Jianmeng Zhu, Yiping Li, Lei Li, Jian Wang, Hongqin Wang, Wenzhong Hong, Ke Hao, Yadan Xue, Bingyu Chen, Zhen Wang

Detection of antigen-specific cytotoxic T lymphocytes (CTLs) is the foundation for understanding hepatocellular carcinoma immune pathology and hepatocellular carcinoma immunotherapy. However, the classical method for labeling CTLs, major histocompatibility complex (MHC)-peptide tetramer, has drawbacks and needs further improvement. Here, as a new detection probe, a graphene-based MHC-peptide multimer was developed for sensitively and selectively identifying hepatocellular carcinoma-specific T-cells. To assess its detection efficiency, reduced graphene oxide (RGO) was functionalized with hemin and streptavidin to prepare a functionalized HRGO-streptavidin complex. Biotinylated MHC-peptide monomer was subsequently constructed onto HRGO to generate a detection probe for CTL labeling. The number of T-cells was detected through the reaction between HRGO and tetramethylbenzidine. Using HRGO/MHC-peptide multimers, the number of T-cells was efficiently detected in both the induction system in vitro and in peripheral blood of patients. HRGO/MHC-peptide multimers methodology has application prospects in the detection of antigen peptide-specific T cells.