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目的建立异型登革热病毒(DENV)感染所产生的抗体依赖感染增强作用(ADE)的体外实验研究模型。方法将梯度稀释的抗-Ⅱ型登革热膜前蛋白prM单克隆抗体与Ⅲ型登革热病毒(MOI=3)于37℃、5%CO2培养箱内孵育90min形成抗体-病毒复合物后感染K562细胞,通过间接免疫荧光染色,real-time PCR检测DENV抗原在细胞内的表达增强。结果抗体依赖增强作用(ADE)的效果取决于抗体浓度。稀释度为1/2560的抗体与DENV结合共同感染K562后,细胞内登革热病毒抗原的免疫荧光染色结果有明显增强,real-time PCR检测的结果也进一步证实了K562细胞内登革热病毒数量显著增加。结论成功建立了登革热病毒的异型抗体依赖感染增强作用的体外研究模拟,为后续的ADE作用机制的研究提供了可靠的研究模型和实验基础。
OBJECTIVE: To establish an in vitro experimental model of ADE induced by DENV infection. Methods K562 cells were infected with gradient dilution anti-dengue membrane preprotein prM monoclonal antibody and dengue virus type III (MOI = 3) at 37 ℃ in 5% CO2 incubator for 90 min to form antibody-virus complex. By indirect immunofluorescence staining, the expression of DENV antigen in cells was detected by real-time PCR. Results The effect of antibody-dependent enhancement (ADE) depends on the antibody concentration. The result of immunofluorescence staining of dengue virus antigens in K562 cells infected with DENV with dilution 1/2560 antibody was significantly enhanced. The result of real-time PCR further confirmed the significant increase of dengue virus in K562 cells. Conclusion The in vitro study of dengue virus-specific antibody-dependent infection enhancement was successfully established in vitro and provided a reliable research model and experimental basis for the subsequent study of ADE mechanism.