Quantum dot/pMHC multimers vs. phycoerythrin/pMHC tetramers for identification of HLA-A*0201-restricted pHBV core antigen18-27-specific T cells.
Mol Med Rep
; 16(6): 8605-8612, 2017 Dec.
Article
in En
| MEDLINE
| ID: mdl-28765875
ABSTRACT
Detection of human leukocyte antigens-A2-restricted p-hepatitis B virus (HBV) core antigenspecific cytotoxic T lymphocytes (CTLs) is important in the study of HBV immunopathogenesis and vaccine design. Currently, major histocompatibility complex (MHC) class I/peptide(p) MHCI tetramers are considered the optimal tools to detect antigenspecific CTLs. However, the MHCtetramer technique also has certain drawbacks and is under continuous development. The quantum dot (QD) bioconjugates nanotechnology with its unique inorganicbiological properties has been developing fast. However, QD/pMHC multimers have seldom been used for the identification of the C1827 epitope, which is important in HBV infection. QD/pMHC multimers were synthesized by metalaffinity coordination and an avidinbiotin system. In the present study they were characterized by transmission electron microscopy, dynamic light scattering and fluorescence spectrophotometry. C1827specific CTLs were obtained by ex vivo expansion of CD8+ T cells. Cultured CTLs were tested for the secretion level of interferon (IFN)γ by ELISA and for cytotoxicity by lactate dehydrogenase release assay. Then, the performance of phycoerythrin (PE)/pMHC tetramers and QD/pMHC multimers were compared by flow cytometry. The synthesized QD/pMHC multimers dispersed well and their emission spectrum exhibited only slight differences compared with original QDs. C1827specific CTLs not only secreted IFNγ but also effectively targeted T2 cells pulsed with peptide C1827. The frequencies of C1827specific CTLs determined by QD/pMHC multimers were higher compared with PE/pMHC tetramers. The present results suggested that QD/pMHC multimers may be able to characterize greater numbers of C1827specific CTLs with increased sensitivity compared to conventional strategies.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Phycoerythrin
/
T-Lymphocytes, Cytotoxic
/
Histocompatibility Antigens Class I
/
HLA-A2 Antigen
/
Hepatitis B virus
/
Quantum Dots
/
Protein Multimerization
/
Antigens, Viral
Type of study:
Diagnostic_studies
Limits:
Humans
Language:
En
Journal:
Mol Med Rep
Year:
2017
Document type:
Article
Affiliation country:
Puerto Rico