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目的观察重组人白细胞介素-2(rhuIL-2)联合弓形虫可溶性速殖子抗原(STAg)滴鼻免疫小鼠诱导的免疫应答,探讨rhuIL-2的佐剂效应及适宜剂量。方法5～6周龄BALB/c小鼠60只,随机分为6组,每组10只。实验组以STAg20μg或分别加rhuIL-2250、500、1000、2000 IU滴鼻免疫,抗原与佐剂溶于20μlPBS中,对照组以PBS滴鼻,免疫2次,间隔2周。末次免疫后30d,颈椎脱臼处死全部小鼠,ELISA法检测血清IgG和粪便sIgA水平;分离脾淋巴细胞、肠上皮内淋巴细胞(iIEL),并计数。结果与PBS组相比,各佐剂组血清IgG水平都有增高,其中STAg+500 IU IL-2组IgG水平增高最显著(P<0.01);STAg+500 IU IL-2组和STAg+1000 IU IL-2组粪便sIgA抗体水平显著高于PBS组和STAg组(P<0.05)。联合IL-2佐剂免疫小鼠脾淋巴细胞和产生了增殖性应答,其中STAg+500 IU IL-2组和STAg+1000 IU IL-2组脾淋巴细胞数显著高于PBS组和STAg组(P<0.05);TAg+500 IU IL-2组iIEL高于PBS组(P<0.05)。结论rhuIL-2作为佐剂联合STAg鼻内免疫小鼠可有效诱导粘膜免疫、系统的细胞免疫和体液免疫应答;500 IU IL-2为鼻内免疫小鼠的适宜剂量。
Objective To observe the immune response induced by recombinant human interleukin-2 (rhuIL-2) combined with intranasal inoculation of Tachyzoites soluble tachyzoite antigen (STAg), and to explore the adjuvant effect and appropriate dose of rhuIL-2. Methods Sixty BALB / c mice aged 5 to 6 weeks were randomly divided into 6 groups with 10 in each group. In the experimental group, intranasal immunization with STAg20μg or rhuIL-2250, 500,1000 and 2000 IU respectively was used. The antigen and adjuvant were dissolved in 20μl PBS, and the control group was intranasally instilled with PBS for 2 weeks at intervals of 2 weeks. Thirty days after the last immunization, all mice were sacrificed by cervical dislocation, serum IgG and feces sIgA were detected by ELISA. Splenic lymphocytes and iIEL were isolated and counted. Results Compared with PBS group, serum IgG levels increased in all adjuvant groups. The highest IgG level in STAg + 500 IU IL-2 group was significantly higher than that in PBS group (P <0.01). The levels of IgG in STAg + 500 IU IL-2 group and STAg + 1000 The level of sIgA antibody in feces of IU IL-2 group was significantly higher than that in PBS group and STAg group (P <0.05). The number of splenic lymphocytes in the group of STAg + 500 IU IL-2 and the group of STAg + 1000 IU IL-2 was significantly higher than that of PBS group and STAg group ( P <0.05). The iIEL in TAg + 500 IU IL-2 group was higher than that in PBS group (P <0.05). Conclusion Mice immunized with rhuIL-2 as an adjuvant and intranasally with STAg can effectively induce mucosal immunity, systemic cellular immunity and humoral immune response. 500 IU IL-2 is the appropriate dose for intranasal immunization in mice.