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目的初步研究登革病毒1、2型单、双价病毒样颗粒(VLP)的免疫原性。方法表达纯化各型VLP,用纯化好的VLP免疫BALB/c小鼠,将BALB/c小鼠随机分PBS组、灭活DENV1、灭活DENV2、纯化VLP1、纯化VLP2、纯化VLP1+2组,以灭活病毒为阳性对照,PBS为阴性对照。ELISA法检测小鼠血清抗VLP抗体效价以及血清IFN-γ、TNF-α细胞因子水平,流式细胞术检测小鼠脾细胞CD4+细胞和CD8+细胞数。结果登革病毒1、2型单、双价VLP均能刺激免疫小鼠产生一定程度的抗血清效价,VLP1的抗体水平较VLP2的低,双价VLP组针对DENV2抗原的抗体水平上升明显,是单价VLP2的2～3倍;攻毒后,灭活DENV1和VLP2可以产生高水平的IFN-γ,分别为60和35pg/ml,双价VLP组的较低;VLP2组的TNF-α一直保持较高水平,双价VLP组中攻DENV2组的TNF-α水平较DENV1组高;三次免疫后,各实验组较之PBS组CD4+细胞的比例都有下降,而CD8+细胞变化不大,除灭活DENV1组有增加,其余各组都有减少。结论无论单、双价VLP均能刺激小鼠产生一定程度的体液免疫和细胞免疫反应,联合免疫有一定程度的协同性。
Objective To study the immunogenicity of dengue virus type 1 and 2 bivalent virus-like particles (VLPs). METHODS: BALB / c mice were immunized with the purified VLP by immunoprecipitation. The BALB / c mice were randomly divided into PBS group, inactivated DENV1, inactivated DENV2, purified VLP1, purified VLP2 and purified VLP1 + To inactivate the virus as a positive control, PBS as a negative control. Serum anti-VLP antibody titers and serum levels of IFN-γ and TNF-α were measured by ELISA. The number of CD4 + cells and CD8 + cells in splenocytes were detected by flow cytometry. Results Both single and double VLP of dengue virus type 1 and 2 could stimulate the mice to produce a certain degree of antiserum titer. The antibody level of VLP1 was lower than that of VLP2. The antibody level of double VLP group against DENV2 antigen increased significantly. Is 2 to 3 times higher than the monovalent VLP2; inactivation of DENV1 and VLP2 can produce high levels of IFN-γ, 60 and 35 pg / ml, respectively, and lower in the bivalent VLP group; TNF-α in the VLP2 group has been The levels of TNF-αin DENV2 group were higher than that of DENV1 group in bivalent VLP group. After three immunizations, the proportion of CD4 + cells in each experimental group decreased compared with that in PBS group, while the changes of CD8 + Inactivated DENV1 group increased, the remaining groups have decreased. Conclusion Both VLP and VLP can stimulate mice to produce a certain degree of humoral and cellular immune responses, and there is a certain degree of synergism in combination immunization.