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棘颚口线虫幼虫部分纯化抗原对人B细胞培养产生抗体的作用。

Roles of partially purified antigens from Gnathostoma spinigerum larvae on antibody production by human B cell culture.

作者信息

Somthana Kasem, Eshita Yuki, Kumsiri Ratchanok, Dekumyoy Paron, Waikagul Jitra, Kalambaheti Thareerat, Maneerat Yaowapa

机构信息

Department of Tropical Pathology, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand.

出版信息

Southeast Asian J Trop Med Public Health. 2011 Jul;42(4):772-81.

Abstract

A 24 kDa protein from advanced third stage Gnathostoma spinigerum larvae (GsAL3) is used for gnathostomiasis serodiagnosis. This study investigated whether partially purified protein antigen (Ag) from GsAL3 (Gnath Ag), prepared by simple gel filtration chromatography, could be used for serodiagnosis. Using DNA microarray analysis, significant gene expression related to immunoreactivity was examined in peripheral blood mononuclear cells (PBMC) cocultured with Gnath Ag. Antigenicity was then determined by its capacity to induce antibody production among purified naive B cells stimulated with Gnath Ag and anti-CD40. Seven and 14 days post-exposure, immunoglobulin levels (Igs) in culture supernatants were determined by enzyme-linked immunosorbent assay. The Gnath Ag stimulated PBMC had a significant increase in gene expression related to an innate immune response and decreased cell mediated immunity, but the expression of gene related antibody production was not markedly increased. The Gnath Ag stimulated naive B cells or lipopolysaccharide primed B cells to produce low levels of specific antibody. Our findings support the assertion that partially purified Gnath Ag possess low antigenicity for Ig induction. Further studies are needed to improve G. spinigerum larva Ag for serodiagnosis.

摘要

来自棘颚口线虫晚期第三期幼虫(GsAL3)的一种24 kDa蛋白用于颚口线虫病的血清学诊断。本研究调查了通过简单凝胶过滤色谱法制备的GsAL3部分纯化蛋白抗原(Ag)(Gnath Ag)是否可用于血清学诊断。使用DNA微阵列分析,在与Gnath Ag共培养的外周血单核细胞(PBMC)中检测了与免疫反应性相关的显著基因表达。然后通过其在Gnath Ag和抗CD40刺激的纯化幼稚B细胞中诱导抗体产生的能力来确定抗原性。暴露后7天和14天,通过酶联免疫吸附测定法测定培养上清液中的免疫球蛋白水平(Igs)。Gnath Ag刺激的PBMC中与先天免疫反应相关的基因表达显著增加,细胞介导的免疫降低,但与抗体产生相关的基因表达没有明显增加。Gnath Ag刺激幼稚B细胞或脂多糖致敏的B细胞产生低水平特异性抗体。我们的研究结果支持以下观点,即部分纯化的Gnath Ag对Ig诱导具有低抗原性。需要进一步研究以改进棘颚口线虫幼虫抗原用于血清学诊断。

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