两种佐剂系统对重组疟疾蛋白疫苗免疫效果的影响

发布时间：2018-03-04 12:20

本文选题：恶性疟原虫融合蛋白-.　切入点：恶性疟原虫环孢子蛋白-　出处：《中国生物制品学杂志》2014年02期 　论文类型：期刊论文

【摘要】：目的比较两种佐剂系统对重组疟疾蛋白疫苗免疫效果的影响。方法将BALB/c小鼠随机分为02、03佐剂组、Pr组和NS组,每组24只。02、03佐剂组、Pr组分别经大腿内侧肌肉注射15μg恶性疟原虫融合蛋白-2.9(combine protein 2.9 of Plasmodium falciparum,PfCP-2.9)+15μg恶性疟原虫环孢子蛋白-2(circumsporozoite protein of Plasmodium falciparum 2,PfCSP-2)+02佐剂系统[75μg BCG-CpG-DNA+0.2 mg A(lOH)3)]、15μg PfCP-2.9+15μg PfCSP-2+03佐剂系统[(50μg PolyI:C+0.2 mg A(lOH)3]和15μg PfCP-2.9+15μg PfCSP-2,NS组注射生理盐水。隔周免疫1次,共5次,于初次免疫2周后每周内眦采血,分离血清,采用ELISA法检测血清IgG值及抗体效价;于2、3、4、5次免疫后2周,无菌取小鼠脾脏,制备脾细胞悬液,采用ELISPOT法,对分泌IFNγ、IL-2、IL-4的特异性淋巴细胞数量进行检测。结果 02佐剂组小鼠血清IgG抗体效价在2次免疫后2周即可达到1∶105,03佐剂组在3次免疫后1周达到1∶105,而Pr组在5次免疫后2周达到1∶105,4次免疫后02、03佐剂组达到最高值。分泌IL-4的特异性淋巴细胞数,03佐剂组均明显高于02佐剂组、Pr组及NS组,02佐剂组3、4、5次免疫后才明显高于NS组,5次免疫后才明显高于Pr组(P均0.05);分泌IL-2的特异性淋巴细胞数,02佐剂组与03佐剂组比较差异无统计学意义(P0.05),3次免疫后02、03佐剂组均明显高于NS组,且03佐剂组也明显高于Pr组,4次免疫后02、03佐剂组、Pr组均明显高于NS组(P均0.05),但三者之间差异无统计学意义(P0.05);分泌IFNγ的特异性淋巴细胞数,02、03佐剂组、Pr组间差异无统计学意义(P0.05),且03佐剂组在3次免疫后明显高于NS组,而02佐剂组、Pr组在5次免疫后才明显高于NS组(P均0.05);02、03佐剂组分别在3、5次免疫后分泌IL-4的特异性淋巴细胞数量达到最高,且分别在5、3次免疫后,分泌IL-2的特异性淋巴细胞数量达到最高,二者均需5次免疫后分泌IFNγ的特异性淋巴细胞数量才可达到最高。结论 03佐剂系统可更加有效地增强重组疟疾蛋白疫苗的免疫原性,诱导机体产生较强的细胞免疫和体液免疫反应,5次免疫后可达到最佳免疫效果。
[Abstract]:Objective to compare the effects of two adjuvant systems on the immune response of recombinant malaria protein vaccine. Methods BALB/c mice were randomly divided into two groups: 0 2n03 adjuvant group and NS group. #number0# 渭 g plasmodium falciparum protein 2.9 of Plasmodium falciparum PfCP-2.9) 15 渭 g plasmodium falciparum cyclosporin protein of Plasmodium falciparum 2PfCSP-2) 02 adjuvant system [75 渭 g BCG-CpG-DNA 0.2 mg AflOH3] 15 渭 g PfCP-2.9 15 渭 g PfCSP-2 03 adjuvant system. The normal saline was injected into the 50 渭 g PolyI:C 0.2 mg A1OH3 and 15 渭 g PfCP-2.9 15 渭 g PfCSP-2Ns group. Blood samples were collected from canthus within 2 weeks after primary immunization, serum samples were isolated, serum IgG values and antibody titers were detected by ELISA method, spleen of mice were collected 2 weeks after 5 immunization, spleen cell suspensions were prepared, and ELISPOT method was used to prepare spleen cell suspensions. Results the antibody titer of serum IgG in the 02 adjuvant group was 1: 105 after 2 weeks of immunization, and 1: 105 in the adjuvant group was 1: 105 one week after 3 immunization, while that in pr group was 5 times. The number of specific lymphocytes secreting IL-4 in the adjuvant group was significantly higher than that in the adjuvant group of 02 adjuvant group and the adjuvant group of NS group after 5 times immunization. The number of specific lymphocytes secreting IL-2 in the adjuvant group was not significantly different from that in the adjuvant group (03), and that in the adjuvant group was significantly higher than that in the NS group. And the adjuvant group 03 was also significantly higher than that of group pr after 4 times immunization, but there was no significant difference among the three groups, but there was no significant difference among the three groups, and there was no significant difference in the number of specific lymphocytes secreting IFN 纬 between the two groups and there was no significant difference between the two groups in the number of specific lymphocytes secreting IFN 纬. The count significance of P0.05A was significantly higher in the 03 adjuvant group than that in NS group after 3 times immunization. The number of specific lymphocytes secreting IL-4 in the 02 adjuvant group was significantly higher than that in the NS group after 5 doses of immunization, and the number of specific lymphocytes secreted by the adjuvant group reached the highest level after 3 or 5 times immunization, and the specific lymphocyte count was higher than that in the NS group after 3 times of immunization, and the number of specific lymphocytes secreted in the adjuvant group was higher than that in the control group. The number of specific lymphocytes secreting IL-2 was the highest, and the number of specific lymphocytes secreting IFN 纬 was the highest after 5 times immunization. Conclusion the adjuvant 03 system can enhance the immunogenicity of recombinant malaria protein vaccine more effectively. The best immune effect can be achieved by inducing strong cellular and humoral immune responses for 5 times.
【作者单位】：河北联合大学;中国食品药品检定研究院;长春工业大学;
【基金】：疫苗效果和质量评价技术研究(2012AA02A402)
【分类号】：R382.31

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