A novel absorption spectrometric method, based on graphene nanomaterials, for detection of hepatocellular carcinoma-specific T lymphocyte cells

Abstract

Jianmeng Zhu,1,* Yiping Li,1,* Lei Li,2 Jian Wang,1 Hongqin Wang,1 Wenzhong Hong,1 Ke Hao,3 Yadan Xue,3 Bingyu Chen,1,3 Zhen Wang1,3 1Department of Clinical Laboratory, Chun’an First People’s Hospital (Zhejiang Provincial People’s Hospital Chun’an Branch), Hangzhou, Zhejiang Province, China; 2Department of Pathophysiology, School of Basic Medical Science, Southern Medical University, Guangzhou, Zhejiang, China; 3Department of Blood Transfusion, Zhejiang Provincial People’s Hospital, People’s Hospital of Hangzhou Medical College, Hangzhou, Zhejiang, China *These authors contributed equally to this work Introduction: 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. Materials and methods: 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. Results: 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. Conclusion: HRGO/MHC-peptide multimers methodology has application prospects in the detection of antigen peptide-specific T cells. Keywords: tetramer, graphene, hemin, major histocompatibility complex multimer, cytotoxic T lymphocytes, hepatocellular carcinoma

Keywords

• tetramer
• graphene
• hemin
• major histocompatibility complex multimer
• cytotoxic T lymphocytes
• hepatocellular carcinoma.