Morphine directs T cells toward T(H2) differentiation.
BACKGROUND
Failure of cell-mediated immunity is thought to increase the morbidity and mortality rates after trauma and major surgical procedures and to be the result, in part, of a redirection of CD4(+) T cells toward T(H2) differentiation. We tested the hypothesis that morphine treatment after injury promotes T(H2) differentiation of precursor T cells through the mu-opioid receptor.


METHODS
Human peripheral blood mononuclear cells (PBMCs) or splenocytes from either wild type or mu-opioid receptor knock-out mice were treated in vitro with either vehicle or morphine and then stimulated with anti-CD3/anti-CD28. The supernatant was assayed for T(H1) (interleukin-2 [IL-2], interferon gamma [IFN gamma]) and T(H2) (IL-4, IL-5) cytokines (enzyme-linked immunosorbent assay). Morphine regulation of IL-4 transcription was investigated in PBMCs (IL-4 messenger RNA, nuclear factor of activated T-cells) and Jurkat T cells transfected with a murine IL-4 promoter-luciferase construct. Morphine-induced nuclear factor of activated T-cell (NFAT) binding was assayed with the electromobility shift assay in Jurkat T cells.


RESULTS
Morphine treatment of PBMCs decreases IL-2 and IFN gamma and increases IL-4 and IL-5 as a function of morphine concentration. Morphine treatment in wild type splenocytes inhibited IFN gamma and stimulated IL-4 protein synthesis. Changes in cytokine synthesis were abolished in mu-opioid receptor knockout mice. Morphine treatment increases IL-4 messenger RNA accumulation in PBMCs and increases IL-4 promoter activity in Jurkat T cells. Morphine increases NFAT nuclear protein binding to an NFAT DNA response element.


CONCLUSIONS
We conclude that morphine treatment promotes T(H2) differentiation through a mu-opioid receptor mechanism and that morphine treatment increases IL-4 transcription, in part, through an NFAT mechanism.
Roy
S
S
Department of Surgery, University of Minnesota, the Minneapolis Veterans Affairs Medical Center, Minneapolis, MN 55417, USA.
Balasubramanian
S
S
Sumandeep
S
S
Charboneau
R
R
Wang
J
J
Melnyk
D
D
Beilman
G J
GJ
Vatassery
R
R
Barke
R A
RA
D000701:Analgesics, Opioid / Q000494:pharmacology*; D000818:Animals; D018106:CD28 Antigens; D017252:CD3 Complex / Q000276:immunology; D002454:Cell Differentiation / Q000187:drug effects / Q000276:immunology; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D007376:Interleukin-2 / Q000378:metabolism; D015847:Interleukin-4 / Q000235:genetics / Q000378:metabolism; D015848:Interleukin-5 / Q000378:metabolism; D019169:Jurkat Cells; D051379:Mice; D018345:Mice, Knockout; D009020:Morphine / Q000494:pharmacology*; D011401:Promoter Regions, Genetic / Q000276:immunology; D017450:Receptors, Opioid, mu / Q000235:genetics / Q000276:immunology; D013154:Spleen / Q000166:cytology; D018418:Th2 Cells / Q000166:cytology* / Q000187:drug effects*; D014158:Transcription, Genetic / Q000187:drug effects / Q000276:immunology; D014162:Transfection
D000701:Analgesics, Opioid; D018106:CD28 Antigens; D017252:CD3 Complex; D007376:Interleukin-2; D015848:Interleukin-5; D017450:Receptors, Opioid, mu; D015847:Interleukin-4; D009020:Morphine; D007371:Interferon-gamma