transforming-growth-factor-beta and Lupus-Nephritis

Although far from complete, the picture of cytokines present in systemic lupus erythematosus (SLE) glomerulonephritis is already complex. Proinflammatory cytokines, such as TNF, IL-6, IL-1, and IL-18 are upregulated, as are both Th1 and Th2 cytokines, with different implications. In many instances, the local effects may be different from the systemic immunoregulatory ones. For some proinflammatory cytokines, and TNF in particular, the local proinflammatory ones may be more relevant to the disease. This may help solve discrepancies between different murine models of the disease and provide a better rationale for targeting certain cytokines in human SLE.

Topics: Animals; Cytokines; Humans; Interferon-gamma; Interleukins; Lupus Nephritis; Mice; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

When last reviewed, bone morphogenetic protein 7 was presented as a potential new renal therapeutic agent, with multiple efficacies in chronic kidney disease. The object of this review is to describe progress from many sources since then in support or denial of the hypothesis.. Bone morphogenetic protein 7 has been shown to be an effective defence in several forms of chronic kidney disease in animal models, and its mechanisms of action have begun to be elucidated. Bone morphogenetic protein 7 inhibits tubular epithelial cell de-differentiation, mesenchymal transformation and apoptosis stimulated by various renal injuries. Bone morphogenetic protein 7 preserves glomerular integrity and inhibits injury-mediated mesangial matrix accumulation. In renal osteodystrophy, bone morphogenetic protein 7 affects osteoblast morphology and number, eliminates peritrabecular fibrosis, decreases bone resorption, and increases bone formation in secondary hyperparathyroidism. Bone morphogenetic protein 7 restores normal rates of bone formation in the adynamic bone disorder. Bone morphogenetic protein 7 is broadly efficacious in renal osteodystrophy, and importantly increases the skeletal deposition of ingested phosphorus and calcium, improving ion homeostasis in chronic kidney disease. Bone morphogenetic protein 7 was shown to prevent vascular calcification in a model of chronic kidney disease associated with the restoration of osteocalcin expression to normal tissue-restricted sites.. Bone morphogenetic protein 7 may be a powerful new therapeutic agent for chronic kidney disease, with the novel attribute of not only treating the kidney disease itself, but also directly inhibiting some of the most important complications of the disease state.

Topics: Bone Morphogenetic Protein 7; Bone Morphogenetic Proteins; Calcinosis; Chronic Kidney Disease-Mineral and Bone Disorder; Diabetic Nephropathies; Fibrosis; Humans; Kidney Diseases; Kidney Failure, Chronic; Lupus Nephritis; Nephritis, Hereditary; Transforming Growth Factor beta; Vascular Diseases

Lupus nephritis results from an acute inflammatory and immunological response to renal immune complex deposition. The acute response is characterized by activation of circulating leukocytes and renal parenchymal cells, triggering the production of pro-inflammatory cytokines and growth factors. In all too many cases, this response is followed by a chronic response, which is characterized by excessive deposition of collagen and other extracellular matrix macromolecules and the development of end-stage renal disease. Mechanisms underlying this chronic response in progressive renal disease are not adequately defined. In this overview, potential roles of reactive oxygen species (ROS) generation and transforming growth factor-beta (TGF-beta) production in the pathogenesis of lupus nephritis are considered. ROS and TGF-beta may be key elements of a pathway leading to persistent and excessive matrix deposition in progressive lupus nephritis. Further studies to define the role of this pathway in lupus nephritis may lead to the development of additional, more specific therapeutic targets to prevent progression of renal disease.

Topics: Animals; Disease Progression; Humans; Kidney; Kidney Diseases; Lupus Nephritis; Reactive Oxygen Species; Transforming Growth Factor beta

This study aims to elucidate the role of Kruppel-like factor (KLF5) and myxovirus resistance 1 (MX1) in the progression of renal fibrosis in lupus nephritis (LN).. First, the expression of KLF5 and MX1 was assessed in the peripheral blood of LN patients and healthy participants. Next, the pathological changes in renal tissues were evaluated and compared in BALB/c and MRL/lpr mice, by detecting the expression of fibrosis marker proteins (transforming growth factor-β [TGF-β] and CTGF) and α-SMA, the content of urine protein, and the levels of serum creatinine, blood urea nitrogen, and serum double-stranded DNA antibody. In TGF-β1-induced HK-2 cells, the messenger RNA levels of KLF5 and MX1 were tested by qRT-PCR, and the protein expression of α-SMA, type I collagen (Col I), fibronectin (FN), and matrix metalloproteinase 9 (MMP9) was measured by western blot analysis. Moreover, the relationship between KLF5 and MX1 was predicted and verified.. In renal tissues of MRL/lpr mice and the peripheral blood of LN patients, KLF5 and MX1 were highly expressed. Pearson analysis revealed that KLF5 was positively correlated with MX1. Furthermore, KLF5 bound to MX1 promoter and promoted its transcription level. MRL/lpr mice showed substantial renal injury, accompanied by increased expression of α-SMA, TGF-β, CTGF, Col I, FN, and MMP9. Injection of sh-KLF5 or sh-MX1 alone in MRL/lpr mice reduced renal fibrosis in LN, while simultaneous injection of sh-KLF5 and ad-MX1 exacerbated renal injury and fibrosis. Furthermore, we obtained the same results in TGF-β1-induced HK-2 cells.. Knockdown of KLF5 alleviated renal fibrosis in LN through repressing the transcription of MX1.

Topics: Animals; Fibrosis; Lupus Nephritis; Matrix Metalloproteinase 9; Mice; Mice, Inbred MRL lpr; Transforming Growth Factor beta; Transforming Growth Factor beta1

Lupus nephritis (LN) is the most common cause of morbidity and mortality in patients with systemic lupus erythematosus (SLE). Currently, immunosuppressive treatments for LN are suboptimal and can induce significant side effects. SB431542 is a selective and potent inhibitor of the TGFβ/Activin/NODAL pathway. Here, we study the effects of SB431542 treatment on LN and discuss the potential mechanisms. SB431542 ameliorated clinical outcomes with a consequent histological improvement in NZB/W mice. A comparative transcriptional profiling analysis revealed 586 differentially expressed genes (247 downregulated genes) in the SB431542 group compared to the control group. We found that the downregulated genes were mainly enriched in the biological processes of B cell activation, B cell proliferation, B cell differentiation, and B cell receptor signaling. Kyoto encyclopedia of genes and genomes pathway analysis revealed that the hematopoietic cell linage pathway was significantly downregulated in the SB431542 group. In addition, we observed that SB431542 reduced the splenic or renal levels of CD20 and the serum levels of anti-dsDNA antibody (IgG) in NZB/W mice. Furthermore, qRT-PCR and immunohistochemistry confirmed that SB431542 inhibits the production of TLR9, TGFβ1, and PDGFB. Thus, due to its immunomodulatory activities, SB431542 could be considered for clinical therapy development for LN.

Topics: Animals; Becaplermin; Lupus Nephritis; Mice; Mice, Inbred NZB; Proto-Oncogene Proteins c-sis; Toll-Like Receptor 9; Transforming Growth Factor beta

Systemic lupus erythematosus (SLE) is a chronic autoimmune inflammatory disease characterized by the involvement of multiple organs. Lupus nephritis (LN) is a major risk factor for overall morbidity and mortality in SLE patients. Hence, designing effective drugs is pivotal for treating individuals with LN. Fisetin plays a senolytic role by specifically eliminating senescent cells, inhibiting cell proliferation, and exerting anti-inflammatory, anti-oxidant, and anti-tumorigenic effects. However, limited research has been conducted on the utility and therapeutic mechanisms of fisetin in chronic inflammation. Similarly, whether the effects of fisetin depend on cell type remains unclear. In this study, we found that LN-prone MRL/lpr mice demonstrated accumulation of Ki-67-positive myofibroblasts and p15

Topics: Animals; Antioxidants; Epithelial Cells; Fibroblasts; Lupus Erythematosus, Systemic; Lupus Nephritis; Mice; Mice, Inbred MRL lpr; Transforming Growth Factor beta

The role of microRNA (miR)-183 has been elucidated in systemic lupus erythematosus, while whether it is also engaged in the lupus nephritis (LN) development remains opaque. The intention of this study is to examine the relevance of miR-183 downregulation in the pathogenesis of LN.. The expression of miR-183 was first detected in MRL/lpr mice at weeks 8 and 12, followed by the assessment the effects of miR-183 on renal fibrosis and inflammatory response after overexpression or silencing of miR-183 in mice with LN. We further overexpressed or knocked-down miR-183 in human renal glomerular endothelial cells (HRGECs), and detected the expression patterns of inflammatory factors and Vimentin and α-SMA in the cells. Differentially expressed genes in HRGECs overexpressing miR-183 by microarrays were intersected with targeting mRNAs of miR-183 predicted by bioinformatics websites. The effects of transforming growth factor beta receptor 1 (Tgfbr1) and TGF-β/Smad/TLR3 pathway on renal damage in mice were verified by rescue experiments.. miR-183 expression was notably lower in MRL/lpr mice, and increased miR-183 expression inhibited renal fibrosis and inflammatory response in mice with LN. Moreover, miR-183 inhibitor in HRGECs remarkably promoted the expression of Vimentin and α-SMA and the secretion of inflammatory factors. miR-183 protected the mouse kidney from pathological damages by targeting and inhibiting Tgfbr1 expression.. miR-183 inhibited the expression of Tgfbr1 by direct targeting to disrupt the TGF-β/Smad/TLR3 pathway, thus repressing renal fibrosis and the secretion of inflammatory factors in LN.

Topics: Animals; Cytokines; Fibrosis; Gene Expression Regulation; Humans; Kidney; Lupus Nephritis; Mice, Inbred C57BL; Mice, Inbred MRL lpr; MicroRNAs; Receptor, Transforming Growth Factor-beta Type I; Signal Transduction; Smad Proteins; Toll-Like Receptor 3; Transforming Growth Factor beta

Many patients with systemic lupus erythematosus (SLE) have lupus nephritis, one of the severe complications of SLE. We previously reported that CD8+CD103+ T regulatory cells induced ex vivo with transforming growth factor β (TGF-β) (iTregs) inhibited immune cells responses to ameliorate excessive autoimmune inflammation. However, the molecular mechanism(s) underlying the role of these CD8+ iTregs is still unclear. Here we identified that CD39, which is highly expressed on CD8+ iTregs, crucially contributes to the immunosuppressive role of the CD8+CD103+ iTregs. We showed that adoptive transfer of CD8+CD103+ iTregs significantly relieves the chronic graft-versus-host disease with lupus nephritis and CD39 inhibitor mostly abolished the functional activities of these CD8+ iTregs in vitro and in vivo. CD39+ cells sorted from CD8+CD103+ iTregs were more effective in treating lupus nephritis than CD39- partner cells in vivo. Furthermore, human CD8+ iTregs displayed increased CD103 and CD39 expressions, and CD39 was involved in the suppressive function of human CD8+ iTregs. Thus, our data implicated a crucial role of CD39 in CD8+CD103+ iTregs in treating lupus nephritis, and CD39 could be a new phenotypic biomarker for the identification of highly qualified CD8+ Tregs. This subpopulation may have therapeutic potential in patients with SLE nephritis and other autoimmune diseases.

Topics: Antigens, CD; Apyrase; CD8-Positive T-Lymphocytes; Cell Differentiation; Cell Proliferation; Chronic Disease; Graft vs Host Disease; Humans; Immune Tolerance; Immunomodulation; Integrin alpha Chains; Lupus Nephritis; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

Lupus nephritis (LN) is a common and severe complication of systemic lupus erythematosus. Without intervention, LN may cause acute kidney injury and end-stage renal disease. This study aims to determine whether microRNA-485 (miR-485) affects renal tubular epithelial cells (RTECs) in LN mice via the TGF-β-MAPK signaling pathway by targeting RhoA. Renal tissue samples were initially extracted from 15 LN and 15 normal mice. RTECs were cultivated in vitro and grouped after transfection of different mimics, inhibitors, or siRNA- RhoA. The target gene of miR-485 was confirmed by dual-luciferase reporter assay. Flow cytometry and MTT assay were applied to detect cell viability and apoptosis. It was determined that RhoA was a target gene of miR-485. We found that urine protein, creatinine, RhoA, interleukin 6 (IL-6), transforming growth factor-β1 (TGF-β1), and p38 mitogen-activated protein kinases (p38MAPK) were highly expressed in renal tissues of LN mice, while poor levels of miR-485 were recorded. The overexpression of miR-485 or siRNA- RhoA or the combination of miR-485 and siRNA- RhoA was demonstrated to lead to a reduction of levels of RhoA, IL-6, TGF-β, and p38MAPK, as well as a promotion of RTECs proliferation and inhibition of RTECs apoptosis. Taken together, these findings indicated that overexpressed miR-485 downregulates RhoA which could promote cell viability and inhibit apoptosis of RTECs by regulating the RhoA-mediated TGF-β-MAPK signaling pathway in LN mice.

Topics: Animals; Apoptosis; Blotting, Western; Cells, Cultured; Flow Cytometry; Lupus Nephritis; MAP Kinase Signaling System; Mice; MicroRNAs; rhoA GTP-Binding Protein; Signal Transduction; Transforming Growth Factor beta

To delineate urine biomarkers that forecast response to therapy of lupus nephritis (LN).. Starting from the time of kidney biopsy, patients with childhood-onset systemic lupus erythematosus who were diagnosed with LN were studied serially. Levels of 15 biomarkers were measured in random spot urine samples, including adiponectin, α-1-acid glycoprotein (AGP), ceruloplasmin, hemopexin, hepcidin, kidney injury molecule 1, monocyte chemotactic protein-1, lipocalin-like prostaglandin D synthase (LPGDS), transforming growth factor-β (TGF-β), transferrin, and vitamin D binding protein (VDBP).. Among 87 patients (mean age 15.6 yrs) with LN, there were 37 treatment responders and 50 nonresponders based on the American College of Rheumatology criteria. At the time of kidney biopsy, levels of TGF-β (p < 0.0001) and ceruloplasmin (p = 0.006) were significantly lower among responders than nonresponders; less pronounced differences were present for AGP, hepcidin, LPGDS, transferrin, and VDBP (all p < 0.05). By Month 3, responders experienced marked decreases of adiponectin, AGP, transferrin, and VDBP (all p < 0.01) and mean levels of these biomarkers were all outstanding (area under the receiver-operating characteristic curve ≥ 0.9) for discriminating responders from nonresponders. Patient demographics and extrarenal disease did not influence differences in biomarker levels between response groups.. Low urine levels of TGF-β and ceruloplasmin at baseline and marked reduction of AGP, LPGDS, transferrin, or VDBP and combinations of other select biomarkers by Month 3 are outstanding predictors for achieving remission of LN. If confirmed, these results can be used to help personalize LN therapy.

Topics: Adolescent; Biomarkers; Ceruloplasmin; Chemokine CCL2; Child; Female; Hepatitis A Virus Cellular Receptor 1; Humans; Intramolecular Oxidoreductases; Lipocalins; Lupus Nephritis; Male; Orosomucoid; Transferrin; Transforming Growth Factor beta; Treatment Outcome; Vitamin D-Binding Protein

To clarify the mechanisms underlying lupus nephritis (LN) amelioration following bortezomib treatment.. Bortezomib was administered subcutaneously every 3 days to NZB/W F1 mice, and the serum anti-double stranded (ds) deoxyribonucleic acid (DNA) antibody titers and proteinuria levels were measured. The renal samples and the splenocytes were examined histologically or used for real-time quantitative reverse transcription-polymerase chain reaction analysis after 18 weeks of treatment. Serum cytokine and anti-dsDNA antibody levels were measured using flow cytometry and enzyme-linked immunoassays every 3 weeks. Transforming growth factor (TGF)-β, angiotensin II type-1 receptor (AT1R), and type I collagen expression levels in the glomeruli were evaluated using immunohistochemistry.. Bortezomib reduced the serum anti-dsDNA antibody titers and the proteinuria levels. It prevented inflammatory cell infiltrations into and the deposition of immunoglobulin G within the glomeruli. Bortezomib reduced the interferon-γ, interleukin (IL)-4, and IL-10 levels in the serum and the ribonucleic acid expression levels for these cytokines within the splenocytes. Bortezomib prevented type I collagen synthesis by downregulating TGF-β and AT1R expression in the glomeruli.. Bortezomib exerts multiple immunosuppressive effects and thus ameliorates LN. Furthermore, bortezomib can prevent glomerulosclerosis formation in NZB/W F1 mice through suppressive effects on the renin-angiotensin system.

Topics: Animals; Bortezomib; Cytokines; Disease Models, Animal; Female; Immune System; Immunoglobulin G; Immunosuppressive Agents; Interferon-gamma; Interleukin-10; Kidney; Kidney Glomerulus; Lupus Nephritis; Mice; Mice, Inbred NZB; Renin-Angiotensin System; Transforming Growth Factor beta

Lupus nephritis is a serious organ involvement with unknown etiology, and glomerulonephritis class IV is one of the most severe forms of the disease which correlates with poor prognosis and death. Immunological abnormalities are implicated in the expression of lupus nephritis. In this study, we examined some T helper 17 and regulatory T-related cytokines and molecules in systemic lupus erythematosus patients with glomerulonephritis class IV.. The study group comprised of 20 glomerulonephritis class IV SLE patients and 20 sex- and age-matched SLE patients without kidney involvement as control group. Blood samples was collected from each participant, lymphocytes were isolated, and RNA was extracted from lymphocytes. Then cDNA was synthesized using reverse transcription enzyme, and finally using specific primers and probes, the expression levels of forkhead box P3 (Foxp3), transforming growth factor (TGF)-β, interferon (IFN)-γ, interleukin (IL)-6, and IL-17 genes were analyzed by real-time polymerase chain reaction based on the TaqMan method.. The expression levels of IL-6, IL-17, IFN-γ, and Foxp3 genes were significantly higher in SLE patients with glomerulonephritis class IV than those with non-nephritis SLE. However, the expression of TGF-β was not significantly different between the SLE patients with and without glomerulonephritis class IV involvement.. According to our results, it seems that in class IV glomerulonephritis patients, increased Foxp3-producing regulatory T cells has an imperfect capacity to control the pathogenic IL-17- and IFN-γ-producing cells.

Topics: Adult; Case-Control Studies; Female; Forkhead Transcription Factors; Gene Expression; Humans; Interferon-gamma; Interleukin-17; Interleukin-6; Lupus Nephritis; Male; Real-Time Polymerase Chain Reaction; T-Lymphocytes, Helper-Inducer; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

Lupus nephritis (LN) is an autoimmune disease that occurs when autoantibodies complex with self-antigen and form immune complexes that accumulate in the glomeruli. These immune complexes initiate an inflammatory response resulting in glomerular injury. LN often concomitantly affects the tubulointerstitial compartment of the kidney, leading first to interstitial inflammation and subsequently to interstitial fibrosis and atrophy of the renal tubules if not appropriately treated. Presently the only way to assess interstitial inflammation and fibrosis is through kidney biopsy, which is invasive and cannot be repeated frequently. Hence, monitoring of disease progression and response to therapy is suboptimal. In this paper we describe a mathematical model of the progress from tubulointerstitial inflammation to fibrosis. We demonstrate how the model can be used to monitor treatments for interstitial fibrosis in LN with drugs currently being developed or used for nonrenal fibrosis.

Topics: Biomarkers; Cell Count; Chemokine CCL2; Disease Progression; Epithelial Cells; Extracellular Matrix; Fibrosis; Humans; Kidney; Kidney Tubules; Lupus Nephritis; Macrophages; Mathematical Concepts; Matrix Metalloproteinases; Models, Biological; Myofibroblasts; Nephritis, Interstitial; Platelet-Derived Growth Factor; Tissue Inhibitor of Metalloproteinases; Transforming Growth Factor beta

Systemic lupus erythematosus (SLE) is an autoimmune systemic disease caused as a result of an imbalance of Th1-/Th2-type cytokines. The soluble form of CD30 (CD30s) released from peripheral blood cells has been described as a marker of active disease in Th2-type immune response as in SLE. However, the expression of CD30 on CD3 T lymphocytes from patients with SLE has not been studied yet. Therefore, we have addressed our study to attempt this issue, studying CD30 expression by flow cytometry on CD3 T lymphocytes and CD4/CD8 subsets in samples from SLE patients mainly with lupus nephritis. In parallel, we have determined the production of the cytokines IL-4 (Th2), IFNγ (Th1), IL-10 and TGFβ by intracellular staining. Differences between positive CD30 T cells in healthy controls and patients with SLE were found, with a higher percentage of CD30-expressing T cells in patients with SLE (P = 0.001). In contrast to healthy controls, CD30 was mainly expressed on CD8 T cells from patients with SLE. The intracellular cytokine staining showed that TGFβ is the main cytokine expressed in CD3 T cells from patients with SLE. In addition to this, we have found a positive correlation between CD30-expressing T cells and IL-4, IFNγ, and immunosuppressive cytokines (IL-10 and TGFβ) (P < 0.05). These results suggest that CD30 could play a role in the pathogenesis of SLE and its expression on CD3 T lymphocytes is not restricted only to Th2-type response.

Topics: Adult; Biomarkers; CD3 Complex; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Female; Humans; Interferon-gamma; Interleukin-10; Interleukin-4; Interleukin-8; Ki-1 Antigen; Lupus Erythematosus, Systemic; Lupus Nephritis; Male; T-Lymphocyte Subsets; Transforming Growth Factor beta

Regulatory T (Treg) cells play an important role in the maintenance of immune tolerance to self and in the pathogenesis of autoimmune disease. Transforming growth factor-beta 1(TGF-β1) is a regulatory cytokine with pleiotropic properties in immune responses. This study was to investigate the role of Treg cells and TGF-β1 in the pathogenesis of patients with lupus nephritis (LN). A total of 42 new-onset systemic lupus erythematosus patients and 22 healthy controls were enrolled. The proportion of Treg cells in peripheral blood mononuclear cells (PBMCs) was evaluated by flow cytometric analysis. The serum and urinary TGF-β1 levels were measured by enzyme-linked immunosorbent assay (ELISA). The results demonstrated a significant decrease in the frequency of CD4(+)CD25(high) and CD4(+)CD25(+)FoxP3(+) T cells in LN patients. The concentration of serum TGF-β1 was found decreased in SLE patients, while urinary TGF-β1 levels were significantly higher in LN patients. Based on our results, decreased Treg cells were accompanied with lower serum TGF-β1 levels and higher urinary TGF-β1 levels in LN patients. TGF-ß1 levels in serum may play a key role in the pathogenesis of renal impairment while the significantly increased urinary TGF-β1 levels may be used as a biological marker in prediction of lupus nephritis.

Topics: Adult; Biomarkers; CD4 Antigens; Cell Count; Cell Separation; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Forkhead Transcription Factors; Humans; Interleukin-2 Receptor alpha Subunit; Kidney; Lupus Erythematosus, Systemic; Lupus Nephritis; Male; Middle Aged; Prognosis; T-Lymphocyte Subsets; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

Anti-double-stranded DNA (dsDNA) antibodies have been indicated to play a major role in the pathogenesis of lupus nephritis (LN), which is characterized by mesangial alterations, including phenotypic changes. To explore the effects of anti-dsDNA antibodies on the phenotype of mesangial cells (MCs), the anti-dsDNA IgG in sera and histological features of glomeruli were analyzed in the mice models of immune-complex glomerulonephritis. The MCs were cultured in vitro with the addition of anti-dsDNA or non-anti-dsDNA IgG. Compared to the anti-dsDNA-negative controls, the serum positive mice had increased extracellular matrix accumulation and higher alpha-smooth muscle actin expression in the mesangial region. The anti-dsDNA IgG enhanced the synthesis of transforming growth factor beta, alpha-smooth muscle actin, and fibronectin, and even induced the myofibroblast-like morphological features in cultured MCs. Our results indicated that anti-dsDNA antibodies contribute to the phenotypic changes in MCs, which suggests another mechanism of renal injuries in LN induced by anti-dsDNA antibodies.

Topics: Actins; Animals; Antibodies, Antinuclear; Cell Shape; Cells, Cultured; Extracellular Matrix; Fibronectins; Gene Expression Regulation; Immunoglobulin G; Lupus Nephritis; Male; Mesangial Cells; Mice; Mice, Inbred C57BL; Myofibroblasts; Signal Transduction; Transforming Growth Factor beta

N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) is an endogenous tetrapeptide which can inhibit the differentiation, migration and activation of macrophages and suppress the proliferation of fibroblast. This study examined the effects of Ac-SDKP on the progression of lupus nephritis (LN). MRL/lpr mice received subcutaneous infusion of Ac-SDKP (1.0 mg kg(-1) d(-1)) or vehicle through implanted osmotic mini-pumps from 12 to 20 weeks until being euthanized. MRL/MpJ mice served as normal controls. The data indicative of renal inflammation and fibrosis were evaluated before and after treatment. Ac-SDKP-treated MRL/lpr mice showed reduced proteinuria and improved renal function compared with vehicle-treated controls. Ac-SDKP-treated mice demonstrated decreased inflammatory infiltrates of T cells and macrophages in the kidneys as compared to vehicle-treated animals. The treatment also inhibited the activation of NF-κB and production of TNF-α. Despite this, immune complex deposition and plasma anti-dsDNA levels were not statistically different between the two groups. In addition, the treatment inhibited renal expression of TGF-β1, α-SMA and fibronectin as well as the phosphorylation of Smad2/3. Ac-SDKP treatment ameliorated LN through exerting anti-inflammatory and anti-fibrotic effects on MRL/lpr mice, providing therapeutic potential for halting the progression of LN.

Topics: Actins; Animals; Antibodies, Antinuclear; Body Weight; Chemokine CCL2; Drug Administration Schedule; Female; Fibronectins; Gene Expression Regulation; Inflammation; Kidney; Leukocytes; Lupus Nephritis; Mice; Mice, Inbred MRL lpr; NF-kappa B; Oligopeptides; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Recent studies suggest that vascular endothelial growth factor (VEGF) plays a crucial role in the preservation of renal function and may also serve as a useful biomarker in monitoring the progression of lupus nephritis (LN). Here we sought to correlate intrarenal VEGF expression with renal histopathology and prognosis of LN. Biopsy specimens from 35 patients with Class III or IV LN (ISN/RPS categorization) were found to have lower levels of intrarenal VEGF than those found in biopsy tissue taken from 10 donor kidneys sampled at the time of allograft reperfusion. This reduced amount of VEGF mRNA in the patients with LN negatively correlated with glomerular endocapillary proliferation, crescent formation, and a high histologic activity index but was positively associated with increased numbers of urinary podocytes. The level of intrarenal VEGF mRNA accurately predicted the deterioration of renal function in these patients within 12 months. Our study shows that expression of VEGF in renal tissue may serve as a molecular marker of renal damage and may be a predictive factor for short-term loss of kidney function in patients with LN.

Topics: Adult; Angiopoietin-1; Biomarkers; Case-Control Studies; Female; Gene Expression; Heme Oxygenase-1; Humans; Kidney; Lupus Nephritis; Male; Podocytes; Prognosis; RNA, Messenger; Time Factors; Transforming Growth Factor beta; Urine; Vascular Endothelial Growth Factor A

The aim of this study was to determine the relationship between histological, immunohistochemical (IHC) and biological data in the assessment of interstitial fibrosis in patients with glomerular diseases. A group of 41 patients with primary and secondary glomerulonephritis was studied. In order to quantify the histological changes and to assess the extent of active-inflammatory and chronic-sclerotic/fibrotic interstitial lesions, we adapted a scoring system, initially used for lupus nephritis, and ANCA-associated vasculitis. IHC labeling procedures with monoclonal antibodies anti-smooth muscle actin (SMA), anti-vimentin and anti-transforming growth factor beta (TGFbeta) were assessed using a semi-quantitative score, correlated with the histological and biological data. Our results showed that interstitial labeling of SMA correlated with scores for sclerotic/fibrotic lesions (chronicity index) and with active-inflammatory lesions (interstitial infiltrate, activity index). Interstitial vimentin correlated with the score for interstitial infiltrate. Both interstitial vimentin and TGFbeta immunopositivity correlated with sclerotic/fibrotic lesions (interstitial fibrosis, tubular atrophies, vascular hyalinosis/fibrosis, chronicity index), and negatively with glomerular filtration rate. An important correlation was found between the interstitial labeling of the two IHC markers of myofibroblasts (SMA and vimentin). We conclude that IHC studies related to clinico-biological and histological data can have an important role in the evaluation of the glomerular diseases, but the classical histological investigation assessed through quantification has still not lost its importance.

Topics: Actins; Adolescent; Adult; Aged; Chronic Disease; Female; Fibrosis; Glomerulonephritis; Humans; Immunohistochemistry; Kidney; Linear Models; Lupus Nephritis; Male; Middle Aged; Transforming Growth Factor beta; Vimentin

B cell depletion may affect T cell activation and costimulation status in rituximab-treated patients with SLE. We examined whether rituximab administration in patients with active lupus nephritis is related to changes in mRNA expression of genes that define regulatory T cells (Tregs) in peripheral blood lymphocytes, measured by real-time PCR. At the early phase of B cell depletion mRNA levels of CD25, CTLA-4, GITR and the bona fide Treg functional marker FOXP3 increased significantly in all 7 patients examined. In contrast, mRNA levels of the costimulatory/activation T cell molecule CD40L were profoundly reduced, while mRNA levels of TGF-beta, a cytokine contributing to Treg induction, increased significantly in all. During follow-up, increased FOXP3 mRNA persisted in those patients in clinical remission, while in those patients with active disease subsequent decreases were noted. Further studies should examine whether modulation of Tregs by therapeutic B cell depletion contributes and/or predicts lupus disease remission.

Topics: Adolescent; Adult; Antibodies, Monoclonal; Antibodies, Monoclonal, Murine-Derived; B-Lymphocytes; CD40 Ligand; Female; Forkhead Transcription Factors; Gene Expression; Humans; Immunologic Factors; Lupus Nephritis; Lymphocyte Activation; Lymphocyte Depletion; Male; Remission Induction; Reverse Transcriptase Polymerase Chain Reaction; Rituximab; RNA, Messenger; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

Lupus nephritis is characterised by intrarenal inflammation and lymphocyte activation.. To examine the profile of cytokine gene expression in glomerulus and tubulointerstitium in patients with lupus nephritis.. 36 consecutive patients with systemic lupus erythematosus having active renal disease were recruited, and they were required to undergo kidney biopsy. Glomerular and tubulointestitial cytokine expression of interleukin (IL)2, 4, 10, 12, 18, interferon gamma (IFN)gamma, T-bet (the Th1 transcription factor), GATA-3 (the Th2 transcription factor), transforming growth factor beta and monocyte chemoattractant protein (MCP)1 were studied by laser microdissection of the renal biopsy specimen, followed by real-time quantitative PCR.. There were 13 patients with World Health Organization class III nephritis, 14 patients with class IV nephritis and 9 patients with class V nephritis. There was a significant correlation between serum C3, C4 and anti-double strand DNA antibody level with glomerular expression of T-bet, IFNgamma and IL2. There was a significant correlation between histological activity index and glomerular expression of IL12, IL18, IL10 and MCP1. In addition, the degree of glomerular leucocyte infiltration significantly correlated with glomerular expression of IFNgamma, IL10, IL12 and IL18. By contrast, histological chronicity index correlated with the tubulointerstitial expression of IL2, MCP1 and GATA-3.. Intraglomerular expression of certain target genes correlate with the severity of systemic as well as histological activity, whereas the tubulointerstitial expression of other target genes correlate with the degree of chronic kidney scarring. This result may shed light on the immunopathogenesis of lupus nephritis.

Topics: Adult; Chemokine CCL2; Cytokines; Female; GATA3 Transcription Factor; Gene Expression; Humans; Interferon-gamma; Interleukins; Kidney; Kidney Glomerulus; Kidney Tubules; Leukocytes; Lupus Nephritis; Male; T-Box Domain Proteins; Transforming Growth Factor beta

Previous studies have shown that messenger RNA (mRNA) expression of target genes is increased in the urinary sediment of patients with active lupus. We study the effect of immunosuppressive therapy on the urinary gene expression profile in patients with active lupus nephritis. Method. We recruited nine patients with active systemic lupus erythematosus (SLE) and renal disease, and required corticosteroid, with or without cytotoxic treatment. They were followed for 6 months, urine samples were collected at 0, 4, 12 and 24 weeks and gene expression profile was determined by polymerase chain reactions. The pattern of gene expression was compared to clinical parameters of therapeutic response.. Amongst the target genes studied, there was a progressive decline in the urinary expression of T-bet, interleukin (IL)-10, transforming growth factor-beta (TGF-beta), monocyte chemoattractant protein-1 (MCP-1), and interferon-gamma (IFN-gamma) after immunosuppressive treatment, although the change of IFN-gamma was not statistically significant. The time course of their urinary expression was parallel to the systemic activity as reflected by the systemic lupus erythematosus disease activity index (SLEDAI). Throughout the study period, the SLEDAI score correlated significantly with the expressions of IFN-gamma (r = 0.43, P = 0.009), T-bet (r = 0.40, P = 0.016), TGF-beta (r = 0.51, P = 0.002) and MCP-1 (r = 0.38, P = 0.022). The anti-double strand(anti-ds)DNA antibody titer correlated significantly with the expressions of IFN-gamma (r = 0.45, P = 0.009), T-bet (r = 0.37, P = 0.034), IL-10 (r = 0.59, P<0.001), TGF-beta (r = 0.44, P = 0.010) and MCP-1 (r = 0.49, P = 0.004). On the other hand, the expression level of IL-2, IL-4, IL-12, IL-18 and GATA-3 remained static throughout the study period.. The mRNA expression of T-bet, IL-10, TGF-beta, MCP-1, and probably IFN-gamma in the urinary sediment of patients with active lupus nephritis improves with successful immunosuppressive therapy, and the change in gene expression profile is in phase with the clinical disease activity. Measurement of urinary mRNA expression of target genes may be a potential non-invasive tool for the monitoring of lupus disease activity.

Topics: Adult; Biomarkers; Chemokine CCL2; Drug Monitoring; Female; Gene Expression Profiling; Gene Expression Regulation; Humans; Immunosuppressive Agents; Interferon-gamma; Interleukin-10; Lupus Nephritis; Middle Aged; RNA, Messenger; T-Box Domain Proteins; Transforming Growth Factor beta

Noninvasive molecular tests of urine cells have been developed to monitor the activity of kidney diseases. We evaluate whether measurement of urinary messenger RNA (mRNA) levels of chemokine and growth factor genes could distinguish between diffuse proliferative lupus nephritis (class IV LN) and others and whether it is able to predict the response to therapy. Prebiopsy urine samples were collected from 26 LN patients. Urine specimens were serially collected over a period of 6 months from class IV LN patients who were receiving standard immunosuppressive treatments. Urinary interferon-producing protein 10 and its CXC chemokine receptor (CXCR)3, transforming growth factor-beta (TGF-beta), and vascular endothelial growth factor (VEGF) mRNA levels were analyzed by quantitative real-time polymerase chain reactions. Levels of chemokine or growth factor mRNAs in urine could distinguish class IV LN from others, with a sensitivity of 85% and a specificity of 94%. The receiver-operative characteristic curve demonstrated that urine mRNA levels of these genes could identify active class IV LN with an accuracy greater than the current available clinical markers, namely systemic lupus erythematosus (SLE) disease activity index, proteinuria, renal function, or urinalysis. A significant reduction of interferon-producing protein 10 (IP-10), CXCR3, TGF-beta, and VEGF mRNA levels from baselines was observed in patients who responded to therapy, whereas the levels tended to increase in those who resisted to treatment. Measurement of urinary chemokine and growth factor mRNAs can precisely distinguish class IV LN from others. Temporal association between these markers and therapeutic response is demonstrated. This noninvasive approach serves as a practical tool in diagnosis and management of LN.

Topics: Adult; Biomarkers; Chemokine CXCL10; Chemokines; Disease Progression; Female; Gene Expression Regulation; Growth Substances; Humans; Immunosuppressive Agents; Longitudinal Studies; Lupus Nephritis; Male; Receptors, Chemokine; Receptors, CXCR3; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sensitivity and Specificity; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A

We induced very low-dose tolerance by injecting lupus prone (SWR x NZB)F1 (SNF1) mice with 1 mug nucleosomal histone peptide autoepitopes s.c. every 2 wk. The subnanomolar peptide therapy diminished autoantibody levels and prolonged life span by delaying nephritis, especially by reducing inflammatory cell reaction and infiltration in kidneys. H4(71-94) was the most effective autoepitope. Low-dose tolerance therapy induced CD8+, as well as CD4+ CD25+ regulatory T (Treg) cell subsets containing autoantigen-specific cells. These adaptive Treg cells suppressed IFN-gamma responses of pathogenic lupus T cells to nucleosomal epitopes at up to a 1:100 ratio and reduced autoantibody production up to 90-100% by inhibiting nucleosome-stimulated T cell help to nuclear autoantigen-specific B cells. Both CD4+ CD25+ and CD8+ Treg cells produced and required TGF-beta1 for immunosuppression, and were effective in suppressing lupus autoimmunity upon adoptive transfer in vivo. The CD4+ CD25+ T cells were partially cell contact dependent, but CD8+ T cells were contact independent. Thus, low-dose tolerance with highly conserved histone autoepitopes repairs a regulatory defect in systemic lupus erythematosus by generating long-lasting, TGF-beta-producing Treg cells, without causing allergic/anaphylactic reactions or generalized immunosuppression.

Topics: Adoptive Transfer; Amino Acid Sequence; Animals; Antibodies, Antinuclear; Autoantigens; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Epitopes; Female; Immune Tolerance; Immunosuppression Therapy; Lupus Erythematosus, Systemic; Lupus Nephritis; Mice; Mice, Inbred NZB; Molecular Sequence Data; Nuclear Proteins; Nucleosomes; Peptides; Receptors, Interleukin-2; T-Lymphocyte Subsets; Transforming Growth Factor beta

Renal biopsies of 69 patients with lupus nephritis were studied according to the WHO classification. The aim of the present study was to correlate the interstitial tryptase-positive mast cells with the interstitial TGF-beta1 and alpha-SM actin expression and clinical outcome of lupus nephritis, and identify the pathological role of the interstitial tryptase-positive mast cells in lupus nephritis. The mean follow-up time was 70.7 +/- 54.4 months. Eight patients were grouped as WHO class II lupus nephritis, 15 patients as class III, 28 patients as class IV and 18 patients as class V. Interstitial tryptase-positive mast cells were not correlated with clinical outcome and interstitial TGF-beta1 expression in lupus nephritis. Interstitial tryptase-positive mast cells were correlated with tubulo-interstitial alpha-SM actin expression for WHO class V lupus nephritis, but not to the other classes. In conclusion, in spite of interstitial tryptase-positive mast cells being related to renal interstitial fibrosis process, their expression according to the clinical outcome of lupus nephritis was not significant.

Topics: Actins; Adult; Biomarkers; Female; Fibroblasts; Humans; Kidney; Lupus Nephritis; Male; Mast Cells; Myocytes, Smooth Muscle; Serine Endopeptidases; Transforming Growth Factor beta; Transforming Growth Factor beta1; Tryptases

The experimental data and the study on human renal tissue in patients with glomerulonephropathies indicate that monocyte chemoattractant protein-1 (MCP-1) plays a main role in progression of inflammatory processes in kidney diseases. Monocytes/macrophages are multifunctional cells that may regulate matrix accumulation by producing transforming growth factor beta-1 (TGF-beta-1), which plays an important role in the progression of renal diseases. The present study was undertaken to evaluate the relationships between the immunoexpression of MCP-1, the number of CD68-positive cells, the immunoexpression of TGF-beta-1 and the extent of renal fibrosis as well serum creatinine level in patients with lupus nephritis. Using immunohistochemistry we analyzed the expression of MCP-1, TGF-beta-1 and the number of CD68+ cells in renal biopsy specimens in 17 patients with IV class of lupus nephritis and in 10 normal kidneys. Statistical analysis revealed significant increase in the tubulointerstitial MCP-beta immunostaining in lupus nephritis as compared to normal controls. In lupus nephritis the amount of glomerular and interstitial CD68+ cells was higher than in control group. None of the control sections have evidence of glomerular or tubulointerstitial immunoexpression of TGF-beta-1. In patients with lupus nephritis TGF-beta-1 was detected in the renal tubular epithelial cells and the interstitium, and to a lesser extent within glomeruli. The tubulointerstitial MCP-1 immunoexpression was significantly correlated with monocyte/macrophage interstitial infiltrates, the immunoexpression of TGF-beta-1 in tubuli and interstitium as well as serum creatinine. Moreover, the tubulointerstitial immunoexpression of TGF-beta-1 was significantly positively correlated with renal interstitial cortical volume and serum creatinine in patients with lupus nephritis. In summary, these data suggest that in lupus nephritis MCP-1 may play a role in modulating interstitial inflammatory process and in tubulointerstitial renal damage via TGF-beta-1 pathway.

Topics: Adolescent; Adult; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Chemokine CCL2; Creatinine; Female; Humans; Immunohistochemistry; Inflammation; Lupus Nephritis; Macrophages; Male; Middle Aged; Transforming Growth Factor beta; Transforming Growth Factor beta1

Angiotensin-converting enzyme (ACE) inhibitors, such as captopril, are used to control hypertension. In patients and animals with primary nephropathies, these agents improve renal function more than that would be expected from their control of hypertension. Here, we examine the effects of treatment with captopril on lupus nephritis and discuss the potential mechanism(s) by which this agent exerts its renoprotective effects.. Lupus-prone, NZB/NZW F1 and MRL-lpr/lpr, mice were treated with captopril or with a control antihypertensive agent, verapamil. Mice were monitored for nephritis, and their sera and tissues analyzed for cytokine and transforming growth factor-beta (TGF-beta) expression.. Captopril treatment delayed the onset of proteinuria when administered to prenephritic mice, whereas verapamil did not. Captopril treatment also retarded disease progression when given to lupus mice that had early disease, and even reversed severe proteinuria in at least some older animals with advanced disease. It reduced chronic renal lesions, but had no effect on autoantibody production. The improvement in renal disease correlated with reduced TGF-beta expression, particularly of the TGF-beta1 and TGF-beta2 isoforms, in the kidneys. Interestingly, in vivo or in vitro exposure to captopril reduced splenic levels of type 2 cytokines, interleukin (IL)-4 and IL-10, suggesting a possible role of the immune system in captopril-mediated disease modulation.. Since type 2 cytokines are known to promote lupus glomerulosclerosis, decreased IL-4 and IL-10 production in captopril-treated mice may be related to this agent's renoprotective effects. We argue here that ACE inhibitors not only act as selective TGF-beta inhibitors, but also as selective immunomodulators, to improve lupus nephritis.

Topics: Angiotensin-Converting Enzyme Inhibitors; Animals; Autoantibodies; Captopril; Chronic Disease; Cytokines; Gene Expression; Interleukin-10; Interleukin-4; Kidney; Lupus Nephritis; Mice; Mice, Inbred BALB C; Mice, Inbred MRL lpr; Mice, Inbred NZB; RNA, Messenger; Spleen; Th2 Cells; Transforming Growth Factor beta; Transforming Growth Factor beta1; Transforming Growth Factor beta2

Slowly progressive renal injury is the major cause for ESRD. The model of progressive immune complex glomerulonephritis in autoimmune MRL(lpr/lpr) mice was used to evaluate whether chemokine receptor CCR1 blockade late in the disease course can affect progression to renal failure. Mice were treated with subcutaneous injections of either vehicle or BX471, a nonpeptide CCR1 antagonist, three times a day from week 20 to 24 of age [corrected]. BX471 improved blood urea nitrogen levels (BX471, 35.1 +/- 5.3; vehicle, 73.1 +/- 39.6 mg/dl; P < 0.05) and reduced the amount of ERHR-3 macrophages, CD3 lymphocytes, Ki-67 positive proliferating cells, and ssDNA positive apoptotic cells in the interstitium but not in glomeruli. Cell transfer studies with fluorescence-labeled T cells that were pretreated with either vehicle or BX471 showed that BX471 blocks macrophage and T cell recruitment to the renal interstitium of MRL(lpr/lpr) mice. This was associated with reduced renal expression of CC chemokines CCL2, CCL3, CCL4, and CCL5 and the chemokine receptors CCR1, CCR2, and CCR5. Furthermore, BX471 reduced the extent of interstitial fibrosis as evaluated by interstitial smooth muscle actin expression and collagen I deposits, as well as mRNA expression for collagen I and TGF-beta. BX471 did not affect serum DNA autoantibodies, proteinuria, or markers of glomerular injury in MRL(lpr/lpr) mice. This is the first evidence that, in advanced chronic renal injury, blockade of CCR1 can halt disease progression and improve renal function by selective inhibition of interstitial leukocyte recruitment and fibrosis.

Topics: Animals; Autoantibodies; Blood Urea Nitrogen; CD3 Complex; CD8-Positive T-Lymphocytes; Chemokines; Disease Progression; DNA; DNA, Single-Stranded; Fibrosis; Glomerulonephritis; In Situ Hybridization; Ki-67 Antigen; Kidney; Leukocytes; Lupus Nephritis; Lymphocytes; Macrophages; Mice; Mice, Inbred MRL lpr; Microscopy, Fluorescence; Phenylurea Compounds; Piperidines; Receptors, CCR1; Receptors, Chemokine; Renal Insufficiency; Reverse Transcriptase Polymerase Chain Reaction; RNA; RNA, Messenger; T-Lymphocytes; Time Factors; Transforming Growth Factor beta; Transforming Growth Factor beta1

Transforming growth factor-beta1 (TGF-beta1) is involved in the generation of CD8+ T suppressor cells, natural killer (NK) cells and regulatory T (Th3) cells for down-regulatory effects on antibody production. We studied TGF-beta1 activity in patients with systemic lupus erythematosus (SLE) to try to clarify whether the dysregulation by TGF-beta1 is genetically determined. Sera from 55 patients with clinically inactive SLE, who were taking minimal steroids and/or hydroxychloroquine, and 40 healthy controls, along with supernatants from concanavalin A-stimulated peripheral blood mononuclear cell (PBMC) cultures from 18 patients with SLE and 10 controls were subjected to TGF-beta1 enzyme-linked immunosorbent assay. A total of 138 patients with SLE and 182 controls were genotyped for 5 single-nucleotide polymorphisms (SNPs) of TGF-beta1: -988C/A, -800G/A, -509C/T, Leu10/Pro10 and Arg25/Pro25. Patients with SLE had lower serum levels of TGF-beta1 compared with controls (p=0.052). The unstimulated and stimulated TGF-beta1 production of PBMCs in patients with SLE was higher than in controls, although these differences did not reach significance (p=0.073 and 0.074, respectively). None of the TGF-beta1 SNPs was strongly associated with SLE in Taiwanese patients or had any prognostic significance in lupus nephritis. Hence these polymorphisms do not represent a genetic predisposition to SLE. The intrinsic capability of immunoregulation for spontaneous B cell hyperactivity through PBMC TGF-beta1 production was presumed to be intact in clinically stable SLE in Taiwanese. Whether the lower serum TGF-beta1 level that causes the defective immune regulation in SLE is primarily under genetic control or secondary to the influence of ongoing cellular interactions in the cytokine context needs to be elucidated.

Topics: Adult; Case-Control Studies; Female; Genetic Predisposition to Disease; Genotype; Humans; Lupus Erythematosus, Systemic; Lupus Nephritis; Male; Polymorphism, Single Nucleotide; Taiwan; Transforming Growth Factor beta; Transforming Growth Factor beta1

Lupus nephritis is characterized by intrarenal inflammation. To assess the extent and severity of disease activity and renal involvement, this study examined the expression of transforming growth factor beta (TGFbeta) and monocyte chemoattractant protein 1 (MCP-1) in the urinary sediment of patients with systemic lupus erythematosus (SLE).. We studied 106 patients with SLE who were classified according to their disease status as those with active disease, those with disease in remission, and those with nonrenal SLE. Ten healthy subjects were used as controls. Lupus activity was assessed by the SLE Disease Activity Index (SLEDAI). If renal biopsy was performed, the histologic activity index and chronicity index were determined, and a morphometry analysis of renal scarring was performed. The urinary expresssion of TGFbeta and MCP-1 messenger RNA (mRNA) was studied by real-time quantitative polymerase chain reaction, and the corresponding protein concentrations of TGFbeta and MCP-1 in the urine were measured by enzyme-linked immunosorbent assay (ELISA).. Expression of TGFbeta and MCP-1 mRNA in the urinary sediment was significantly elevated in the active disease group (P < 0.001 for both). These expression levels of TGFbeta and MCP-1 mRNA correlated with the SLEDAI score (TGFbeta r = 0.71, P < 0.001; MCP-1 r = 0.72, P < 0.001), and also significantly correlated with the histologic activity index (TGFbeta r = 0.487, P = 0.004; MCP-1 r = 0.357, P = 0.038). The urinary protein concentration of MCP-1, but not of TGFbeta, correlated with the SLEDAI score (r = 0.66, P < 0.001). However, neither the protein concentration of TGFbeta nor that of MCP-1 as measured by ELISA in the urine correlated with the histologic activity index.. The measurement of urinary mRNA expression may be a noninvasive method for the assessment of lupus disease activity and the severity of renal involvement in patients with lupus nephritis.

Topics: Adult; Biopsy; Chemokine CCL2; Female; Gene Expression; Humans; Kidney; Lupus Nephritis; Male; Middle Aged; RNA, Messenger; Severity of Illness Index; Transforming Growth Factor beta; Urine

Inducible, high-output nitric oxide (NO) production has been identified as a central mediator of cell injury in immune-mediated renal disease. In acute anti-thy-1 glomerulonephritis prefeeding with the NO precursor L-arginine increases mesangial cell injury and the subsequent fibrosis. The present study tested the hypothesis that L-arginine supplementation may also be detrimental in chronic, NO-mediated murine lupus nephritis.. Groups (N = 18) of female MRL/lpr mice with lupus nephritis were fed the following diets: (1) normal protein (22% casein); (2) normal protein and 1.0% L-arginine in the drinking water; (3) low protein (6% casein); (4) low protein + 0.4%l-arginine; and (5) low protein + 1.0% L-arginine. After 40 days mouse survival, albuminuria, matrix accumulation, inflammatory cell infiltration, immunoglobulin G (IgG) deposition, expression of transforming growth factor-beta 1 (TGF-beta 1), fibronectin and plasminogen activator inhibitor-1 (PAI-1) mRNA and protein, anti-DNA antibody titer, inducible nitric oxide synthase (iNOS) mRNA expression, blood amino acid levels, blood urea nitrogen (BUN) concentrations and blood and urinary NOx (nitrite + nitrate) levels were assessed.. L-Arginine supplementation increased mortality significantly (P < 0.02). The death rate increased from 0% in the lowest to 50% in the highest L-arginine intake group (normal protein + 1.0% L-arginine). L-Arginine administration increased albuminuria, renal matrix accumulation, TGF-beta 1, fibronectin, PAI-1, blood L-arginine, L-citrulline, BUN and blood and urine NOx levels, while protein restriction reduced these parameters. Renal cell infiltration and iNOS mRNA expression were decreased in the low protein group only. Anti-ds DNA-IgG and renal IgG deposition were comparable in all groups. Increasing L-arginine intake increases the severity of renal fibrosis and the likelihood of death in MRL/lpr mice. The results appear to be at least in part mediated through enhanced cytotoxic NO generation via iNOS. The data suggest that L-arginine restriction should be considered in human immune-mediated renal diseases.

Topics: Albuminuria; Amino Acids; Animals; Arginine; Autoantibodies; Blood Urea Nitrogen; DNA; Extracellular Matrix; Female; Fibronectins; Fibrosis; Immunoglobulin G; Leukocytes, Mononuclear; Lupus Nephritis; Mice; Mice, Inbred MRL lpr; Nitrates; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Nitrites; Plasminogen Activator Inhibitor 1; RNA, Messenger; Transforming Growth Factor beta; Transforming Growth Factor beta1

Tubulointerstitial fibrosis is a hallmark feature of chronic renal injury. Specific therapies to control the progression of renal fibrosis toward end-stage renal failure are limited. Previous studies have demonstrated that expression of endogenous bone morphogenic protein-7 (BMP-7) is reduced in the kidneys of several inducible mouse models of acute and chronic renal disease and that administration of exogenous recombinant human BMP-7 (rhBMP-7) has a beneficial effect on kidney function. Here we report that treatment with rhBMP-7 leads to improved renal function, histology, and survival in mice deficient in the alpha3-chain of type IV collagen and MRL/MpJlpr/lpr lupus mice, two genetic models for chronic renal injury and fibrosis. Such therapeutic benefit is also associated with a significant decrease in the expression of profibrotic molecules, such as type I collagen and fibronectin, in renal fibroblasts. Additionally, rhBMP-7 induces expression of active matrix metalloproteinase-2, which is potentially important for removal of fibrotic matrix. Collectively, these studies provide further evidence for rhBMP-7 as an important bone-associated protein with protective function against renal pathology.

Topics: Animals; Autoantigens; Bone Morphogenetic Protein 7; Bone Morphogenetic Proteins; Cells, Cultured; Collagen Type IV; Disease Models, Animal; Female; Fibroblasts; Fibrosis; Humans; Kidney Failure, Chronic; Lupus Nephritis; Mice; Mice, Inbred MRL lpr; Mice, Mutant Strains; Neuroprotective Agents; Transforming Growth Factor beta

To study the protective effects of Tripterine on experimental lupus nephritis glomerulosclerosis.. Different doses of Tripterine were injected peritoneally to BW F1 mice at different stages. 24-hour urine protein excretion, serum anti-dsDNA antibodies, renal pathology and RT-nested PCR were analyzed to study the preventive effects of Tripterine on LN glomerulosclerosis and its mechanisms.. (1) Tripterine suppressed the development of proteinuria, decreased the level of serum anti-dsDNA antibodies, reduced the expressions of collagen type IV, fibronectin, TIMP-1, TIMP-2, TGF-beta(1) and improved the expressions of MMP-1, -2 in the murine kidney. (2) The use of Tripterine before occurrence of proteinuria had more obvious protective effects than its use after the occurrence of proteinuria. (3) No significant difference was found between the 3 mg/kg/week Tripterine-treated-group and the 6 mg/kg/week Tripterine-treated-group. (4) No obvious change was observed in the expression of MMP-3.. Tripterine has a definite protective effect on glomerulosclerosis of the lupus murine model. The decrease of renal collagen type IV and fibronectin is probably due to its suppressive effect on the expressions of local TGF-beta(1) and TIMP-1, -2, and its improvement effect on the local expressions of MMP-1, -2. Tripterine may have no obvious influence on MMP-3 in this model.

Topics: Animals; Antibodies, Antinuclear; Female; Glomerulosclerosis, Focal Segmental; Immunohistochemistry; Kidney; Lupus Nephritis; Mice; Pentacyclic Triterpenes; Proteinuria; Reverse Transcriptase Polymerase Chain Reaction; Time Factors; Transforming Growth Factor beta; Transforming Growth Factor beta1; Triterpenes

Smad6 and Smad7 are inhibitory SMADs with putative functional roles at the intersection of major intracellular signaling networks, including TGF-beta, receptor tyrosine kinase (RTK), JAK/STAT, and NF-kappaB pathways. This study reports differential functional roles and regulation of Smad6 and Smad7 in TGF-beta signaling in renal cells, in murine models of renal disease and in human glomerular diseases. Smad7 is upregulated in podocytes in all examined glomerular diseases (focal segmental glomerulosclerosis [FSGS], minimal-change disease [MCD], membranous nephropathy [MNP], lupus nephritis [LN], and diabetic nephropathy [DN]) with a statistically significant upregulation in "classical" podocyte-diseases such as FSGS and MCD. TGF-beta induces Smad7 synthesis in cultured podocytes and Smad6 synthesis in cultured mesangial cells. Although Smad7 expression inhibited both Smad2- and Smad3-mediated TGF-beta signaling in podocytes, it inhibited only Smad3 but not Smad2 signaling in mesangial cells. In contrast, Smad6 had no effect on TGF-beta/Smad signaling in podocytes and enhanced Smad3 signaling in mesangial cells. These data suggest that Smad7 is activated in injured podocytes in vitro and in human glomerular disease and participates in negative control of TGF-beta/Smad signaling in addition to its pro-apoptotic activity, whereas Smad6 has no role in TGF-beta response and injury in podocytes. In contrast, Smad6 is upregulated in the mesangium in human glomerular diseases and may be involved in functions independent of TGF-beta/Smad signaling. These data indicate an important role for Smad6 and Smad7 in glomerular cells in vivo that could be important for the cell homeostasis in physiologic and pathologic conditions.

Topics: Adaptor Proteins, Signal Transducing; Animals; Cells, Cultured; Cytoskeletal Proteins; Diabetic Nephropathies; DNA-Binding Proteins; Glomerular Mesangium; Glomerulosclerosis, Focal Segmental; Humans; Kidney Diseases; Kidney Glomerulus; Lupus Nephritis; Mice; Mice, Inbred C57BL; Mice, Inbred NZB; Mice, Knockout; Nephrosis, Lipoid; Proteins; Reference Values; Signal Transduction; Smad2 Protein; Smad3 Protein; Smad6 Protein; Smad7 Protein; Trans-Activators; Transforming Growth Factor beta

Although elevated levels of transforming growth factor-beta (TGF-beta) and tumor necrosis factor-alpha (TNF-alpha) have been implicated in renal disease, the tissue distribution and cellular localization of the induced cytokines is not well established. In this study, we investigated the expression of these cytokines during the progression of lupus nephritis in MRL lpr/lpr mice. The concentration of both cytokines increased in the plasma of these animals in an age-dependent manner, and there was an age-dependent induction of TGF-beta and TNF-alpha mRNAs in their kidneys. Although the increase in TGF-beta mRNA was specific for the kidney, the increase in TNF-alpha mRNA was widespread and also could be demonstrated in the liver, lung, and heart. In situ hybridization analysis of renal tissues from the lupus-prone mice localized TGF-beta mRNA to the glomerulus, and more specifically, to resident glomerular cells and inflammatory cells infiltrating periglomerular spaces in the nephritic lesions. The signals for TNF-alpha mRNA were detected only in inflammatory cells and were distributed throughout the nephritic kidney. Plasminogen activator inhibitor-1 (PAI-1) is known to be elevated in the glomeruli of MRL lpr/lpr mice, and intraperitoneal administration of either TGF-beta or TNF-alpha into normal mice markedly induced the expression of this potent inhibitor of fibrinolysis in renal glomerular or tubular cells in vivo. These results suggest that the increased renal expression of both cytokines may contribute to the development of lupus nephritis in this model and raise the possibility that PAI-1 may be involved. The fact that TGF-beta is specifically induced in the kidney whereas TNF-alpha increases in a variety of tissues, supports the hypothesis that the renal specificity of this disorder reflects the abnormal expression of TGF-beta.

Topics: Animals; Female; Gene Expression Regulation; Kidney; Lupus Nephritis; Mice; Mice, Inbred MRL lpr; Plasminogen Activator Inhibitor 1; RNA, Messenger; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

To understand better the function of endothelin-1 (ET-1) in renal physiology, we examined vascular and glomerular expression of ET-1 in normal human kidney and in lupus nephritis. Immunohistochemical analysis revealed that renal endothelium of glomeruli, arteries, veins, and capillaries expressed ET-1. Endothelial cells were the principal source of glomerular ET-1; positive immunostaining was detected only rarely in mesangial cells and vascular smooth muscle cells from normal kidney. However, mesangial staining for ET-1 was elevated in patients with lupus nephritis, suggesting that under certain conditions mesangial cells elaborate ET-1. Indeed cultured human mesangial cells from normal subjects secreted ET-1 peptide. ET-1 secretion was augmented by the protein kinase C activator phorbol ester and by transforming growth factor-beta1 (TGF-beta1), a cytokine implicated in the development of glomerulosclerosis. Transient transfection of cultured mesangial cells with a preproET-1 reporter construct showed that the preproET-1 promoter is transcriptionally active in mesangial cells and is stimulated by TGF-beta1, phorbol ester, or ectopic expression of protein kinase beta1. Cultured human mesangial cells have both ETA and ETB receptors that contribute to ET-1-stimulated mitogenesis. Taken together, these results demonstrate that ET-1 is expressed at sites where paracrine or autocrine signaling by ET-1 might control renal vasoconstriction, glomerular filtration rate, and remodeling of the glomerulus in renal disease.

Topics: Cells, Cultured; Endothelin-1; Endothelium, Vascular; Gene Expression Regulation; Glomerular Mesangium; Humans; Kidney Glomerulus; Lupus Nephritis; Muscle, Smooth, Vascular; Polymerase Chain Reaction; Protein Kinase C; Recombinant Proteins; Renal Circulation; RNA, Messenger; Tetradecanoylphorbol Acetate; Transcription, Genetic; Transfection; Transforming Growth Factor beta

Over-expression of iNOS is implicated in the pathogenesis of glomerulonephritis in animal models of systemic lupus erythematosus. The aim of this study was to evaluate the effect of aminoguanidine, a selective inhibitor of iNOS, for the protection from glomerulosclerosis in NZB/W F1 mice. Female NZB/W F1 mice (n = 8) were treated with aminoguanidine (1 g/l) in drinking water for 4 months starting at age 2 months before the onset of glomerulonephritis. Controls were age- and sex-matched mice (n = 10) without aminoguanidine treatment. By glomerular microdissection and reverse-transcription competitive polymerase chain reaction, we found that glomerular iNOS/beta-actin and TGF-beta1/beta-actin mRNA ratios were reduced 15.1% (P<0.05) and 61.3% (P<0.01), respectively, in aminoguanidine-treated mice. Aminoguanidine significantly reduced the glomerular iNOS staining, urinary nitrite production and degree of glomerulosclerosis. In addition, the glomerular volume and mean glomerular cell number were reduced 33.2% (P<0.01) and 32.8% (P<0.01), respectively. Likewise, the urinary proteinuria was also significantly reduced by aminoguanidine. These results indicate that administration of aminoguanidine may reduce the progression of glomerulosclerosis in NZB/W F1 mice, possibly through inhibition of glomerular nitric oxide production.

Topics: Animals; Body Weight; Cell Count; Creatine; Crosses, Genetic; Enzyme Inhibitors; Female; Guanidines; Hypertrophy; Kidney Glomerulus; Lupus Nephritis; Male; Mice; Mice, Inbred NZB; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Nitrites; Organ Size; Proteinuria; RNA, Messenger; Transforming Growth Factor beta

Studies from our laboratory indicate that n-3 (fish oil, FO) lipids at 10% (w/w) in a nutritionally adequate, semipurified diet, and supplemented with equal levels of antioxidants, extended the life span of lupus-prone (NZB/NZW)F1 (B/W) female mice as compared to n-6 (corn oil, CO) lipids. The early rise of autoimmune disease in CO-fed mice was closely linked to the loss of T-cell function. Both IL-2 production and IL-2 receptor expression were reduced due to the loss of naive T-cells and a rise in memory T-cells. Proliferative response to both mitogens and superantigens (staphylococcal enterotoxins A and B) was higher in FO-fed 6.5-mon-old mice. These changes paralleled decreased PGE2 production by splenic cells from FO-fed mice. Analysis of mRNA expression in different organs revealed differential effects of dietary lipids. In FO-fed mice, transforming growth factor beta 1 (TGF beta 1) expression was decreased in kidneys, but splenic tissues had higher expression of TGF beta mRNA. As TGF beta promotes programmed cell death (PCD), we studied the effects of CO and FO on PCD rates in lymphocytes. Both propidium iodide staining and DNA fragmentation were elevated in lymphocytes of FO-fed mice when compared to CO-fed mice of similar age. Also, increased PCD correlated closely with increased Fas gene expression. Thus, in addition to various other antiinflammatory effects, dietary FO appears to increase PCD and prevent accumulation of self-reactive immune cells in lymphoid organs. Further studies are required to dissect the pro- and antiinflammatory mechanisms associated with dietary n-3 and n-6 lipids in modulating autoimmune disorders or malignancy during aging.

Topics: Animals; Antioxidants; Apoptosis; Autoimmune Diseases; Autoimmunity; Fatty Acids, Omega-3; Female; Gene Expression Regulation, Enzymologic; Lupus Nephritis; Male; Mice; Mice, Inbred NZB; Transforming Growth Factor beta

The present study was carried out to test whether transforming growth factor beta (TGF beta) plays a pathological role in the induction or progression of glomerulonephritis in a murine model of systemic lupus erythematosus (SLE), and whether dietary supplementation with fish oil (FO) can modulate the expression of TGF beta. Weanling female (NZB x NZW) F1 (B/W) mice were divided into three groups. One group was fed an unmanipulated diet (lab. chow; LC) and the other two groups were fed a nutritionally adequate semipurified diet supplemented with 10% CO or FO. Both water and food were provided ad libitum. Proteinuria and serum anti-dsDNA antibody levels were measured to assess disease progression. Mice were killed at 3.5 and 6.5 months of age and renal mRNA levels for TGF beta isoforms, fibronectin-1 (FN-1) and intercellular adhesion molecule-1 (ICAM-1) were studied by Northern blot analysis. TGF beta 1 protein levels were also examined in kidneys by Western blot analysis. Our results indicate that at 3.5 months of age, when urinary protein levels were undetectable and very low levels of anti-dsDNA were detected, no mRNA signal could be detected for TGF beta isoforms, ICAM-1 and FN-1 in either dietary group. However, at 6.5 months, the FO-fed mice, compared to LC and CO, had [1] greatly reduced proteinuria (LC: 2-3+, CO: 2-3+; FO: trace -1+) and serum anti-dsDNA antibodies; [2] improved survival (CO: 100% death (15/15) occurred by 8 months; FO: 50% were alive at 12 months (8/15) and [3] reduced renal TGF beta 1 mRNA and protein levels. TGF beta 2 and beta 3 were not significantly affected by FO diet. Similarly, lower levels of renal FN-1 and ICAM-1 mRNA were observed in FO fed mice. These data indicate that in B/W mice on a FO diet, prolonged survival and amelioration of renal disease may be attributed at least in part to lower levels of TGF beta 1 mRNA and protein in the kidneys.

Topics: Adjuvants, Immunologic; Animals; Autoimmune Diseases; Blotting, Northern; Blotting, Western; Body Weight; Fatty Acids, Omega-3; Female; Kidney; Lupus Nephritis; Mice; Mice, Inbred NZB; Proteinuria; RNA, Messenger; Transforming Growth Factor beta

Topics: Biopsy; Diabetic Nephropathies; Glomerulonephritis, IGA; Humans; In Situ Hybridization; Kidney Glomerulus; Lupus Nephritis; Macrophages; Nephrosis, Lipoid; RNA, Messenger; Transforming Growth Factor beta

Three major components of the plasminogen activators (PA)/plasmin system are synthesized physiologically in glomeruli, and can be involved in glomerular proteolysis and extracellular matrix metabolism: tissue-type PA (tPA), urokinase (uPA) and PA inhibitor type 1 (PAI-1). To explore the possible role of a dysregulation of the plasmin protease system in the development and progression of lupus-like glomerulonephritis, we studied the expression of the renal plasmin protease components during the course of the disease, either acute, induced by IgG3 monoclonal cryoglobulins, or chronic, occurring spontaneously in three different lupus-prone mice: (NZBxNZW)F1, BXSB and MRL-lpr/lpr. RNase protection assays and in situ hybridizations revealed a marked glomerular induction of PAI-1 mRNA abundance without any significant changes in renal tPA and uPA mRNA levels in the two different types of lupus-like glomerulonephritis. The overexpression of PAI-1 mRNA occurred in parallel with a significant decrease in glomerular tPA-catalyzed enzymatic activity as determined by zymographic analysis. In addition, a concomitant increase in glomerular expression of transforming growth factor beta 1 (TGF-beta 1) mRNA was observed. The demonstration of a close correlation between the PAI-1 and TGF-beta 1 mRNA levels and the severity of lupus-like glomerular lesions suggests that a pertubation of the glomerular PA/PAI balance, resulting from a marked TGF-beta 1-mediated induction of PAI-1 gene expression, plays an important role in the progression of lupus-like glomerular lesions, leading to glomerulosclerosis.

Topics: Animals; Antibodies, Monoclonal; Chronic Disease; Cryoglobulins; Immunoglobulin G; Kidney Glomerulus; Lupus Nephritis; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Plasminogen Activator Inhibitor 1; Rheumatoid Factor; RNA, Messenger; Tissue Distribution; Transforming Growth Factor beta