摘要
目的利用重组亮氨酸氨基肽酶(rSjLAP)和果糖二磷酸醛缩酶(rSjFBPA)辅以佐剂免疫BALB/c小鼠,观察基于2-DE和血清蛋白质组筛选的上述二种重组抗原的免疫保护效果;通过福氏佐剂和纳米C60佐剂在增强免疫效果方面的比较,探讨纳米C60佐剂在血吸虫分子疫苗接种中的价值。方法从重组质粒转化的E.coli中诱导表达rSjLAP和rSjFBPA,纯化并检测含量。将6周龄BALB/c小鼠随机分为7组,生理盐水对照组(A组)、rSjLAP+福氏佐剂组(B组)、rSjFBPA+福氏佐剂组(C组)、rSjLAP+rSjFBPA+福氏佐剂组(D组)、rSjLAP+纳米C60佐剂组(E组)、rSjFBPA+纳米C60佐剂组(F组)、rSjLAP+rSjFBPA+纳米C60佐剂组(G组),每组6只。除A组外,B～G组小鼠皮下多点注射rSjLAP或rSjFBPA100μg,共免疫3次,每次间隔2周。用尾蚴感染小鼠6周后,处死小鼠,收集成虫,计算减虫率;收集肝组织虫卵,计算减卵率。结果 B～G组的减虫率分别为:35.59%、33.05%、37.98%、38.99%、43.21%和45.75%,与A组比较,差异均有统计学意义(P<0.05);B～G组的减卵率分别为:40.25%、42.57%、48.88%、41.64%、45.44%和46.88%,与A组比较,差异均有统计学意义(P<0.05)。结论 BALB/c小鼠经rSjLAP和rSjFBPA辅以福氏佐剂或纳米C60佐剂免疫,获得了一定的免疫保护力;福氏佐剂和纳米C60佐剂均增强了重组疫苗的免疫效果,但两种佐剂的免疫增强效果差异无统计学意义。
Objective To observe the immune protective effects of two recombinant antigens of rSjLAP and rSjFBPA based on the 2-DE oriented screening of the sera proteomics with adjuvants in BALB/c mice.The immune enhancing effect of Freund＇s adjuvant and Nano-C60 adjuvant was compared for their potential use in development of molecular vaccine for Schistosomiasis japonica.Methods rSjLAP and rSjFBPA,induced and expressed from the transformed E.coli,were purified with affinity chromatography.The 6-week-old BALB/c mice were randomly divided into seven groups,saline control group （group A） ,rSjLAP ＋ Freund＇s adjuvant group（group B） ,rSjFBPA ＋ Freund＇s adjuvant group（group C） ,rSjLAP ＋ rSjFBPA ＋ Freund＇ s adjuvant group（group D） ,rSjLAP ＋ Nano-C60 adjuvant group（group E） ,rSjFBPA ＋ Nano-C60 adjuvant group（group F） ,rSjLAP ＋ rSjFBPA ＋ Nano-C60 adjuvant group（group G） ,with 6 mice each.All mice（from group B to group G） ,except for the group A,were immunized three times at 2-week＇s intervals by subcutaneous multi-point injection with rSjLAP or rSjFBPA in the dosage of 100 μg.The mice were infected with schistosome cercariae.After 6 weeks,the mice were sacrificed under anesthesia for calculating the reduction of worm burden and hepatic tissue eggs.Results From group B to group G,the worm reduction rates were found to be 35.59% ,33.05% ,37.98% ,38.99% ,43.21% ,and 45.75% ,respectively;compared with the group A,statistical significance was noted（P 0.05） .From group B to group G,the eggs reduction rates were 40.25% ,42.57% ,48.88% ,41.64% ,45.44% ,and 46.88% ,respectively;compared with the group A,statistical significance was noted（P 0.05） .Conclusion The BALB/c mice,which is immunized with rSjLAP and rSjFBPA with Freud＇s adjuvant or Nano-C60 adjuvant show a certain immune protection.No statistical significance notes between Freud＇s and Nano-C60 adjuvents although both of them may give rise to a relative immune enhancement.
出处
《安徽医科大学学报》
CAS
北大核心
2010年第6期740-743,共4页
Acta Universitatis Medicinalis Anhui
基金
安徽省自然科学基金(编号:090413089)
