Procalcitonin expression in human peripheral blood mononuclear cells and its modulation by lipopolysaccharides and sepsis-related cytokines in vitro.
Procalcitonin (PCT), the precursor of calcitonin, was recently put forward as a diagnostic marker of systemic bacterial infection and sepsis. The major PCT production site in sepsis still remains unclear. Because of a certain association between increased levels of PCT and leukocyte-derived cytokines during sepsis, we assessed the possible expression of PCT in human peripheral blood mononuclear cells (PBMCs) and the modulation of PCT by lipopolysaccharides (LPS) and various sepsis-related cytokines by reverse transcriptase-polymerase chain reaction (RT-PCR) by using a novel primer set and flow cytometric analysis with intracellular staining with antibodies to the PCT components calcitonin and katacalcin. RT-PCR and flow cytometric analysis demonstrated that PBMCs express PCT both on mRNA and on protein levels. LPS and various proinflammatory cytokines (interleukin-1beta (IL-1beta), IL-6, tumor necrosis factor-alpha (TNF-alpha), IL-2) had pronounced stimulatory effects on the expression of PCT mRNA. Under identical experimental conditions the anti-inflammatory cytokine IL-10 had no effect on the expression of mRNA for PCT. Flow cytometric analysis demonstrated increased intracellular amounts of PCT components after LPS stimulation. Thus we demonstrate for the first time that PCT is expressed in PBMCs. This expression is modulated by bacterial LPS and sepsis-related cytokines. Therefore PBMCs may be among the sources of elevated PCT levels in patients with sepsis.
Department of Anesthesiology and Intensive Care Medicine and the Institute of Clinical Immunology, Friedrich Schiller University, Jena, Germany.
M
M
Oberhoffer
I
I
Stonans
S
S
Russwurm
E
E
Stonane
H
H
Vogelsang
U
U
Junker
L
L
Jäger
K
K
Reinhart
D001483:Base Sequence; D002116:Calcitonin / Q000097:blood* / Q000235:genetics; D015740:Calcitonin Gene-Related Peptide; D016207:Cytokines / Q000494:pharmacology*; D004247:DNA; D017931:DNA Primers; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D066298:In Vitro Techniques; D018950:Lipopolysaccharide Receptors / Q000032:analysis; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009000:Monocytes / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D010446:Peptide Fragments / Q000097:blood; D010835:Phytohemagglutinins / Q000494:pharmacology; D011498:Protein Precursors / Q000097:blood* / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018805:Sepsis / Q000378:metabolism*
C000606589:CALCA protein, human; D016207:Cytokines; D017931:DNA Primers; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D010446:Peptide Fragments; D010835:Phytohemagglutinins; D011498:Protein Precursors; D012333:RNA, Messenger; C037553:katacalcin; D002116:Calcitonin; D004247:DNA; D015740:Calcitonin Gene-Related Peptide