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Richard Horuk - One of the best experts on this subject based on the ideXlab platform.

  • expression purification and in vitro functional reconstitution of the Chemokine Receptor CCR1
    Protein Expression and Purification, 2009
    Co-Authors: Samantha J Allen, Richard Horuk, Sofia Ribeiro, Tracy M. Handel
    Abstract:

    Chemokine Receptors are a specific class of G-protein-coupled Receptors (GPCRs) that control cell migration associated with routine immune surveillance, inflammation and development. In addition to their roles in normal physiology, these Receptors and their ligands are involved in a large number of inflammatory diseases, cancer and AIDS, making them prime therapeutic targets in the pharmaceutical industry. Like other GPCRs, a significant obstacle in determining structures and characterizing mechanisms of activation has been the difficulty in obtaining high levels of pure, functional Receptor. Here we describe a systematic effort to express the Chemokine Receptor CCR1 in mammalian cells, and to purify and reconstitute it in functional form. The highest expression levels were obtained using an inducible HEK293 system. The Receptor was purified using a combination of N- (StrepII or Hemagglutinin) and C-terminal (His8) affinity tags. Function was assessed by ligand binding using a novel fluorescence polarization assay with fluorescein-labeled Chemokine. A strict dependence of function on the detergent composition was observed, as solubilization of CCR1 in n-dodecyl-beta-D-maltopyranoside/cholesteryl hemisuccinate yielded functional Receptor with a K(d) of 21 nM for the Chemokine CCL14, whereas it was non-functional in phosphocholine detergents. Differences in function were observed despite the fact that both these detergent types maintained the Receptor in a state characterized by monomers and small oligomers, but not large aggregates. While optimization is still warranted, yields of approximately 0.1-0.2mg of pure functional Receptor per 10(9) cells will permit biophysical studies of this medically important Receptor.

  • Chemokine Receptor CCR1 regulates inflammatory cell infiltration after renal ischemia reperfusion injury
    Journal of Immunology, 2008
    Co-Authors: Richard Horuk, Kengo Furuichi, Takashi Wada, Shuichi Kaneko, Philip M Murphy
    Abstract:

    Neutrophils and macrophages rapidly infiltrate the kidney after renal ischemia-reperfusion injury, however specific molecular recruitment mechanisms have not been fully delineated for these cell types. Here we provide genetic and pharmacologic evidence supporting a positive role for the Chemokine Receptor CCR1 in macrophage and neutrophil infiltration in a 7 day mouse model of renal ischemia-reperfusion injury. By day 7, injured kidneys from mice lacking CCR1 contained 35% fewer neutrophils and 45% fewer macrophages than injured kidneys from wild-type control mice. Pretreatment of wild-type mice with the specific CCR1 antagonist BX471 also suppressed neutrophil and macrophage infiltration in the model. Injured kidneys from mice lacking CCR1 also had reduced content of the CCR1 ligands CCL3 (MIP-1α) and CCL5 (RANTES) compared with injured kidneys from wild-type controls, suggesting a leukocyte source for these inflammatory Chemokines and existence of a CCR1-dependent positive feedback loop for leukocyte infiltration in the model. Local leukocyte proliferation and apoptosis were detected after injury, but were not dependent on CCR1. Also, the extent of necrotic and fibrotic damage and decline in renal function in injured kidneys was similar in wild-type and CCR1-deficient mice. Thus, CCR1 appears to regulate trafficking of macrophages and neutrophils to kidney in a mouse model of renal ischemia-reperfusion injury, however this activity does not appear to affect tissue injury.

  • CCR1 blockade reduces interstitial inflammation and fibrosis in mice with glomerulosclerosis and nephrotic syndrome
    Kidney International, 2004
    Co-Authors: Volker Vielhauer, Stephan Segerer, Richard Horuk, Hermann Josef Gröne, Detlef Schlondorff, Frank Strutz, Matthias Kretzler, Elias Berning, Hans-joachim Anders
    Abstract:

    CCR1 blockade reduces interstitial inflammation and fibrosis in mice with glomerulosclerosis and nephrotic syndrome. Background CC Chemokines mediate leukocyte infiltration into inflamed tissue. We have recently shown that blockade of the CC Chemokine Receptor CCR1 reduces interstitial inflammation and fibrosis in murine obstructive nephropathy. However, it is not known whether CCR 1 blockade is protective in progressive renal injury associated with severe proteinuria. We therefore studied the effect of the small-molecule CCR1 antagonist BX471 in a murine model of adriamycin-induced focal segmental glomerulosclerosis (FSGS) with nephrotic syndrome and progressive interstitial inflammation and fibrosis. Methods Adriamycin nephropathy with persistent proteinuria was induced in male BALB/c mice by two intravenous injections of adriamycin (13 mg/kg) at day 0 and 14. BX471 treatment was started at day 14 when proteinuria and interstitial inflammation had developed. At 6 weeks, renal histology was studied by morphometry and immunohistochemistry. Results At week 6, adriamycin-treated mice showed FSGS, associated with tubulointerstitial injury consisting of tubular dilation and atrophy, interstitial leukocyte infiltration, and fibrosis. The mRNA expression of CCR1 and CC Chemokines, including the CCR1 ligands CCL3 (MIP-1α) and CCL5 (RANTES), was up-regulated in diseased kidneys, with a prominent interstitial expression of CCL5. Compared to vehicle-treated controls BX471 significantly reduced the amount of macrophages and T lymphocytes in interstitial lesions by 51% and 22%, respectively. Markers of renal fibrosis such as interstitial fibroblasts (48%) and interstitial volume (23%) were significantly reduced by BX471 treatment. In contrast, the extent of proteinuria and glomerular sclerosis was not affected by BX471 treatment. Conclusion Blockade of CCR1 substantially reduced interstitial leukocyte accumulation and the subsequent renal fibrosis in a murine model of nephrotic syndrome and FSGS. These findings support a role for CCR1 in interstitial leukocyte recruitment and suggest that CCR1 blockade might be a new therapeutic strategy in progressive nephropathies such as FSGS.

  • late onset of treatment with a Chemokine Receptor CCR1 antagonist prevents progression of lupus nephritis in mrl fas lpr mice
    Journal of The American Society of Nephrology, 2004
    Co-Authors: Hans-joachim Anders, Stephan Segerer, Guillermo Perez De Lema, Emilia Belemezova, Volker Vielhauer, Clemens D Cohen, Richard Horuk, Michael Frink, Matthias Kretzler, Kelly L Hudkins
    Abstract:

    ABSTRACT. 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 week from week 20 to 24 of age. BX471 improved blood urea nitrogen levels (BX471, 35.1 ± 5.3; vehicle, 73.1 ± 39.6 mg/dl; P 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-β. 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.

  • late onset of treatment with a Chemokine Receptor CCR1 antagonist prevents progression of lupus nephritis in mrl fas lpr mice
    Journal of The American Society of Nephrology, 2004
    Co-Authors: Hans-joachim Anders, Stephan Segerer, Guillermo Perez De Lema, Emilia Belemezova, Volker Vielhauer, Clemens D Cohen, Michael Frink, Matthias Kretzler, Vaclav Eis, Richard Horuk
    Abstract:

    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.

Matthias Kretzler - One of the best experts on this subject based on the ideXlab platform.

  • Expression of the Chemokine Receptor CCR1 in human renal allografts
    Nephrology Dialysis Transplantation, 2007
    Co-Authors: Verena Mayer, Kelly L Hudkins, Peter J Nelson, Hans-joachim Anders, Matthias Kretzler, Florian Heller, Holger Schmid, Ulrike Brandt, Heinz Regele, Charles E. Alpers
    Abstract:

    Background. Chemokines are involved in the recruitment of leukocytes to vascularized allografts. CCR1 is a Receptor for various proinflammatory Chemokines and CCR1 blockade reduces renal allograft injury in rabbits. The purpose of the study was to characterize CCR1-positive cells in human renal allografts. Methods. Formalin-fixed, paraffin-embedded allograft nephrectomies (n ¼ 9) and non-involved parts of tumour nephrectomies (n ¼ 10) were studied. Immunohistochemistry for CCR1, CD3 and CD68 was performed on consecutive sections. Double immunofluorescence for CCR1 and CD3, CD20, CD68, DC-SIGN and S100 was used on selected cases. Expression of CCR1 mRNA and the ligands CCL3 and CCL5 was studied in renal allograft biopsies with acute rejection (n ¼ 10), with chronic allograft nephropathy (n ¼ 8) and controls (n ¼ 8). Results. CCR1 protein was expressed by circulating cells in glomerular and peritubular capillaries, colocalizing with CD68. In renal allografts CCR1-positive cells were present within glomerular tufts, but only scattered CCR1-positive cells were found in tubulointerstitial infiltrates. CCR1 did not colocalize with the majority of CD68-positive cells in the interstitium. The small number of CCR1-positive interstitial cells were identified as CD20- or DC-SIGN-positive by double immunofluorescence. CCR1 mRNA was significantly increased in renal biopsies with acute allograft rejection (P < 0.001), and with chronic allograft nephropathy (P < 0.05), it correlated with the expression of CCL3 and CCL5, and with serum-creatinine. Conclusions. CCR1 mRNA expression was associated with renal function in allografts. CCR1 protein expression was restricted to monocytes, CD20-positive B cells and DC-SIGN-positive dendritic cells. Thus most interstitial macrophages were CCR1 negative, which may relate to down-regulation after migration into the interstitium in human renal allografts.

  • CCR1 blockade reduces interstitial inflammation and fibrosis in mice with glomerulosclerosis and nephrotic syndrome
    Kidney International, 2004
    Co-Authors: Volker Vielhauer, Stephan Segerer, Richard Horuk, Hermann Josef Gröne, Detlef Schlondorff, Frank Strutz, Matthias Kretzler, Elias Berning, Hans-joachim Anders
    Abstract:

    CCR1 blockade reduces interstitial inflammation and fibrosis in mice with glomerulosclerosis and nephrotic syndrome. Background CC Chemokines mediate leukocyte infiltration into inflamed tissue. We have recently shown that blockade of the CC Chemokine Receptor CCR1 reduces interstitial inflammation and fibrosis in murine obstructive nephropathy. However, it is not known whether CCR 1 blockade is protective in progressive renal injury associated with severe proteinuria. We therefore studied the effect of the small-molecule CCR1 antagonist BX471 in a murine model of adriamycin-induced focal segmental glomerulosclerosis (FSGS) with nephrotic syndrome and progressive interstitial inflammation and fibrosis. Methods Adriamycin nephropathy with persistent proteinuria was induced in male BALB/c mice by two intravenous injections of adriamycin (13 mg/kg) at day 0 and 14. BX471 treatment was started at day 14 when proteinuria and interstitial inflammation had developed. At 6 weeks, renal histology was studied by morphometry and immunohistochemistry. Results At week 6, adriamycin-treated mice showed FSGS, associated with tubulointerstitial injury consisting of tubular dilation and atrophy, interstitial leukocyte infiltration, and fibrosis. The mRNA expression of CCR1 and CC Chemokines, including the CCR1 ligands CCL3 (MIP-1α) and CCL5 (RANTES), was up-regulated in diseased kidneys, with a prominent interstitial expression of CCL5. Compared to vehicle-treated controls BX471 significantly reduced the amount of macrophages and T lymphocytes in interstitial lesions by 51% and 22%, respectively. Markers of renal fibrosis such as interstitial fibroblasts (48%) and interstitial volume (23%) were significantly reduced by BX471 treatment. In contrast, the extent of proteinuria and glomerular sclerosis was not affected by BX471 treatment. Conclusion Blockade of CCR1 substantially reduced interstitial leukocyte accumulation and the subsequent renal fibrosis in a murine model of nephrotic syndrome and FSGS. These findings support a role for CCR1 in interstitial leukocyte recruitment and suggest that CCR1 blockade might be a new therapeutic strategy in progressive nephropathies such as FSGS.

  • late onset of treatment with a Chemokine Receptor CCR1 antagonist prevents progression of lupus nephritis in mrl fas lpr mice
    Journal of The American Society of Nephrology, 2004
    Co-Authors: Hans-joachim Anders, Stephan Segerer, Guillermo Perez De Lema, Emilia Belemezova, Volker Vielhauer, Clemens D Cohen, Richard Horuk, Michael Frink, Matthias Kretzler, Kelly L Hudkins
    Abstract:

    ABSTRACT. 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 week from week 20 to 24 of age. BX471 improved blood urea nitrogen levels (BX471, 35.1 ± 5.3; vehicle, 73.1 ± 39.6 mg/dl; P 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-β. 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.

  • late onset of treatment with a Chemokine Receptor CCR1 antagonist prevents progression of lupus nephritis in mrl fas lpr mice
    Journal of The American Society of Nephrology, 2004
    Co-Authors: Hans-joachim Anders, Stephan Segerer, Guillermo Perez De Lema, Emilia Belemezova, Volker Vielhauer, Clemens D Cohen, Michael Frink, Matthias Kretzler, Vaclav Eis, Richard Horuk
    Abstract:

    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.

  • Chemokine Receptor CCR1 but not ccr5 mediates leukocyte recruitment and subsequent renal fibrosis after unilateral ureteral obstruction
    Journal of The American Society of Nephrology, 2004
    Co-Authors: Bruno Luckow, Yvonne Linde, Jens T Siveke, Stephan Segerer, Guillermo Perez De Lema, Volker Vielhauer, Clemens D Cohen, Matthias Mack, Matthias Kretzler, Richard Horuk
    Abstract:

    ABSTRACT. As Chemokine Receptor CCR1 and CCR5 expression on circulating leukocytes is thought to contribute to leukocyte recruitment during renal fibrosis, the authors examined the effects of unilateral ureteral obstruction (UUO) in mice deficient for CCR1 or CCR5. Analysis of UUO kidneys from CCR1-deficient mice revealed a reduction of interstitial macrophages and lymphocytes (35% and 55%, respectively) compared with wild-type controls. CCR1-deficient mice had reduced CCR5 mRNA levels in UUO kidneys, which correlated with a reduction of CCR5+ T cell infiltrate as determined by flow cytometry. Interstitial fibroblasts, renal TGF-β1 mRNA expression, interstitial volume, and collagen I deposits were all significantly reduced in CCR1-deficient mice. In contrast, renal leukocytes and fibrosis were unaffected in CCR5-deficient mice with UUO. However, if treated with the CCR1 antagonist BX471, CCR5-deficient mice showed a similar reduction of renal leukocytes and fibrosis as CCR1-deficient mice. To determine the underlying mechanism labeled macrophages and T cells isolated from either wild-type, CCR1-deficient, or CCR5-deficient mice were injected into wild-type mice with UUO. Three hours later, renal cell recruitment was reduced for CCR1-deficient cells or cells pretreated with BX471 compared with CCR5-deficient or wild-type cells. Thus, CCR1 but not CCR5 is required for leukocyte recruitment and fibrosis after UUO in mice. Therefore, CCR1 is a promising target for therapeutic intervention in leukocyte-mediated fibrotic tissue injury, e.g. progressive renal fibrosis.

Hans-joachim Anders - One of the best experts on this subject based on the ideXlab platform.

  • Expression of the Chemokine Receptor CCR1 in human renal allografts
    Nephrology Dialysis Transplantation, 2007
    Co-Authors: Verena Mayer, Kelly L Hudkins, Peter J Nelson, Hans-joachim Anders, Matthias Kretzler, Florian Heller, Holger Schmid, Ulrike Brandt, Heinz Regele, Charles E. Alpers
    Abstract:

    Background. Chemokines are involved in the recruitment of leukocytes to vascularized allografts. CCR1 is a Receptor for various proinflammatory Chemokines and CCR1 blockade reduces renal allograft injury in rabbits. The purpose of the study was to characterize CCR1-positive cells in human renal allografts. Methods. Formalin-fixed, paraffin-embedded allograft nephrectomies (n ¼ 9) and non-involved parts of tumour nephrectomies (n ¼ 10) were studied. Immunohistochemistry for CCR1, CD3 and CD68 was performed on consecutive sections. Double immunofluorescence for CCR1 and CD3, CD20, CD68, DC-SIGN and S100 was used on selected cases. Expression of CCR1 mRNA and the ligands CCL3 and CCL5 was studied in renal allograft biopsies with acute rejection (n ¼ 10), with chronic allograft nephropathy (n ¼ 8) and controls (n ¼ 8). Results. CCR1 protein was expressed by circulating cells in glomerular and peritubular capillaries, colocalizing with CD68. In renal allografts CCR1-positive cells were present within glomerular tufts, but only scattered CCR1-positive cells were found in tubulointerstitial infiltrates. CCR1 did not colocalize with the majority of CD68-positive cells in the interstitium. The small number of CCR1-positive interstitial cells were identified as CD20- or DC-SIGN-positive by double immunofluorescence. CCR1 mRNA was significantly increased in renal biopsies with acute allograft rejection (P < 0.001), and with chronic allograft nephropathy (P < 0.05), it correlated with the expression of CCL3 and CCL5, and with serum-creatinine. Conclusions. CCR1 mRNA expression was associated with renal function in allografts. CCR1 protein expression was restricted to monocytes, CD20-positive B cells and DC-SIGN-positive dendritic cells. Thus most interstitial macrophages were CCR1 negative, which may relate to down-regulation after migration into the interstitium in human renal allografts.

  • Progression of kidney disease: blocking leukocyte recruitment with Chemokine Receptor CCR1 antagonists.
    Kidney International, 2006
    Co-Authors: Hans-joachim Anders, Volha Ninichuk, Detlef Schlondorff
    Abstract:

    Chronic kidney disease (CKD) is usually associated with interstitial leukocytic cell infiltrates, which may contribute to disease progression by production of proinflammatory, proapoptotic, and profibrotic mediators. Recruiting leukocytes into the kidney involves local expression of chemotactic cytokines, that is, Chemokines, that interact with respective Chemokine Receptors on the leukocyte's outer surface. Thus, specific Chemokine Receptor antagonists may represent an attractive therapeutic concept to interfere with renal leukocyte recruitment. Among the proinflammatory Chemokine Receptors, Chemokine Receptor (CCR)-1 has nonredundant roles for leukocyte adhesion to activated vascular endothelium and for transendothelial migration. In fact, blocking CCR-1 with specific small-molecule antagonists was shown to retard progression in various types of rodent CKD models. Here we discuss the perspective of CCR-1 as a new potential target for the treatment of CKD.

  • Chemokine Receptor CCR1: a new target for progressive kidney disease.
    American Journal of Nephrology, 2005
    Co-Authors: Volha Ninichuk, Hans-joachim Anders
    Abstract:

    Infiltrating leukocytes are thought to contribute to the progression of kidney disease. Locally produced Chemokines guide circulating leukocytes into the kidney, which renders therapeutic blockade of respective Chemokine Receptors on the leukocyte surface as potential targets for the inhibition of renal leukocyte recruitment. By using mutant mice and specific antagonists, we found that Chemokine Receptor CCR1 has non-redundant functions for leukocyte adhesion to activated vascular endothelium and for transendothelial diapedesis. Most importantly, CCR1 blockade with a specific small molecule antagonist can improve injury in several types of progressive kidney disease models, even if treatment is initiated in advanced disease states. Identification of new targets may add to the therapeutic options in chronic kidney disease.

  • CCR1 blockade reduces interstitial inflammation and fibrosis in mice with glomerulosclerosis and nephrotic syndrome
    Kidney International, 2004
    Co-Authors: Volker Vielhauer, Stephan Segerer, Richard Horuk, Hermann Josef Gröne, Detlef Schlondorff, Frank Strutz, Matthias Kretzler, Elias Berning, Hans-joachim Anders
    Abstract:

    CCR1 blockade reduces interstitial inflammation and fibrosis in mice with glomerulosclerosis and nephrotic syndrome. Background CC Chemokines mediate leukocyte infiltration into inflamed tissue. We have recently shown that blockade of the CC Chemokine Receptor CCR1 reduces interstitial inflammation and fibrosis in murine obstructive nephropathy. However, it is not known whether CCR 1 blockade is protective in progressive renal injury associated with severe proteinuria. We therefore studied the effect of the small-molecule CCR1 antagonist BX471 in a murine model of adriamycin-induced focal segmental glomerulosclerosis (FSGS) with nephrotic syndrome and progressive interstitial inflammation and fibrosis. Methods Adriamycin nephropathy with persistent proteinuria was induced in male BALB/c mice by two intravenous injections of adriamycin (13 mg/kg) at day 0 and 14. BX471 treatment was started at day 14 when proteinuria and interstitial inflammation had developed. At 6 weeks, renal histology was studied by morphometry and immunohistochemistry. Results At week 6, adriamycin-treated mice showed FSGS, associated with tubulointerstitial injury consisting of tubular dilation and atrophy, interstitial leukocyte infiltration, and fibrosis. The mRNA expression of CCR1 and CC Chemokines, including the CCR1 ligands CCL3 (MIP-1α) and CCL5 (RANTES), was up-regulated in diseased kidneys, with a prominent interstitial expression of CCL5. Compared to vehicle-treated controls BX471 significantly reduced the amount of macrophages and T lymphocytes in interstitial lesions by 51% and 22%, respectively. Markers of renal fibrosis such as interstitial fibroblasts (48%) and interstitial volume (23%) were significantly reduced by BX471 treatment. In contrast, the extent of proteinuria and glomerular sclerosis was not affected by BX471 treatment. Conclusion Blockade of CCR1 substantially reduced interstitial leukocyte accumulation and the subsequent renal fibrosis in a murine model of nephrotic syndrome and FSGS. These findings support a role for CCR1 in interstitial leukocyte recruitment and suggest that CCR1 blockade might be a new therapeutic strategy in progressive nephropathies such as FSGS.

  • late onset of treatment with a Chemokine Receptor CCR1 antagonist prevents progression of lupus nephritis in mrl fas lpr mice
    Journal of The American Society of Nephrology, 2004
    Co-Authors: Hans-joachim Anders, Stephan Segerer, Guillermo Perez De Lema, Emilia Belemezova, Volker Vielhauer, Clemens D Cohen, Richard Horuk, Michael Frink, Matthias Kretzler, Kelly L Hudkins
    Abstract:

    ABSTRACT. 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 week from week 20 to 24 of age. BX471 improved blood urea nitrogen levels (BX471, 35.1 ± 5.3; vehicle, 73.1 ± 39.6 mg/dl; P 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-β. 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.

Volker Vielhauer - One of the best experts on this subject based on the ideXlab platform.

  • CCR1 blockade reduces interstitial inflammation and fibrosis in mice with glomerulosclerosis and nephrotic syndrome
    Kidney International, 2004
    Co-Authors: Volker Vielhauer, Stephan Segerer, Richard Horuk, Hermann Josef Gröne, Detlef Schlondorff, Frank Strutz, Matthias Kretzler, Elias Berning, Hans-joachim Anders
    Abstract:

    CCR1 blockade reduces interstitial inflammation and fibrosis in mice with glomerulosclerosis and nephrotic syndrome. Background CC Chemokines mediate leukocyte infiltration into inflamed tissue. We have recently shown that blockade of the CC Chemokine Receptor CCR1 reduces interstitial inflammation and fibrosis in murine obstructive nephropathy. However, it is not known whether CCR 1 blockade is protective in progressive renal injury associated with severe proteinuria. We therefore studied the effect of the small-molecule CCR1 antagonist BX471 in a murine model of adriamycin-induced focal segmental glomerulosclerosis (FSGS) with nephrotic syndrome and progressive interstitial inflammation and fibrosis. Methods Adriamycin nephropathy with persistent proteinuria was induced in male BALB/c mice by two intravenous injections of adriamycin (13 mg/kg) at day 0 and 14. BX471 treatment was started at day 14 when proteinuria and interstitial inflammation had developed. At 6 weeks, renal histology was studied by morphometry and immunohistochemistry. Results At week 6, adriamycin-treated mice showed FSGS, associated with tubulointerstitial injury consisting of tubular dilation and atrophy, interstitial leukocyte infiltration, and fibrosis. The mRNA expression of CCR1 and CC Chemokines, including the CCR1 ligands CCL3 (MIP-1α) and CCL5 (RANTES), was up-regulated in diseased kidneys, with a prominent interstitial expression of CCL5. Compared to vehicle-treated controls BX471 significantly reduced the amount of macrophages and T lymphocytes in interstitial lesions by 51% and 22%, respectively. Markers of renal fibrosis such as interstitial fibroblasts (48%) and interstitial volume (23%) were significantly reduced by BX471 treatment. In contrast, the extent of proteinuria and glomerular sclerosis was not affected by BX471 treatment. Conclusion Blockade of CCR1 substantially reduced interstitial leukocyte accumulation and the subsequent renal fibrosis in a murine model of nephrotic syndrome and FSGS. These findings support a role for CCR1 in interstitial leukocyte recruitment and suggest that CCR1 blockade might be a new therapeutic strategy in progressive nephropathies such as FSGS.

  • late onset of treatment with a Chemokine Receptor CCR1 antagonist prevents progression of lupus nephritis in mrl fas lpr mice
    Journal of The American Society of Nephrology, 2004
    Co-Authors: Hans-joachim Anders, Stephan Segerer, Guillermo Perez De Lema, Emilia Belemezova, Volker Vielhauer, Clemens D Cohen, Richard Horuk, Michael Frink, Matthias Kretzler, Kelly L Hudkins
    Abstract:

    ABSTRACT. 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 week from week 20 to 24 of age. BX471 improved blood urea nitrogen levels (BX471, 35.1 ± 5.3; vehicle, 73.1 ± 39.6 mg/dl; P 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-β. 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.

  • late onset of treatment with a Chemokine Receptor CCR1 antagonist prevents progression of lupus nephritis in mrl fas lpr mice
    Journal of The American Society of Nephrology, 2004
    Co-Authors: Hans-joachim Anders, Stephan Segerer, Guillermo Perez De Lema, Emilia Belemezova, Volker Vielhauer, Clemens D Cohen, Michael Frink, Matthias Kretzler, Vaclav Eis, Richard Horuk
    Abstract:

    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.

  • Chemokine Receptor CCR1 but not ccr5 mediates leukocyte recruitment and subsequent renal fibrosis after unilateral ureteral obstruction
    Journal of The American Society of Nephrology, 2004
    Co-Authors: Bruno Luckow, Yvonne Linde, Jens T Siveke, Stephan Segerer, Guillermo Perez De Lema, Volker Vielhauer, Clemens D Cohen, Matthias Mack, Matthias Kretzler, Richard Horuk
    Abstract:

    ABSTRACT. As Chemokine Receptor CCR1 and CCR5 expression on circulating leukocytes is thought to contribute to leukocyte recruitment during renal fibrosis, the authors examined the effects of unilateral ureteral obstruction (UUO) in mice deficient for CCR1 or CCR5. Analysis of UUO kidneys from CCR1-deficient mice revealed a reduction of interstitial macrophages and lymphocytes (35% and 55%, respectively) compared with wild-type controls. CCR1-deficient mice had reduced CCR5 mRNA levels in UUO kidneys, which correlated with a reduction of CCR5+ T cell infiltrate as determined by flow cytometry. Interstitial fibroblasts, renal TGF-β1 mRNA expression, interstitial volume, and collagen I deposits were all significantly reduced in CCR1-deficient mice. In contrast, renal leukocytes and fibrosis were unaffected in CCR5-deficient mice with UUO. However, if treated with the CCR1 antagonist BX471, CCR5-deficient mice showed a similar reduction of renal leukocytes and fibrosis as CCR1-deficient mice. To determine the underlying mechanism labeled macrophages and T cells isolated from either wild-type, CCR1-deficient, or CCR5-deficient mice were injected into wild-type mice with UUO. Three hours later, renal cell recruitment was reduced for CCR1-deficient cells or cells pretreated with BX471 compared with CCR5-deficient or wild-type cells. Thus, CCR1 but not CCR5 is required for leukocyte recruitment and fibrosis after UUO in mice. Therefore, CCR1 is a promising target for therapeutic intervention in leukocyte-mediated fibrotic tissue injury, e.g. progressive renal fibrosis.

  • Chemokine Receptor CCR1 but not ccr5 mediates leukocyte recruitment and subsequent renal fibrosis after unilateral ureteral obstruction
    Journal of The American Society of Nephrology, 2004
    Co-Authors: Vaclav Eis, Yvonne Linde, Jens T Siveke, Stephan Segerer, Guillermo Perez De Lema, Volker Vielhauer, Clemens D Cohen, Bruno Luckow, Matthias Kretzler, Matthias Mack
    Abstract:

    As Chemokine Receptor CCR1 and CCR5 expression on circulating leukocytes is thought to contribute to leukocyte recruitment during renal fibrosis, the authors examined the effects of unilateral ureteral obstruction (UUO) in mice deficient for CCR1 or CCR5. Analysis of UUO kidneys from CCR1-deficient mice revealed a reduction of interstitial macrophages and lymphocytes (35% and 55%, respectively) compared with wild-type controls. CCR1-deficient mice had reduced CCR5 mRNA levels in UUO kidneys, which correlated with a reduction of CCR5+ T cell infiltrate as determined by flow cytometry. Interstitial fibroblasts, renal TGF-beta1 mRNA expression, interstitial volume, and collagen I deposits were all significantly reduced in CCR1-deficient mice. In contrast, renal leukocytes and fibrosis were unaffected in CCR5-deficient mice with UUO. However, if treated with the CCR1 antagonist BX471, CCR5-deficient mice showed a similar reduction of renal leukocytes and fibrosis as CCR1-deficient mice. To determine the underlying mechanism labeled macrophages and T cells isolated from either wild-type, CCR1-deficient, or CCR5-deficient mice were injected into wild-type mice with UUO. Three hours later, renal cell recruitment was reduced for CCR1-deficient cells or cells pretreated with BX471 compared with CCR5-deficient or wild-type cells. Thus, CCR1 but not CCR5 is required for leukocyte recruitment and fibrosis after UUO in mice. Therefore, CCR1 is a promising target for therapeutic intervention in leukocyte-mediated fibrotic tissue injury, e.g. progressive renal fibrosis.

Clemens D Cohen - One of the best experts on this subject based on the ideXlab platform.

  • late onset of treatment with a Chemokine Receptor CCR1 antagonist prevents progression of lupus nephritis in mrl fas lpr mice
    Journal of The American Society of Nephrology, 2004
    Co-Authors: Hans-joachim Anders, Stephan Segerer, Guillermo Perez De Lema, Emilia Belemezova, Volker Vielhauer, Clemens D Cohen, Richard Horuk, Michael Frink, Matthias Kretzler, Kelly L Hudkins
    Abstract:

    ABSTRACT. 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 week from week 20 to 24 of age. BX471 improved blood urea nitrogen levels (BX471, 35.1 ± 5.3; vehicle, 73.1 ± 39.6 mg/dl; P 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-β. 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.

  • late onset of treatment with a Chemokine Receptor CCR1 antagonist prevents progression of lupus nephritis in mrl fas lpr mice
    Journal of The American Society of Nephrology, 2004
    Co-Authors: Hans-joachim Anders, Stephan Segerer, Guillermo Perez De Lema, Emilia Belemezova, Volker Vielhauer, Clemens D Cohen, Michael Frink, Matthias Kretzler, Vaclav Eis, Richard Horuk
    Abstract:

    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.

  • Chemokine Receptor CCR1 but not ccr5 mediates leukocyte recruitment and subsequent renal fibrosis after unilateral ureteral obstruction
    Journal of The American Society of Nephrology, 2004
    Co-Authors: Bruno Luckow, Yvonne Linde, Jens T Siveke, Stephan Segerer, Guillermo Perez De Lema, Volker Vielhauer, Clemens D Cohen, Matthias Mack, Matthias Kretzler, Richard Horuk
    Abstract:

    ABSTRACT. As Chemokine Receptor CCR1 and CCR5 expression on circulating leukocytes is thought to contribute to leukocyte recruitment during renal fibrosis, the authors examined the effects of unilateral ureteral obstruction (UUO) in mice deficient for CCR1 or CCR5. Analysis of UUO kidneys from CCR1-deficient mice revealed a reduction of interstitial macrophages and lymphocytes (35% and 55%, respectively) compared with wild-type controls. CCR1-deficient mice had reduced CCR5 mRNA levels in UUO kidneys, which correlated with a reduction of CCR5+ T cell infiltrate as determined by flow cytometry. Interstitial fibroblasts, renal TGF-β1 mRNA expression, interstitial volume, and collagen I deposits were all significantly reduced in CCR1-deficient mice. In contrast, renal leukocytes and fibrosis were unaffected in CCR5-deficient mice with UUO. However, if treated with the CCR1 antagonist BX471, CCR5-deficient mice showed a similar reduction of renal leukocytes and fibrosis as CCR1-deficient mice. To determine the underlying mechanism labeled macrophages and T cells isolated from either wild-type, CCR1-deficient, or CCR5-deficient mice were injected into wild-type mice with UUO. Three hours later, renal cell recruitment was reduced for CCR1-deficient cells or cells pretreated with BX471 compared with CCR5-deficient or wild-type cells. Thus, CCR1 but not CCR5 is required for leukocyte recruitment and fibrosis after UUO in mice. Therefore, CCR1 is a promising target for therapeutic intervention in leukocyte-mediated fibrotic tissue injury, e.g. progressive renal fibrosis.

  • Chemokine Receptor CCR1 but not ccr5 mediates leukocyte recruitment and subsequent renal fibrosis after unilateral ureteral obstruction
    Journal of The American Society of Nephrology, 2004
    Co-Authors: Vaclav Eis, Yvonne Linde, Jens T Siveke, Stephan Segerer, Guillermo Perez De Lema, Volker Vielhauer, Clemens D Cohen, Bruno Luckow, Matthias Kretzler, Matthias Mack
    Abstract:

    As Chemokine Receptor CCR1 and CCR5 expression on circulating leukocytes is thought to contribute to leukocyte recruitment during renal fibrosis, the authors examined the effects of unilateral ureteral obstruction (UUO) in mice deficient for CCR1 or CCR5. Analysis of UUO kidneys from CCR1-deficient mice revealed a reduction of interstitial macrophages and lymphocytes (35% and 55%, respectively) compared with wild-type controls. CCR1-deficient mice had reduced CCR5 mRNA levels in UUO kidneys, which correlated with a reduction of CCR5+ T cell infiltrate as determined by flow cytometry. Interstitial fibroblasts, renal TGF-beta1 mRNA expression, interstitial volume, and collagen I deposits were all significantly reduced in CCR1-deficient mice. In contrast, renal leukocytes and fibrosis were unaffected in CCR5-deficient mice with UUO. However, if treated with the CCR1 antagonist BX471, CCR5-deficient mice showed a similar reduction of renal leukocytes and fibrosis as CCR1-deficient mice. To determine the underlying mechanism labeled macrophages and T cells isolated from either wild-type, CCR1-deficient, or CCR5-deficient mice were injected into wild-type mice with UUO. Three hours later, renal cell recruitment was reduced for CCR1-deficient cells or cells pretreated with BX471 compared with CCR5-deficient or wild-type cells. Thus, CCR1 but not CCR5 is required for leukocyte recruitment and fibrosis after UUO in mice. Therefore, CCR1 is a promising target for therapeutic intervention in leukocyte-mediated fibrotic tissue injury, e.g. progressive renal fibrosis.

  • a Chemokine Receptor ccr 1 antagonist reduces renal fibrosis after unilateral ureter ligation
    Journal of Clinical Investigation, 2002
    Co-Authors: Hans-joachim Anders, Yvonne Linde, Simone M Blattner, Volker Vielhauer, Clemens D Cohen, Frank Strutz, Matthias Mack, Michael Frink, Matthias Kretzler, Hermann Josef Gröne
    Abstract:

    The expression of Chemokines and their Receptors is thought to contribute to leukocyte infiltration and progressive renal fibrosis after unilateral ureter obstruction (UUO). We hypothesized that blocking the Chemokine Receptor CCR1 using the nonpeptide antagonist BX471 could reduce leukocyte infiltration and renal fibrosis after UUO. UUO kidneys from BX471-treated mice (day 0–10 and day 6–10) revealed a 40–60% reduction of interstitial macrophage and lymphocyte infiltrate compared with controls. Treated mice also showed a marked reduction of CCR1 and CCR5 mRNA levels, and FACS analysis showed a comparable reduction of CD8+/CCR5+ T cells. Markers of renal fibrosis, such as interstitial fibroblasts, interstitial volume, mRNA and protein expression for collagen I, were all significantly reduced by BX471-treatment compared with vehicle controls. By contrast treatment was ineffective when the drug was supplied only from days 0 to 5. In summary, blockade of CCR1 substantially reduces cell accumulation and renal fibrosis after UUO. Most interestingly, late onset of treatment is also effective. We therefore conclude that CCR1 blockade may represent a new therapeutic strategy for reducing cellular infiltration and renal fibrosis as major factors in the progression to end-stage renal failure.