Patients with acute pancreatitis complicated by organ failure show highly aberrant monocyte signaling profiles assessed by phospho-specific flow cytometry.
OBJECTIVE
To outline signaling profiles and transmigration capacity of monocytes of patients with severe acute pancreatitis.


METHODS
Prospective study.


METHODS
University hospital intensive care unit.


METHODS
Thirteen patients with severe acute pancreatitis. All patients had organ dysfunction (acute respiratory distress syndrome in 12, renal dysfunction in eight). Healthy volunteers served as reference subjects.


METHODS
Blood samples were collected after admission to the intensive care unit.


RESULTS
Phosphorylation of nuclear factor-kappaB and p38, signal transducers and activators of transcription (STATs) 1, 3, 5, and extracellular signal-regulated kinases 1/2 in appropriately stimulated and nonstimulated samples were studied using phospho-specific whole-blood flow cytometry. Monocyte chemotactic protein-1-induced transmigration of monocytes among mononuclear cells obtained by density gradient centrifugation was studied using Transwell cell culture inserts covered with confluent layer of endothelial EA-HY cells. Phosphorylation levels of nuclear factor-kappaB induced by tumor necrosis factor, bacterial lipopolysaccharide, muramyl dipeptide, Escherichia coli, Staphylococcus aureus, and Staphylococcus epidermidis were significantly lower in patients' monocytes than monocytes of healthy reference subjects, whereas mitogen-activated protein kinase p38 phosphorylation levels were normal. Phosphorylation levels induced by interleukin-6 in STAT1 and STAT3 and by combination of phorbol 12-myristate 13-acetate and calcium ionophore A23187 in extracellular signal-regulated kinases 1/2, members of a mitogen-activated protein kinase family, were depressed in patients' monocytes, whereas phosphorylation levels induced by granulocyte-macrophage colony-stimulating factor in STAT5 was normal. In nonstimulated samples, phosphorylation levels were normal. The transmigration percentage of patients' monocytes was significantly lower than that of reference monocytes.


CONCLUSIONS
In severe acute pancreatitis, monocytes show impaired nuclear factor kappaB and STAT1 activation, which may increase susceptibility to secondary infections. p38 activation is normal and STAT3 activation is depressed, which may contribute to maintenance of systemic inflammation. Extracellular signal-regulated kinases 1/2 activation is impaired, which may depress monocytes' transmigration and may consequently increase risk of infection. Monitoring of monocyte signaling profiles may aid in finding new therapeutic approaches and predictors of outcome of severe acute pancreatitis.
Oiva
Jani
J
Department of Surgery, Helsinki University Central Hospital, University of Helsinki, Helsinki, Finland.
Mustonen
Harri
H
Kylänpää
Marja-Leena
ML
Kyhälä
Lea
L
Alanärä
Tiina
T
Aittomäki
Saara
S
Siitonen
Sanna
S
Kemppainen
Esko
E
Puolakkainen
Pauli
P
Repo
Heikki
H
D000328:Adult; D015415:Biomarkers / Q000097:blood; D016022:Case-Control Studies; D003422:Critical Care / Q000379:methods; D016638:Critical Illness / Q000628:therapy; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005434:Flow Cytometry; D006785:Hospitals, University; D006801:Humans; D007362:Intensive Care Units; D008297:Male; D008875:Middle Aged; D009000:Monocytes / Q000502:physiology*; D009102:Multiple Organ Failure / Q000097:blood* / Q000150:complications / Q000175:diagnosis; D019283:Pancreatitis, Acute Necrotizing / Q000097:blood* / Q000150:complications / Q000175:diagnosis / Q000628:therapy; D010766:Phosphorylation; D011237:Predictive Value of Tests; D011379:Prognosis; D011446:Prospective Studies; D012016:Reference Values; D018570:Risk Assessment; D050799:STAT5 Transcription Factor / Q000378:metabolism; D012680:Sensitivity and Specificity; D015398:Signal Transduction / Q000502:physiology*; D018709:Statistics, Nonparametric; D025521:Tumor Suppressor Proteins / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
D015415:Biomarkers; D050799:STAT5 Transcription Factor; C494092:STAT5A protein, human; D025521:Tumor Suppressor Proteins; D048049:Extracellular Signal-Regulated MAP Kinases; D048051:p38 Mitogen-Activated Protein Kinases