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.

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

COSENTINO, MARCO;FERRARI, MARCO;MARINO, FRANCA;LECCHINI, SERGIO;FRIGO, GIANMARIO
2000-01-01

Abstract

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.
2000
Cosentino, Marco; Bombelli, R; Ferrari, Marco; Marino, Franca; Rasini, E; Maestroni, Gjm; Conti, A; Boveri, M; Lecchini, Sergio; Frigo, Gianmario
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11383/1707211
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