title: [Expression profiling and immunofluorescence localization of the major egg antigen p40 of Schistosoma japonicum in the liver of infected New Zealand white rabbits].
abstract: OBJECTIVE
  To examine the expression profile and immunofluorescence localization of the major egg antigen p40 of Schistosoma japonicum (Sjp40) during granuloma formation in the liver of infected New Zealand white rabbits.
  
  
  METHODS
  New Zealand white rabbits were infected with S. japonicum cercariae, and the livers were harvested at 29 and 45 days post-infection (dpi). The total RNA of the liver tissues was extracted for expression profiling of Sjp40 by quantitative reverse transcription-PCR (qRT-PCR) with GAPDH of S. japonicum as the endogenous reference gene. The expression of Sjp40 in the liver were detected by Western blotting using anti-Sjp40 monoclonal antibody (mAb) 9G7 or anti-Toxoplasma gondii tSAG1 mAb Y3A8 (control) as the primary antibody. Paraffin sections of the liver were prepared for observing egg granuloma formation using HE staining and for indirect immunofluorescence assay of Sjp40 location in the trapped eggs and egg granulomas.
  
  
  RESULTS
  The level of Sjp40 mRNA in the eggs trapped in rabbit livers was significantly higher at 45 dpi than that at 29 dpi (P<0.05), and Western blotting confirmed the presence of Sjp40 protein in the rabbit livers at both 29 and 45 dpi. Immunofluorescence assay demonstrated localized expression of Sjp40 in the immature eggs in the rabbit liver at 29 dpi, but at 45 dpi fluorescence was detected in clusters of mature eggs containing miracidium and in the surrounding egg granulomas.
  
  
  CONCLUSIONS
  The transcriptional levels of Sjp40 significantly increased with the maturation of eggs trapped in the rabbit livers. Sjp40 protein spread from the eggs to the surrounding egg granuloma at 45 dpi when acute liver granulomatous lesions occur, suggesting that Sjp40 plays a key role in egg granulomas formation in the livers of infected New Zealand white rabbits.
authors.affiliation: Key Laboratory of Prevention and Control for Emerging Infectious Diseases of Guangdong Higher Institutes, Department of Pathogen Biology, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, China. E-mail: zixia-wang@outlook.com.
authors.forename: Dan
authors.initials: D
authors.lastname: Xia
authors.forename: Ganming
authors.initials: G
authors.lastname: Deng
authors.forename: Pingying
authors.initials: P
authors.lastname: Teng
authors.forename: Yu
authors.initials: Y
authors.lastname: Xie
authors.forename: Yaomin
authors.initials: Y
authors.lastname: Li
authors.forename: Chunmei
authors.initials: C
authors.lastname: Wang
authors.forename: Shujie
authors.initials: S
authors.lastname: Chen
authors.forename: Minfang
authors.initials: M
authors.lastname: Chen
authors.forename: Rongjia
authors.initials: R
authors.lastname: Mai
authors.forename: Haiyan
authors.initials: H
authors.lastname: Liao
authors.forename: Lingyu
authors.initials: L
authors.lastname: Shi
authors.forename: Liyan
authors.initials: L
authors.lastname: Ou
authors.forename: Qiwei
authors.initials: Q
authors.lastname: Chen
authors.forename: Xiaoguang
authors.initials: X
authors.lastname: Chen
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Zhou
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D020869:Gene Expression Profiling; D006099:Granuloma / Q000469:parasitology; D015801:Helminth Proteins / Q000378:metabolism*; D008099:Liver / Q000469:parasitology*; D012333:RNA, Messenger; D011817:Rabbits; D012549:Schistosoma japonicum; D012554:Schistosomiasis japonica
chemical_list: D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth; D015801:Helminth Proteins; D012333:RNA, Messenger; C055560:p40 egg antigen, Schistosoma