Adenoviral gene therapy leads to rapid induction of multiple chemokines and acute neutrophil-dependent hepatic injury in vivo
Adenoviral gene therapy leads to rapid induction of multiple chemokines and acute neutrophil-dependent hepatic injury in vivo
复制标题
DOI：
10.1089/10430349950018364
复制
发表时间：
1999-04-10
影响因子：
4.2
通讯作者：
Libermann, TA
中科院分区：
文献类型：
--
作者：
Muruve, DA;Barnes, MJ;Libermann, TA
众所周知，复制缺陷腺病毒会引起感染组织的急性损伤和炎症，因此限制了它们在人类基因治疗中的应用。然而，触发这种反应的分子机制尚未完全确定。为了表征这种反应，在静脉注射各种腺病毒载体后，对DBA/2小鼠的趋化因子表达进行了评估。静脉注射adCMV β - gal、adCMV- gfp或FG140可迅速诱导小鼠肝脏中C-X-C和C-C趋化因子以剂量依赖的方式表达。10(10) PFU adCMV β - gal感染1小时后，肝脏MIP-2 mRNA水平比基线增加60倍。MCP-1和IF-10 mRNA水平在感染各种腺病毒载体后也立即升高，在6小时达到峰值，>分别表达25倍和>100倍。腺病毒载体也可以早期诱导RANTES和MIP-1 β mRNA，但诱导程度较低。趋化因子的诱导与病毒基因表达无关，因为补骨脂素失活的腺病毒颗粒在给药后的前16小时内产生了相同的趋化因子基因转录模式。趋化因子的表达与中性粒细胞和CD11b（+）细胞流入感染动物肝脏的预期相关。在高滴度下，所有腺病毒载体在系统给药DBA/2小鼠后引起明显的肝坏死和凋亡。为了研究中性粒细胞在腺病毒诱导的肝损伤中的作用，用中和的抗mip -2抗体或去除中性粒细胞对动物进行预处理。MIP-2拮抗和中性粒细胞耗竭均导致血清ALT/AST水平降低和腺病毒诱导的肝损伤的组织学减弱，证实这种早期损伤主要是由于趋化因子的产生和中性粒细胞的募集。我们的研究结果进一步阐明了针对复制缺陷腺病毒载体的早期免疫反应，并提出了一种通过干扰趋化因子或中性粒细胞功能来预防腺病毒介导的炎症和组织损伤的策略。
Replication-deficient adenoviruses are known to induce acute injury and inflammation of infected tissues, thus limiting their use for human gene therapy. However, molecular mechanisms triggering this response have not been fully defined. To characterize this response, chemokine expression was evaluated in DBA/2 mice following the intravenous administration of various adenoviral vectors. Administration of adCMV beta gal, adCMV-GFP, or FG140 intravenously rapidly induced a consistent pattern of C-X-C and C-C chemokine expression in mouse liver in a dose-dependent fashion. One hour following infection with 10(10) PFU of adCMV beta gal, hepatic levels of MIP-2 mRNA were increased >60-fold over baseline. MCP-1 and IF-10 mRNA levels were also increased immediately following infection with various adenoviral vectors, peaking at 6 hr with >25- and >100-fold expression, respectively. Early induction of RANTES and MIP-1 beta mRNA by adenoviral vectors also occurred, but to a lesser degree. The induction of chemokines occurred independently of viral gene expression since psoralen-inactivated adenoviral particles produced an identical pattern of chemokine gene transcription within the first 16 hr of administration. The expression of chemokines correlated as expected with the influx of neutrophils and CD11b(+) cells into the livers of infected animals. At high titers, all adenoviral vectors caused significant hepatic necrosis and apoptosis following systemic administration to DBA/2 mice. To investigate the role of neutrophils in this adenovirus-induced hepatic injury, animals were pretreated with neutralizing anti-MIP-2 antibodies or depleted of neutrophils. MIP-2 antagonism and neutrophil depletion both resulted in reduced serum ALT/AST levels and attenuation of the adenovirus-induced hepatic injury histologically, confirming that this early injury is largely due to chemokine production and neutrophil recruitment. Our findings further clarify the early immune response against replication-deficient adenoviral vectors and suggest a strategy to prevent adenovirus-mediated inflammation and tissue injury by interfering with chemokine or neutrophil function.