DO11.10小鼠卵清蛋白免疫耐受模型的建立

为建立D011.10小鼠的卵清蛋白(OVA)免疫耐受模型,试验组小鼠尾静脉注射5 μg/只OVA,对照组注射PBS,共3次,每次间隔5 d.最后一次注射后的第3天、第12天分别用OVA进行免疫,第18天颈椎脱臼处死小鼠.利用MTT法检测小鼠OVA、OVA323-339肽段特异性脾淋巴细胞增殖情况,利用流式细胞术检测CD...

Establishment of Ovalbumin Immunotolerance Model in DO11.10 Mice

To induce immunotolerance to ovalbumin in DO11.10 mice,the mice in the experimental and immunized control groups were injected i.v.with 5 μg of OVA or PBS for three times at 5 days intervals,respectively.Experimental and immunized control mice were challenged with 200 μg of OVA at 3 and 12 days post injection.All mice were sacrificed at 6 days after the final immunization.The state of immunotolerance to ovalbumin in DO11.10 mice was evaluated using MTT,flow cytometry,and ELISA to determine the lymphocyte proliferation,the amout of CD4+CD25+T cell subset,and the content of suppressive cytokines(TGF-β and IL-10).The results showed that cell proliferation in mice of experimental groups were significantly suppressed to specific antigen OVA or OVA323-339 peptide(P<0.05),and the ratio of CD4+CD25+ to CD4+ cells in mice of this group was significantly increased(P<0.05).The suppressive cytokine of TGF-β but not IL-10 levels were also significantly elevated in co-cultures of OVA323-339 peptide and lymphocytes from experimental mice(P<0.05),which indicated that OVA-specific immunotolerance could be induced in DO11.10 mice with injection i.v.of low-dose OVA.