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通过cDNA文库的λ展示对人类针对弓形虫感染的B细胞反应进行分子剖析。

Molecular dissection of the human B-cell response against Toxoplasma gondii infection by lambda display of cDNA libraries.

作者信息

Beghetto Elisa, Spadoni Andrea, Buffolano Wilma, Del Pezzo Mariassunta, Minenkova Olga, Pavoni Emiliano, Pucci Andrea, Cortese Riccardo, Felici Franco, Gargano Nicola

机构信息

Kenton Laboratories, Via Pontina km 30.400, 00040 Rome, Italy.

出版信息

Int J Parasitol. 2003 Feb;33(2):163-73. doi: 10.1016/s0020-7519(02)00256-4.

Abstract

The disorders generated by Toxoplasma gondii infection are closely associated with the competence of the host immune system and both humoral and cell mediated immunity are involved in response to parasite invasion. To identify antigens implicated in human B-cell responses, we screened a phage-display library of T. gondii cDNA fragments with sera of infected individuals. This approach identified a panel of recombinant phage clones carrying B-cell epitopes. All the peptide sequences selected by this procedure are regions of T. gondii gene products. These regions contain epitopes of the T. gondii antigens SAG1, GRA1, GRA7, GRA8 and MIC5, which are recognised by human immunoglobulins. Moreover, we report the isolation and characterisation of two additional immunodominant regions encoded by GRA3 and MIC3 genes, whose products have never been described as antigens of the human B-cell response against T. gondii infection. These results demonstrate potential of lambda-display technology for antigen discovery and for the study of the human antibody response against infectious agents.

摘要

由弓形虫感染引发的疾病与宿主免疫系统的能力密切相关,体液免疫和细胞介导免疫均参与对寄生虫入侵的反应。为了鉴定与人类B细胞反应相关的抗原,我们用感染个体的血清筛选了一个弓形虫cDNA片段的噬菌体展示文库。这种方法鉴定出一组携带B细胞表位的重组噬菌体克隆。通过该程序选择的所有肽序列都是弓形虫基因产物的区域。这些区域包含弓形虫抗原SAG1、GRA1、GRA7、GRA8和MIC5的表位,它们可被人类免疫球蛋白识别。此外,我们报告了由GRA3和MIC3基因编码的另外两个免疫显性区域的分离和特性,其产物从未被描述为人类针对弓形虫感染的B细胞反应的抗原。这些结果证明了λ展示技术在抗原发现以及研究人类针对感染因子的抗体反应方面的潜力。

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