甲乙肝联合疫苗中不同配比对小鼠Th1/Th2型细胞因子变化的影响

Changes of Th1/Th2 cytokine in mouse immunized by the different concentrations of combined HAB vaccine

目的通过注射甲乙肝联合疫苗和单价甲肝、乙肝疫苗来观察小鼠在免疫过程中细胞因子的变化情况,衡量应答的性质,探讨两种抗原对小鼠细胞因子变化的不同作用以及乙肝抗原对甲肝抗原的作用。方法选用150只Balb/c小鼠随机分成注射联苗640Eu/mL:10μg/mL组、640Eu/mL:20μg/mL组、单价甲肝抗原640Eu/mL组、单价乙肝抗原10μg/mL组、单价乙肝抗原20μg/mL组和佐剂AL(OH)3对照组,每组25只。于第4周加强免疫1次,分别在初次免疫注射后第1、2、3、4周及加强免疫后第2周,眼眶采血,分离血清,每组各5只小鼠,上流式细胞仪检测小鼠血清5种细胞因子(TNF、IFN-γ、IL-5、IL-4、IL-2)含量。结果疫苗诱导产生的Th1/Th2型细胞因子均随时间的变化而变化,5种细胞因子均约在第3周达最大值,其中小鼠产生最多的是TNF;加强免疫后,640Eu/mL:20μg/mL联苗组刺激小鼠产生细胞因子水平高于640Eu/mL:10μg/mL联苗组。结论甲乙肝联合疫苗能刺激机体产生特异性细胞免疫应答,甲乙肝联合疫苗能诱导机体产生与单价相同或略强的Th1/Th2型细胞因子。

Objective To approach different contribution of cytokine in mouse serum induced by hepatitis A and hepatitis B antigen, and to investigate the effects hepatitis B surface antigen on hepatitis A antigen in the course of immunization by observing the changes of Th1/ Th2 cytokine in mouse immunized respectively by the two different concentrations of combined HAB vaccine, monovalent hepatitis A vaccine, and monovalent hepatitis B vaccine. Methods A total of 150 female Balb/c mice were divided into 6 groups randomly （25 mice/group） ： control group[ 1 mg/mL AL （OH）3 ], univalent hepatitis A group （640 Eu/mL）, univalent hepatitis B group 1 （10μg/mL）, univalent hepatitis B group 2 （20μg/mL）, combined HAB vaccine group 1 （640 Eu/mL： 10μg/mL）, combined HAB vaccine group 2 （640 Eu/mL：20 μg/mL）. Prime-boost immunization was carried out on the forth week. The blood in fossa orbitalis of 5 mice from every group were collected at week 1, week 2, week 3, week 4, week 6 after the first injecting. The content of cytokines （TNF, IFN-γ, IL-5, IL-4, and IL-2） were counted by the flow cytometry after the serum were separated. Results TNF was the highest cytokine in five cytokines detected. The cytokines in 640 Eu/ml： 20 μg/mL combined HAB vaccine group is higher than that of 640 Eu/ml： 10 μg/mL group. Th1/Th2 cytokine is maximal on week 3 after first immunization. Conclusion Combined HAB vaccine can boost specific immune response on the initial and boost immunity, and also can induce Th1/Th2 cytokine which is same or higher as compared with that induced by monovalent hepatitis A vaccine and monovalent hepatitis B vaccine.