动物实验,虫卵抗原Sjp40 z-bo 2020-08-22 589

　　OBJECTIVE: To observe the expression of Sjp40 in the formation of liver eggs and granulomas in infected New Zealand white rabbits, and to perform immunolocalization.

　　Method: Infect New Zealand white rabbits with Schistosoma schistosome tail c, collect livers of uninfected group, 29dpi group and 45dpi group, and extract the total RNA of each group by Trizol method. Taqman probe qRT-PCR was used to detect the expression of Sjp40 mRNA; total liver protein was extracted from each group; anti-Sjp40-McAb9G7 and anti-Toxoplasma tSAG1-McAbY3A8 were extracted by ammonium sulfate salting out method and Protein G immunoaffinity chromatography (control antibody). Western blot was used to detect the expression of Sjp40 protein. Prepare liver paraffin sections, observe the formation and development of liver egg granuloma by HE staining, and use immunofluorescence technology to further place Sjp40 in the liver deposits of egg and granuloma tissue.

　　Result: The level of Sjp40 mRNA in 45dpi liver deposited eggs with acute hepatic granulomatous lesions infected by Schistosoma japonicum was significantly higher than that of immature liver eggs without egg granuloma nodules deposited at 29dpi. u003c0.05). Westernbblot confirmed the expression of Sjp40 protein in rabbit liver at 29 and 45 dpi. Immunofluorescence showed that Sjp40 was present in immature eggs of rabbit liver at 29 dpi, but at 45 dpi, many mature eggs were deposited in the liver of infected rabbits, including the surrounding areas of trichomes. Clear fluorescence is seen in the granuloma tissue.

　　Conclusion: As the eggs mature, the transcription level of Sjp40 in the eggs deposited in the liver of schistosomiasis-infected rabbits increases significantly. During acute granulomatous lesions (45 dpi) in infected rabbits, the eggs spread to the surrounding granulomatous tissue.