Improved preparation of class I HLA tetramers and their use in detecting CMV-specific CTL.
Different methods were used to prepare HLA tetramers and the yields of each method were compared. Our results indicate that preliminary refolding of the heavy chain (Hc) and light chain (beta 2m) yields more monomer than the typical conventional method with urea-solubilized Hc and beta 2m. We then used the corresponding tetramers to detect cytomegalovirus (CMV)-specific cytotoxic T lymphocytes (CTL). Increasing data suggest that the adoptive transfer of CMV-specific CTL constitutes an effective strategy against CMV infections. We designed a method that efficiently induces CMV-specific CTL to a higher frequency in vitro than is currently achieved. This method increased the percentage of CMV-specific CTL from below 1% to 20% of PBL, accounting for more than 40% of CD8+ T cells. Successful HLA tetramer preparation provides the basis for the subsequent detection of CMV-specific CTL in clinical applications.
Ruan
Guang-Ping
GP
Institute of Molecular Immunology, Southern Medical University, Guangzhou 510515, China.
Ma
Li
L
Meng
Min-Jie
MJ
Zhu
Yong
Y
Chen
Ze-Hong
ZH
Lin
Ying
Y
Wu
Zheng-Qiang
ZQ
He
Xiao-Wei
XW
Wang
Ju-Fang
JF
Wang
Xiao-Ning
XN