Studies on gene analysis and immunological function of 47KD polypeptide of Plasmodium falciparum.

恶性疟原虫47KD多肽基因分析及免疫功能研究

• 批准号: 03404024
• 负责人: SUZUKI Mamoru
• 金额: $ 16.51万
• 依托单位: Gunma University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (A)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03404024/
• 关键词: Malaria; Plasmodium falciparum; Gene analysis; Immunology; Vaccine; Epidemiology; 分子生物学; 遺伝子解所

Comparative studies on immunue status of individuals living in the endemic areas will elucidate the antigen epitopes related to protective immunity. Present study was performed by using anti-P.falicparum polyclonal antibodies taken from individuals in the endemic tropical areas and also from patients who developed falciparum malaria in Japan. By an immunoblotting, electrophoresed 47kD molecule (pf47kD) was presented by the sera taken from the acute phase of the infection, thus it was presumed that pf47kD was a parasite epitope inducing immune reactions at an early stage of malaria infection. One monoclonal antibody was developed to study localization of pf47kD in the parasite. A lazer-scanning fluorescent microscopy rvealed that pf47kD was on the surface of developing schizont. Studies on parasite gene which encodes the pf47kD sequence was attempted. cDNA library from the clutured falciparum parasite was prepared and were ligated to vectors and were introduced into host cells. Screening was processed by using high titered polyclonal antibodies and the monoclonal antibody. the resulted pf47kD sequence was coincidental with an sequence of human gene. The reason is still on the study. Simultaneously, the parasite antigen was purified by using HPLC to analyze the sequence and results will be comparatively studied with the results shown above. In the field studies, a 23kD parasite moleculte was highlighted by the use of the sera from chronic type of P.falciparum infection. The molecule seemed to induce another type of protective immune reaction. In addition, roles of cytokines in the protective immunity was studied using animal models of inbred mice and monkeys. INF-gamma and TNF-alpha worked in the protection and probably in the detrimental reaction in malaria infection.

通过对疫区个体免疫状况的比较研究，将有助于阐明与保护性免疫相关的抗原表位。本研究采用anti-P。从热带流行地区的个体以及日本恶性疟疾患者身上提取的恶性疟原虫多克隆抗体。通过免疫印迹法，在感染急性期血清中发现了电泳的47kD分子（pf47kD），推测pf47kD是疟疾感染早期诱导免疫反应的寄生虫表位。制备了一种单克隆抗体，用于研究pf47kD在寄生虫中的定位。激光扫描荧光显微镜显示，pf47kD存在于发育中的分裂体表面。对编码pf47kD序列的寄生虫基因进行了研究。从培养的恶性疟原虫中制备cDNA文库，并连接到载体上，导入宿主细胞。采用高滴度多克隆抗体和单克隆抗体进行筛选。pf47kD序列与人类基因序列一致。原因还在研究中。同时，采用高效液相色谱法纯化寄生虫抗原，分析其序列，并将结果与上述结果进行对比研究。在现场研究中，利用慢性恶性疟原虫感染的血清，发现了一个23kD的寄生虫分子。这种分子似乎诱导了另一种保护性免疫反应。此外，我们还利用近交系小鼠和猴子的动物模型研究了细胞因子在保护性免疫中的作用。inf - γ和tnf - α在疟疾感染的保护和可能的有害反应中起作用。

项目成果

狩野 繁之,鈴木 守: "マラリアDNA診断法の進歩と利点" 日本臨床. 50. 458-463 (1992)

Shigeyuki Kano、Mamoru Suzuki：“疟疾 DNA 诊断方法的进展和优点”日本临床研究 50. 458-463 (1992)。

Waki, S.Nakane, K.Zhu, M.et al.: "A DNA hybridation assay for use in drug sensitivity test in vitro for P.falciparum under field cindition" Trans.Roy.Soc.Trop.Med.Hyg.86. 227-228 (1992)

Waki, S.Nakane, K.Zhu, M.et al.：“在野外条件下用于恶性疟原虫体外药物敏感性测试的 DNA 杂交测定” Trans.Roy.Soc.Trop.Med.Hyg.86

Akanmori,B.D.,Waki,S.,Suzuki,M.: "Early secretion of INF-gamma and TNF-alpha by spleen cells for the protection against Plasmodium bergehi NK65 infection in CBA mice" Parasitology Research. (submitted).

Akanmori,B.D.、Waki,S.、Suzuki,M.：“脾细胞早期分泌 INF-γ 和 TNF-α，以保护 CBA 小鼠免受伯氏疟原虫 NK65 感染”寄生虫学研究。

Kano, S.El Safi, S.H.Omer et al.: "Antibody frequency distribution curve for risk assessment of a malaria epidemic in the Sudan" Japan.J.Trop.Med.Hyg.21. 207-211 (1993)

Kano、S.El Safi、S.H.Omer 等人：“苏丹疟疾流行风险评估的抗体频率分布曲线”Japan.J.Trop.Med.Hyg.21。

Y.Sugioka,M.Suzuki: "The Chemical basis for the ferriprotoporphyrin IXーchloroquine complex induced lipid peroxidation" Biochemica et Biophysica Acta. 1074. 19-24 (1991)

Y.Sugioka，M.Suzuki：“铁原卟啉 IX-氯喹复合物诱导脂质过氧化的化学基础”Biochemica et Biophysical Acta. 1074. 19-24 (1991)