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HPLC-ED measurement of endogenous catecholamines in human immune cells and hematopoietic cell lines

机译：HPLC-ED测定人体免疫细胞和造血细胞系中内源性儿茶酚胺

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A rapid and simple HPLC-ED method is described to identify and measure catecholamines (CTs) and their major metabolites in immune cells. Using this method, intracellular CTs were quantified in human peripheral blood mononuclear cells (PBMCs), T and B lymphocytes, monocytes and granulocytes. Immune cell subsets were separated by density gradient centrifugation and immunomagnetic cell sorting. CTs were also found in the human hematopoietic cell lines NALM-6 (pre-B) and (in smaller amounts) in Jurkat (T lymphoblastoid) and U937 (promonocytic). In cultured PBMCs, intracellular CTs were reduced by both the tyrosine hydroxylase inhibitor alpha -methyl-p-tyrosine and the chromaffin granule depletant reserpine. In NALM-6 cells, both alpha -methyl-p-tyrosine and the dopamine-beta-hydroxylase inhibitor disulfiram reduced intracellular CTs, supporting the presence of active synthetic pathways in these cells. Since sympathoadrenergic mechanisms play a key role in the interactions between the immune system and the nervous system, these findings may be relevant for a better understanding of the neuro-immune network. (C) 2000 Elsevier Science Inc. All rights reserved. [References: 25]
机译：描述了一种快速简单的HPLC-ED方法来鉴定和测量免疫细胞中的儿茶酚胺（CT）及其主要代谢产物。使用这种方法，可以对人外周血单个核细胞（PBMC），T和B淋巴细胞，单核细胞和粒细胞中的细胞内CT进行定量。通过密度梯度离心和免疫磁性细胞分选来分离免疫细胞亚群。在人类造血细胞系NALM-6（前B型）和Jurkat（T淋巴母细胞）和U937（原单核细胞）中也发现了CT。在培养的PBMC中，酪氨酸羟化酶抑制剂α-甲基-p-酪氨酸和嗜铬粒蛋白颗粒利血平均降低了细胞内CT。在NALM-6细胞中，α-甲基-对-酪氨酸和多巴胺-β-羟化酶抑制剂双硫仑都降低了细胞内CT，从而支持了这些细胞中活性合成途径的存在。由于交感神经功能机制在免疫系统和神经系统之间的相互作用中起关键作用，因此这些发现可能与更好地理解神经免疫网络有关。 （C）2000 Elsevier Science Inc.保留所有权利。 [参考：25]

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