CAJ | 학술논문

目的研究构建的沙眼衣原体E型DNA疫苗所产生的细胞免疫效应.方法以BALB/c小鼠为实验动物,采用肌内接种免疫的方式,将小鼠分为接种空质粒的阴性对照组、DNA疫苗免疫组和接种灭活沙眼衣原体E型原体的阳性对照组.通过观察小鼠后足垫局部的迟发型超敏反应、沙眼衣原体原体刺激后脾淋巴细胞增殖试验、ELISA检测血清IFN-γ水平、实验动物经生殖道进行同型沙眼衣原体攻击后局部的衣原体培养情况以及局部组织病理学变化来研究所构建疫苗的细胞免疫效果.结果 DNA疫苗组和灭活原体组小鼠的迟发型超敏反应水平高于阴性对照组.空质粒组、DNA疫苗组和灭活原体组的脾淋巴细胞刺激指数分别为1.48±0.15、2.35±0.25、3.81±0.30,后两组与空质粒组比较差异有统计学意义,且灭活原体组增殖更明显.三组的血清IFN-γ水平分别为(309.2±157.9)μg/L、(593.3±342.6)μg/L、(2891.7±1048.8)μg/L;DNA疫苗组和灭活原体组与空质粒组比较,差异有统计学意义,且灭活原体组IFN-γ水平更高.沙眼衣原体E型原体攻击后,空质粒组小鼠生殖道局部能够培养出衣原体,生殖道局部组织有破坏.另两组衣原体培养阴性,生殖道局部组织完好.结论沙眼衣原体E型DNA疫苗免疫小鼠后能够诱导产生一定的沙眼衣原体特异性细胞免疫,该疫苗对同型沙眼衣原体的生殖道攻击具有保护作用.
목적 연구구건적사안의원체E형DNA역묘소산생적세포면역효응.방법 이BALB/c소서위실험동물,채용기내접충면역적방식,장소서분위접충공질립적음성대조조、DNA역묘면역조화접충멸활사안의원체E형원체적양성대조조.통과관찰소서후족점국부적지발형초민반응、사안의원체원체자격후비림파세포증식시험、ELISA검측혈청IFN-γ수평、실험동물경생식도진행동형사안의원체공격후국부적의원체배양정황이급국부조직병이학변화래연구소구건역묘적세포면역효과.결과 DNA역묘조화멸활원체조소서적지발형초민반응수평고우음성대조조.공질립조、DNA역묘조화멸활원체조적비림파세포자격지수분별위1.48±0.15、2.35±0.25、3.81±0.30,후량조여공질립조비교차이유통계학의의,차멸활원체조증식경명현.삼조적혈청IFN-γ수평분별위(309.2±157.9)μg/L、(593.3±342.6)μg/L、(2891.7±1048.8)μg/L;DNA역묘조화멸활원체조여공질립조비교,차이유통계학의의,차멸활원체조IFN-γ수평경고.사안의원체E형원체공격후,공질립조소서생식도국부능구배양출의원체,생식도국부조직유파배.령량조의원체배양음성,생식도국부조직완호.결론 사안의원체E형DNA역묘면역소서후능구유도산생일정적사안의원체특이성세포면역,해역묘대동형사안의원체적생식도공격구유보호작용.
Objective To study cellular immune responses induced by DNA vaccine against Chlamydia trachomatis (Ct) serotype E. Methods BALB/c mice were divided into three groups to be intramuscularly immunized by blank plasmid (negative control group), DNA vaccine against Ct serotype E (vaccine group), and inactivated Ct elementary body (positive control group), respectively. Two weeks after the last immunization,delayed-type hypersensitivity (DTH) response was evaluated; MTT assay was performed to detect the proliferation of spleen lymphocytes, ELISA to measure the serum level of interferon-γin mice. Some immunized mice underwent a genital challenge with Ct elementary body followed by isolation of Ct from exfoliated epithelial cells in genital tract and pathological examination of cervical tissue from the challenged mice. Results Compared to negative control group, vaccine group and positive control group experienced a stronger DTH response.The lymphocyte stimulating index and serum level of IFN-γwere highest in the positive control group (3.81 ±0.30, 2891.7 ± 1048.8 μg/L), followed by vaccine group (2.35 ± 0.25, 593.3 ± 342.6 μg/L) and negative control group (1.48 ± 0.15, 309.2 ± 157.9 μg/L), and significant difference was observed between the three groups (P ＜ 0.05 or 0.01 ). After Ct challenge, Ct was isolated from exfoliated epithelial cells and cervical tissue was damaged in the negative control group, while in the other two groups, Ct was undetected and genital tract tissue was intact. Conclusions The DNA vaccine against Ct serotype E could induce Ct-specific cellular immune responses to some extent, and offer a protection against vaginal challenge with Ct.