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首页> 外文期刊>Infection and immunity >Characterization of Entamoeba histolytica Intermediate Subunit Lectin-Specific Human Monoclonal Antibodies Generated in Transgenic Mice Expressing Human Immunoglobulin Loci
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Characterization of Entamoeba histolytica Intermediate Subunit Lectin-Specific Human Monoclonal Antibodies Generated in Transgenic Mice Expressing Human Immunoglobulin Loci

机译：在表达人类免疫球蛋白基因座的转基因小鼠中产生的变形虫变形虫中间亚单位凝集素特异性人类单克隆抗体的表征。

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Four fully human monoclonal antibodies (MAbs) to Entamoeba histolytica intermediate subunit lectin (Igl) were prepared in XenoMouse mice, which are transgenic mice expressing human immunoglobulin loci. Examination of the reactivities of these MAbs to recombinant Igl1 and Igl2 of E. histolytica showed that XEhI-20 {immunoglobulin G2(κ) [IgG2(κ)]} and XEhI-28 [IgG2(κ)] were specific to Igl1, XEhI-B5 [IgG2(κ)] was specific to Igl2, and XEhI-H2 [IgM(κ)] was reactive with both Igls. Gene analyses revealed that the VH and VL germ lines were VH3-48 and L2 for XEhI-20, VH3-21 and L2 for XEhI-28, VH3-33 and B3 for XEhI-B5, and VH4-4 and A19 for XEhI-H2, respectively. Flow cytometry analyses showed that the epitopes recognized by all of these MAbs were located on the surfaces of living trophozoites. Confocal microscopy demonstrated that most Igl1 and Igl2 proteins were colocalized on the surface and in the cytoplasm, but different localization patterns in intracellular vacuoles were also present. The preincubation of trophozoites with XEhI-20, XEhI-B5, and XEhI-H2 caused significant inhibition of the adherence of trophozoites to Chinese hamster ovary cells, whereas preincubation with XEhI-28 did not do so. XEhI-20, XEhI-B5, and XEhI-H2 were injected intraperitoneally into hamsters 24 h prior to intrahepatic challenge with E. histolytica trophozoites. One week later, the mean abscess size in groups injected with one of the three MAbs was significantly smaller than that in controls injected with polyclonal IgG or IgM isolated from healthy humans. These results demonstrate that human MAbs to Igls may be applicable for immunoprophylaxis of amebiasis.
机译：在XenoMouse小鼠中制备了四种抗溶组织性阿米巴凝集素中间亚基凝集素（Igl）的四种完全人单克隆抗体（MAb），它们是表达人免疫球蛋白基因座的转基因小鼠。检查这些单克隆抗体对 E的重组Igl1和Igl2的反应性。 histolytica 显示XEhI-20 {免疫球蛋白G2（κ）[IgG2（κ）]}和XEhI-28 [IgG2（κ）]对Igl1具有特异性，XEhI-B5 [IgG2（κ）]对Igl1具有特异性Igl2和XEhI-H2 [IgM（κ）]与两个Igls都具有反应性。基因分析显示，XEhI-20的V H 和V L 种系分别为VH3-48和L2，XEhI-28和VH3-33分别为VH3-48和L2。对于XEhI-B5，分别为B3和B3;对于XEhI-H2，分别为VH4-4和A19。流式细胞仪分析表明，所有这些单克隆抗体识别的表位位于活体滋养体的表面。共聚焦显微镜显示，大多数Igl1和Igl2蛋白共定位在表面和细胞质中，但是在细胞内液泡中也存在不同的定位方式。将滋养体与XEhI-20，XEhI-B5和XEhI-H2一起预孵育会显着抑制滋养体与中国仓鼠卵巢细胞的粘附，而与XEhI-28一起预孵育则不会。 XEhI-20，XEhI-B5和XEhI-H2在肝内用 E攻击前24小时腹膜内注射到仓鼠中。组织溶菌滋养体。一周后，注射了三种单克隆抗体之一的组的平均脓肿大小显着小于注射了从健康人体内分离出的多克隆IgG或IgM的对照组。这些结果证明，人单克隆抗体至Igs可能适用于免疫预防阿米巴病。

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