Adenovirus-mediated gene transfer in the transplant setting - Early events after orthotopic transplantation of liver allografts expressing TGF-beta 1
Adenovirus-mediated gene transfer in the transplant setting - Early events after orthotopic transplantation of liver allografts expressing TGF-beta 1
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DOI：
10.1097/00007890-199610270-00010
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发表时间：
1996-10-27
期刊：
影响因子：
6.2
通讯作者：
Shaked, A
中科院分区：
文献类型：
--
作者：
Drazan, KE;Olthoff, KM;Shaked, A
我们假设腺病毒介导的TGF-β 1基因转移到肝移植物中会增强这种重组蛋白的局部表达，并下调炎症和同种异体免疫反应。使用编码鼠TGF-β 1的全长DNA替换5型腺病毒（AdmTGF-β 1）的E1区。在AdmTGF-β 1转导的Hep G2细胞和TGF-β敏感的MV 1细胞中检测了生物活性鼠TGF-β 1的表达和蛋白质产生。在移植环境中，复制缺陷型载体用于灌注冷保存的ACI肝同种异体移植物，然后移植到刘易斯受体中。对照组肝脏同样灌注冷乳酸林格氏液，随后无免疫抑制。在移植后1、3和5天处死动物。采用ELISA和定量PCR测定移植物内TNF α和IFN γ的细胞因子水平。AdmTGF-β 1转导的肝细胞系Hep G2的培养物上清液的TGF-β 1 ELISA分泌的TGF-β 1的量与感染复数（MOI，载体：肝细胞比）直接相关。通过MV 1 TGF-β敏感细胞的生长抑制证实了分泌的重组蛋白的生物活性。与非免疫抑制对照组相比，转导同种异体移植物中TGF-β 1的产生增加与TNF α和IFN γ水平降低相关。腺病毒介导的鼠TGF-β 1基因转移到肝细胞中导致生物活性蛋白的表达。同种异体移植物转导TGF-β 1下调原位移植后早期TNF α和IFN γ的产生TGF-β 1的移植物转导为研究单一免疫调节剂对同种异体反应性免疫应答、T细胞功能和细胞因子级联反应的影响提供了一种新的方法。
We hypothesized that adenovirus mediated gene transfer of TGF-beta 1 into liver grafts would enhanced local expression of this recombinant protein and down-regulate inflammatory and alloreactive immune response. A full length DNA encoding the murine TGF-beta 1 was used to replaced the E1 region of adenovirus type 5 (AdmTGF-beta 1). Expression and protein production of biologically active murine TGF-beta 1 was tested in AdmTGF-beta 1-transduced Hep G2 cells and TGF-beta-sensitive MV1 cells. In the transplant setting, the replication-defective vector was used to perfused cold preserved ACI liver allograft prior to transplantation into Lewis recipients. Control livers were similarly perfused with cold lactated Ringer's solution and were followed without immunosuppression. Animals were sacrificed at 1, 3, and 5 days after transplantation. Intragraft cytokine levels of TNF alpha, and IFN gamma were determined using ELISA and quantitative PCR. TGF-beta 1 ELISA of culture supernatants from AdmTGF-beta 1 transduced hepatocyte cell line Hep G2 excreted TGF-beta 1 in quantities directly correlated with multiplicity of infection (MOI, vector:hepatic cell ratio). The biological activity of the excreted recombinant protein was confirmed by growth inhibition of MV1 TGF-beta-sensitive cells. Enhanced production of TGF-beta 1 in transduced allografts was associated with decreased levels of TNF alpha and IFN gamma when compared with nonimmunosuppressed controls. Adenovirus-mediated gene transfer of murine TGF-beta 1 into hepatic cells results in the expression of biologically active protein. Transduction of allografts with TGF-beta 1 down-regulates TNF alpha and IFN gamma production early after orthotopic transplantation. Graft transduction with TGF-beta 1 offers a novel approach to study the effects of single immune modulator on alloreactive immune response, T cell function, and cytokine cascade.