We purchased C1R (B lymphoblasts lacking endogenous human leukocyte antigen (HLA)-A and HLA-B expression), Jiyoye, and EB-3 cells from the American Type Culture Collection (Rockville, MD, USA) and cultured them in RPMI1640 supplemented with 10% FBS. We developed C1R cells stably expressing HLA-A*02:01 or HLA-A*24:02 (C1R-A0201 or C1R-A2402, respectively) in our previous study [4 (link)]. We generated C1R-A0206, C1R-A1101, C1R-A3101, and C1R-A3303 by the nucleofection of pCAGGS vectors encoding HLA-A*02:06, HLA-A*11:01, HLA-A*31:01, and HLA-A*33:03 cDNAs, respectively.
We used the following anti-human antibodies for cell surface staining by flow cytometry analyses: We purchased CD3-APC (cat# 300311), CD3-APC-Cy7 (clone HIT3a, cat# 300317), CD4-PE-Cy7 (clone RPA-T4, cat# 300511), CD8-FITC (cat# 560960), and CD8-PE-Cy7 (clone HIT8a, cat# 301012) from BioLegend (San Diego, CA, USA). We also obtained an anti-mouse TCRβ chain-APC antibody (clone H57-597, cat# 109212) from BioLegend (San Diego, CA, USA). We purchased peptide-HLA tetramers labeled with PE from MBL (Tokyo, Japan). We analyzed all flow cytometry data using FlowJo software (ver.10, BD Biosciences).
We used the following anti-human antibodies for cell surface staining by flow cytometry analyses: We purchased CD3-APC (cat# 300311), CD3-APC-Cy7 (clone HIT3a, cat# 300317), CD4-PE-Cy7 (clone RPA-T4, cat# 300511), CD8-FITC (cat# 560960), and CD8-PE-Cy7 (clone HIT8a, cat# 301012) from BioLegend (San Diego, CA, USA). We also obtained an anti-mouse TCRβ chain-APC antibody (clone H57-597, cat# 109212) from BioLegend (San Diego, CA, USA). We purchased peptide-HLA tetramers labeled with PE from MBL (Tokyo, Japan). We analyzed all flow cytometry data using FlowJo software (ver.10, BD Biosciences).