HPLC-ED measurement of endogenous catecholamines in human immune cells and hematopoietic cell lines
HPLC-ED measurement of endogenous catecholamines in human immune cells and hematopoietic cell lines
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DOI：
10.1016/s0024-3205(00)00937-1
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发表时间：
2000-12-08
期刊：
影响因子：
6.1
通讯作者：
Frigo, G
中科院分区：
文献类型：
--
作者：
Cosentino, M;Bombelli, R;Frigo, G
描述了一种快速、简单的 HPLC-ED 方法来识别和测量儿茶酚胺 (CT) 及其在免疫细胞中的主要代谢物。使用这种方法，对人外周血单核细胞 (PBMC)、T 和 B 淋巴细胞、单核细胞和粒细胞的细胞内 CT 进行定量。通过密度梯度离心和免疫磁性细胞分选分离免疫细胞亚群。在人类造血细胞系 NALM-6（前 B）和（少量）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-酪氨酸和嗜铬颗粒消耗剂利血平均降低了细胞内 CT。在 NALM-6 细胞中，α-甲基-p-酪氨酸和多巴胺-β-羟化酶抑制剂双硫仑均可降低细胞内 CT，支持这些细胞中存在活性合成途径。由于交感肾上腺素能机制在免疫系统和神经系统之间的相互作用中发挥着关键作用，这些发现可能有助于更好地理解神经免疫网络。 (C) 2000 Elsevier Science Inc. 保留所有权利。
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.