CAJ | 학술논문

目的对小鼠血清抗体滴度检测和对小鼠的保护力观察,评价鼠疫菌荚膜抗原(F1抗原)和重组rV270抗原对鼠疫的免疫保护效果.方法 40只6～8周雌性Balb/c小鼠,按体质量随机分成4个实验组(F1-10 μg+铝佐剂、F1-20μg+铝佐剂、rV-10 μg+铝佐剂、rV-20 μg+铝佐剂)和1个对照组,每组8只.将天然F1抗原和重组rV270抗原分别吸附到25％铝佐剂中免疫实验组小鼠,对照组小鼠以免疫等量的铝佐剂.每只动物每次后腿肌肉免疫100 μ1,初次免疫后,第21天加强免疫1次.所有动物分别于第1次免疫后第8周采血,酶联免疫吸附试验(ELISA)测定抗体滴度.同时用2000倍半数致死量(LD50)的鼠疫菌141强毒株皮下攻毒,观察14d,以观察免疫后保护效果.结果对照组未产生抗体,F1-10 μg+铝佐剂组、F1-20 μg+铝佐剂组的抗体几何平均滴度(GMT)分别为1∶30443.9、1∶21527.8,组间比较差异无统计学意义(t=1.1282,P＞0.05)；rV-10 μg+铝佐剂组和rV-20μg+铝佐剂组的GMT分别为1∶13957.3、1∶18100.9,组间比较差异无统计学意义(t=0.9408,P＞0.05).用141强毒株皮下攻毒后,实验组小鼠全部存活,对照组8只小鼠全部死亡.结论实验用天然F1抗原及重组rV270抗原具有较高的免疫活性,可作为鼠疫亚单位疫苗的主要组分用于鼠疫亚单位疫苗的研究.
목적 대소서혈청항체적도검측화대소서적보호력관찰,평개서역균협막항원(F1항원)화중조rV270항원대서역적면역보호효과.방법 40지6～8주자성Balb/c소서,안체질량수궤분성4개실험조(F1-10 μg+려좌제、F1-20μg+려좌제、rV-10 μg+려좌제、rV-20 μg+려좌제)화1개대조조,매조8지.장천연F1항원화중조rV270항원분별흡부도25％려좌제중면역실험조소서,대조조소서이면역등량적려좌제.매지동물매차후퇴기육면역100 μ1,초차면역후,제21천가강면역1차.소유동물분별우제1차면역후제8주채혈,매련면역흡부시험(ELISA)측정항체적도.동시용2000배반수치사량(LD50)적서역균141강독주피하공독,관찰14d,이관찰면역후보호효과.결과 대조조미산생항체,F1-10 μg+려좌제조、F1-20 μg+려좌제조적항체궤하평균적도(GMT)분별위1∶30443.9、1∶21527.8,조간비교차이무통계학의의(t=1.1282,P＞0.05)；rV-10 μg+려좌제조화rV-20μg+려좌제조적GMT분별위1∶13957.3、1∶18100.9,조간비교차이무통계학의의(t=0.9408,P＞0.05).용141강독주피하공독후,실험조소서전부존활,대조조8지소서전부사망.결론 실험용천연F1항원급중조rV270항원구유교고적면역활성,가작위서역아단위역묘적주요조분용우서역아단위역묘적연구.
Objective To evaluate the protective effect of Yersinia pestis capsular antigen F1 and recombinant rV270 on mice after immunization with them.Methods According to body weight,40 female Balb/c mice aged 6 to 8 weeks were randomly divided into four experimental groups(Fl-10 μg + aluminum adjuvant,F1-20 μg + aluminum adjuvant,rV-10 μg + aluminum adjuvant,and rV-20 μg + aluminum adjuvant) and a control group,8 in each group.Mice in experimental groups were immunized with the natural antigen F1 and recombinant antigen rV270 adsorbed to 25％ aluminum adjuvant and the control group was immunized with the same amount of aluminum adjuvant.Each mouse was immunized at the hind leg muscle with 100 ml immunizing agent,then a booster immunization was done once on the 21st day after the first immunization.The blood of all mice was collected on the 8th week after the first immunization,serum antibody titers were detected by ELISA and the data of antibody titers were analyzed by t test for comparison between groups.At the same time the mice were injected subcutaneously with 2000-fold LD50 of Yersinia pestis virulent strain 141,after 14 days,the protective effect of immunization was analyzed.Results The control group did not produce antibody.Antibody geometric mean titers (GMT) of the F1-10 mg + aluminum adjuvant and F1-20 mg + aluminum adjuvant groups were 1 ∶ 30443.9,and 1 ∶21527.8,respectively,and compared between the two groups,the difference was not statistically significant (t =1.1282,P ＞ 0.05).The GMTs of the rV-10 μg + aluminum adjuvant and rV-20 μg + aluminum adjuvant groups were 1 ∶ 13957.3 and 1 ∶18100.9,respectively,and compared between the two groups,the difference was not statistically significant(t =0.9408,P ＞ 0.05 ).After subcutaneous injection with Yersinia pestis virulent strain 141,all mice died in the control group but all survived in the experimental group.Conclusion The immune activity of natural antigen F1 and recombinant rV270 is high,which can be used as the main component of subunit vaccine in the plague subunit vaccine study.