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. 2019 Mar 20;49(5):737–746. doi: 10.1002/eji.201847949

Figure 5.

Figure 5

CD1b‐BbGL‐II‐binding T cells are autoreactive and recognize diacylglycerol. (A) IFN‐γ production was measured by co‐culture of BC24A T cells and C1R cells transduced to express human CD1a or CD1b. The cells were incubated in the presence of BbGL‐II lipid, sonicated Borrelia burgdorferi (B.b.), or without antigen (–). A blocking antibody or isotype control was added. Data is mean value of triplicates with error bars showing SD. Data shown is representative of three independent experiments. (B) Staining of BC24A with the indicated tetramers. (C) Detection of C36:2 DAG standard or lipid extracts of human cell lines C1R, K562, and 293T cells by high mass resolution mass HPLC time of flight MS. (D) Mass spectrum between 13.5 and 13.7 min shows the presence of C34:1 DAG and C36:2 DAG in cells. (E) Cultured primary T cells or J76 cells transduced with the BC24A TCR were stained with CD1b tetramers loaded with the indicated antigens. Pregating strategy is shown in Figures 2A and 3C, respectively. BC24A staining was performed twice independently, J76‐BC24A staining was performed three times independently. One representative staining is shown.