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重组鸡γ干扰素在体外介导对柔嫩艾美耳球虫发育的抑制作用,并在堆型艾美耳球虫攻击感染后减少卵囊产生和体重减轻。

Recombinant chicken interferon-gamma-mediated inhibition of Eimeria tenella development in vitro and reduction of oocyst production and body weight loss following Eimeria acervulina challenge infection.

作者信息

Lillehoj H S, Choi K D

机构信息

USDA-ARS, Livestock and Poultry Sciences Institute, Immunology and Disease Resistance Laboratory, Beltsville, MD 20705-2350, USA.

出版信息

Avian Dis. 1998 Apr-Jun;42(2):307-14.

PMID:9645322
Abstract

Recombinant chicken interferon-gamma (chIFN-gamma) was produced in CHO-K1 or Spodoptera frugiperda (SF9) insect cells by transfection with a pcDNA vector or recombinant baculovirus (SF9-interferon-gamma [IFN-gamma] carrying the chIFN-gamma gene. A rabbit antibody against a synthetic peptide corresponding to an immunogenic portion of chIFN-gamma recognized a 22-23-kDa band in SF9-IFN-gamma cell extracts by western blot analysis. Biological activity of recombinant chIFN-gamma was shown by its inhibition of vesicular stomatitis virus-induced cytotoxicity of chicken embryonic fibroblast cells in vitro. To investigate the role of chIFN-gamma during Eimeria infection, CHCC-OU2 chicken cells either pretreated with chIFN-gamma or stably transfected with the chIFN-gamma gene were infected with Eimeria tenella sporozoites. IFN-gamma demonstrated significant reductions in intracellular sporozoite development without affecting sporozoite invasion of host cells. Furthermore, chickens treated with recombinant chIFN-gamma showed decreased oocyst production and significant improvement in body weight gain following Eimeria acervulina challenge infection. These results provide the first direct evidence that chIFN-gamma exerts an inhibitory effect against Eimeria and provides a rational basis for use of this cytokine as a vaccine adjuvant against coccidiosis.

摘要

通过用pcDNA载体或重组杆状病毒(携带chIFN-γ基因的SF9-干扰素-γ[IFN-γ])转染,在CHO-K1或草地贪夜蛾(SF9)昆虫细胞中生产重组鸡γ干扰素(chIFN-γ)。一种针对与chIFN-γ免疫原性部分相对应的合成肽的兔抗体,通过蛋白质印迹分析在SF9-IFN-γ细胞提取物中识别出一条22-23 kDa的条带。重组chIFN-γ的生物活性通过其在体外抑制水疱性口炎病毒诱导的鸡胚成纤维细胞的细胞毒性得以体现。为了研究chIFN-γ在艾美耳球虫感染过程中的作用,用chIFN-γ预处理或稳定转染chIFN-γ基因的CHCC-OU2鸡细胞被柔嫩艾美耳球虫子孢子感染。IFN-γ显示细胞内子孢子发育显著减少,而不影响子孢子对宿主细胞的侵袭。此外,用重组chIFN-γ处理的鸡在堆型艾美耳球虫攻击感染后,卵囊产量降低,体重增加显著改善。这些结果提供了首个直接证据,表明chIFN-γ对艾美耳球虫发挥抑制作用,并为使用这种细胞因子作为抗球虫病疫苗佐剂提供了合理依据。

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