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目的:探讨重组IL-12腺病毒载体(AdCMV IL-12)对肺炎小鼠NK细胞功能的影响。方法:90只BALB/c小鼠随机分为4组,A组:空白对照组15只,不给予病毒,滴鼻生理盐水。B组:对照组25只,每只鼻腔滴入105TCID50/0.1mL的Ad3悬液100μL。C组:AdCMV IL-12组25只,每只鼻腔滴入105TCID50的Ad3/0.1mL悬液100μL,3d后滴鼻给药吸入AdCMV IL-12(5×108PFU/只)100μL。D组:AdCMVLacZ组25只,每只鼻腔滴入105TCID50/0.1mL的Ad3悬液100μL,3d后滴鼻给药吸入AdCMVLacZ(5×108PFU/只)100μL。开始滴入病毒定为0d。于第5天,摘眼球取血,颈椎脱臼处死,无菌条件下取肺,用于转染及病毒毒力检测。无菌条件下取脾脏做NK细胞活性检测。测定血清IL-12及FIN-γ含量。结果:含有目的基因的腺病毒载体能有效转染支气管及肺组织并在局部表达目的基因。血清IL-12浓度为C组>A组>B组、D组,NK、FIN-γ浓度为C组>B组、D组>A组。病毒滴度:A组细胞生长良好。B、D组于10-4出现细胞病变,C组于原液中出现病变。结论:重组IL-12腺病毒载体可通过滴鼻给药成功转染至肺组织,升高血液IL-12浓度,通过增加NK细胞活性而增加了IFN-γ的释放,从而导致病毒滴度下降,限制CVB病毒复制的作用。
Objective: To investigate the effect of recombinant IL-12 adenovirus vector (AdCMV IL-12) on NK cell function in pneumonia mice. Methods: Ninety BALB / c mice were randomly divided into 4 groups. Group A: 15 rabbits in control group, without virus and nasal saline. Group B: control group of 25, each nasal drop 105TCID50 / 0.1mL Ad3 suspension 100μL. Group C: AdCMV IL-12 group, 25 mice, each with 105TCID50 Ad3 / 0.1mL nasal instillation 100μL, nasal administration inhaled AdCMV IL-12 (5 × 108PFU / only) 100μL. Group D: AdCMVLacZ group 25, each nasal drop 105TCID50 / 0.1mL Ad3 suspension 100μL, nasal administration after 3 days inhalation AdCMVLacZ (5 × 108PFU / only) 100μL. Start dropping the virus at 0d. On the fifth day, the eyeball was taken for blood, the cervical dislocation was sacrificed and the lungs were removed under aseptic conditions for transfection and virulence detection. Aseptic conditions to take spleen NK cell activity test. Serum IL-12 and FIN-γ levels were measured. Results: The adenoviral vector containing the target gene could effectively transfect the bronchus and lung tissue and express the target gene locally. Serum levels of IL-12 in group C> group A> group B, group D, group NK and group FIN-γ were group C> group B, group D> group A. Virus titer: A group of cells grow well. B, D group appeared in 10-4 cytopathic lesions in group C in the original solution. CONCLUSIONS: Recombinant IL-12 adenovirus vector can be successfully transfected into the lung tissue by intranasal administration to increase the concentration of blood IL-12 and increase the release of IFN-γ by increasing the activity of NK cells, leading to the decrease of virus titer , Limiting the role of CVB virus replication.