Reed W A, Elzer P H, Enright F M, Jaynes J M, Morrey J D, White K L
Department of Animal, Dairy, and Veterinary Sciences, Logan, Utah State University 84322-4700, USA.
Transgenic Res. 1997 Sep;6(5):337-47. doi: 10.1023/a:1018423015014.
The addition of an antimicrobial that can be synthesized by the mammalian immune system at the point of challenge may enhance disease resistance. A possible group of agents are cecropins, broad-spectrum antimicrobial peptides, which have been described and characterized. They are relatively non-toxic to normal cells from multicellular organisms but are toxic to a wide range of bacteria, protozoa and fungi, as well as infected and abnormal cells. Twenty-six lines of transgenic mice were produced by pronuclear injection of DNA consisting of the 5'-flanking region from -593 to +110 of the mouse interleukin 2 (IL-2) gene, Shiva 1a (a synthetic cecropinclass lytic peptide), and the SV40 polyadenylation/splice signal. A reverse-transcription PCR assay determined that two lines of transgenic mice were produced whose spleen-derived lymphocytes could be induced to transcribe and mature mRNA for Shiva 1a by exposure to 3.25 mg ml-1 of Con A. Two lines were challenged with an inoculation of 5 x 10(4) Brucella abortus strain 2308. After four weeks, there were significantly fewer B. abortus organisms in the spleens of transgenic mice than in non-transgenic control mice of the same strain (p < 0.05). Since the controlling regions of the IL-2 enhancer and the amino acid sequence of the signal peptide are highly conserved among several species, it is likely that this recombinant gene will function in other mammals.
在受到感染时添加一种可由哺乳动物免疫系统合成的抗菌物质,可能会增强抗病能力。一类可能的物质是杀菌肽,即已被描述和鉴定的广谱抗菌肽。它们对多细胞生物的正常细胞相对无毒,但对多种细菌、原生动物和真菌以及受感染和异常的细胞有毒性。通过原核注射由小鼠白细胞介素2(IL-2)基因从-593至+110的5'侧翼区域、Shiva 1a(一种合成的杀菌肽类裂解肽)以及SV40聚腺苷酸化/剪接信号组成的DNA,产生了26株转基因小鼠。逆转录聚合酶链反应测定表明,产生了两株转基因小鼠,其脾脏来源的淋巴细胞在暴露于3.25 mg/ml的刀豆球蛋白A时可被诱导转录并成熟Shiva 1a的信使核糖核酸。给两株小鼠接种5×10⁴流产布鲁氏菌2308菌株进行攻毒。四周后,转基因小鼠脾脏中的流产布鲁氏菌数量明显少于同一品系的非转基因对照小鼠(p<0.05)。由于IL-2增强子的调控区域和信号肽的氨基酸序列在几个物种中高度保守,这种重组基因很可能在其他哺乳动物中发挥作用。