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The Research on the BTV and Its dsRNA Inducing Cells to Produce IFN and to Get into Apoptosis

Author: WangHaiZuo
Tutor: DongChangYuan
School: Wuhan University
Course: Pathogen Biology
Keywords: BTV dsRNA Interferon Transfection Apoptosis
CLC: R346
Type: Master's thesis
Year: 2005
Downloads: 51
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Abstract

IFNs are one group of extensive anti-viral substance, which are induced through the interferon inducer that could incite cultivated cell and human or animal body to defeat virus and tumor. IFN inducer are another substance that could induce cell to produce IFN, but currently it’s thought that Poly[I:C] are the most ideal and important cell IFN inducer, which could directly apply in vitro to induce IFN. However, it’s also found that Poly[I:C] was amiable to be destroyed in human and primates body by nucleic acid enzyme in blood serum, and its ability and efficiency were a little low. According this, this research was ready to explore and discuss that whether the animal BTV and its genome dsRNA which could not infect human normal cell and arouse any human diseases had the capability of inducing cell to produce IFN and tumor cell to enter into apoptosis. BTV belongs to respiratory enteric orphan virus branch; genus orbivirus category, it’s able to arouse sheep and wild ruminant to suffer from the bluetongue disease, the cattle, goat and deer are the viral storage of BTV. The whole inherit information is composed of ten gene segements, the whole genome has 19218 bp sequences, its whole molecular weight is 13 × 10~6Da, and so far it’s thought the biggest RNA virus. The complete inherit information of BTV genome dsRNA bases its possibility and infinitude potential as a new kind of endogenesis IFN inducer. When the exacted viral nucleic acid segments are entered into cell, the dsRNA would be lack of infection ability because of the defect of RNA polymerize enzyme and the non-integrality of BTV whole genome, but its biological function of inducing IFN is remain, moreover the dsRNA would be safety and effective than its BTV granule to body.It’s studied in our research that the genome daRNA was exacted through Trizol, and it was directly wrapped by LP and transfected into Hela cell and HEL cell differently. At the same time, we took BTV and bare dsRNA as comparison, BTV infected the two kinds of cell; bare dsRNA was also added into the two cells’ plant. We compared and sdudied that the differences among three treatments, and observed two cells’ morphological change, detected the IFN content in two cells’ suspension through β -IFN ELISA kit, tested the cells apoptosis rate through flow cytometor. Finally, all of the results were analyzed through statistics method. It’s found that no matter Hela cell or HEL cell, BTV and bare dsRNA and dsRNA wrapped LP were all able to induce the two kinds of cell to produce IFN, which had significant differences, the IFN output induced under different content of dsRNA wrappedlipofactamine comopound was higher than that of different content of simple dsRNA, which however was higher than that of different dosage of BTV. According to the analysis of statistic variance, the differences among them possessed significance (P<0.05) ; and the ability to produce IFN of HEL cell was higher than that of Hela, which also was proved to be possessed significance (P<0.05) , and cell apoptosis of Hela cell induced under three treatment were all higher than that of HEL cells, the apoptosis rate of HEL cell almost had not any change , which was accord with the formal results of the experiment of our laboratory.Conclusion: The result of our research approved that BTV and its dsRNA molecule possessed the ability to induce cell to produce IFN and tumor cell to enter into apoptosisk, and bare BTV dsRNA molecule was most likely to be developed into a new latent kind of endogenesis IFN inducer, the study afford a great experimental basis for the theory research of endogenesisi IFN inducer, and it’s also established stable theory and experimental basis for our laboratory to develop deep into its animal experiment and clinical trial, our research would embody its important merit among treatment to kinds of viral infection and tumor.

The Research on the BTV and Its dsRNA Inducing Cells to Produce IFN and to Get into Apoptosis

Abstract

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