ABSTRACT
Objective To explore the distribution of dendritic cells (DCs) and the expression of adhesion molecules in rat kidney with unilateral ureteral obstruction (UUO),as well as the regulatory effect of anti-P-selectin lectin-EGF domain monoelonal antibody (PsL-EGFmAb) on adhesion,maturation and function of human DCs cultured in vitro.Methods UUO rat models were established,which were divided into sham group (n=6),untreated group (n=18)and treated group with PsL-EGFmAb(n=18).DCs were analyzed with Axioplan 2 microscopy,while P-selectin being observed by immunohistochemistry.CD34~+ stem cells were isolated from cord blood and cultured in 20%IMDM medium with SCF,GM-CSF,TGF-?1,Flt-3L and TNF-?in vitro.During development, PsL-EGFmAb was added and IL-10 served as control.FACS was performed to detect the expression of HLA-DR,CD1a,CD11c,CD54,CD83,CD80,CD86,CD209 (DC-SIGN) and CD62-P,-E,-L (P-,E-,L-selectin) on DCs.RT-PCR was performed to detect the expression of NF-_KB P50, P65 mRNA.MLR was performed to detect the stimulatory effect of DCs on T cell proliferation and ELISA to determine IL-12p70 amount.Results Comparing with Sham group,the expression of P- selectin was up-regulated among tubulointerstitium mainly on renal tubular epithelial cell after unilateral ureteral obstruction on day 1,while CD1a~+CD80~+DCs being also found in renal interstitium. The expression of P-seleetin and CD1a~+CD80~+DCs was increased evidently on day 7,and correlated with the degree of renal tubulointerstitial fibrosis closely.However,these changes became less conspicuous in rat treated with PsL-EGFmAb.In vitro experiment showed on day 5 after cultured with the induction of TNF-?,immature DCs highly expressed C-type lectin DC-SIGN of pattern recognition receptors;the expression of co-stimulatory molecules such as CD11c,CD83,CD80 and CD86 on mature DCs was up-regulated in paralleling with the mRNA level of NF-_KB;the secretion of IL-12 was enhanced,as well as displaying the features of antigen-presenting cells with a higher ability to induce proliferation of T lymphocytes in vitro.In addition,L-selectin expressed highly on immature DCs,but lowly on mature DCs,neither of two DCs expressed P- and E-selectin.Compared with the IL-10 treated group,PsL-EGFmAb had an inhibitory effect on DC-SIGN of DCs with down- regulating the mRNA level of NF-_KB.PsL-EGFmAb could also inhibited CD11c,CD83,CD80, CD86 expression,reduced secretion of IL-12,and inhibited T cell proliferation stimulated by DCs in vitro.Conclusion DCs may play a critical role on initiating the inflammatory injury of renal tubulointerstitium,and the inhibitory effect of PsL-EGFmAb on DC maturation and function correlated with the inhibition of DC-SIGN,which is mainly mediated through NF-_KB signaling pathway.