Xu, Yuexin http://orcid.org/0000-0002-7140-6505
Morales, Alicia J.
Cargill, Michael J.
Towlerton, Andrea M. H.
Coffey, David G.
Warren, Edus H.
Tykodi, Scott S.
Article History
Received: 25 March 2019
Accepted: 18 October 2019
First Online: 4 November 2019
Compliance with ethical standards
:
: Aprovisional patent application was filed by Yuexin Xu, Edus H. Warren and Scott S. Tykodi regarding TCRs described in this study. The other authors declare that they have no conflict of interest.
: All study protocols and donor consent documents were approved by the Institutional Review Board of Fred Hutchinson Cancer Research Center, Seattle, Washington, USA and included protocols 1495 (skin biopsy), 999.209 (skin biopsy), 868 (healthy donor leukapheresis), 1246 (cancer patient leukapheresis), and 1810 (RCC tumor, normal kidney, peripheral blood).
: Written informed consent was obtained from all individual donors of biological samples for research use included in this study. Kidney cancer donor D consent for the collection of RCC tumor tissue for study 1810 also authorized collection of clinical data from the donor’s electronic medical record, including treatment history and disease status.
: The cell lines T2, A498, MDA-231, BT-20, and SW480 were obtained from American Type Culture Collection (Manassas, VA). The cell line lenti-X 293T virus packaging cells was obtained from Clontech Laboratories (Mountain View, CA). The RCC tumor lines LB1828, BB65, DOBSKI, and the BB65-LCL line were a generous gift from Dr. Benoît Van den Eynde (Ludwig Institute for Cancer Research, Brussels, Belgium). The TM-LCL line was a generous gift from Dr. Phillip D. Greenberg and Dr. Tom B. Schmitt (Fred Hutchinson Cancer Research Center, Seattle, WA, USA). The phenotypes of these lines were verified by the source before arriving at our facility. The phenotypic characterization of RCC tumor line SST548 was previously verified [CitationRef removed]. Paired RCC tumor and PTEC cultures from four clear-cell RCC tumors and adjacent normal kidney cortex, as well as three fibroblast lines derived from skin biopsies of three additional donors were established following specific cell culture protocols to ensure propagation of the targeted cell types [CitationRef removed–CitationRef removed]. Expression of the 5T4 tumor marker on the four primary RCC lines was verified by flow cytometry, as shown in Supplementary Fig. 6. No 5T4 expression was detected on any fibroblast or PTEC lines [CitationRef removed]. PCR-based HLA-typing of genomic DNA was performed for all cell lines to select for HLA-A2 expression [CitationRef removed]. Mycoplasma was routinely tested. Phenotypes for cell morphology and growth rate were monitored regularly and did not change over time. All the experiments were performed with cells at a low passage number (≤ 10).