Detection Of Eosinophils Ex Vivo And In Vivo By Silencing Murine CCR3Gene Via RNA Interference

Objective：Using RNA interference for silencing mice CCR3gene to evaluate the effect oneosinophils and to provide the basis for the pathogenesis of allergic rhinitis.Methods：An ex vivo culture system was adopted to purified eosinophils from mice bonemarrow progenitors. The lentiviral vector expressing shRNA for RNA interference ofmice CCR3gene was transfected into eosinophils by calcium phosphate precipitationmethod. The interference effects of interfering virions were detected by RT-PCR andWestern Blot. Using flow cytometric to analyze the apoptosis of eosinophil. TheCCR3gene lentiviral vector was adopted in the murine allergic rhinitis by the use ofnasal topical administration， the application of RT-PCR and Western Blot weredetected by mouse bone marrow，peripheral blood and nasal mucosa eosinophilsCCR3mRNA and protein expression level. Analyzed numbers of CD34+cells of micesin each group by Flow cytometric.Results：（1）The RT-PCR and Western Blot results indicate that lentiviral vector of CCR3gene has interference effect on the level of mRNA and protein inhibition.（2）Interfering virions promote the apoptosis of eosinophil.（3） The expression of CCR3mRNA and protein of eosinophils from bonemarrow，peripheral blood and nasal mucosa was lower in the CCR3shRNA treatmentgroup than the PBS treatment control and shRNA treatment control group (p<005).（4）The numbers of CD34+cells had no statistic difference (p＞005).Conclusion：This study suggests a therapeutic approach the genes CCR3through intranasalmucosal administration of target gene shRNA can inhibit in development，migrationand infiltration of eosinophils in the animal models with allergic rhinitis.