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目的建立以人原代树突状细胞(dendritic cells,DCs)为宿主的登革病毒(dengue virus,DENV)感染模型。方法采集基因型为CD209,-139A/-336A的人抗凝血标本,分离获得DCs前体细胞,利用rh GM-CSF及rh IL-4诱导,获得未成熟DCs。瑞氏染色鉴定DCs的形态,流式细胞术鉴定DCs的分化效率,淋巴细胞增殖实验鉴定DCs的生物学功能。以I型登革病毒GZ2002株为模式病毒进行病毒感染实验,建立并评估以人原代DCs为宿主的登革病毒感染模型。结果分离获得的DCs前体细胞经细胞因子刺激后成功分化为未成熟DCs,分化效率高达90%。获得的DCs具有典型的细胞形态及生物学功能,可有效刺激淋巴细胞增殖。登革病毒GZ2002株能成功感染诱导获得的原代DCs,促进DCs成熟,并复制产生感染性子代病毒,证明细胞感染模型成功建立。结论成功建立以DCs为宿主的登革病毒感染模型。
Objective To establish a dengue virus (DENV) infection model using human primary dendritic cells (DCs) as the host. Methods Human anticoagulant samples of CD209, -139A / -336A genotypes were collected. DCs precursor cells were isolated and induced by rh GM-CSF and rhIL-4 to obtain immature DCs. The morphology of DCs was identified by Wright’s staining, the differentiation efficiency of DCs was identified by flow cytometry, and the biological function of DCs was identified by lymphocyte proliferation assay. The dengue virus type I dengue virus strain GZ2002 was used as a model virus for virus infection experiments to establish and evaluate a dengue virus infection model using human primary DCs as a host. Results The isolated DCs precursor cells successfully differentiated into immature DCs stimulated by cytokines, the differentiation efficiency of up to 90%. The obtained DCs have typical cell morphology and biological functions, which can effectively stimulate lymphocyte proliferation. The dengue virus GZ2002 strain can successfully infect the induced primary DCs, promote the maturation of DCs and replicate the infectious progeny virus to prove that the cell infection model is successfully established. Conclusion The model of dengue virus infection with DCs was successfully established.