Raz E, Carson D A, Parker S E, Parr T B, Abai A M, Aichinger G, Gromkowski S H, Singh M, Lew D, Yankauckas M A
Department of Medicine, University of California, San Diego, La Jolla 92093-0663.
Proc Natl Acad Sci U S A. 1994 Sep 27;91(20):9519-23. doi: 10.1073/pnas.91.20.9519.
The skin and mucous membranes are the anatomical sites were most viruses are first encountered by the immune system. Previous experiments have suggested that striated muscle cells are unique among mammalian cell types in their capacity to take up and express free DNA in the absence of a viral vector or physical carrier. However, we have found that mice injected into the superficial skin with free (naked) plasmid DNA encoding the influenza nucleoprotein gene had discrete foci of epidermal and dermal cells, including cells with dendritic morphology, that contained immunoreactive nucleoprotein antigen. A single intradermal administration of 0.3-15 micrograms of free plasmid DNA induced anti-nucleoprotein-specific antibody and cytotoxic T lymphocytes that persisted for at least 68-70 weeks after vaccination. Intradermal gene administration induced higher antibody titers than did direct gene injection into skeletal muscle and did not cause local inflammation or necrosis. Compared with control animals, the gene-injected mice were resistant to challenge with a heterologous strain of influenza virus. These results indicate that the cells of the skin can take up and express free foreign DNA and induce cellular and humoral immune responses against the encoded protein. We suggest that DNA uptake by the skin-associated lymphoid tissues may play a role in the induction of cytotoxic T cells against viruses and other intracellular pathogens.
皮肤和黏膜是免疫系统最先接触大多数病毒的解剖部位。先前的实验表明,横纹肌细胞在哺乳动物细胞类型中独一无二,它能够在没有病毒载体或物理载体的情况下摄取并表达游离DNA。然而,我们发现,向浅表皮肤注射编码流感核蛋白基因的游离(裸露)质粒DNA的小鼠,其表皮和真皮细胞出现离散病灶,包括具有树突形态的细胞,这些细胞含有免疫反应性核蛋白抗原。单次皮内注射0.3 - 15微克游离质粒DNA可诱导产生抗核蛋白特异性抗体和细胞毒性T淋巴细胞,疫苗接种后至少持续68 - 70周。皮内基因给药诱导产生的抗体滴度高于直接向骨骼肌注射基因,且不会引起局部炎症或坏死。与对照动物相比,基因注射小鼠对异源流感病毒株的攻击具有抵抗力。这些结果表明,皮肤细胞能够摄取并表达游离的外源DNA,并诱导针对编码蛋白的细胞免疫和体液免疫反应。我们认为,皮肤相关淋巴组织摄取DNA可能在诱导针对病毒和其他细胞内病原体的细胞毒性T细胞中发挥作用。