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目的通过过继转移日本血吸虫可溶性虫卵抗原(SEA)免疫的小鼠树突状细胞(DC)亚群,探讨SEA免疫的DC亚群在抑制过敏性哮喘中的作用及可能的作用机制。方法用CD8α和CD11c磁珠分离纯化日本血吸虫SEA免疫小鼠CD8α+DC、CD8α-DC亚群。另取24只BALB/c小鼠,随机分为4组:正常对照组、单纯哮喘组、过继转移SEA免疫CD8α+DC(SEA-CD8α+DC)组和过继转移SEA免疫CD8α-DC(SEA-CD8α-DC)组。SEA-CD8α+DC组、SEA-CD8α-DC组每只小鼠分别经尾静脉过继转移SEA免疫CD8α+DC或CD8α-DC 5×105个。1h后单纯哮喘组、SEA-CD8α+DC组、SEA-CD8α-DC组小鼠同时用卵白蛋白(OVA)诱发哮喘,4周后剖杀,取肺组织做病理切片和免疫组织化学染色,观察炎症变化,同时检测肺组织CCL-11和IL-13Rα2表达情况。结果与单纯哮喘组和SEA-CD8α+DC组比较,SEACD8α-DC组小鼠肺部炎症显著减轻。免疫组化染色后小鼠肺组织CCL-11平均吸光度值分别为:正常对照组0.1496±0.0009,单纯哮喘组1.7121±0.4994,SEA-CD8α+DC组0.9631±0.1201,SEA-CD8α-DC组0.3458±0.0543,SEACD8α-DC组与单纯哮喘组和SEA-CD8α+DC比较差异有统计学意义(P<0.05);小鼠肺组织IL-13Rα2平均吸光度值分别为:正常对照组0.1029±0.0103,单纯哮喘组0.3136±0.0174,SEA-CD8α+DC组0.3263±0.0128,SEA-CD8α-DC组0.1859±0.0294,SEA-CD8α-DC组与单纯哮喘组和SEA-CD8α+DC组比较差异有统计学意义(P<0.05)。结论日本血吸虫SEA免疫的DC亚群对过敏性哮喘有抑制作用,且CD8α-DC亚群起主要作用。
Objective To investigate the role of SEA-immunized DC subsets in the inhibition of allergic asthma and its possible mechanism by adoptive transfer of DCs subcloned by soluble Schistosoma japonicum soluble egg antigen (SEA). Methods The CD8α + DC and CD8α-DC subsets of mice immunized with SEA of Schistosoma japonicum were isolated and purified by CD8α and CD11c magnetic beads. Another 24 BALB / c mice were randomly divided into 4 groups: control group, asthma group, SEA-CD8α + DC group and adoptive transfer of SEA immunized CD8α-DC (SEA- CD8α-DC) group. Each mouse in SEA-CD8α + DC group and SEA-CD8α-DC group was immunized with 5 × 105 CD8α + DC or CD8α-DC by adoptive transfer of SEA via tail vein. After 1h, the mice in the asthma group, the SEA-CD8α + DC group and the SEA-CD8α-DC group were ovalbumin (OVA) -induced asthma at the same time. After 4 weeks, the mice were sacrificed and the lungs were taken for histopathological examination and immunohistochemical staining Inflammatory changes were detected simultaneously with the detection of lung tissue CCL-11 and IL-13Rα2 expression. Results Compared with the asthma group and the SEA-CD8α + DC group, the lung inflammation in SEACD8α-DC group was significantly reduced. The average absorbance of CCL-11 in lung tissue of mice after immunohistochemical staining were 0.1496 ± 0.0009 in normal control group, 1.7121 ± 0.4994 in simple asthma group, 0.9631 ± 0.1201 in SEA-CD8α + DC group and 0.3458 ± in SEA-CD8α-DC group 0.0543. There was significant difference between SEACD8α-DC group and asthmatic group and SEA-CD8α + DC (P <0.05). The mean IL-13Rα2 absorbance in lung tissue of the mice in the SEACD8α-DC group were 0.1029 ± 0.0103 in the normal control group, CD8α-DC group and SEA-CD8α-DC group were significantly higher than SEA-CD8α + DC group (P <0.01) <0.05). Conclusion The DC subgroup of SEA immunized by Schistosoma japonicum has an inhibitory effect on allergic asthma, and the CD8α-DC subsets play a major role.