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目的观察120μg重组peroxiredoxin蛋白(rTgPrx)联合弗氏佐剂皮下免疫小鼠诱导的特异性体液免疫应答和细胞免疫应答。方法 105只6周龄BALB/c小鼠随机分为3组,分别用100μl PBS(PBS组)、120μg rTgPrx(120P组)或120μg rTgPrx+50μl佐剂(120PA组)皮下免疫3次,间隔2周。rTgPrx溶于50μl PBS,首次免疫和加强免疫分别加入弗氏完全佐剂和不完全佐剂。分别在末次免疫后第1、2、3、4、5、6、7周,每组处死5只小鼠,ELISA法检测血清IgG和小肠冲洗液sIgA;分离并计数小肠上皮内淋巴细胞(intestinal intraepithelial lymphocytes,IEL)和脾淋巴细胞。结果 120PA组小鼠免疫后第2~5周血清IgG水平显著高于120P组(F=16.875,P<0.05),120P组免疫后第2、3周显著高于PBS组(F=19.417,P<0.05);120PA组免疫后第4周小肠冲洗液sIgA水平显著高于120P组(F=14.638,P<0.05),于第5周有所下降并保持平稳,但仍显著高于120P组(F=15.316,P<0.05);120PA组免疫后第1~4周IEL计数显著高于120P组(F=23.634,P<0.05),第2~6周脾淋巴细胞数显著高于120P组(F=15.279,P<0.05)。结论 120μg rTgPrx联合弗氏佐剂皮下免疫小鼠可诱导高水平的体液免疫应答及细胞免疫应答。
Objective To observe the specific humoral and cellular immune responses induced by subcutaneous immunization with 120 μg recombinant peroxiredoxin protein (rTgPrx) and Freund’s adjuvant. Methods 105 6-week-old BALB / c mice were randomly divided into three groups and immunized subcutaneously with 100μl PBS (PBS group), 120μg rTgPrx (120P group) or 120μg rTgPrx + 50μl adjuvant (120PA group) week. rTgPrx was dissolved in 50μl PBS, the first immunization and booster immunization were added Freund’s complete adjuvant and incomplete adjuvant. At the 1st, 2nd, 3rd, 4th, 5th, 6th and 7th week after the last immunization, 5 mice in each group were sacrificed and the serum IgG and intestinal sIgA were detected by ELISA. The intestinal epithelial lymphocytes intraepithelial lymphocytes, IEL) and splenic lymphocytes. Results The level of serum IgG in 120PA group was significantly higher than that in 120P group (F = 16.875, P <0.05) at 2 ~ 5 weeks after immunization, <0.05). The level of sIgA in the intestinal fluid of the 120PA group was significantly higher than that of the 120P group (F = 14.638, P <0.05) in the 4th week after immunization. It decreased at the 5th week and remained stable but still significantly higher than that of the 120P group F = 15.316, P <0.05). The number of IELs in the first to fourth week after immunization in 120PA group was significantly higher than that in 120P group (F = 23.634, P <0.05), and the number of spleen lymphocytes in the second to sixth weeks was significantly higher than that in 120P group F = 15.279, P <0.05). Conclusion 120μg rTgPrx combined with Freund’s adjuvant can induce high humoral and cellular immune responses in mice.