HPLC-ED measurement of endogenous catecholamines in human immune cells and hematopoietic cell lines.
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.
Cosentino
M
M
Department of Internal Medicine and Therapeutics, University of Insubria, Varese, Italy.
Bombelli
R
R
Ferrari
M
M
Marino
F
F
Rasini
E
E
Maestroni
G J
GJ
Conti
A
A
Boveri
M
M
Lecchini
S
S
Frigo
G
G
D001402:B-Lymphocytes / Q000737:chemistry; D002395:Catecholamines / Q000032:analysis*; D002469:Cell Separation; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004221:Disulfiram / Q000494:pharmacology; D004563:Electrochemistry / Q000379:methods*; D006098:Granulocytes / Q000737:chemistry; D006412:Hematopoietic Stem Cells / Q000737:chemistry*; D006801:Humans; D019169:Jurkat Cells; D007963:Leukocytes, Mononuclear / Q000737:chemistry*; D009000:Monocytes / Q000737:chemistry; D012110:Reserpine / Q000494:pharmacology; D013601:T-Lymphocytes / Q000737:chemistry; D013997:Time Factors; D020298:U937 Cells; D019805:alpha-Methyltyrosine / Q000494:pharmacology
D002395:Catecholamines; D019805:alpha-Methyltyrosine; D012110:Reserpine; D004221:Disulfiram