![Figure 1. Recombinant HLA-A2/peptide monomers can stimulate allo-restricted, peptide-specific CTLs. (A-E) PBMCs of an HLA-A2- donor were stimulated 4 times in vitro with autologous B lymphocytes coated with A2/pMelA monomers. Responding T lymphocytes were stained with FITC-labeled antihuman CD8 antibodies and PE-labeled A2 tetramers containing the peptide epitopes indicated in each panel. The percentage of tetramer-binding CD8+ T lymphocytes is also indicated in each panel. (F) The killing activity of the same cultures stained in panels A to E was tested in chromium-release assays using the human TAP-deficient, HLA-A2+ T2 cells coated with the indicated peptides as targets. Black bars (▪) show the killing activity (effector-target ratio [E/T], 10:1) of cultures that were stimulated with A2/pMelA monomers in the presence of IL-2 and IL-7; white bars (□), the killing activity of control cultures stimulated in parallel with cytokines only. (G-K) PBMCs of an HLA-A2- donor were stimulated 5 times in vitro with autologous B lymphocytes coated with A2/pWT126 monomers and stained with FITC-labeled antihuman CD8 antibodies and PE-labeled A2 tetramers containing the peptide epitopes indicated in each panel. The percentage of tetramer-binding CD8+ T lymphocytes is also indicated in each panel. (L) The killing activity of the same cultures stained in panels G to K was tested against T2 cells coated with the indicated peptides. Filled bars show the killing activity (E/T, 10:1) of cultures that were stimulated with A2/pWT126 monomers in the presence of IL-2 and IL-7; open bars show the killing activity of control cultures stimulated in parallel with cytokines only.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/103/12/10.1182_blood-2003-11-3903/5/zh80120462980001.jpeg?Expires=1722567849&Signature=kIZVl8aLvKD31VpScb8ZGYbDdj7vWPietrywJt~3XIcEMkM0ZnctvmKlzuKd9NNedrtxQibo1UxiV92zjDKIap5Rh2z2kvd9EOvHtE1sCODQ4c8m9mLd8KWRkLWD1EAWUUCS9uK8vRQ3G8Z~UdYZ91t~8r~F73N1Hq-llfE96kgQwMy0rOZvmn3Rz7y2lcoOlx0Wl-hkUASEKHeFp9dA1bbsexNZn5bHgOgAXXqz88ehAw2HuPWKp8WYPLg0agWRwjvpxcBJ-O4JKYiI0~aCbu0toR~D4HfhMnhSFVGYE4EPgk7afES-5oirrcgV--D91XSHnfdd8~yCl1Gyg1T~2g__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Recombinant HLA-A2/peptide monomers can stimulate allo-restricted, peptide-specific CTLs. (A-E) PBMCs of an HLA-A2- donor were stimulated 4 times in vitro with autologous B lymphocytes coated with A2/pMelA monomers. Responding T lymphocytes were stained with FITC-labeled antihuman CD8 antibodies and PE-labeled A2 tetramers containing the peptide epitopes indicated in each panel. The percentage of tetramer-binding CD8+ T lymphocytes is also indicated in each panel. (F) The killing activity of the same cultures stained in panels A to E was tested in chromium-release assays using the human TAP-deficient, HLA-A2+ T2 cells coated with the indicated peptides as targets. Black bars (▪) show the killing activity (effector-target ratio [E/T], 10:1) of cultures that were stimulated with A2/pMelA monomers in the presence of IL-2 and IL-7; white bars (□), the killing activity of control cultures stimulated in parallel with cytokines only. (G-K) PBMCs of an HLA-A2- donor were stimulated 5 times in vitro with autologous B lymphocytes coated with A2/pWT126 monomers and stained with FITC-labeled antihuman CD8 antibodies and PE-labeled A2 tetramers containing the peptide epitopes indicated in each panel. The percentage of tetramer-binding CD8+ T lymphocytes is also indicated in each panel. (L) The killing activity of the same cultures stained in panels G to K was tested against T2 cells coated with the indicated peptides. Filled bars show the killing activity (E/T, 10:1) of cultures that were stimulated with A2/pWT126 monomers in the presence of IL-2 and IL-7; open bars show the killing activity of control cultures stimulated in parallel with cytokines only.
