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目的:观察用毒蕈碱3型受体(M3R)胞外区第二环多肽免疫NOD-scid小鼠(nonobese diabetic mice CB17.Prkdcscid/J,非肥胖糖尿病/重症联合免疫缺陷小鼠)后,小鼠体内IL-17和IFN-γ分泌水平的变化。方法:200μg的M3R胞外区第二环多肽和不完全弗氏佐剂(IFA)以1∶1的比例配制而成后,皮下免疫NOD-scid小鼠。免疫后1、7、14、21 d尾部采血检测血清中的抗M3R第二环多肽抗体,血清中IL-17和IFN-γ含量的变化,同时测定每星期每只小鼠的饮水量。21 d给予小鼠安乐死,取小鼠脾脏细胞与抗M3R第二环多肽共同培养,观察细胞上清中IL-17和IFN-γ含量的变化。同时取小鼠泪腺做免疫荧光观察IL-17和IFN-γ的变化。结果:用M3R胞外区第二环多肽免疫NOD-scid小鼠14 d后,小鼠体内的抗M3R第二环多肽抗体显著高于对照肽组和PBS组(P<0.05),血清中IL-17和IFN-γ含量在第14天和第7天显著升高,具有统计学意义(P<0.01)。脾细胞多肽共培养后细胞上清中的IL-17的含量维持在一定的水平,而对照肽组和PBS组则显著下降(P<0.01)。组织免疫荧光显示泪腺中的IL-17和IFN-γ的含量也显著提高。每只小鼠的饮水量3组没有显著差别(P>0.05)。结论:用M3R胞外区第二环多肽免疫NOD-scid小鼠后成功诱导出抗M3R第二环多肽抗体,并且促进了NOD-scid小鼠体内IL-17和IFN-γ的分泌。
OBJECTIVE: To observe whether NOD-scid mice (nonobese diabetic mice CB17.Prkdcscid / J, non-obese diabetic / severe combined immunodeficient mice) were immunized with the second polypeptide in the extracellular domain of muscarinic type 3 receptor (M3R) Changes of IL-17 and IFN-γ secretion in mice. METHODS: NOD-scid mice were immunized subcutaneously after 200 μg of extracellular domain of M3R and incomplete Freund’s adjuvant (IFA) were formulated in a 1: 1 ratio. Blood samples were collected at the end of 1, 7, 14, and 21 days after immunization to detect the change of serum anti-M3R second-ring polypeptide antibody and IL-17 and IFN-γ in serum. Meanwhile, the drinking amount per mouse per week was measured. The mice were euthanized on day 21 and co-cultured with anti-M3R second-ring polypeptide. The changes of IL-17 and IFN-γ in the supernatant were observed. At the same time, the changes of IL-17 and IFN-γ in lacrimal gland of mice were observed by immunofluorescence. Results: After 14 days of NOD-scid mice immunized with the second-ring polypeptide of M3R, the anti-M3R second-ring antibody in mice was significantly higher than that of the control peptide group and PBS group (P <0.05) -17 and IFN-γ levels increased significantly on the 14th and 7th days (P <0.01). The content of IL-17 in the supernatant of the spleen cell polypeptide co-cultured was maintained at a certain level, while the control peptide group and the PBS group were significantly decreased (P <0.01). Tissue immunofluorescence showed a significant increase in the levels of IL-17 and IFN-γ in the lacrimal gland. There was no significant difference in water intake per mouse between the three groups (P> 0.05). CONCLUSION: NOD-scid mice immunized with the second-ring polypeptide of extracellular domain of M3R successfully induced anti-M3R second-ring polypeptide antibody and promoted the secretion of IL-17 and IFN-γ in NOD-scid mice.