Figure 1

hCARΔcyt transgene map and expression in lymph node cells and thymocytes of hCAR transgenic mice. A. An 807 bp fragment (hatched box) encoding the extracellular and transmembrane region with a truncated cytoplasmic tail (TM-RKKKRR^*) of the human CAR was cloned into the EcoRI/SmaI site of VA hCD2. The hCARΔcyt coding region was inserted between intron 1 of hCD2 and the poly A1 site from the hCD2 3' untranslated region. All potential ATG start codons in exon 1 of hCD2 have been mutated [16]. B. Lymphoid cells were isolated from three different transgenic lines (6, 13 and 18) and stained with RmcB anti-human CAR mAb and anti-mouse CD4, CD8 or CD19 before analysis on a FACSscan. The quadrants were set based on the staining of a hCARΔcyt negative littermate control (data not shown). The graphs represent two-color dot plots of lymph node cells gated on viable lymphoid gate; the percentage of cells in each quadrant is indicated. C. Two-color dot plots of thymocytes stained with anti-CD4 and anti-CD8 mAbs. The staining with the anti-hCAR RmcB mAb is represented as a histogram for each corresponding quadrant with the percentage of positive cells.