Wu H L, Chen P J, Lin H K, Lee R S, Lin H L, Liu C J, Lee P J, Lee J J, Chen D S
Hepatitis Research Center, National Taiwan University Hospital, 7 Chung-Shan South Road, Taipei, Taiwan.
J Med Virol. 2001 Nov;65(3):567-75.
Granulocyte-macrophage colony stimulating factor (GM-CSF) has immunoregulatory and antiviral effects, and may thus be promising for the treatment of chronic hepatitis B. Using woodchuck hepatitis virus (WHV)-infected woodchuck as an animal model to test the efficacy and safety of GM-CSF on the therapy of chronic hepatitis B, woodchuck GM-CSF will be required due to the apparent species-specific activity of GM-CSF. The cDNA of woodchuck GM-CSF was cloned using reverse transcription-polymerase chain reaction (RT-PCR) with primers deriving from highly conserved regions of GM-CSF genes from other species. The deduced amino acids, including the signal peptide, is 138 in length and its identities to human, murine, canine and bovine GM-CSFs are 63, 49, 63, and 63% respectively. The genomic DNA of woodchuck GM-CSF was also cloned by PCR. Its organization is highly homologous to that of human and murine GM-CSF genes, consisting of four exons and three introns. Cloned woodchuck GM-CSF was expressed transiently in 293T cells. The recombinant protein expressed was found to stimulate the growth and differentiation of woodchuck bone marrow cells, indicating the protein expressed by the cloned gene is functional. These results pave the way for future studies on the potential role of GM-CSF for the treatment of chronic hepatitis B by using this animal model.
粒细胞巨噬细胞集落刺激因子（GM-CSF）具有免疫调节和抗病毒作用，因此可能有望用于治疗慢性乙型肝炎。由于GM-CSF明显的种属特异性活性，需使用感染土拨鼠肝炎病毒（WHV）的土拨鼠作为动物模型来测试GM-CSF治疗慢性乙型肝炎的疗效和安全性。利用逆转录聚合酶链反应（RT-PCR），以源自其他物种GM-CSF基因高度保守区域的引物克隆了土拨鼠GM-CSF的cDNA。推导的氨基酸（包括信号肽）长度为138个，其与人类、小鼠、犬和牛GM-CSF的同一性分别为63%、49%、63%和63%。还通过PCR克隆了土拨鼠GM-CSF的基因组DNA。其结构与人类和小鼠GM-CSF基因高度同源，由四个外显子和三个内含子组成。克隆的土拨鼠GM-CSF在293T细胞中瞬时表达。发现表达的重组蛋白可刺激土拨鼠骨髓细胞的生长和分化，表明克隆基因表达的蛋白具有功能。这些结果为未来利用该动物模型研究GM-CSF在治疗慢性乙型肝炎中的潜在作用铺平了道路。
