Figure 1.

Proof of the HLA-C*06:02-restricted autoimmune response against melanocytes in psoriasis. Following separation of epidermis and dermis of lesional biopsies from HLA-C*06:02⁺ psoriasis patients CD8⁺ T cells (A) were isolated from epidermal cell suspensions using magnetic beads coated with CD8 antibodies. The arrow “A” points to a CD8⁺ T cells (encircled) isolated from lesional psoriatic epidermis which is rosetted by magnetic beads with a melanocyte attached. T-cell receptor (TCR) α and β chain mRNA of single T cells was transcribed into cDNA and sequenced by a newly developed method for single cell TCR analysis (54), cloned into expression plasmids and (B) expressed in a TCR^α−β− mouse reporter T-hybridoma cell line stably transfected with a plasmid for super green fluorescent protein under control of NFAT (57–59). The TCR hybridoma cells were cocultured with various primary cell types or cell lines either positive or negative for HLA-C*06:02 (C). Upon TCR ligation, the hybridoma cells produce green fluorescent protein which was visualized by UV-fluorescence microscopy or FACS analysis (D). (E) Shows the activation of the Vα3S1/Vβ13S1-TCR hybridoma by spindle-shaped HLA-C*06:02⁺ primary melanocytes in a coculture experiment.