Elucidation the mechanism of the immunological changes during anesthesia and abdominal surgery

本研究的主要目的之一是建立新的方法来分析辅助性T淋巴细胞和自然杀伤细胞的细胞因子发育模式。具有挑战性的新方法是：1)利用流式细胞术直接从单核细胞亚群中提取辅助性T淋巴细胞和自然杀伤细胞。2)用ELIS POT法对单个核细胞亚群中发育细胞因子的细胞数量进行计数。3)细胞因子向Th1或Th2分化及发育比例变化的定量分析。结果如下：1)利用LAG-3和cd30双染色的流式细胞术无法获得完全的重现性。2)评价TNFgamma、lL-4等细胞因子发育变化的方法不足以捕捉辅助性T淋巴细胞的微妙变化。3)我们正在开发一种新的方法来评估辅助T淋巴细胞的变化，利用另一种标志物，如tnf - β， IL-5, il - 10，IL-13与cd4。这种方法尚未确定，但我们很快就会成功地发展这种方法。4)采用CD3和TCR双染色的流式细胞术，观察热休克蛋白对细胞凋亡的影响，以评价作为应答细胞和参与细胞凋亡的γ δ - t细胞率的变化。5)认识到上腹部手术过程中外周血γ - t细胞率的变化。

One of the main purpose of this study was to develop the new method to analyze cytokine development pattern of helper T lymphocyte and natural killer cell. The challenging new method was following,1) Extract helper T lymphocyte and natural killer cell from subpopulations of mononuclear cell directly utilizing flow cytometry.2) Count the cell number of developing cytokine from subpopulations of mononuclear cell by ELIS POT method.3) Quantification of the differentiation to Th1 or Th2 and the change of the developmental ratio of the cytokines.The results were following,1 )We could not develop the methods to gain the complete reproducibility with flow cytometry utilizing double stain with LAG-3 and CD3O.2)The method to evaluate the change of cytokine development such as TNFgamma and lL-4 was not enough to catch the delicate change of helper T lymphocuytes.3)We are developing the new method to evaluate the change of helper T lymphocuytes utilizimg another markers such as TNFbeta, IL-5, IL-1O,IL-13 with cd4. The method has not be established yet but we will soon succeed in the develop of the method.4)To evaluate the change of the rates of gammadelta-T cell which is quite important as responding cell and participate to apotosis depended on the development of heat shock protein, we used the flow cytometry by the double stain with CD3 and TCR gamma/delta.5)The change of the rates of gammadelta-T cell in the peripheral blood during upper abdominal surgery was recognize.