Stage-specific excretory-secretory small heat shock proteins from the parasitic nematode Strongyloides ratti--putative links to host's intestinal mucosal defense system.
Stage-specific excretory-secretory small heat shock proteins from the parasitic nematode Strongyloides ratti--putative links to host's intestinal mucosal defense system.
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DOI：
10.1111/j.1742-4658.2011.08248.x
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发表时间：
2011-09
期刊：
影响因子：
--
通讯作者：
Brattig NW
中科院分区：
文献类型：
--
作者：
Younis AE;Geisinger F;Ajonina-Ekoti I;Soblik H;Steen H;Mitreva M;Erttmann KD;Perbandt M;Liebau E;Brattig NW
为了寻找肠道线虫与哺乳动物粘膜宿主细胞相互作用的相关分子，我们采用质谱法鉴定了鼠圆线虫的排泄/分泌蛋白（ESP）。除其他肽外，我们在寄生雌虫阶段的ESP中检测到与秀丽隐杆线虫热休克蛋白-17同源的肽，命名为Sra-HSP-17.1 （~19 kDa）和Sra-HSP-17.2 (~ 18 kDa)，氨基酸同源性为49%。鉴定出全长cdna（分别为483 bp和474 bp），并进行基因组组织分析。为了进一步鉴定，对蛋白进行重组表达和纯化。利用qRT-PCR和ELISA分析了不同发育阶段寄生蜂的特异性表达。DNA序列分析和氨基酸序列分析表明，两个基因共享一个保守的α -结晶蛋白结构域和可变的n端。Sra-HSP-17蛋白与推断的人粪球菌热休克小蛋白序列同源性最高。我们观察到两种蛋白的强免疫原性，导致大鼠感染后的高IgG反应。流式细胞分析表明，Sra-HSP-17s与单核/巨噬细胞系结合，但不与外周淋巴细胞或中性粒细胞结合。大鼠肠上皮细胞系与Sra-HSP-17.1的结合呈剂量依赖性，而与Sra-HSP-17.2的结合无剂量依赖性。暴露的单核细胞在对Sra-HSP-17s的反应中释放IL-10，但不释放tnf - α，这表明可能参与了宿主免疫反应中分泌的雌性蛋白。
In search of molecules involved in the interaction of intestinal nematodes and mammalian mucosal host cells, we performed mass spectrometry to identify excretory/secretory proteins (ESP) from Strongyloides ratti. In addition to other peptides, we detected in the ESP of parasitic female stage peptides homologous to the Caenorhabditis elegans heat shock protein-17, named Sra-HSP-17.1 (~19 kDa) and Sra-HSP-17.2 (~ 18 kDa) with 49% amino acid identity. The full-length cDNAs (483 bp and 474 bp, respectively) were identified and the genomic organization analyzed. To allow further characterization, the proteins were recombinantly expressed and purified. Profiling of transcription by qRT-PCR and of protein by ELISA in various developmental stages revealed parasitic female-specific expression. The sequence analysis of both DNA and amino acid sequence showed two genes share a conserved alpha-crystallin domain and variable N-terminals. The Sra-HSP-17 proteins showed the highest homology to the deduced small heat-shock protein sequence of the human pathogen S. stercoralis. We observed strong immunogenicity of both proteins, leading to high IgG responses following infection of rats. Flow cytometric analysis indicated the binding of Sra-HSP-17s to the monocytes/macrophage lineage but not to peripheral lymphocytes or neutrophils. A rat intestinal epithelial cell line showed dose dependent binding to Sra-HSP-17.1, but not to Sra-HSP-17.2. Exposed monocytes released IL-10 but not TNF-alpha in response to Sra-HSP-17s, suggesting a possible involvement of secreted female proteins in host immune responses.
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影响因子：
1.5
作者：
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通讯作者：
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影响因子：
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作者：
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作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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发表时间：
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影响因子：
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作者：
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