[Using flow cytometry to detect peripheral blood eosinophils and their related molecules].
OBJECTIVE
To set up an accurate and rapid method to detect the phenotype of peripheral blood eosinophils using flow cytometry.


METHODS
10(-4) mol/L of theophylline, 10(-4) of dexamethasone and 10(-8) mol/L of rhIL-5 were added to peripheral blood sampled from normal human subjects. Anti-CD16-PE mAb and FITC-labelled mAbs to eosinophil's molecules were used to perform double labeling staining. Simultaneously, CD16-FL2 was used to install gating so as to accurately locate eosinophils. And then their molecules were examined and analyzed.


RESULTS
The eosinophils were accurately located. Theophylline and dexamethasone effectively inhibited activation of eosinophils and shedding of CD62L on eosinophils caused by IL-5.


CONCLUSIONS
Our method can accurately detect eosinophils in peripheral blood samples and their molecules. In addition, only a tiny amount of blood sample is needed and few artificial factors are involved in the detection. Therefore, this method may be an ideal both in basic immunological research and clinical laboratory examination.
Cao
Yun-xin
YX
Department of Immunology, Fourth Military Medical University, Xi'an 710032, China.
Shi
Jie-ran
JR
Han
Wei-ning
WN
D003907:Dexamethasone / Q000494:pharmacology*; D047091:Eosinophil Granule Proteins / Q000378:metabolism; D004804:Eosinophils / Q000166:cytology / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D015848:Interleukin-5 / Q000037:antagonists & inhibitors; D019041:L-Selectin / Q000378:metabolism; D010641:Phenotype; D011994:Recombinant Proteins / Q000494:pharmacology; D013806:Theophylline / Q000494:pharmacology*
D047091:Eosinophil Granule Proteins; D015848:Interleukin-5; D011994:Recombinant Proteins; D019041:L-Selectin; D003907:Dexamethasone; D013806:Theophylline