Viral IL-10 gene therapy inhibits TNF-alpha and IL-1 beta, not IL-6, in the newborn endotoxemic mouse
Viral IL-10 gene therapy inhibits TNF-alpha and IL-1 beta, not IL-6, in the newborn endotoxemic mouse
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DOI：
10.1016/s0022-3468(96)90749-6
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发表时间：
1996-03-01
影响因子：
2.4
通讯作者：
Shaked, A
中科院分区：
文献类型：
--
作者：
Drazan, KE;Wu, L;Shaked, A
引言：重症婴儿肝脏内炎症级联反应的调节可能会提高生存机会。使用基因治疗，作者假设，增强的反调节细胞因子病毒白细胞介素-10（IL-10）在体内的局部生产将调节炎症反应中的关键细胞因子。本研究的目的是确定是否复制缺陷型腺病毒介导的病毒IL-10（vIL-10）基因转移和表达的肝脏内可以实现这一目标，在新生小鼠。材料与方法：给4周龄Balb/c小鼠腹腔注射每毫升1 × 109空斑形成单位（pfu）的腺病毒载体（E1 a/b缺失），该载体编码sv 40启动子和BCRF 1 cDNA，或对照载体dl 434不表达外源基因。48小时后，用50 μ g/kg的脂多糖（LPS）攻击小鼠; 1、2、6或24小时后将它们杀死（每个时间点6只）。对从肝脏、肺和肾脏中分离的基因组DNA进行Southern印迹分析，以评估BCRF 1的基因转移。分析匀浆肝蛋白中的肿瘤坏死因子-α（TNF-α）、IL-1 β、IL-6和重组vIL-10。结果如下：Southern印迹分析证实，在接受腺病毒载体的小鼠中，成功地将基因转移到肝脏，但不转移到肺、肾或dl 434转导的肝脏。肝脏内的病毒IL-10水平范围为14 - 18 ng/mL。在对照组中，TNF-α的产生在早期时间点升高至18，000 pg/mL，但在LPS攻击后24小时迅速下降。在整个研究过程中，用Ad 5svBCRF 1治疗的动物的TNF-α水平显著低于对照组（P
Introduction: Modulation of the inflammatory cascade within the liver of critically ill infants may improve the chance of survival. Using gene therapy, the authors hypothesized that augmented local production of the counter-regulatory cytokine viral interleukin-10 (IL-10) in vivo will modulate the critical cytokines in the inflammatory response. The purpose of the present study was to determine whether replication-defective adenovirus-mediated viral IL-10 (vIL-10) gene transfer and expression within the liver can achieve this goal in newborn mice. Materials and Methods: Four-week-old Balb/c mice were administered (intraperitoneally) 1 x 10(9) plaque-forming units (pfu) per milliliter of an adenovirus vector (E1a/b-deleted) that encodes the sv40 promoter and the BCRF1 cDNA, or of control vector dl434 that expresses no foreign gene. Forty-eight hours later the mice were challenged with 50 mu g/kg of lipopolysaccharide; (LPS) they were killed 1, 2, 6, or 24 hours later (six at each time point). Southern blot analysis was performed on genomic DNA isolated from the liver, lung, and kidney to assess gene transfer of BCRF1. Homogenized liver protein was analyzed for tumor necrosis factor-alpha (TNF-alpha), IL-1 beta, IL-6, and recombinant vIL-10. Results: Southern blot analysis confirmed successful gene transfer to the liver but not to the lung, kidney, or dl434-transduced liver in mice that received adenovectors. Viral IL-10 levels within the liver ranged from 14 to 18 ng/mL. In controls, TNF-alpha production was elevated at early time points, to 18,000 pg/mL, but decreased rapidly by 24 hours after LPS challenge. The TNF-alpha levels of animals treated with Ad5svBCRF1 were significantly lower than those of controls throughout the course of study (P