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PHA体外诱导草鱼白细胞产生γ-干扰素的研究
邵健忠, 项黎新
浙江大学生命科学学院 杭州310012
摘要:
于1997年10月–1999年3月,采用半数细胞病变抑制等方法,进行植物凝集素(Phytoagglutinin,PHA)体外诱导草鱼白细胞产生γ-干扰素的研究。结果表明,在PHA诱导的草鱼白细胞培养液中出现了一种抗病毒因子,经理化和生物学性质鉴定证明,该因子是一种干扰素活性物质。但它不同于病毒诱导的α/β-干扰素,主要表现为:对56℃、pH=2和0.1%SDS敏感;其活性不能被病毒诱导的干扰素抗体所中和;其诱生剂为PHA,来源于白细胞;在诱导条件上,佛波酯(PMA)和白细胞介素-2(IL-2)能显著地促进其诱生,这些特性与人类和高等脊椎动物中报道的γ-干扰素相符,表明是一种γ性质的干扰素
关键词:  植物凝集素  草鱼白细胞  γ-干扰素  体外诱生
DOI:
分类号:
基金项目:国家自然科学基金资助项目,39400100号,39970589号;浙江省自然科学基金资助项目,397300号
IN VITRO INDUCTION OF INTERFERON-GAMMA FROM CTENOPHARYNGODON IDELLUSS LEUCOCYTES BY PHYTOAGGLUTININ
SHAO Jian Zhong, XIANG Li Xin
College of Life Science, Zhejiang University, Hangzhou, 310012
Abstract:
An antiviral factor was detected in grass carp (Ctenopharyngodon idelluss) leucocyte culture supernatant following induction with phytoagglutinin during October, 1997 to March, 1999 in Hangzhou City, Zhejiang Province, China. Biochemical study showed that the factor could not be dialyzed and sedimented at 10×104g, exhibited stability to treatment of DNase, RNase and lipid solvent, but was sensitive to trypsin, glucosidase and NaIO4. According to the above characteristics it should be a glycoprotein. Its inhibitory effect on virus replication was dependent on the process of cellular RNA and protein syntheses. It could inhibit different kinds of virus in the cells of grass carp, but failed to de so in human, mammalian, avian and shellfish cells. The result indicated the antiviral factor was specific to fish cells and viruses. Its biological characteristics were consistent with those of high vertebrate interferon. Furthermore, it was induced from the leucocytes by phytoagglutinin, sensitive to heat (56°C), pH=2 and 0.1% SDS. Its antigenic characteristic was different from that of interferon-α/β induced by the virus, and its production could be promoted significantly by the phorbolester and interleukin-2. These characteristics of the antiviral factor accorded with those of interferon-gamma. The results revealed that the antiviral factor was an interferon-gamma and there were different types of interferon in fish.
Key words:  Phytoagglutinin, Leucocytes of grass carp, Interferon-Gamma, Induction in vitro
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