Joucher Franck, Mazmanian Guy-Michel, German-Fattal Michele
CNRS UMR 8078, I.P.S.C., Université Paris XI, Centre Chirurgical Marie-Lannelongue, Le Plessis-Robinson, France.
Transplantation. 2004 Nov 15;78(9):1283-9. doi: 10.1097/01.tp.0000137324.87116.41.
The interaction between host lymphocytes and graft endothelial cells plays an important role in graft rejection.
Using our model of isolated ventilated lung from female mouse perfused with fresh blood from either isogeneic or allogeneic male mouse for 3 hours without noticeable ischemia, we have investigated the kinetics of the early events after endothelial cell triggering by E-selectin engagement.
Isogeneic perfusion induced nonspecific endothelial cell activation, which was characterized by up-regulation of E-selectin, intercellular adhesion molecule (ICAM)-1, and of the pro-inflammatory cytokines tumor necrosis factor (TNF)-alpha, interleukin (IL)-2, and lymphotoxin-alpha (mRNAs by real-time polymerase chain reaction). Allogeneic perfusion was characterized after 3 hours by an additional loose adhesion of lymphocytes mediated by the E-selectin and related to the allogeneic activation of endothelial cells. These in turn expressed the I-A molecule (immunostaining). ICAM-1 and lymphocyte function-associated antigen (LFA)-3 mRNA levels were significantly increased in lung extracts after 2 hours, then vascular cell adhesion molecule (VCAM)-1 and TNF-alpha mRNAs after 3 hours without evidence of TNF-alpha production (enzyme-linked immunoadsorbent assay). The major participation of the E-selectin in early allogeneic activation by way of the protein kinase (PK)C pathway was confirmed by using a neutralizing anti-CD62E monoclonal antibody or the inhibitory PKC 19-31 fragment.
Altogether, our results demonstrate that E-selectin expression (1) is not a consequence of TNF-alpha triggering, (2) up-regulates its own expression and expression of I-A, VCAM-1, TNF-alpha, and lymphotoxin-alpha mRNAs, and (3) down-regulates expression of LFA-3 and ICAM-1 mRNAs. In conclusion, in our physiologic model, the E-selectin highly participates in the loose adhesion of allogeneic lymphocytes and in the early activation of endothelial cell and therefore in structural and functional lung alterations.
宿主淋巴细胞与移植血管内皮细胞之间的相互作用在移植排斥反应中起重要作用。
利用我们的模型，将雌性小鼠分离的通气肺用同基因或异基因雄性小鼠的新鲜血液灌注3小时，无明显缺血，我们研究了E-选择素结合触发内皮细胞后早期事件的动力学。
同基因灌注诱导非特异性内皮细胞活化，其特征在于E-选择素、细胞间粘附分子（ICAM）-1以及促炎细胞因子肿瘤坏死因子（TNF）-α、白细胞介素（IL）-2和淋巴毒素-α（通过实时聚合酶链反应检测mRNA）上调。异基因灌注3小时后的特征是，由E-选择素介导的淋巴细胞额外的松散粘附，这与内皮细胞的异基因活化有关。这些内皮细胞进而表达I-A分子（免疫染色）。2小时后肺提取物中ICAM-1和淋巴细胞功能相关抗原（LFA）-3 mRNA水平显著升高，3小时后血管细胞粘附分子（VCAM）-1和TNF-α mRNA水平显著升高，但无TNF-α产生的证据（酶联免疫吸附测定）。使用中和抗CD62E单克隆抗体或抑制性PKC 19-31片段证实了E-选择素通过蛋白激酶（PK）C途径在早期异基因活化中的主要作用。
总之，我们的结果表明，E-选择素表达（1）不是TNF-α触发的结果，（2）上调其自身表达以及I-A、VCAM-1、TNF-α和淋巴毒素-α mRNA的表达，（3）下调LFA-3和ICAM-1 mRNA的表达。总之，在我们的生理模型中，E-选择素高度参与异基因淋巴细胞的松散粘附以及内皮细胞的早期活化，因此参与肺的结构和功能改变。
