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Insights into the proteomic profile and gene expression of Lutzomyia longipalpis-derived Lulo cell line
Memórias do Instituto Oswaldo Cruz ( IF 2.8 ) Pub Date : 2020-10-26 , DOI: 10.1590/0074-02760200113
Luzia Monteiro de Castro Côrtes 1 , Daniela de Pita-Pereira 1 , Priscila Silva Grijó Farani 1 , Bernardo Acácio Santini Pereira 1 , Geovane Dias-Lopes 1 , Franklin Souza da Silva 1 , Paloma Resende Corrêa 1 , Roger Magno Macedo Silva 1 , Suzana Côrte-Real 1 , Felio Jesus Bello 2 , Leila Mendonça-Lima 1 , Otacilio da Cruz Moreira 1 , Mariana Caldas Waghabi 1 , Carlos Roberto Alves 1
Affiliation  

BACKGROUND Lutzomyia longipalpis-derived cell line (Lulo) has been suggested as a model for studies of interaction between sandflies and Leishmania. OBJECTIVES Here, we present data of proteomic and gene expression analyses of Lulo cell related to interactions with Leishmania (Viannia) braziliensis. METHODS Lulo cell protein extracts were analysed through a combination of two-dimensional gel electrophoresis and mass spectrometry and resulting spots were further investigated in silico to identify proteins using Mascot search and, afterwards, resulting sequences were applied for analysis with VectorBase. RESULTS Sixty-four spots were identified showing similarities to other proteins registered in the databases and could be classified according to their biological function, such as ion-binding proteins (23%), proteases (14%), cytoskeletal proteins (11%) and interactive membrane proteins (9.5%). Effects of interaction with L. (V.) braziliensis with the expression of three genes (enolase, tubulin and vacuolar transport protein) were observed after an eight-hour timeframe and compared to culture without parasites, and demonstrated the impact of parasite interaction with the expression of the following genes: LLOJ000219 (1.69-fold), LLOJ000326 (1.43-fold) and LLOJ006663 (2.41-fold). CONCLUSIONS This set of results adds relevant information regarding the usefulness of the Lulo cell line for studies with Leishmania parasites that indicate variations of protein expression.

中文翻译:

深入研究Lutzomyia longipalpis衍生的Lulo细胞系的蛋白质组学特征和基因表达

背景技术长期以来,人们认为Lutzomyia longipalpis衍生的细胞系(Lulo)是研究沙蝇和利什曼原虫之间相互作用的模型。目的在这里,我们介绍与巴西利什曼原虫(Viannia)的相互作用相关的Lulo细胞的蛋白质组学和基因表达分析数据。方法通过二维凝胶电泳和质谱相结合的方法对Lulo细胞蛋白提取物进行分析,并通过Mascot搜索在计算机上进一步研究所得斑点,以鉴定蛋白质,然后将所得序列用于VectorBase分析。结果确定了64个点,这些点与数据库中注册的其他蛋白质相似,可以根据其生物学功能进行分类,例如离子结合蛋白(23%),蛋白酶(14%),细胞骨架蛋白(11%)和相互作用膜蛋白(9.5%)。在八个小时的时间框架内,观察了与巴西利亚L.(V.)的相互作用与三个基因(烯醇酶,微管蛋白和液泡转运蛋白)表达的影响,并将其与无寄生虫的培养进行了比较,并证明了寄生虫与巴西寄生虫相互作用的影响表达以下基因:LLOJ000219(1.69倍),LLOJ000326(1.43倍)和LLOJ006663(2.41倍)。结论这组结果增加了有关Lulo细胞系在利什曼原虫寄生虫研究中的有用性的相关信息,该研究表明蛋白质表达的变化。8小时后观察到微管蛋白和液泡转运蛋白),并将其与不含寄生虫的培养进行比较,并证明了寄生虫相互作用对以下基因表达的影响:LLOJ000219(1.69倍),LLOJ000326(1.43倍)和LLOJ006663(2.41倍)。结论这组结果增加了有关Lulo细胞系在利什曼原虫寄生虫研究中的有用性的相关信息,该研究表明蛋白质表达的变化。8小时后观察到微管蛋白和液泡转运蛋白),并将其与不含寄生虫的培养进行比较,并证明了寄生虫相互作用对以下基因表达的影响:LLOJ000219(1.69倍),LLOJ000326(1.43倍)和LLOJ006663(2.41倍)。结论这组结果增加了有关Lulo细胞系在利什曼原虫寄生虫研究中的有用性的相关信息,该研究表明蛋白质表达的变化。
更新日期:2020-10-27
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