Optimizing the conditions for in vitro expansion of HBV protein epitope-specific CTLs
HAO Youhua
CHEN Ling
YANG Xinxing
DING Honghui
FENG Xuemei
ZHU Fan
LIU Zhi
YANG Dongliang
Abstract：Objective To optimize the in vitro expansion conditions HBV protein epitope-specific CTL. Methods PBMC from the acute self-limited HBV patients was stimulated by peptide under the conditions of different concentrations, different stimulating intervals, and different cytokines. The cells were harvested and the frequencies of the HBV specific CTLs were obtain by FACS using FITC-marked CD8 antibody and HBV peptide -MHC-I molecular tetramers. Then the obtained frequencies of HBV specific CTLs in dif-ferent culture conditions were compared. Results 1) When the peptide concentration was 20 μg/μL, the peak of specific CTLa was the highest compared with other concentration; 2) when the stimulating interval was 10 days, the peak of specific CTLs was the highest compared with other intervals; 3) when the cocuhure eytokines were IL-2, IL-7, and IL-15, the peak of specific CTLs was the highest compared with other cytokines. Conclusion The highest frequency of specific CTLs is appeared when PBMC is in vitro stimulated by the stimulating peptide of 20 μ/mL, for 10 days, and with addition of IL-2, IL-7, IL-15 cytokines.
Keywords：HBV proteinepitope-specific CTLexpanding
Publication Date：2009-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 413-417 )