ABSTRACT
Objective To explore the expression of DC?SIGN, the phenotype of dendritic cells (DCs), on podocytes, and its role in immune and inflammatory responses of lupus nephritis (LN). Methods DC?SIGN and IgG1 expression in renal tissues of lupus nephritis patients were observed by immunohistochemistry and immunofluorescence. The 4?week old LN mice were randomly divided into the experimental group and the intervention group. C57BL/6J mice were used as normal control group. Mice of the intervention group were injected anti?DC?SIGN antibody at 6?week old. Mice were sacrificed at 16, 20, 24, 28?week old respectively, to observe the mice renal function and pathological changes. And DC?SIGN and IgG1 expression in renal tissue were observed by immunohistochemistry and immunofluorescence. In addition, mice podocytes were treated with serum of LN mice. Flow cytometry was used to investigate the expression of MHC II, CD80 and DC?SIGN expression on podocytes. Mixed lymphocyte reaction was used to detect the ability of stimulating T cells proliferation. IFN?gamma and IL?4 in supernatant were determined by ELISA. Results (1) Expression of DC?SIGN and IgG1 was found in glomeruli of lupus nephritis patients. (2) Accompanied by increased proteinuria of LN mice from 20?week old (P<0.01), DC?SIGN and IgG1 expression was found in glomeruli, and the renal function deteriorated up to 24 week?old (P<0.01). Mice with anti?DC?SIGN antibody intervention appeared reduced proteinuria and remission of renal function (P<0.01). (3) After stimulated by serum of LN mice, the expression of DC?SIGN, MHC II and CD80 was up?regulated, stimulation of T cell proliferation was enhanced (P<0.01), and IFN?gamma/IL?4 ratio increased (P<0.01). Anti?DC?SIGN antibody treatment down?regulated the expressions of DC?SIGN, MHC II and CD80 on podocytes, decreased the ability of stimulating T cell proliferation and lowered the ratio of IFN?gamma/IL?4 (P<0.01). Conclusions Podocytes in lupus nephritis can play DC?like function through the expression of DC?SIGN, which may be involved in immune and inflammatory responses of renal tissue. However, inhibiton of DC?SIGN can depress immune function of podocytes and have prevention and treatment effect.
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Objective To investigate the expression and distribution of parathyroid hormone (PTH) in renal tissues of early stage chronic kidney disease (CKD),and to elucidate its potential role in renal lesion.Methods Eighty-two patients of early stage CKD (stage 1 and 2) diagnosed as glomerulonephritis (GN) with different pathologic types by renal biopsy in our department between 2009 and 2012 were enrolled in the study.Renal tissues of eight patients with mismatched HLA haplotype or the normal part of renal cancer were chosen as controls.Scr,BUN,serum calcium,phosphorus,PTH and 25(OH)VitD3 were measured.Creatinine clearance (Ccr) was calculated by Cockcroft-Gault (CG)formula.99mTc-DTPA clearance rate was used to detect GFR.Patients were divided into mild,moderate and severe groups according to the renal interstitial extent of inflammatory cells infiltration.Immunohistochemistry was used to observe the expression and distribution of PTH in renal tissues.Image-Pro Plus software was used to calculate A value of PTH in renal tissues and compare the extent of PTH expression.Results The levels of calcium,phosphorus,25(OH)VitD3 and PTH in peripheral blood from GN patients of CKD stage Ⅰ and 2 were normal.PTH had no correlation with the above indexes.PTH expression could be seen in renal tissues of all the GN patients with different pathologic types,and it mainly located in renal tubular,only a few in glomeruli and interstitium.The expression of PTH in renal tissues of GN increased compared with the controls (P ＜ 0.01).Furthermore,PTH expression elevated with the increase of inflammatory cells infiltration in interstitium.However the expression of PTH was not significantly different among different pathologic types of GN.Conclusions In the early stage CKD,PTH expression in patients of GN increases,which occurs earlier as compared to PTH elevation in peripheral blood and the imbalance of minerals and bone metabolism.The intensity of PTH expression is associated with the local inflammation.
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Objective To explore the role of dendritic cell-specific intercellular adhesion molecule-3-grabbing nonintegrin (DC-SIGN) in the tubulointerstitial lesions of immune-mediated nephrotoxic nephritis (NTN) and the intervention regulation by anti-P-selectin lectin-EGF domain monoclonal antibody (PsL-EGFmAb). Methods WKY rats were randomly divided into control,NTN and PsL-EGFmAb-treated groups. The mrs in NTN group were injected with 1 ml nephrotoxic rabbit serum per kilogram of rat body weight; the ones in PsL-EGFmAb-treated group were injected with 2 mg PsL-EGFmAb per kilogram of rat body weight simultaneously and 2 h later after nephrotoxic rabbit serum injection; and those in control group were injected with equal volume of 0.9% saline. Renal function and pathology were observed at day 4, 7 and 14 after the induction of NTN. Distribution of DC-SIGN + dendritic cells (DCs) in renal tissues was measured by immunofluorescence. Real-time PCR was performed to examine the expression of P-selectin,RANTES, TNF-α, IL-10, IFN-γ and IL-4. Expression of MHC Ⅱ , CD80 and DC-SIGN on dendritic cells was analyzed by flow cytometry. Transendothelial migration was used to detect the ability of DCs migration. DCs ability to activate T cells was determined by mixed lymphocyte reaction (MLR). ELISA was used to detect the concentration of IFN-γ and IL-4 in the supernatant of MLR. Results At day 4, immature DC-SIGN+ DCs infiltrated the rat renal tubulointerstium of NTN group, matured at day 14, and enhanced the ability to migrate and activate T cells. The distribution of DC-SIGN + DCs was significantly related to the form of crescent, tubulointerstial lesions and renal function. In addition, expression of chemokine RANTES and proinflammatory cytokine TNF-α continuously augmented since day 4, while anti-inflammatory eytokine IL-10 decreased after markedly increased at day 4. At day 14, IFN-γ/IL-4 mRNA increased, which was obviously related to DCs maturation. The intervention of PsL-EGFmAb supressed the expression of DC-SIGN and CD80 on DCs, depressed DCs maturation, migration and ability to activate T cells,down-regulated proinflammatory cytokines and up-regulated anti-inflammatory cytokines in kidney,and thus regulated Th1/Th2 bias. At the same time, kidneys showed the decrease of crescents,improvement of tnbulointerstium damage and renal function. Conclusions DC-SIGN may mediate DCs tubulointerstitial infiltration. It may be also a potent regulator of local immune reaction imbalance and pathology of tubulointerstium. PsL-EGFmAb may depress DCs migration and downregulate DCs maturation and function through DC-SIGN, and thus having a role in prevention and treatment.