Disappearance of GFP-positive hepatocytes transplanted into the liver of syngeneic wild-type rats pretreated with retrorsine.
Disappearance of GFP-positive hepatocytes transplanted into the liver of syngeneic wild-type rats pretreated with retrorsine.
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DOI：
10.1371/journal.pone.0095880
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Sun Z
中科院分区：
文献类型：
--
作者：
Maeda H;Shigoka M;Wang Y;Fu Y;Wesson RN;Lin Q;Montgomery RA;Enzan H;Sun Z
绿色荧光蛋白（GFP）是一种广泛使用的分子标签，用于在啮齿动物肝损伤模型中追踪移植细胞。先前报道的各种使用 GFP 的研究的不同结果可能归因于 GFP 的免疫原性。肝细胞取自表达 GFP 的转基因 (Tg) Lewis 大鼠，并在野生型 Lewis 大鼠接受部分肝切除后移植到其肝脏中。逆转录酶预处理可抑制受体大鼠内源性肝细胞的增殖，从而增强移植肝细胞的增殖。还将野生型肝细胞移植到 GFP-Tg 大鼠肝脏中进行比较。移植后7天采集的所有活检标本均显示移植肝细胞定植，直到第14天移植肝细胞数量不断增加。野生型大鼠肝脏中的GFP阳性肝细胞在第28天减少，到第42天检测不到，而GFP-Tg大鼠肝脏中野生型肝细胞数量稳定增加。组织学检查显示第28天GFP阳性肝细胞发生退行性改变以及浸润细胞的积累。GFP转基因的PCR分析表明移植的肝细胞被消除而不是随着GFP表达的丧失而被保留。使用他克莫司和骨髓移植改变免疫反应均延长了 GFP 阳性肝细胞的存活。相反，用 GFP 阳性肝细胞进行宿主免疫导致 GFP 阳性肝细胞在第 14 天完全丧失。从 GFP-Tg Lewis 大鼠分离的 GFP 阳性肝细胞在经逆转录碱预处理的野生型 Lewis 大鼠的肝脏中不能长期存活。这种现象的机制很可能涉及针对 GFP 的免疫反应。在计划使用 GFP 的体内实验并解释其结果时，应考虑 GFP 免疫原性对细胞移植模型的影响。
Green fluorescent protein (GFP) is a widely used molecular tag to trace transplanted cells in rodent liver injury models. The differing results from various previously reported studies using GFP could be attributed to the immunogenicity of GFP. Hepatocytes were obtained from GFP-expressing transgenic (Tg) Lewis rats and were transplanted into the livers of wild-type Lewis rats after they had undergone a partial hepatectomy. The proliferation of endogenous hepatocytes in recipient rats was inhibited by pretreatment with retrorsine to enhance the proliferation of the transplanted hepatocytes. Transplantation of wild-type hepatocytes into GFP-Tg rat liver was also performed for comparison. All biopsy specimens taken seven days after transplantation showed engraftment of transplanted hepatocytes, with the numbers of transplanted hepatocytes increasing until day 14. GFP-positive hepatocytes in wild-type rat livers were decreased by day 28 and could not be detected on day 42, whereas the number of wild-type hepatocytes steadily increased in GFP-Tg rat liver. Histological examination showed degenerative change of GFP-positive hepatocytes and the accumulation of infiltrating cells on day 28. PCR analysis for the GFP transgene suggested that transplanted hepatocytes were eliminated rather than being retained along with the loss of GFP expression. Both modification of the immunological response using tacrolimus and bone marrow transplantation prolonged the survival of GFP-positive hepatocytes. In contrast, host immunization with GFP-positive hepatocytes led to complete loss of GFP-positive hepatocytes by day 14. GFP-positive hepatocytes isolated from GFP-Tg Lewis rats did not survive long term in the livers of retrorsine-pretreated wild-type Lewis rats. The mechanism underlying this phenomenon most likely involves an immunological reaction against GFP. The influence of GFP immunogenicity on cell transplantation models should be considered in planning in vivo experiments using GFP and in interpreting their results.
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