JCI - Identification of heme oxygenase-1–specific regulatory CD8+ T cells in cancer patients

Identification of heme oxygenase-1–specific regulatory CD8⁺ T cells in cancer patients

Mads Hald Andersen, Rikke Bæk Sørensen, Marie K. Brimnes, Inge Marie Svane, Jürgen C. Becker, Per thor Straten

Published in Volume 119, Issue 8

J Clin Invest. 2009; 119(8):2245–2256 doi:10.1172/JCI38739

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Figure 7

The inhibitory activity of supernatant from HO-1–reactive cells.

(A) 200 HLA-A2/HO212–pentamer⁺ T cells were directly isolated by FACS sorting from PBLs from an MM patient and cultured overnight in 100 μl medium. The supernatant was removed and added to an ELISPOT with PBLs from either an RCC patient or another MM patient. The TAA (Survivin_96–104) was added to the wells. The same ELISPOT was performed with and without TAA in regular medium with no supernatant. Numbers represent counted spots. (B) 300 HLA-A2/HO212 pentamer⁺ T cells were directly isolated by FACS sorting from PBLs from an RCC patient and cultured overnight in 100 μl medium. The supernatant was removed and added to a chrome release assay. The effector cells were a RhoC-specific T cell clone; target cells were T2-A3 cells pulsed with RhoC (RAGLQVRKNK) peptide. The effector/target ratio was 3:1. (C) The supernatants from either directly isolated HO-1–specific T cells, CMV-specific T cells, or bulk CD8⁺ cells were added to an ELISPOT with PBLs from 4 different cancer patients; 2 MM patients (MM50, MM48) and 2 RCC patients (Ur27, Ur25), as well as TAA (MART-1_27–35 or Survivin_96–104). All measurements were made in duplicate. Error bars indicate SD.