本文選題：惡性瘧原蟲融合蛋白-.　切入點：惡性瘧原蟲環(huán)孢子蛋白-　出處：《中國生物制品學雜志》2014年02期 　論文類型：期刊論文

【摘要】：目的比較兩種佐劑系統對重組瘧疾蛋白疫苗免疫效果的影響。方法將BALB/c小鼠隨機分為02、03佐劑組、Pr組和NS組,每組24只。02、03佐劑組、Pr組分別經大腿內側肌肉注射15μg惡性瘧原蟲融合蛋白-2.9(combine protein 2.9 of Plasmodium falciparum,PfCP-2.9)+15μg惡性瘧原蟲環(huán)孢子蛋白-2(circumsporozoite protein of Plasmodium falciparum 2,PfCSP-2)+02佐劑系統[75μg BCG-CpG-DNA+0.2 mg A(lOH)3)]、15μg PfCP-2.9+15μg PfCSP-2+03佐劑系統[(50μg PolyI:C+0.2 mg A(lOH)3]和15μg PfCP-2.9+15μg PfCSP-2,NS組注射生理鹽水。隔周免疫1次,共5次,于初次免疫2周后每周內眥采血,分離血清,采用ELISA法檢測血清IgG值及抗體效價;于2、3、4、5次免疫后2周,無菌取小鼠脾臟,制備脾細胞懸液,采用ELISPOT法,對分泌IFNγ、IL-2、IL-4的特異性淋巴細胞數量進行檢測。結果 02佐劑組小鼠血清IgG抗體效價在2次免疫后2周即可達到1∶105,03佐劑組在3次免疫后1周達到1∶105,而Pr組在5次免疫后2周達到1∶105,4次免疫后02、03佐劑組達到最高值。分泌IL-4的特異性淋巴細胞數,03佐劑組均明顯高于02佐劑組、Pr組及NS組,02佐劑組3、4、5次免疫后才明顯高于NS組,5次免疫后才明顯高于Pr組(P均0.05);分泌IL-2的特異性淋巴細胞數,02佐劑組與03佐劑組比較差異無統計學意義(P0.05),3次免疫后02、03佐劑組均明顯高于NS組,且03佐劑組也明顯高于Pr組,4次免疫后02、03佐劑組、Pr組均明顯高于NS組(P均0.05),但三者之間差異無統計學意義(P0.05);分泌IFNγ的特異性淋巴細胞數,02、03佐劑組、Pr組間差異無統計學意義(P0.05),且03佐劑組在3次免疫后明顯高于NS組,而02佐劑組、Pr組在5次免疫后才明顯高于NS組(P均0.05);02、03佐劑組分別在3、5次免疫后分泌IL-4的特異性淋巴細胞數量達到最高,且分別在5、3次免疫后,分泌IL-2的特異性淋巴細胞數量達到最高,二者均需5次免疫后分泌IFNγ的特異性淋巴細胞數量才可達到最高。結論 03佐劑系統可更加有效地增強重組瘧疾蛋白疫苗的免疫原性,誘導機體產生較強的細胞免疫和體液免疫反應,5次免疫后可達到最佳免疫效果。
[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 緯 between the two groups and there was no significant difference between the two groups in the number of specific lymphocytes secreting IFN 緯. 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 緯 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.
【作者單位】： 河北聯合大學;中國食品藥品檢定研究院;長春工業(yè)大學;
【基金】：疫苗效果和質量評價技術研究(2012AA02A402)
【分類號】：R382.31

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