Figure 3.

Stimulation of CTL 121 by COS-7 cells transiently cotransfected with cDNA clone 668 and with a sequence encoding an HLA-B35 molecule. 1,500 CTL were added into microwells containing 10,000 BB49-SCCHN or 20,000 transfected COS-7 cells. The production of TNF was measured after 24 h of coculture by testing the toxicity of the supernatants for the TNF-sensitive WEHI-164.13 cells. (A) COS-7 cells were cotransfected with cDNA 668 and a cDNA coding for HLA-B35 obtained from the RNA of patient LB1047. The autologous BB49-SCCHN cells were used as a positive control for stimulation. COS-7 cells, cotransfected with HLA-B35 and nonrelevant cDNA clone F10, were used as a negative control. (B) COS-7 cells were cotransfected with cDNA 668 and either a cDNA coding for the autologous HLA-B*3503 molecule or with a cDNA coding for the allogeneic HLA-B*3501 molecule. Stimulator cells included the autologous BB49-SCCHN cells as a positive control and COS-7 cells transfected with either cDNA 668, the HLA-B*3503 cDNA, or the HLA-B*3501 cDNA alone as negative controls.