Joucher Franck, Mazmanian Guy-Michel, German-Fattal Michele
Laboratory of Thymic Physiology, CNRS UMR 8078, IPSC, Université Paris-Sud, Centre Chirurgical Marie-Lannelongue, Le Plessis Robinson, France.
Transplantation. 2002 Nov 27;74(10):1461-9. doi: 10.1097/00007890-200211270-00020.
The early inflammatory response induced following vascularized organ allotransplantation is mediated by adhesin-ligand interactions between blood leukocytes and allogeneic endothelial cells. In the present study, we focused on the role of allogeneic blood lymphocytes in early immune alterations following lung reperfusion.
We developed an experimental model of isolated and ventilated lung in female C57BL/6 mouse perfused with either isogeneic (C57BL/6) or allogeneic (C3H/He) male mouse fresh blood for 3 hours. SRY DNA quantification by polymerase chain reaction (PCR) was used to assess the presence of perfusate cells in lung extract. Using quantitative reverse transcriptase (RT)-PCR, we measured mRNA expression both of the adhesion molecules--intercellular adhesion molecule (ICAM)-1, lymphocyte function-associated antigen (LFA)-3, vascular cell adhesion molecule (VCAM)-1, and endothelial leukocyte adhesion molecule (ELAM)-1--and of tumor necrosis factor (TNF)-alpha in lung tissue.
The number of blood lymphocytes is significantly decreased in allogeneic versus isogeneic condition at 3 hours of perfusion, without evidence of increased apoptosis or necrosis. In parallel, SRY DNA quantity recovered in the lung was 2.5 times higher in allogeneic condition, which was related to cells loosely adherent to endothelial cells. Lastly, the levels of mRNAs of all adhesion molecules and of TNF-alpha were significantly increased in allogeneic versus isogeneic conditions.
These results demonstrate that an early interaction between allogeneic blood lymphocytes and vascular endothelial cells is correlated with a high mRNA expression both of adhesion molecules and of TNF-alpha in the perfused lung. Our model of a mouse lung perfused with fresh blood appears to be a useful clinical assessment system for in-depth investigation of early cell activation and the resulting intragraft immune alterations.
血管化器官同种异体移植后诱导的早期炎症反应是由血液白细胞与同种异体内皮细胞之间的粘附素-配体相互作用介导的。在本研究中，我们重点关注同种异体血淋巴细胞在肺再灌注后早期免疫改变中的作用。
我们建立了一个实验模型，用同基因（C57BL/6）或异基因（C3H/He）雄性小鼠的新鲜血液灌注雌性C57BL/6小鼠的离体通气肺3小时。通过聚合酶链反应（PCR）对SRY DNA进行定量，以评估肺提取物中灌注细胞的存在情况。使用定量逆转录（RT）-PCR，我们测量了肺组织中粘附分子——细胞间粘附分子（ICAM）-1、淋巴细胞功能相关抗原（LFA）-3、血管细胞粘附分子（VCAM）-1和内皮白细胞粘附分子（ELAM）-1——以及肿瘤坏死因子（TNF）-α的mRNA表达。
在灌注3小时时，异基因条件下的血淋巴细胞数量与同基因条件相比显著减少，且没有凋亡或坏死增加的证据。同时，在异基因条件下肺中回收的SRY DNA量高出2.5倍，这与松散粘附在内皮细胞上的细胞有关。最后，与同基因条件相比，异基因条件下所有粘附分子和TNF-α的mRNA水平均显著升高。
这些结果表明，同种异体血淋巴细胞与血管内皮细胞之间的早期相互作用与灌注肺中粘附分子和TNF-α的高mRNA表达相关。我们用新鲜血液灌注小鼠肺的模型似乎是一个有用的临床评估系统，可用于深入研究早期细胞活化及由此产生的移植物内免疫改变。
