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Updated: Aug 17, 2026

Measuring TCR-pMHC Binding In Situ using a FRET-based Microscopy Assay
Published on: October 30, 2015
T-cell receptor (TCR)/CD3 is involved in sulfated polymannuroguluronate (SPMG)-induced T lymphocyte activation
Benchun Miao1, Jing Li, Xueyan Fu
1Department of Pharmacology, Marine Drug and Food Institute, Ocean University of China, Qingdao 266003, P.R. China.
Abstract:
Sulfated polymannuroguluronate (SPMG) has entered the Phase II clinical trial as the first anti-acquired immune deficiency syndrome (AIDS) drug candidate in China. Proliferation assays showed that SPMG was effective at enhancing the proliferative response of T lymphocytes either with or without concanavalin A (ConA) stimulation. Flow cytometry (FCM) and fluorescence microscope examination revealed the significant binding of SPMG to T lymphocytes. The significant engagement of SPMG with TCR/CD3 complex was verified by competitive inhibition assay and one of the SPMG binding proteins purified by affinity chromatography from thymocyte membrane preparations was further confirmed to be CD3 component of TCR/CD3 complex via Western blotting analysis. In addition, SPMG was demonstrated to dramatically interact with ConA in a multivalent manner by surface plasmon resonance (SPR) assay. Notably, the concomitant presence of ConA and SPMG facilitated each other's binding to T cells. Together, the simultaneous interactions of SPMG with TCR/CD3 and with ConA can be highly proposed to facilitate the cross-linking of these molecules, and thus favoring costimulatory signaling, which serves to well explain the immunopotentiation and anti-human immunodeficiency virus (HIV) activities of SPMG.
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