Coagulation cascade activation causes CC chemokine receptor-2 gene expression and mononuclear cell activation in hemodialysis patients.
Priming of the coagulation cascade during hemodialysis (HD) leads to the release of activated factor X (FXa). The binding of FXa to its specific receptors, effector protease receptor-1 (EPR-1) and protease-activated receptor-2 (PAR-2), may induce the activation of peripheral blood mononuclear cells (PBMC) and promote a chronic inflammatory state that is responsible for several HD-related morbidities. In the attempt to elucidate the mechanisms underlying the coagulation-associated inflammation in HD, 10 HD patients were randomized to be treated subsequently with a cellulose acetate membrane (CA) and Ethylen-vinyl-alcohol (EVAL), a synthetic membrane that has been shown to reduce FXa generation. At the end of each experimental period, surface FXa and thrombin receptors (EPR-1 and PAR-1, -2, and -4) and CCR2 (monocyte chemoattractant protein-1 receptor) gene expression in isolated PBMC were examined. the ability of dialytic membranes to activate protein-tyrosine kinases and the stress-activated kinase JNK and to modulate the generation of terminal complement complex (TCC) was also investigated. EPR-1 and PAR-2 and -4 mRNA expression, barely detectable in normal PBMC, were significantly upregulated in HD patients, particularly in those who were treated with CA. A striking increase of tyrosine-phosphorylated proteins and JNK activation was observed at the end of HD only in CA-treated patients. Simultaneously, an increased gene expression for both splicing isoforms of CCR2, A and B, only in PBMC from CA-treated patients was demonstrated. The increased CCR-2 mRNA abundance was followed by a significant increase in its protein synthesis. The high expression of CCR2 was associated with an increased generation of plasma TCC and a significant drop in leukocyte and monocyte count. By contrast, EVAL treatment slightly lowered TCC generation and normalized leukocyte count. In vitro FXa induced CCR2 A and B expression and JNK activation in freshly isolated PBMC. FXa-induced CCR2 mRNA expression was completely abolished by JNK and tyrosine kinase inhibition. In conclusion, these data suggest that subclinical clotting activation may cause an increased CCR2 gene and protein expression on uremic PBMC, contributing to HD-related chronic microinflammation. The use of the less coagulation-activating membrane, EVAL, may reduce PBMC activation through the modulation of the stress-activated kinase JNK.
Division of Nephrology, Department of Emergency and Transplantation, University of Bari, Policlinico, Piazza Giulio Cesare 11, Bari 70124, Italy.
Giovanni
G
Pertosa
Simona
S
Simone
Michela
M
Soccio
Daniela
D
Marrone
Loreto
L
Gesualdo
Francesco Paolo
FP
Schena
Giuseppe
G
Grandaliano
D000328:Adult; D000368:Aged; D051017:Apoptosis Regulatory Proteins / Q000096:biosynthesis; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism; D002482:Cellulose / Q000031:analogs & derivatives / Q000737:chemistry; D015938:Complement Membrane Attack Complex / Q000096:biosynthesis; D004268:DNA-Binding Proteins / Q000096:biosynthesis; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015951:Factor Xa / Q000096:biosynthesis / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D007249:Inflammation; D007962:Leukocytes / Q000166:cytology; D007963:Leukocytes, Mononuclear / Q000166:cytology* / Q000378:metabolism; D048670:MAP Kinase Kinase 4 / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D009000:Monocytes / Q000166:cytology; D010766:Phosphorylation; D011145:Polyvinyls / Q000737:chemistry; D011485:Protein Binding; D020033:Protein Isoforms; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis; D050783:Proto-Oncogene Proteins c-ets; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D044463:Receptor, PAR-1 / Q000096:biosynthesis; D044464:Receptor, PAR-2 / Q000096:biosynthesis; D054390:Receptors, CCR2; D019707:Receptors, Chemokine / Q000096:biosynthesis*; D006435:Renal Dialysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014157:Transcription Factors / Q000096:biosynthesis
D051017:Apoptosis Regulatory Proteins; C515334:CCR2 protein, human; D015938:Complement Membrane Attack Complex; D004268:DNA-Binding Proteins; C105529:ELF3 protein, human; D004791:Enzyme Inhibitors; D011145:Polyvinyls; D020033:Protein Isoforms; D011518:Proto-Oncogene Proteins; D050783:Proto-Oncogene Proteins c-ets; D012333:RNA, Messenger; D044463:Receptor, PAR-1; D044464:Receptor, PAR-2; D054390:Receptors, CCR2; D019707:Receptors, Chemokine; D014157:Transcription Factors; C101296:prostate apoptosis response-4 protein; C020320:ethylene-vinyl alcohol copolymer; C005062:acetylcellulose; D012313:RNA; D002482:Cellulose; D011505:Protein-Tyrosine Kinases; D048670:MAP Kinase Kinase 4; D015951:Factor Xa