, shown in panel A, exhibited less than 25% variation. Forty-six sera demonstrated more that 25% variation in MFI values and are not shown to simplify analysis. To determine if residual antibodies to TKO swine PBMC-targeted class I SLA molecules, absorbed sera from panel A were incubated with either GGTA1/CMAH/B4GalNT2 knockout (x axis, class I SLA+) or GGTA1/SLA1 knockout (y axis, class I SLA−) swine PBMC. MFI of IgG binding was determined by flow cytometry. Sera exhibiting greater (B) or less (C) than 25% variation of binding in the presence or absence of class I SLA are shown. Nine sera with high levels of binding to GGTA1/CMAH/B4GalNT2 knockout caused by class I SLA are highlighted (red dots, panels A and B).