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首页> 外文期刊>Parasitology >Dissimilar peptidase production by avirulent and virulent promastigotes of Leishmania braziliensis: inference on the parasite proliferation and interaction with macrophages.
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Dissimilar peptidase production by avirulent and virulent promastigotes of Leishmania braziliensis: inference on the parasite proliferation and interaction with macrophages.

机译：巴西利什曼原虫无毒和有毒力的前鞭毛体产生不同的肽酶：推断寄生虫的增殖和与巨噬细胞的相互作用。

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摘要

In the present paper, we have analysed the cellular and extracellular proteolytic activity profiles in 2 distinct Leishmania braziliensis strains: a recently isolated (virulent) and a laboratory-adapted (avirulent) strain. Quantitative and qualitative differences on the peptidase expression were observed in both strains. For instance, low-molecular mass acidic cysteine peptidase activities were detected exclusively in the virulent strain. Similarly, metallopeptidase activities were mainly produced by L. braziliensis virulent promastigotes. Interestingly, metallo- and cysteine peptidase activities were drastically reduced after several in vitro passages of the virulent strain. Western blotting, flow cytometry and fluorescence microscopy analyses were performed to detect homologous of the major leishmania metallopeptidase (gp63) and cysteine peptidase (cpb) in virulent and avirulent strains of L. braziliensis. Our results revealed that the virulent strain produced higher amounts of gp63 and cpb molecules, detected both in the surface and cytoplasm regions, than the avirulent counterpart. Metallo- (1,10-phenanthroline and EGTA) and cysteine peptidase (E-64) inhibitors arrested the growth of L. braziliensis virulent strain in a dose-dependent manner, as well as the association index with peritoneal murine macrophages. Conversely, these peptidase inhibitors did not affect either the proliferation or the cellular interaction of the avirulent strain. Corroborating these findings, the pre-treatment of the virulent strain with both anti-peptidase antibodies promoted a prominent reduction in the interaction with macrophages, while the association index of the avirulent strain to macrophage was only slightly diminished. Moreover, the spent culture medium from virulent strain significantly enhanced the association index between avirulent strain and macrophages, and this effect was reversed by 1,10-phenanthroline. Collectively, the results presented herein suggest that peptidases participate in several crucial processes of L. braziliensis.
机译：在本文中，我们分析了两种巴西利什曼原虫菌株的细胞和细胞外蛋白水解活性谱：最近分离的（有毒）菌株和实验室适应的（无毒）菌株。在两种菌株中均观察到肽酶表达的数量和质量上的差异。例如，仅在强毒株中检测到低分子酸性半胱氨酸肽酶活性。同样，金属肽酶活性主要由巴西乳杆菌强毒前鞭毛体产生。有趣的是，在毒性菌株经过数次体外传代后，金属和半胱氨酸肽酶的活性急剧下降。进行了蛋白质印迹，流式细胞仪和荧光显微镜分析，以检测巴西利兹氏菌有毒力和无毒力菌株中主要的利什曼原虫金属肽酶（gp63）和半胱氨酸肽酶（cpb）的同源性。我们的结果表明，与无毒菌株相比，在表面和细胞质区域均检测出有毒力的菌株产生了大量的gp63和cpb分子。金属（1,10-菲咯啉和EGTA）和半胱氨酸肽酶（E-64）抑制剂以剂量依赖性以及与腹膜鼠巨噬细胞的结合指数抑制了巴西乳杆菌的强毒株的生长。相反，这些肽酶抑制剂不影响无毒力菌株的增殖或细胞相互作用。证实了这些发现，用两种抗肽酶抗体对有毒力的菌株进行的预处理均促进了与巨噬细胞相互作用的显着降低，而无毒力的菌株与巨噬细胞的缔合指数仅略有降低。此外，来自强毒株的废培养基显着提高了无毒株与巨噬细胞之间的结合指数，这种作用被1,10-菲咯啉所逆转。总体而言，本文呈现的结果表明肽酶参与了巴西乳杆菌的几个关键过程。

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