Establishment of a flow cytometry-based assay for the detection of monocyte-mediated antibody-de-pendent cell-mediated cytotoxicity
10.3760/cma.j.issn.0254-5101.2015.01.004
- VernacularTitle:基于流式细胞术评价单核细胞介导的抗体依赖性细胞介导的细胞毒效应的方法学建立和应用
- Author:
Hua LIANG
;
Xiangbo HUANG
;
Tao SHEN
;
Yiming SHAO
- Publication Type:Journal Article
- Keywords:
Monocytes;
Antibody-dependent cell-mediated cytotoxicity;
Flow cytometry
- From:Chinese Journal of Microbiology and Immunology
2015;(1):18-22
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a flow cytometry-based assay for the detection of monocyte-me-diated antibody-dependent cell-mediated cytotoxicity ( ADCC ) .Methods P815 cells double stained with PKH26 and carboxyfluorescein succinimidyl ester ( CFSE ) were used as target cells and coated with P 815 specific antibodies to form antigen-antibody complexes .The peripheral blood mononuclear cells were isolated as effector cells and co-cultured with the antigen-antibody complexes .The CD3-CD14+PKH26+CFSE-cell population were gated by flow cytometry .Optimized effector/target cell ratio and incubation time for killing assay were identified .Monocyte-mediated ADCC in 23 patients with chronic HCV infection and 22 healthy subjects were analyzed .Results The monocyte-mediated ADCC could be evaluated through analyzing the CD3-CD14+PKH26+CFSE-cells with flow cytometry .The optimized effector/target cell ratio was 10 ∶1 and the optimized time for incubation was 4 h.Monocyte-mediated ADCC was inhibited in patients with chronic HCV infection as compared with healthy subjects (P=0.009).Conclusion A flow cytometry-based assay for the detection of monocyte-mediated ADCC was established , which could be used as a fast , sensitive and safety method for the evaluation of monocyte-mediated ADCC during viral infections and the research and de-velopment of drugs .
