Related Experiment Video
Updated: Aug 20, 2026
Imaging of In Situ Interferon Gamma Production in the Mouse Spleen following Listeria monocytogenes Infection
Published on: July 16, 2019
[Detection of IFN-gamma level in single CD8+ T cell]
Zhi-Jie Wang1, Hua Zhang, Xue-Liang Chen
1Department of Hematology, Qilu Hospital,Shandong University, Jinan 250012, China. jie_969@163.com
Aim:
To establish a new method capillary electrophoretic immunoassay laser-induced fluorescence (CEIA-LIF), to detect IFN-gamma level in single CD8(+) T cell.
Methods:
CD8(+) T cells were isolated from peripheral blood of two patients with severe aplastic anemia(SAA) and one normal person by Ficoll-Hypaque gradient centrifugation and then were purified by immunomagnetic microbead separation. Then purified CD8(+) T cells were incubated with digitonin for 15 min followed by FITC-anti-IFN-gamma mAb for 20 min. The single cell was detected continuously by CEIA-LIF. The feasibility of the method was confirmed by inverted microscopy and laser scanning confocal fluorescence microscopy.
Results:
The IFN-gamma content in purified CD8(+) T cells was detected under the condition of cell-membrane integrity. The IFN-gamma level in single CD8(+) T cell from 2 SAA patients was (151.53+/-28.92)zmol and (223.72+/-45.23)zmol, respectively, and much higher that from normal control (47.47+/-17.97)zmol ( P=0.001).
Conclusion:
It is feasible to quantitate IFN-gamma in single CD8(+) T cell by CEIA-LIF. CEIA-LIF might be useful in the clinical detection of intracellular cytokines.
