In vitro expansion of Wilms' tumor protein 1 epitope- specific primary T cells from healthy human peripheral blood mononuclear cells

被引:1
|
作者
Heijden, Sanne van der [1 ,2 ]
Flumens, Donovan [1 ]
Versteven, Maarten [1 ]
Peeters, Stefanie [1 ]
De Reu, Hans [1 ]
Campillo-Davo, Diana [1 ]
Willemen, Yannick [1 ]
Ogunjimi, Benson [4 ]
Van Tendeloo, Viggo [1 ]
Berneman, Zwi N. [1 ]
Anguille, Sebastien [1 ,3 ]
Smits, Evelien [2 ]
Lion, Eva [1 ]
机构
[1] Univ Antwerp, Vaccine & Infect Dis Inst VAXINFECTIO, Fac Med & Hlth Sci, Lab Expt Hematol LEH, Univplein 1, B-2610 Antwerp, Belgium
[2] Univ Antwerp, Ctr Oncol Res CORE, Integrated Personalized & Precis Oncol Network IPP, Univplein 1, B-2610 Antwerp, Belgium
[3] Antwerp Univ Hosp UZA, Div Hematol, Drie Eikenstr 655, B-2650 Edegem, Belgium
[4] Univ Antwerp, Ctr Hlth Econ Res & Modeling Infect Dis CHERMID, VAXINFECTIO, Univplein 1, B-2610 Antwerp, Belgium
来源
STAR PROTOCOLS | 2023年 / 4卷 / 01期
关键词
Cancer; Cell Biology; Cell Culture; Cell Isolation; Flow Cytometry/Mass Cytometry; Health Sciences; Immunology;
D O I
10.1016/j.xpro.2023.102053
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页数:18
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