Investigation of Vaccination Effect by Hsp 70-Malaria Antigen Peptide Complex

热休克蛋白70-疟疾抗原肽复合物的疫苗接种效果研究

• 批准号: 06670259
• 负责人: UDONO Heiichiro
• 金额: $ 1.28万
• 依托单位: Okayama University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06670259/
• 关键词: Mice Malaria; Plasmodium berghei; CS protein; Killer T cell; Heat Shock Protein 70; マラリア; P.bergfei; 熱ショック蛋白; CTLエピトープ; ワクチン

For establishment of preerythrocytic vaccination model against mice malaria Plasmodium berghei, we tried to induce cellular immunity to CS protein of plasmodium berghei.1.Cytotoxic T cell epitope, NDDSYIPSAEKI which is recognized in a context with H-2 K^d by CTL lines against CS protein of Plasmodium berghei was biochemically synthesized. This 12mer peptide and heat shock protein 70 were mixed and incubated for 1 hour ar 37ﾟC for making the hsp70-peptide.2.BALB/c mice were immunized with 10mg (per mouse) of this hsp70-peptide complex twice at a week interval and spleens were removed and suspended cells were incubated with the peptide for five days in 5% CO_2,37ﾟC.Cytotoxic activity of the effector cells was assayd with ^<51>Cr labelled P815 target cells pulsed with the peptide.3.By in vitro stimulation with the different dose, 1 x 10^<-4>,1 x 10^<-5>,1 x 10^<-6>,1 x 10^<-7>,1 x 10^<-8> mol of the peptide, optimal concentrations of the peptide were identified as 1 x 10^<-6> and 1 x 10^<-7> mol for induction of primary CTLs.4.Incubation of spleen cells with the peptide in the presence of IL2 revealed no mounting effect of cytotoxic activity was observed, rather IL2 decreased the activity.5.Since repeated stimulation of effector cells with the peptide apparently increased the cytolytic activity, establishment of long term cultured cell lines were on going now.

为了建立针对小鼠疟疾伯氏疟原虫的红细胞前疫苗接种模型，我们尝试诱导针对伯氏疟原虫CS蛋白的细胞免疫。 1.针对伯氏疟原虫CS蛋白的CTL系在H-2 K^d的背景下识别细胞毒性T细胞表位NDDSYIPSAEKI，该表位是生化的 合成的。将此 12mer 肽和热休克蛋白 70 混合并在 37°C 下孵育 1 小时，以制备 hsp70-肽。 2. 用 10mg（每只小鼠）此 hsp70-肽复合物每周两次免疫 BALB/c 小鼠，取出脾脏，将悬浮细胞与该肽在 5% CO_2,37°C 中孵育 5 天。的 效应细胞用^<51>Cr标记的P815靶细胞进行肽脉冲分析。3.通过不同剂量的体外刺激，1 x 10^<-4>,1 x 10^<-5>,1 x 10^<-6>,1 x 10^<-7>,1 x 10^<-8>mol肽，确定肽的最佳浓度为1 x 10^<-6> 和 1 x 10 ^ -7 mol用于诱导初级CTL。4.在IL2存在下用该肽孵育脾细胞显示没有观察到细胞毒性活性的增加效应，相反IL2降低了活性。5.由于用肽重复刺激效应细胞明显增加了细胞溶解活性，所以长期培养细胞系的建立正在进行中。

项目成果

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Udono,H.,Srivastava,P.K.: "Comparison of Tumor-specific immunogenicities of stress-induced proteins gp96,hsp90,and hap 70." J.Immunol.152. 5398-5403 (1994)

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Udono H.,et al.: "Heat shock proteins in cancer immunotherapy." Medical Intelligence Unit.in press.(1996)

Udono H.等人：“癌症免疫疗法中的热休克蛋白”。

Udono, H.: "Heat shock protein and cancer immunology" Blood Immunity Cancer. 1. 36-40 (1996)

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