Studies on the development of protective immunity with the 150-kDa lectin of Entamoeba histolytica

溶组织内阿米巴 150-kDa 凝集素发展保护性免疫的研究

• 批准号: 14570223
• 负责人: TACHIBANA Hiroshi
• 金额: $ 2.24万
• 依托单位: Tokai University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570223/
• 关键词: Entamoeba histolytica; adherence; lectin; monoclonal antibody; recombinant protein; epitope; Entamoeba dispar; モノクロナール抗体

We have recently identified a 150-kDa surface antigen of Entamoeba histolytica as an intermediate subunit (Igl) of galactose-and N-acetyl-D-galactosamine-inhibitable lectin. In the present study, full-length Igl except for the signal sequences [amino acid (aa) 14-1,088] and three fragments of Igl, the N-terminal part (aa 14-382), middle part (aa 294-753), and C-terminal part (aa 603-1,088), were prepared in Escherichia coli and the reactivity of these recombinant proteins with sera from patients with amebiasis was examined by means of enzyme-linked immunosorbent assay (ELISA). Sera from symptomatic patients with amebic liver abscess or amebic colitis, sera from asymptomatic cyst passers, sera from individuals with other protozoan infections, and sera from healthy controls were used. Sensitivity and specificity of the recombinant full-length Igl in the ELISA were 90% and 94% respectively. When three fragments were used as antigens in the ELISA, sensitivities were 56% in the N-terminus, … More 92% in the middle part, and 97% in the C-terminus. Specificities of the three antigens were 96% in the N-terminus and 99% in both the middle and C-terminus fragments. None of the sera from asymptomatic cyst passers was negative when F-Igl, M-Igl and C-Igl were used as the antigen. On the other hand, in the cases of ELISA using M-Igl and C-Igl, false-negative results were detected in only the colitis cases. These observations suggest that the antibodies which recognized the epitopes located in M-Igl and C-Jgl may function to prevent the invasion of trophozoites into host tissues and that the carboxyl terminus of Igl is an especially useful antigen for the serodiagnosis of amebiasis. When the reactivity of adherence-inhibitory monoclonal antibodies to various recombinant fragments was also examined, one of the epitope was localized in the part of aa 989-1, 088. Liver abscess formation of hamsters was significantly inhibited by the immunization with native Igl, but not with the F-Igl, suggesting that the epitope (s) on sugar chain(s) of Igi may also be important for the induction of protective immunity. Two Igl genes, coding 1, 110 and 1, 106 aa, were also cloned from Entamoeba dispar and were further analyzed. Less

我们最近已经确定了一个150 kDa的溶组织内阿米巴表面抗原的半乳糖和N-乙酰基-D-半乳糖胺-凝集素的中间亚基（Igl）。在本研究中，全长Igl除了信号序列[氨基酸（aa）14- 1，088]和Igl的三个片段，即N-末端部分，（aa 14-382），中间部分（aa 294-753）和C-末端部分（aa 603- 1，088），在大肠杆菌中制备了重组蛋白，并通过酶联免疫吸附试验检测了这些重组蛋白与阿米巴病患者血清的反应性。免疫吸附试验（ELISA）。使用来自阿米巴肝脓肿或阿米巴结肠炎的症状性患者的血清、来自无症状囊肿传者的血清、来自其他原生动物感染的个体的血清和来自健康对照的血清。ELISA检测的灵敏度和特异性分别为90%和94%。当三个片段用作ELISA中的抗原时，N-末端的灵敏度为56%， ...更多信息 92%在中间部分，97%在C-末端。三种抗原的特异性在N-末端为96%，在中间和C-末端片段均为99%。以F-Igl、M-Igl和C-Igl为抗原，无症状传囊者血清均为阴性。另一方面，在使用M-Igl和C-Igl的ELISA的情况下，仅在结肠炎病例中检测到假阴性结果。这些观察结果表明，识别位于M-Igl和C-Jgl中的表位的抗体可能具有防止滋养体侵入宿主组织的功能，并且Igl的羧基端是用于阿米巴病血清学诊断的特别有用的抗原。当还检测粘附抑制性单克隆抗体对各种重组片段的反应性时，表位之一位于aa 989-1，088部分。用天然Igl免疫显著抑制仓鼠的肝脓肿形成，但用F-Igl免疫则没有，这表明Igi糖链上的表位对于诱导保护性免疫也可能是重要的。另外还从舞帕内阿米巴（Entamoeba dispar）中克隆了编码1，110和1，106个氨基酸的Ig 1基因，并对其进行了分析。少

项目成果

Y.Tsutsumi et al.: "Acanthamebic meningoencephalitis associated with alcoholic liver cirrhosis"Pathology Case Reviews. 7(6). 273-277 (2002)

Y.Tsutsumi 等人：“与酒精性肝硬化相关的棘阿米巴性脑膜脑炎”病理学病例评论。

Tsutsumi, Y., et al.: "Acanthamebic meningoencephalitis associated with alcoholic liver cirrhosis"Pathol.Case Rev.. 7. 273-277 (2002)

Tsutsumi, Y. 等人：“与酒精性肝硬化相关的棘阿米巴性脑膜脑炎”Pathol.Case Rev.. 7. 273-277 (2002)

Tachibana, H., et al.: "VH3 gene usage in neutralizing human antibodies specific for the histolytica Gal/GalNAc lectin heavy subunit"Infect.Immun.. 71(8). 4313-4319 (2003)

Tachibana, H. 等人：“VH3 基因在中和对溶组织性 Gal/GalNAc 凝集素重亚基特异的人抗体中的应用”Infect.Immun.. 71(8)。

橘 裕司: "いま日常診療で注目すべき原虫症・寄生虫症 トリパノソーマ症・リーシュマニア症"JIM. 13. 251-253 (2003)

Yuji Tachibana：“日常临床实践中需要注意的锥虫病和利什曼病、原虫和寄生虫病”JIM 13. 251-253 (2003)。

Tachibana, H., et al.: "Evaluation of recombinant fragments of Entamoeba histolytica Gal/GalNAc lectin intermediate subunit for serodiagnosis of amebiasis"J.Clin.Microbiol.. 42(3)(印刷中). (2004)

Tachibana, H., et al.：“用于阿米巴病血清诊断的溶组织内阿米巴 Gal/GalNAc 凝集素中间亚基的重组片段的评估”J.Clin.Microbiol.. 42(3)（出版中）。