In vitro expansion of Wilms’ tumor protein 1 epitope-specific primary T cells from healthy human peripheral blood mononuclear cells
oleh: Sanne van der Heijden, Donovan Flumens, Maarten Versteven, Stefanie Peeters, Hans De Reu, Diana Campillo-Davo, Yannick Willemen, Benson Ogunjimi, Viggo Van Tendeloo, Zwi N. Berneman, Sébastien Anguille, Evelien Smits, Eva Lion
| Format: | Article |
|---|---|
| Diterbitkan: | Elsevier 2023-03-01 |
Deskripsi
Summary: Wilms’ tumor protein 1 (WT1) is a tumor-associated antigen overexpressed in various cancers. As a self-antigen, negative selection reduces the number of WT1-specific T cell receptors (TCRs). Here, we provide a protocol to generate WT137-45-specific TCRs using healthy human peripheral blood mononuclear cells. We describe the expansion of WT1-specific T cell clones by two consecutive in vitro stimulations with autologous WT137-45-pulsed dendritic cells and peripheral blood lymphocytes. We then detail the detection with human leukocyte antigen/WT137-45 tetramers. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.