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.

为建立伯氏疟原虫红内期前免疫小鼠模型，我们尝试诱导针对伯氏疟原虫CS蛋白的细胞免疫。1.生物化学合成了抗伯氏疟原虫CS蛋白CTL细胞株在H-2 K^d背景下识别的细胞毒性T细胞表位NDDSYIPSAEKI。将此12肽与热休克蛋白70混合，37 ℃孵育1小时，制备hsp 70-肽（每只小鼠）以一周间隔两次注射该hsp 70-肽复合物，取出脾脏，将悬浮细胞与肽在5%CO_2中孵育5天，3.<51>用不同浓度的多肽（1 × 10 ~（-1）、1 × 10 ~（-1）、1 × 10 ~（-1）<-4>、1 × 10 <-5>~（-1）、<-6>1 × 10 ~（-1<-7>）mol）体外刺激小鼠脾细胞，<-8>发现1 × 10 ~（-1）和1 × 10 ~（-1）mol的多肽诱导小鼠脾细胞产生原代CTL的最佳浓度为1 × 10 ~<-6>（-1）和1 × 10 ~（-1）mol<-7>。5.由于用该肽反复刺激效应细胞明显增加了细胞溶解活性，因此目前正在建立长期培养的细胞系。

项目成果

鵜殿 平一郎: "腫瘍細胞熱ショック蛋白の抗腫瘍効果における役割-腫瘍拒絶抗原としての熱ショック蛋白-" 別冊 医学の歩み 免疫疾患-state of arts-. 239-242 (1995)

Heiichiro Udono：“肿瘤细胞热休克蛋白在抗肿瘤作用中的作用 - 热休克蛋白作为肿瘤排斥抗原”单独卷，State of Arts，239-242（1995）

Srivastava P.K.,Udono H.: "Heat shock protein-peptide complexes in cancer immunotherapy." Curr.Opin.Immunol.6. 728-732 (1994)

Srivastava P.K.、Udono H.：“癌症免疫治疗中的热休克蛋白-肽复合物。”

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)

Udono,H.,Srivastava,P.K.：“应激诱导蛋白 gp96、hsp90 和 hap 70 的肿瘤特异性免疫原性的比较。”

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)

Udono, H.：“热休克蛋白和癌症免疫学”血液免疫癌症。