DEVELOPMENT OF SEROLOGICAL DIAGNOSIS OF BANCROFTIAN FILARIASIS BY A MONOCLONAL ANTIBODY

单克隆抗体班氏丝虫病血清学诊断方法的研究进展

• 批准号: 04670224
• 负责人: KOBAYASHI Masashi
• 金额: $ 1.34万
• 依托单位: CHIBA 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: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04670224/
• 关键词: Wuchereria bancrofti; Filariasis; Monoclonal antibody; Microfilaria; Enzyme Linked Immunoassay; 糸状虫症; ミクロフィラリア; 血清抗体価

Hybridoma cell lines, which secreted antibodies directed against either the soluble fraction of microfilaria or surface of sheath of microfilaria, were produced by fusion of spleen cells of W.bancrofti microfilariae-immunized mice with NS-1 myeloma cells. Fourteen cloned antibody-secreting cell lines were successfully established. Two of these clones were characterized. One clone (1G10) secreted an IgM antibody that had a high stage specificity with recognizing the microfilaria but did not react against adult worm. Another clone (2G01) directed against the sheath of microfilaria was found to produce an IgG_1, but did not show a binding to adult filaria.The evaluation of monoclonal-based enzyme immunoassay for detecting soluble microfilarial antigen in sera collected in an endemic area of filariasis was carried out. Filarial antigen was detected in sera from 135 of 140 microfilaremic patients, 1 of 25 amicrofilaremic patients with clinical filariasis, and 5 of 30 endemic controls. Antigen was not detected in sera form nonendemic areas s who had a variety of other helminth infections. Parasite antigen titer were significantly correlated with microfilrial counts in night blood smears. Antibodies to circulating W.bancrofti antigen were detected 16 of 23 antigen-negative sera from patients with clinical filariasis. Despite this limitation, detecting parasite antigen by enzyme immunoassay provides significant advantages over previously available methods for diagnosing active W.bancrofti infection.

将班氏丝虫免疫小鼠的脾细胞与NS-1骨髓瘤细胞融合，获得了分泌抗微丝蚴可溶性部分或微丝蚴鞘表面抗体的杂交瘤细胞系。成功建立了14株克隆化抗体分泌细胞系。对其中两个克隆进行了表征。1个克隆（1G 10）分泌的IgM抗体具有高度的阶段特异性，可识别微丝蚴，但不与成虫反应。另一个抗微丝蚴鞘的克隆（2G 01）能产生IgG_1，但不与成虫结合。140例微丝蚴血症者中135例检出丝虫抗原，25例无微丝蚴血症者中1例检出丝虫抗原，30例地方病对照中5例检出丝虫抗原。在患有各种其他蠕虫感染的非流行地区的血清中未检测到抗原。寄生虫抗原滴度与夜间血涂片中微丝蚴计数显著相关。对23例班氏丝虫病患者血清中16例进行了抗班氏丝虫循环抗原抗体检测。尽管存在这种局限性，但通过酶免疫测定法检测寄生虫抗原提供了优于先前可用的用于诊断活动性班氏瓦氏体感染的方法的显著优势。

项目成果

Masashi Kobayashi,Munetoshi Niimura,Elizabeth M.Santana: "A monoclonal antibody-based Enzyme linked immunoassay for detecting parasite antigen Bancroftian Filariasis." The Journal of Infectious Diseases. (準備中).

Masashi Kobayashi、Munetoshi Niimura、Elizabeth M.Santana：“用于检测寄生虫抗原班克罗夫特丝虫病的单克隆抗体酶联免疫测定法。”传染病杂志（正在准备中）。

小林 仁、小島 荘明、E.Malagueno,T.Miranda: "バンクロフト糸状虫ミクロフィラリアの特異的なモノクローナル抗体の産性とその性状" 日本寄生虫学雑誌. 39. 67 (1990)

Hitoshi Kobayashi、Shoaki Kojima、E.Malagueno、T.Miranda：“班克罗夫特微丝蚴特异性单克隆抗体的生产力和特性”日本寄生虫学杂志 39. 67 (1990)。

Masashi Kobayashi, Somei Kojima Elizabeth M.Santana, Luciano T.Montenegro and Paulo J.Miranda: "Production and properties of Mouse Monoclonal antibodies to Wuchereria Bancrofti microfilaria." Jpn.J.Parasitol.39. 67 (1990)

Masashi Kobayashi、Somei Kojima Elizabeth M.Santana、Luciano T.Montenegro 和 Paulo J.Miranda：“针对 Wuchereria Bancrofti 微丝蚴的小鼠单克隆抗体的生产和特性。”

Masashi Kobayashi, Munetoshi Niimura and Elizabeth M.Santana: "A monoclonal antibody-based Enzyme linked immunoassay for detecting parasite antigen of Bancroftian Filariasis." J.Inf.Diseases.(in press).

Masashi Kobayashi、Munetoshi Niimura 和 Elizabeth M.Santana：“一种基于单克隆抗体的酶联免疫测定法，用于检测班克罗夫特丝虫病的寄生虫抗原。”

小林 仁、小島 荘明、E.Malagueno,P.Miranda: "バンクロフト糸状虫ミクロフィラリアに特異的なモノクローナル抗体の産性とその性状" 日本寄生虫学雑誌. 39. 67 (1990)

Hitoshi Kobayashi、Shoaki Kojima、E.Malagueno、P.Miranda：“班克罗夫特微丝蚴特异性单克隆抗体的生产力和特性”日本寄生虫学杂志 39. 67 (1990)。