Li Jun, Zheng Chanyu, Zhang Xin, Liu Xiaoyong, Zhang Chengmei, Goldsmith Corinne M, Baum Bruce J, Wang Songlin
Salivary Gland Disease Center and the Molecular Laboratory for Gene Therapy, Faculty of Stomatology, Capital University of Medical Sciences, Beijing, P. R. China.
J Gene Med. 2004 Jan;6(1):55-63. doi: 10.1002/jgm.476.
Localized gene transfer to salivary glands has great potential for the treatment of salivary gland, systemic, and oral diseases. The minipig parotid gland, given its volume and morphological similarities to the human parotid gland, may be useful as a large animal model for pre-clinical gene transfer experiments. The purpose of this study was to perform an initial assessment of the efficacy and safety of adenoviral-vector-mediated gene transfer to parotid glands of miniature pigs.
AdCMVluc, a recombinant type 5 adenoviral (rAd5) vector containing a luciferase reporter gene, was administered to miniature pig parotid glands by intraductal cannulation. Five regions of gland tissue were obtained to measure the distribution of luciferase activity. The effects of time, viral dose, infusate buffer volume, and gland anatomical region on transgene expression were determined. Detailed serum chemistry and hematological analyses were performed. In addition, AdCMVlacZ, a similar rAd5 vector encoding beta-galactosidase, was also delivered to determine the parotid gland cell types transduced.
Luciferase assays indicated that gene transfer to miniature pig salivary glands could be readily accomplished using rAd5 vectors. Highest transgene expression was found in the center of glands, which was > posterior > inferior > anterior > superior tissue regions. Expression was maximal on day 2 and declined to background by day 14, and observed in both acinar and ductal cells. Several serum chemistry and hematology parameters were transiently changed following rAd5 administration.
Transgene expression by, and inflammatory response to, rAd5 vectors in minipig parotid glands are similar to results seen earlier in rodent studies. This suggests that results of salivary gland gene transfer from rodent studies can be extended to a larger animal model, and supports the value of using minipigs for pre-clinical applications of gene transfer to these tissues. Published in 2004 by John Wiley & Sons, Ltd.
向唾液腺进行局部基因转移在治疗唾液腺疾病、全身性疾病和口腔疾病方面具有巨大潜力。小型猪的腮腺因其体积和形态与人类腮腺相似，可能作为临床前基因转移实验的大型动物模型。本研究的目的是对腺病毒载体介导的基因转移至小型猪腮腺的有效性和安全性进行初步评估。
通过导管插管将AdCMVluc（一种含有荧光素酶报告基因的重组5型腺病毒（rAd5）载体）注入小型猪腮腺。获取五个区域的腺组织以测量荧光素酶活性的分布。确定时间、病毒剂量、灌注缓冲液体积和腺体解剖区域对转基因表达的影响。进行详细的血清化学和血液学分析。此外，还递送了AdCMVlacZ（一种编码β-半乳糖苷酶的类似rAd5载体）以确定转导的腮腺细胞类型。
荧光素酶测定表明，使用rAd5载体可轻松实现向小型猪唾液腺的基因转移。在腺体中心发现最高的转基因表达，其顺序为中心>后部>下部>前部>上部组织区域。表达在第2天达到最大值，并在第14天降至背景水平，在腺泡细胞和导管细胞中均有观察到。给予rAd5后，一些血清化学和血液学参数发生短暂变化。
rAd5载体在小型猪腮腺中的转基因表达和炎症反应与早期在啮齿动物研究中观察到的结果相似。这表明啮齿动物唾液腺基因转移的结果可以扩展到更大的动物模型，并支持使用小型猪进行这些组织基因转移临床前应用的价值。2004年由John Wiley & Sons, Ltd.出版
