Analysis of leukocyte binding to depletion filters: role of passive binding, interaction with platelets, and plasma components
Analysis of leukocyte binding to depletion filters: role of passive binding, interaction with platelets, and plasma components
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白细胞与消耗过滤器结合的分析：被动结合的作用、与血小板的相互作用以及血浆成分
DOI：
10.1007/s00277-004-0994-0
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发表时间：
2005
影响因子：
3.5
通讯作者：
E. Seifried
中科院分区：
文献类型：
--
作者：
R. Henschler;B. Rüster;A. Steimle;H. L. Hansmann;W. Walker;T. Montag;E. Seifried
由于对调节细胞与白细胞去除过滤器表面结合的机制了解有限，我们研究了白细胞与白细胞去除过滤器各层的结合模式。在通过1单位全血后，在载玻片上对分离的过滤层进行印迹或从过滤层洗脱细胞显示，大多数白细胞位于总共28个过滤层的前10个内，在第6至8层达到峰值，粒细胞平均比淋巴细胞更早结合到过滤层。与肌动蛋白激活抑制剂预孵育的白细胞显示不变的分布之间的过滤层，这表明细胞骨架激活并不显着有助于其结合。当白细胞直接与单个过滤层孵育时，在不存在Ca2+的情况下记录了高达30%的输入细胞的结合。免疫组织学分析显示血小板和白细胞共定位，血小板和白细胞共聚集。单核细胞和某种程度上的淋巴细胞（但不是粒细胞）与血小板竞争滤器结合。用单个血浆成分预涂过滤层表明，与白蛋白涂层相比，透明质酸、血浆型纤连蛋白和纤维蛋白原均增加了白细胞的结合。总之，白细胞可以被动结合过滤器的过程中，不需要Ca2+，这是独立的细胞骨架激活，这可能取决于个别血浆成分。当设想通过特定过滤器的新的选择性细胞富集或耗尽策略时，这些结果是重要的。
Since limited knowledge exists on the mechanisms which regulate cell binding to leukocyte removal filter surfaces, we investigated the binding patterns of leukocytes to individual layers of leukocyte depletion filters. After passage of 1 unit of whole blood, blotting of isolated filter layers on glass slides or elution of cells from filter layers revealed that most leukocytes were located within the first 10 of a total of 28 filter layers, peaking at layers 6 to 8, with granulocytes binding on average to earlier filter layers than lymphocytes. Leukocytes preincubated with inhibitors of actin activation showed unchanged distribution between filter layers, suggesting that cytoskeletal activation does not significantly contribute to their binding. When leukocytes were directly incubated with single filter layers, binding of up to 30% of input cells was recorded in the absence of Ca2+. Immunohistological analyses showed colocalization of platelets and leukocytes, with co-clustering of platelets and leukocytes. Monocytes and to some degree lymphocytes but not granulocytes competed with platelets for filter binding. Precoating of filter layers with individual plasma components showed that hyaluronic acid, plasma type fibronectin, and fibrinogen all increased the binding of leukocytes compared with albumin coating. In conclusion, leukocytes can bind passively to filters in a process which does not require Ca2+, which is independent of cytoskeletal activation and which may depend on individual plasma components. These results are of importance when new selective cell enrichment or depletion strategies through specific filters are envisaged.