Targeting the CD134-CD134L interaction using anti-CD134 and/or rhCD134 fusion protein as a possible strategy to prevent lupus nephritis.

Lupus nephritis (LN) is characterized by an increased upregulation of Th1. This study was undertaken to evaluate the role of CD134 in cytokine production in peripheral blood mononuclear cells (PBMCs) from subjects with LN. Percentages of IFN-gamma- (Th1), IL-4-, and IL-10- (Th2) producing cells within the PBMC CD4+ T cell population of LN subjects were found to be higher than those of healthy subjects. Stimulation of PBMC from LN subjects with anti-CD3 epsilon mAb/rIL-2 resulted in further increases in cytokine production. Stimulation in the presence of anti-CD134 mAb resulted in reduced IL-4 and IL-10 production; however, it also resulted in increased IFN-gamma production. Stimulation in the presence of the fusion protein rhCD134:Fc resulted in decreased production of all three cytokines. The possibilities that anti-CD134 therapy may control the extent of IL-4- and IL-10-mediated damage in active LN and that rhCD134:Fc therapy may prevent occurrence of LN are discussed.

Effect of anti-CD134L mAb and CTLA4Ig on ConA-induced proliferation, Th cytokine secretion, and anti-dsDNA antibody production in spleen cells from lupus-prone BXSB mice.

We sought to evaluate the effects of combined downregulation of CD134 and cytotoxic T lymphocyte-associated antigen 4 (CTLA4) on the autoimmune process of lupus. Concanavalin A (ConA)-induced proliferation, T helper cell cytokine secretion, and anti-double stranded DNA (dsDNA) antibody production were measured in cultures of splenic lymphocytes derived from lupus-prone BXSB mice. Splenocytes from six prednisone-treated and six untreated male lupus-prone BXSB mice, as well as from six syngeneically normal C57BL/6 male mice, were stimulated with ConA. BXSB splenocytes from untreated mice were exposed to anti-CD134L mAb, CTLA4 linked to the Fc portion of IgG1 (CTLA4Ig), or both. The magnitude of splenocyte proliferation and the levels of IFN-gamma, IL-6, and anti-dsDNA antibody were: (1) significantly higher in cultures of ConA-stimulated control and other cells than in unstimulated cells, (2) similar in cultures of normal and BXSB cells treated with anti-CD134 and CTLA4Ig or prednisone and (3) significantly reduced in cultures of ConA-stimulated and unstimulated cells treated with anti-CD134L and CTLA4Ig or prednisone compared with cells treated with CD134L or CTLA4Ig alone. Like corticosteroids, anti-CD134L mAb or CTLA4Ig can inhibit T- and B-cell activation by blocking the CD134-CD134L or CD28/CTLA4-B7 co-stimulatory pathway. The combined immune intervention described herein may prove useful for the treatment of autoimmune diseases such as systemic lupus erythematosus.

[Risk factors affecting the long-term outcome of IgA nephropathy].

OBJECTIVE: To study the risk factors predicting long-term renal survival of IgA nephropathy in Chinese. METHODS: Clinical and pathological data of 317 patients (124 males and 193 females) with IgA nephropathy confirmed by renal biopsy in our center from January 1987 to February 2003 were reviewed retrospectively and were correlated with outcomes. A semiquantitative scoring system was used to evaluate individual pathological lesion of the kidney. Patients were followed for at least 6 months and doubling of serum creatinine level was defined as endpoint of follow-up. Renal survival was calculated by Kaplan-Meier survival analysis and risk factors of progression were analyzed by using univariate and multi-variate Cox regression models. RESULTS: The average age at renal biopsy was (30.1 +/- 10.9) years and the average duration from onset of disease to the time of biopsy was (20.1 +/- 33.7) months. Thirty-two percent of the patients had 24 h-urinary protein excretion greater than 1.0 g at the time of biopsy. Thirty-two percent of the patients had hypertension and 20.8% had renal insufficiency. Thirty-five percent of the patients were of Lee's grade IV or above and 20.5% presented with small proportion of crescent formation (usually less than 20%). Patients were followed for an average duration of (43.5 +/- 32.2) months with 39 patients (12.3%) reaching the endpoint. The 1-, 3-, 5- and 10-year renal survival was 99.5%, 93.1%, 84.5% and 60.1% respectively. Univariate Cox regression analysis revealed that longer duration of the disease before biopsy, serum creatinine > 115 micromol/L, proteinuria > 1.0 g/d, hypertension, Lee's grading of IV-V, moderate-severe glomerulosclerosis, crescent formation, moderate-severe interstitial fibrosis and renal arteriolar lesion were risk factors of disease progression, with an odds ratio of 1.007, 9.61, 7.31, 3.97, 5.41, 5.78, 4.65, 14.05 and 2.28 respectively (P < 0.001). Episodic macro-hematuria had an odds ratio of 0.194 (P < 0.05). Age, sex, serum cholesterol and triglyceride level had no significant impact on prognosis. Proteinuria, elevated serum creatinine, glomerulosclerosis, crescent formation and interstitial fibrosis were confirmed to be independent risk factors by multi-variate Cox regression model while the remaining variables were not statistically significant. Patients with both renal insufficiency and proteinuria greater that 1.0 g/24 h at the time of biopsy had a very poor 5-year renal survival (41.8%). CONCLUSIONS: Proteinuria, renal insufficiency, glomerulosclerosis, crescent formation and interstitial fibrosis were independent risk factors predicting the renal survival. IgA nephropathy presented with proteinuria, hypertension and crescent formation may need intervention.

[Role of Akt/NF-kappa B signal transduction pathway in murine glomerular mesangial cell proliferation induced by immune complex].

AIM: To explore whether immune complex (IC) can directly induce glomerular mesangial cells(MCs) proliferation and the role of Akt/NF-kappa B signal pathway in the proliferation. METHODS: The mice were divided into control, stimulation and oligodeoxynucleotide(ODN) groups. In ODN group, MCs isolated from mice were transfected with Akt1 sense, mismatched or antisense ODN for 8 h, respectively, by using lipofectin, control and stimulation groups were incubated with lipofectin for 8 h. Then stimulation and ODN groups were incubated with aggregated IgG(AIgG)(a standard IC model), while the control group with monomeric IgG. MTT colorimetry was used to detect MCs proliferation. Distribution of MCs in cell cycle was analyzed by flow cytometry. Cyclin D1 mRNA and its protein expression were determined by RT-PCR and Western blot, respectively.The activity of NF-kappa B in MCs was determined by EMSA. RESULTS: AIgG activated NF-kappa B, upregulated cyclin D1 mRNA and its protein expression, and induced majority of MCs to enter S-phase in cell cycle. Akt1 antisense ODN specifically decreased AIgG-induced NF-kappa B activation, cyclin D1 mRNA and its protein expression, and then inhibited MCs to progress to S-phase and cell proliferation. Sense ODN and mismatched ODN had no such effects. CONCLUSION: IC can directly stimulate MCs proliferation through Akt/NF-kappa B signal pathway, suggesting that NF-kappa B probably be a useful molecule for targeted therapy in IC-mediated MC overproliferation.

CD40 is expressed on rat peritoneal mesothelial cells and upregulates ICAM-1 production.

BACKGROUND: CD40 has been identified on a variety of cell types, and it plays an important role in adaptive immunity and inflammation. Peritoneal mesothelial cells (PMCs) are the main cell layer that line the peritoneal membrane. Previously we found that CD40 ligand (CD154) is functionally expressed on peritoneal macrophages during continuous ambulatory peritoneal dialysis peritonitis. However, there are few studies that have examined both CD40 expression on PMCs and the function of CD40 signalling in peritoneal local defence. The purpose of this study was to determine whether PMCs express CD40 and to investigate potential mechanisms of CD40-CD154 interactions that may be involved in the inflammation of the peritoneal membrane. METHODS: Rat PMCs were harvested from the peritoneal cavity and maintained under defined in vitro conditions. We examined expression of CD40 on PMCs under normal culture or stimulation with interferon-gamma (IFN-gamma), tumour necrosis factor-alpha (TNF-alpha) or interleukin (IL)-1 by reverse transcription-polymerase chain reaction and fluorescence-activated cell sorting (FACS) analysis. After activation with CD40 monoclonal antibody (mAb), the expression of intercellular adhesion molecule-1 (ICAM-1) on PMCs was analysed by FACS. RESULTS: A portion of rat PMCs cultured in vitro expressed CD40 constitutively. The expression of CD40 mRNA and protein was upregulated markedly following stimulation with IFN-gamma, but not following IL-1 or TNF-alpha. The expression of ICAM-1 on PMCs was significantly increased after activation of CD40 with IFN-gamma and with CD40 mAb. CONCLUSION: PMCs functionally express CD40. The interaction between CD40 on PMCs and CD154-positive cells in the peritoneal cavity may play an important role in peritoneal local defence and may be involved in the inflammation process of the peritoneum.

IL-17 induces autoantibody overproduction and peripheral blood mononuclear cell overexpression of IL-6 in lupus nephritis patients.

OBJECTIVE: To investigate the role of IL-17 in the overproduction of autoantibodies and IL-6 overexpression by peripheral blood mononuclear cells (PBMC) of lupus nephritis (LN) patients. METHODS: Fifteen consecutively hospitalized LN patients were selected as subjects and 15 healthy adults as normal controls. PBMC were obtained by Ficoll density gradient centrifugation. IgG, anti-dsDNA antibody and IL-6 protein levels were assessed using enzyme-linked immunosorbent assays (ELISA) on the supernatant of cultured PBMC of LN patients or normal controls. IL-6 mRNA levels in PBMC were measured using reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: In medium culture, IgG, anti-dsDNA and IL-6 protein levels of the supernatant of PBMC from LN patients were significantly higher than those from normal controls (1492.1 +/- 73.2 ng/ml vs 636.7 +/- 51.9 ng/ml for IgG, 306.6 +/- 53.7 IU/ml vs 95.8 +/- 11.6 IU/ml for anti-dsDNA and 50.92 +/- 15.92 ng/ml vs 1.77 +/- 0.73 ng/ml for IL-6, all P < 0.001). In LN patients, IgG, anti-dsDNA and IL-6 protein levels were higher in the supernatants of PBMC in the IL-17-stimulated culture than the medium culture, but in normal controls, only the IL-6 protein levels were significantly higher. The increase in IgG, anti-dsDNA and IL-6 protein levels induced by IL-17 was dose-dependent and could be completely blocked by IL-17 monoclonal antibody mIgG(28) and partially blocked by dexamethasone. Similarly, IL-6 mRNA overexpression of PBMC in LN patients or normal controls induced by IL-17 was both dose- and time-dependent. During medium culture, IL-6 mRNA levels in LN patients were significantly higher than those in normal controls (1.80 +/- 0.11 vs 0.36 +/- 0.07). During stimulation with IL-17, IL-6 mRNA levels in LN patients were higher than those in normal controls (3.21 +/- 0.24 vs 1.30 +/- 0.14, P < 0.05) and also significantly higher when comparing the stimulated culture with the medium culture either in LN patients or normal control. CONCLUSIONS: IL-17 may play an important role in the pathogenesis of LN through the induction of IgG, anti-dsDNA overproduction and IL-6 overexpression of PBMC in LN patients.

[Effects of LFA-1 costimulation on proliferation and IgG production of PBMCs from lupus nephritis patients].

AIM: To study the effect of lymphocyte function associated antigen-1(LFA-1) costimulation on proliferation and immunoglobin production of peripheral blood mononuclear cells(PBMCs) form lupus nephritis(LN) patients. METHODS: 29 LN patients were enrolled in this study, and 12 healthy persons served as control. PBMCs from LN patients and healthy persons were obtained by Ficoll density gradient centrifugation, and cell proliferation was detected by (3)H-TdR incorporation. IgG content in cultural supermatant was detected by ELISA. RESULTS: Stimulation of anti-CD3 mAb alone could enhance the proliferation and IgG production of PBMCs from 29 LN patients,while the effects on PBMCs from patients in active phase were stronger than those from the patients in the inactive phase (P<0.01). But anti-CD3 mAb had no influenence on PBMCs from healthy persons.The costimulation of anti-CD3 mAb and LFA-1 could increase proliferation and IgG production of PBMCs from LN patients and healthy persons. The effects of this costimulation decreased in turn from active and inactive LN patients to normal persons (P<0.01). The costimulant effects of LFA-1 was inhibited by anti-LFA-1 mAb. CONCLUSION: The effects of enhancing PBMC proliferation and IgG production by LFA-1 may be a mechanism of LN pathogenesis.

Progress of intervention of renal interstitial fibrosis with Chinese traditional herbal medicine.

This article reviews the current status of progress in the research of renal interstitial fibrosis therapy using traditional Chinese herbal medicine, which exerts its therapeutic effect through inhibiting cytokine expression and fibroblast proliferation, inducing apoptosis of the renal myofibroblasts and other mechanisms.

Expression of interleukin-12 and its signaling molecules in peripheral blood mononuclear cells in systemic lupus erythematosus patients.

OBJECTIVE: To determine the in vitro expression of interleukin-12 (IL-12) and its effect on signal transducers and activators of transcription (STAT) signaling molecules in peripheral blood mononuclear cells (PBMCs) in patients with systemic lupus erythematosus (SLE). METHODS: Peripheral blood mononuclear cells in 39 patients with definite systemic lupus erythematosus and 11 healthy volunteers were collected. Expression of IL-12 P40mRNA in PBMCs was determined with reverse transcription-polymerase chain reaction (RT-PCR). Quantity of IL-12 protein supernatant was measured by enzyme-linked immunosorbent assay (ELISA). The levels of phosphorylated STAT3 and STAT4 signaling molecules in PBMCs were detected by immunoblot. RESULTS: Levels of IL-12 protein and mRNA expression in patients with active or inactive SLE were significantly higher than those in controls. Phytohemagglutinin (PHA ) may promote the expression of IL-12. IL-12 alone induced the phosphorylation of STAT3 and STAT4 in PBMCs from patients with SLE, especially in active SLE. However it had no obvious effect on normal PBMCs. Phosphorylated STAT3 and STAT4 might be observed in normal PBMCs treated with IL-12 plus PHA. CONCLUSION: IL-12 is produced aberrantly in patients with SLE. IL-12 might exert its biological role in SLE via the aberrantly phosphorylated STAT3 and STAT4 signaling molecules.

Long-term effect of TCM decoctions in treatment of nephrotic syndrome.

Fifty-seven cases of nephrotic syndrome were treated with TCM decoctions as accessory treatment for prednisone and cyclophosphamide, and the effects were observed in a follow-up period of 5-15 years. The long-term complete remission rate of 68.4% and recurrence rate of 26.3% in the treatment group were respectively higher and lower than those in the control group (P < 0.01, and P < 0.01). The results suggested that the TCM decoctions were very helpful in treating this condition.