FIGURE 6.

L363 Fab binds similarly to mCD1d and CD1d mutants loaded with KRN7000 compared with NKT hybridoma cells. A coated plate ELISA assay was used to measure L363 binding, and an APC-free NKT cell stimulation assay was used to assess TCR recognition based on IL-2 secretion. Similar patterns of recognition were observed of various CD1d mutants by both L363 Fab and iNKT cells hybridomas expressing TCRs containing Vβ8.2 (2C12 and 1.2) or Vβ10 (1.4). With the exception of the G155W mutation of mCD1d that blocks the binding of L363 Fab, the observed binding pattern correlates with the shared footprint illustrated in Fig. 3. The data represent the means ± range of two independent experiments.