Beta defensin-3 engineered epidermis shows highly protective effect for bacterial infection
Beta defensin-3 engineered epidermis shows highly protective effect for bacterial infection
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DOI：
10.1038/sj.gt.3302472
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发表时间：
2005-05-01
期刊：
影响因子：
5.1
通讯作者：
Shimizu, H
中科院分区：
文献类型：
--
作者：
Sawamura, D;Goto, M;Shimizu, H
防御素是一种小分子阳离子蛋白质，具有广谱的杀菌活性，可抵抗细菌、真菌和病毒。本研究检测了表皮工程化表达人类β-防御素3（HBD 3）以对抗细菌感染对病原体的影响。首先，我们研究了HBD 3在表皮中的定位，并观察到HBD 3在上表皮层的细胞间隙和板层体中。这一结果表明，HBD 3在表皮外层表达和组装，被怀疑是对抗入侵微生物的。接下来，我们建立了稳定表达HBD 3的角质形成细胞系，并发现培养基显示出抗菌活性。此外，我们制备了这些细胞与HBD 3基因的表皮片，并将其移植到裸大鼠的皮肤伤口上。HBD 3工程表皮显示出显著的抗微生物活性。没有表皮的皮肤溃疡经常暴露于入侵的微生物。收集皮肤溃疡患者表皮再上皮化的活检样品，并测量表皮中HBD 3 mRNA水平。来自溃疡皮肤的表皮样品表达的HBD 3转录水平比对照皮肤中的HBD 3转录水平高2.5倍。这些结果表明，HBD 3基因治疗性导入体细胞可能为难治性感染性疾病提供一种新的基因治疗策略。
Defensins are small cationic proteins that harbor broad-spectrum microbicidal activity against bacteria, fungi and viruses. This study examines the effects on pathogens of the epidermis engineered to express human beta-defensin 3 (HBD3) to combat bacterial infections. First, we examined the localization of HBD3 in the epidermis and observed HBD3 in the intercellular spaces and lamellar bodies of the upper epidermal layers. This result showed HBD3 expressed and assembled in the outer layers of the epidermis was suspected to counter the invading microorganisms. Next, we established a keratinocyte cell line that stably expressed HBD3 and found that the culture medium showed antibacterial activity. Furthermore, we prepared an epidermal sheet of these cells with the HBD3 gene and grafted this onto a dermal wound on a nude rat. The HBD3 engineered epidermis demonstrated significant antimicrobial activity. Skin ulcers without epidermis are constantly exposed to invading microorganisms. Biopsy samples of re-epithelizing epidermis from patients with skin ulcers were collected, and HBD3 mRNA level measured in the epidermis. The epidermal samples from the ulcer skin expressed 2.5 times higher levels of HBD3 transcript than those in the control skin. These results, taken together, indicate that the therapeutic introduction of the HBD3 gene into somatic cells may provide a new gene therapy strategy for intractable infectious diseases.