Isolation,purification and functional identification of LAP +CD4 + regulatory T ceIIs from human peripheral blood of patients with coIorectal carcinoma
YANG Zuqing
JIANG Zhiyuan
ZHONG Wu
WANG Shijun
CAO Yunfei
Abstract：Objective To explore a method for isolating and purifying LAP +CD4 +regulatory T (LAP +CD4 +T)cells from human peripheral blood of patients with colorectal carcinoma (CRC)by magnetic activated cell sorting (MACS)and to investigate the function of these T cells in vitro.Methods LAP +CD4 +T cells were isolated by MACS two-step method (negative sorting and positive sorting).The purity and the survival of these T cells were identified by flow cytometry (FACS)and Typan blue staining.The inhibition effect of LAP +CD4 +T cells on the proliferation of LAP -CD4 +T cells was determined by CCK-8.The expression of IL-10 and TGF-βwas measured by ELISA.Results The purity of LAP +CD4 +T cells isolated by MACS was 95.16% ±2.11%.The cell viability was 93.64% ±1.33%.The proliferation of LAP -CD4 +T was obviously inhibited by intervention of LAP +CD4 +T cells.The suppressor cytokine,TGF-βand IL-10,which were secreted by LAP +CD4 +T cells was more than that of LAP -CD4 +T cells.Conclusion MACS used for isolating LAP +CD4 +T cells from peripheral blood of patients withe CRC has advantages of high efficiency,high speed,and high purity and no damage to cell activity.
Keywords：colorectal carcinomamagnetic activated cell sortingLAP +CD4 + regulatory T cells
Publication Date：2016-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 26-29 )