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目的研究微小隐孢子虫表面抗原CP23 DNA疫苗产生的免疫反应及对小鼠的免疫保护作用。方法选BALB/c小鼠60只,随机分为3组,即疫苗免疫组、PBS对照组及空质粒对照组。疫苗免疫组用构建的真核表达质粒pcDNA 3.0-23每隔2周免疫1次小鼠,共免疫3次。PBS对照组肌注等量PBS,空质粒对照组肌注pcDNA3.0,免疫方法及次数同疫苗免疫组。末次免疫2周后,分别取小鼠脾脏、血清,检测CD4+和CD8+T细胞、细胞因子IFN-γ及抗CP23特异性抗体IgG和IgA滴度;用微小隐孢子虫攻击感染被免疫小鼠,收集小鼠粪便,计算小鼠排出的卵囊量。结果 PBS对照组及空质粒组比较,疫苗免疫组小鼠的CD4+T细胞、CD4+/CD8+比值及脾细胞培养上清中IFN-γ滴度均显著升高(P<0.05),小鼠血清抗CP23特异性抗体IgG及IgA滴度随免疫次数增加显著升高(P<0.05),攻击感染后小鼠排卵囊量显著减少(P<0.05),且排出时间缩短。结论构建的微小隐孢子虫表面抗原CP23真核表达质粒能够诱导小鼠产生细胞及体液免疫反应,对小鼠有较好的免疫保护作用。
Objective To study the immune response induced by Cryptosporidium parvum surface antigen CP23 DNA vaccine and its immunoprotective effect on mice. Methods Sixty BALB / c mice were randomly divided into three groups: vaccine immunized group, PBS control group and empty plasmid control group. Vaccine group immunized mice constructed with the eukaryotic expression plasmid pcDNA 3.0-23 immunized once every 2 weeks, co-immunization 3 times. PBS control group, intramuscular injection of PBS, empty plasmid control group intramuscular pcDNA3.0, immunization methods and the number of immunized group with the vaccine. Two weeks after the last immunization, the spleens and serums of mice were taken, and the CD4 and CD8 + T cells, IFN-γ and anti-CP23 specific antibodies IgG and IgA titers were detected. The infected mice were challenged with Cryptosporidium parvum , Collect mouse feces, and calculate the amount of oocysts discharged from mice. Results Compared with the control group and empty plasmid group, the CD4 + T cells, CD4 + / CD8 + ratio and the IFN-γ titer in the splenocyte culture supernatants were significantly increased (P <0.05) The titers of anti-CP23 IgG and IgA were significantly increased with the increase of immunization times (P <0.05). The number of ovulation cysts in mice was significantly decreased (P <0.05) and the discharge time was shortened. CONCLUSION: The constructed CP23 eukaryotic expression plasmid of Cryptosporidium parvum can induce cellular and humoral immune responses in mice, and has good immunoprotective effect on mice.