Anti-dsDNA antibody up-regulates interleukin 6, but not cyclo-oxygenase, gene expression in glomerular mesangial cells: a marker of immune-mediated renal damage?
OBJECTIVE
To determine whether anti-double stranded DNA antibody (anti-dsDNA) can affect the synthesis of eicosanoids and cytokines in rat glomerular mesangial cells (RMC).


METHODS
Glomerular mesangial cells were isolated and subcultured from Sprague-Dawley rats. Monoclonal anti-dsDNA (12B3 clone) was derived from autoimmune MRL-lpr/lpr mouse by hybridoma technology.


METHODS
The mRNA expression of cyclo-oxygenase type 1 (COX-1), type 2 (COX-2), Th1 (IL-2 and IFN-gamma)/Th2 (IL-4 and IL-10) and proinflammatory (IL-6 and TNF-alpha) and anti-inflammatory (TGF-beta) cytokines of RMC +/- anti-dsDNA was detected by RT-PCR. The PGE2 production by RMC +/- anti-dsDNA was measured by ELISA. The statistical significance was assessed by non-parametric Wilcoxon signed rank test.


RESULTS
We found RMC spontaneously expressed COX-1, but not COX-2. The incubation of RMC with anti-dsDNA (50 ng/ml) did not affect COX expression and PGE2 production by RMC. RMC also spontaneously expressed IL-6, TNF-alpha and TGF-beta mRNA. However, only IL-6 was up-regulated by anti-dsDNA.


CONCLUSIONS
Increased IL-6 expression in RMC may become a marker of anti-dsDNA-mediated immune damage of mesangial cells.
Department of Medicine & Institute of Molecular Medicine, National Taiwan University College of Medicine, Taipei. clyu@ha.mc.ntu.edu.tw
C L
CL
Yu
K H
KH
Sun
C Y
CY
Tsai
S C
SC
Hsieh
H S
HS
Yu
D000818:Animals; D000974:Antibodies, Antinuclear / Q000494:pharmacology*; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D015415:Biomarkers; D051545:Cyclooxygenase 1; D051546:Cyclooxygenase 2; D015232:Dinoprostone / Q000096:biosynthesis; D005786:Gene Expression Regulation; D005920:Glomerular Mesangium / Q000737:chemistry / Q000378:metabolism*; D007371:Interferon-gamma / Q000235:genetics; D016753:Interleukin-10 / Q000235:genetics; D007376:Interleukin-2 / Q000235:genetics; D015847:Interleukin-4 / Q000235:genetics; D015850:Interleukin-6 / Q000235:genetics*; D007527:Isoenzymes / Q000235:genetics; D007674:Kidney Diseases / Q000276:immunology*; D008565:Membrane Proteins; D011451:Prostaglandin-Endoperoxide Synthases / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000974:Antibodies, Antinuclear; D000911:Antibodies, Monoclonal; D015415:Biomarkers; D007376:Interleukin-2; D015850:Interleukin-6; D007527:Isoenzymes; D008565:Membrane Proteins; D012333:RNA, Messenger; D016753:Interleukin-10; D015847:Interleukin-4; D007371:Interferon-gamma; D051545:Cyclooxygenase 1; D051546:Cyclooxygenase 2; D011451:Prostaglandin-Endoperoxide Synthases; C496513:Ptgs1 protein, rat; D015232:Dinoprostone