CHARACTERIZATION OF B-CELL EPITOPES ON IPAB, AN INVASION-ASSOCIATED ANTIGEN OF SHIGELLA-FLEXNERI - IDENTIFICATION OF AN IMMUNODOMINANT DOMAIN RECOGNIZED DURING NATURAL INFECTION
CHARACTERIZATION OF B-CELL EPITOPES ON IPAB, AN INVASION-ASSOCIATED ANTIGEN OF SHIGELLA-FLEXNERI - IDENTIFICATION OF AN IMMUNODOMINANT DOMAIN RECOGNIZED DURING NATURAL INFECTION
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DOI：
10.1128/iai.61.9.3825-3831.1993
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发表时间：
1993-09-01
影响因子：
3.1
通讯作者：
PHALIPON, A
中科院分区：
文献类型：
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作者：
BARZU, S;NATO, F;PHALIPON, A
侵袭质粒抗原B（Ipa B）是一种与志贺氏菌侵袭上皮细胞的能力相关的62-kDa质粒编码的蛋白质，是在感染期间最强烈和一致地被宿主识别的细菌抗原。针对这种侵袭素观察到的强烈的全身和粘膜免疫应答促使我们绘制其B细胞表位。为此目的，IpaB首先在福氏志贺菌中过表达，并用于产生兔多克隆抗血清和鼠单克隆抗体，随后用于筛选IgDagt 11 ipaB文库。对被抗血清和抗体特异性识别的重组DNA克隆进行测序，并鉴定出三个不同的决定簇。这些决定因素的进一步表征表明，它们被志贺氏菌病恢复期患者的血清所识别，这表明它们与自然感染期间的体液反应有关。此外，IpaB区域包括三个决定因素被系统地识别的所有血清从感染的患者，我们测试，而其他地区的蛋白质没有。这些数据表明，该区域，位于氨基酸残基147和258之间，是在自然感染过程中的侵袭素的主要免疫原性结构域。
The invasion plasmid antigen B (IpaB), a 62-kDa plasmid-encoded protein associated with the ability of shigellae to invade epithelial cells, is the bacterial antigen most strongly and consistently recognized by the host during infection. The strong systemic and mucosal immune responses observed against this invasin prompted us to map its B-cell epitopes. For this purpose, IpaB was first overexpressed in Shigella flexneri and used to raise rabbit polyclonal antiserum and murine monoclonal antibodies, which were subsequently used to screen a lambdagt11 ipaB library. Inserts of recombinant DNA clones that were specifically recognized by the antisera and antibodies were sequenced, and three distinct determinants were identified. Further characterization of these determinants showed that they were recognized by sera from patients convalescent from shigellosis, suggesting that they are relevant to the humoral response during natural infection. Moreover, the IpaB region comprising the three determinants was systematically recognized by all sera from infected patients that we tested, whereas other regions of the protein were not. These data suggest that this region, located between amino acid residues 147 and 258, is the major immunogenic domain of the invasin in the course of natural infection.