Hense M, Domann E, Krusch S, Wachholz P, Dittmar K E, Rohde M, Wehland J, Chakraborty T, Weiss S
Department of Cell Biology and Immunology, GBF, German Research Centre for Biotechnology, Mascheroder Weg 1, D-38124 Braunschweig, Germany.
Cell Microbiol. 2001 Sep;3(9):599-609. doi: 10.1046/j.1462-5822.2001.00138.x.
The facultative intracellular, Gram-positive bacterium Listeria monocytogenes invades phagocytic and non-phagocytic cells from the tissues and organs of a wide variety of animals and humans. Here, we report the use of these bacteria as vehicles for gene transfer. Eukaryotic expression plasmids were introduced into the nucleus of host cells following lysis of the intracytosolic, plasmid-carrying bacteria with antibiotics. Cell lines of different tissues and species could be transfected in this way. We examined bacterial properties required for delivery of the expression plasmids and found that this was strictly dependent on the ability of these bacteria to both invade eukaryotic cells and egress from the vacuole into the cytosol of the infected host cells. Macrophage-like cell lines or primary, peritoneal macrophages proved to be almost refractory to Listeria-mediated gene transfer. Thus, attenuated L. monocytogenes represents a serious candidate for consideration as a DNA-transfer vehicle for in vivo somatic gene therapy. The potential for oral administration of L. monocytogenes and the ease in producing and cultivating recombinant strains are further attributes that make its use as a gene transfer vehicle attractive.
兼性胞内革兰氏阳性菌单核细胞增生李斯特菌可侵入多种动物和人类组织及器官中的吞噬细胞和非吞噬细胞。在此，我们报告了利用这些细菌作为基因转移载体的情况。在用抗生素裂解携带质粒的胞内细菌后，真核表达质粒被导入宿主细胞的细胞核。不同组织和物种的细胞系都可以通过这种方式进行转染。我们研究了表达质粒传递所需的细菌特性，发现这严格依赖于这些细菌侵入真核细胞并从液泡进入受感染宿主细胞胞质溶胶的能力。巨噬细胞样细胞系或原代腹膜巨噬细胞对李斯特菌介导的基因转移几乎具有抗性。因此，减毒单核细胞增生李斯特菌是体内体细胞基因治疗中作为DNA转移载体的一个值得认真考虑的候选者。单核细胞增生李斯特菌可口服给药，且易于生产和培养重组菌株，这些进一步特性使其作为基因转移载体具有吸引力。
