LEUKOCYTE DEFECTS ASSOCIATED WITH THERMAL INJURY

热损伤后白细胞的研究已经确定， 吞噬细胞功能出现显著缺陷。 这 观察可以用两种可能性之一来解释： 热损伤，白细胞生长和分化发生在 这样，内在缺陷的发展阻止了一些重要的 机制运行。 或者，与 热损伤影响白细胞功能的继发性 方式 本建议旨在评估这两种可能性 通过使用流式细胞术技术， 深度分析采用热损伤模型，烧伤大鼠。 流式细胞术将允许研究多种白细胞 对微升量（100 μ l）的全血起作用。 因此，在本发明中， 对单个大鼠的动力学研究现已实用。 而 研究这种热损伤模型，一些新的 将应用流式细胞术的应用，即使用 二氯荧光素细胞内H2O2测定，评价 膜流动性作为早期检测缺陷的手段 激活事件和荧光方法用于 同时测量吞噬作用和杀伤作用。 因为 流式细胞术利用白细胞制剂的能力 混合人群，这些研究将在 单核细胞和中性粒细胞吞噬细胞。 在显影之后 和应用所需的技术，并在燃烧后， 更好地理解大鼠模型，热损伤的血液 患者将使用。 这些病人将来自一种特殊的烧伤 该机构的一个部门非常热衷于协助 合作学习 这些将成为重要的研究， 了解免疫反应性 和热损伤后败血症。

Studies of leukocytes after thermal injury have determined that significant defects occur in phagocytic cell function. This observation is explained by one of two possibilities: That after thermal injury, leukocyte growth and differentiation occurs in such a way that intrinsic defects develop preventing some vital mechanism from operating. Alternatively, factors related to the thermal injury affect the leukocyte function in a secondary manner. This proposal aims at evaluating both these possibilities by utilizing flow cytometric techniques which will allow an in- depth analysis using a model of thermal injury, the burned rat. Flow cytometry will allow investigation of multiple leukocyte functions on microliter quantities (100 ul) of whole blood. Thus, kinetic studies on single rats are now practical. While investigating this model of thermal injury, a number of new applications of flow cytometry will be applied, namely the use of the dichlorofluorescein intracellular H2O2 assay, evaluation of membrane fluidity as a means of detecting defects in early activation events and the use of a fluorescence method for simultaneously measuring phagocytosis and killing. Because of the ability of flow cytometry to utilize leukocyte preparations of mixed populations, these investigations will be performed on both the monocyte and neutrophil phagocytic cells. After development and application of the required techniques, and after the burned rat model is better understood, blood from thermally injured patients will be used. These patients will be from a special burn unit at this institution which is very keen to assist in a cooperative study. These will become significant studies in understanding the relationship between immune responsiveness and sepsis after thermal injury.