IMMUNOPROTECTION AND DIAGNOSTIC POTENTIAL OF SIGNALING PROTEIN 14-3-3 OF SCHISTOSOMA JAPONICUM×

刘庆中; 沈继龙; 李德发; 李锋; 汪学龙; 江宝玲

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IMMUNOPROTECTION AND DIAGNOSTIC POTENTIAL OF SIGNALING PROTEIN 14-3-3 OF SCHISTOSOMA JAPONICUM×

首发时间：2006-02-06

Liu Qingzhong ¹ Shen Jilong ¹ Li Defa ¹ Li Feng ¹ Wang Xuelong ¹ JIang Baoling ¹

1、Department of Microbiology and Parasitology, Anhui Medical University

Abstract：The 14-3-3 protein is a key player in signal transduction processes in various species of animals and plants. Here, we cloned and expressed the 14-3-3 of Schistosoma japonicum (Sj14-3-3, Chinese mainland strain) in pET28a/E.coli. And the protein can be strongly recognized by polyclonal anti-14-3-3εand sera from animals infected with S. japonicum. Monoclonal antibodies against Sj14-3-3 were raised using a highly specific, affinity-purified protein. The antibodies preparation by ammonium sulfate precipitation was employed for the localization of the native 14-3-3 protein in the parasite by immunofluorescence microscopy. The results demonstrate a wide distribution of this protein in tegument, subtegument, muscle and parenchyma of male, female adult parasites and 15-day-old schistosomulum. The potential use of this protein or its DNA as a vaccine candidate against infection with S. japonicum in BALB/c mice and the recombinant Sj14-3-3 (rSj14-3-3) antigen for diagnosis of schistosomiasis was evaluated. Challenge infection with S. japonicum, immunization with rSj14-3-3, rSjGST, and rSj14-3-3+rSjGST proteins using Freund’s adjuvant or Mycobacterium tuberculosis (Mtb) low molecular peptide as adjuvant respectively, led to 26.0-32.2％ protection, as determined by reduction of adult worm burden; 50.4-58.6％ protection as determined by reduction of the eggs in liver tissue. The mice were directly inoculated with rSj14-3-3 and rSj14-3-3+rSjGST cDNAs through quadriceps femoris muscle for immunization. As the consequence, the worm reduction was found to be 34.20％ in rSj14-3-3 group, 32.40％in rSj14-3-3+rSjGST group; the number of eggs in liver tissue was reduced by 50.74％ and 55.23％, respectively. Indirect ELISA suggested that the rSj14-3-3 antigen is highly sensitive and specific for the diagnosis of schitosomiasis.

keywords： Schistosome recombinant vaccine immunodiagnosis

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IMMUNOPROTECTION AND DIAGNOSTIC POTENTIAL OF SIGNALING PROTEIN 14-3-3 OF SCHISTOSOMA JAPONICUM×

刘庆中 ¹ 沈继龙 ¹ 李德发 ¹ 李锋 ¹ 汪学龙 ¹ 江宝玲 ¹

1、安徽医科大学基础医学院

摘要：The 14-3-3 protein is a key player in signal transduction processes in various species of animals and plants. Here, we cloned and expressed the 14-3-3 of Schistosoma japonicum (Sj14-3-3, Chinese mainland strain) in pET28a/E.coli. And the protein can be strongly recognized by polyclonal anti-14-3-3εand sera from animals infected with S. japonicum. Monoclonal antibodies against Sj14-3-3 were raised using a highly specific, affinity-purified protein. The antibodies preparation by ammonium sulfate precipitation was employed for the localization of the native 14-3-3 protein in the parasite by immunofluorescence microscopy. The results demonstrate a wide distribution of this protein in tegument, subtegument, muscle and parenchyma of male, female adult parasites and 15-day-old schistosomulum. The potential use of this protein or its DNA as a vaccine candidate against infection with S. japonicum in BALB/c mice and the recombinant Sj14-3-3 (rSj14-3-3) antigen for diagnosis of schistosomiasis was evaluated. Challenge infection with S. japonicum, immunization with rSj14-3-3, rSjGST, and rSj14-3-3+rSjGST proteins using Freund’s adjuvant or Mycobacterium tuberculosis (Mtb) low molecular peptide as adjuvant respectively, led to 26.0-32.2％ protection, as determined by reduction of adult worm burden; 50.4-58.6％ protection as determined by reduction of the eggs in liver tissue. The mice were directly inoculated with rSj14-3-3 and rSj14-3-3+rSjGST cDNAs through quadriceps femoris muscle for immunization. As the consequence, the worm reduction was found to be 34.20％ in rSj14-3-3 group, 32.40％in rSj14-3-3+rSjGST group; the number of eggs in liver tissue was reduced by 50.74％ and 55.23％, respectively. Indirect ELISA suggested that the rSj14-3-3 antigen is highly sensitive and specific for the diagnosis of schitosomiasis.

关键词：血吸虫重组疫苗免疫诊断

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1. 国家自然基金（30571631）

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Liu Qingzhong,Shen Jilong,Li Defa,et al. IMMUNOPROTECTION AND DIAGNOSTIC POTENTIAL OF SIGNALING PROTEIN 14-3-3 OF SCHISTOSOMA JAPONICUM×[EB/OL]. Beijing:Sciencepaper Online[2006-02-06]. https://www.paper.edu.cn/releasepaper/content/200602-15.

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论文编号	200602-15
论文题目	IMMUNOPROTECTION AND DIAGNOSTIC POTENTIAL OF SIGNALING PROTEIN 14-3-3 OF SCHISTOSOMA JAPONICUM×
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