Recognition and clearance of apoptotic cells: a role for complement and pentraxins.
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Apoptosis↗
Biomedical subjects
Cees van Kooten
Publications and source records attributed to Cees van Kooten.
Glucose-induced fibronectin and collagen type III expression in renal fibroblasts can occur independent of TGF-beta1.
BACKGROUND: Various renal cell types have been shown to contribute to the excessive matrix deposition observed in diabetic nephropathy. The present study examined the effect of high ambient glucose and transforming growth factor-beta1 (TGF-beta1) on matrix production by human renal fibroblasts. METHODS: Human renal fibroblasts (TK173) were used to examine the effects of high glucose and TGF-beta1 on fibronectin and collagen type III expression. Stable transfectants were generated of TK173 cells expressing a dominant negative TGF-beta type II receptor. Matrix components were measured in enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Fibronectin secretion by renal fibroblasts was increased upon exposure to high glucose, but with delayed kinetics compared to TGF-beta1-induced fibronectin. Exposure to high glucose resulted in an increased secretion of latent TGF-beta1. However, treatment with neutralizing pan-specific anti-TGF-beta antibodies could not attenuate the effects of glucose. Furthermore, collagen type III was up-regulated by high glucose, but not by TGF-beta1. Importantly, fibroblasts expressing a dominant negative TGF-beta type II receptor were defective in TGF-beta1-induced fibronectin production, whereas glucose-induced fibronectin and collagen type III were unaffected. CONCLUSIONS: These data show that in renal fibroblasts exposure to high glucose can increase matrix production independent of endogenous TGF-beta1. Although glucose activation is accompanied by an increased production of latent TGF-beta1, which can have an important role in vivo, the data suggest involvement of alternative growth factors in the mechanism by which hyperglycemic conditions can modulate matrix accumulation in diabetic nephropathy.
Antibodies↗
Functional modulation of dendritic cells to suppress adaptive immune responses.
In recent years, dendritic cells (DCs) have entered the center court of immune regulation. Dependent on their ontogeny, state of differentiation, and maturation and thereby a variable expression of membrane-bound and soluble molecules, DCs can induce immunostimulatory as well as immunoregulatory responses. This dual function has made them potential targets in vaccine development in cancer and infections as well as for the prevention and treatment of allograft rejection and autoimmune diseases. The present review is focused on the effect of immune-modulatory factors, such as cytokines and immunosuppressive drugs, and on the survival, differentiation, migration, and maturation of DC human subsets. A better understanding of DC immunobiology may lead to the development of specific therapies to prevent or dampen immune responses.
Animals↗
Connective tissue growth factor and igf-I are produced by human renal fibroblasts and cooperate in the induction of collagen production by high glucose.
Tubulointerstitial fibrosis is an important component in the development of diabetic nephropathy. Various renal cell types, including fibroblasts, contribute to the excessive matrix deposition in the kidney. Although transforming growth factor-beta (TGF-beta) has been thought to play a major role during fibrosis, other growth factors are also involved. Here we examined the effects of connective tissue growth factor (CTGF) and IGF-I on collagen type I and III production by human renal fibroblasts and their involvement in glucose-induced matrix accumulation. We have demonstrated that both CTGF and IGF-I expressions were increased in renal fibroblasts under hyperglycemic conditions, also in the absence of TGF-beta signaling. Although CTGF alone had no effect on collagen secretion, combined stimulation with IGF-I enhanced collagen accumulation. Furthermore, IGF-I also had a synergistic effect with glucose on the induction of collagens. Moreover, we observed a partial inhibition in glucose-induced collagen secretion with neutralizing anti-CTGF antibodies, thereby demonstrating for the first time the involvement of endogenous CTGF in glucose-induced effects in human renal fibroblasts. Therefore, the cooperation between CTGF and IGF-I might be involved in glucose-induced matrix accumulation in tubulointerstitial fibrosis and might contribute to the pathogenesis of diabetic nephropathy.
Cells, Cultured↗
Steroid responsiveness of renal epithelial cells. Dissociation of transrepression and transactivation.
Glucocorticoids modulate cellular and inflammatory responses via stimulation or inhibition of gene transcription. Inhibition of cytokine gene expression is mediated via repression of transcription factors, including NF-kappaB. Previously we have shown that cytokine production by renal epithelial cells is insensitive to the inhibitory action of dexamethasone. In this study we demonstrate that dexamethasone is unable to inhibit NF-kappaB activation in the renal epithelial cell line HK-2, as measured by IkappaB-alpha degradation and DNA binding activity. Transfection of an NF-kappaB-inducible reporter gene demonstrated that non-stimulated HK-2 cells contain a high level of constitutively active NF-kappaB compared with the steroid-sensitive airway epithelial cell line A549, which was not blocked by dexamethasone. Expression and nuclear translocation of the glucocorticoid receptor (GR) was comparable in both cell types. In HK-2 cells, dexamethasone stimulated expression of two glucocorticoid-responsive genes, beta(2)-adrenoreceptors and angiotensinogen. The capacity of GR to transactivate the native angiotensinogen glucocorticoid-responsive element (GRE) using chromatin-IP was not impaired. Moreover, dexamethasone activation of a GRE-driven reporter construct appeared to be equally effective, although less sensitive compared with A549 cells. In conclusion, we provide evidence that glucocorticoids are unable to repress the activity of NF-kappaB in renal epithelial cells in the presence of an intact stimulatory pathway.
Cell Line↗
Rapamycin specifically interferes with GM-CSF signaling in human dendritic cells, leading to apoptosis via increased p27KIP1 expression.
The longevity of dendritic cells (DCs) is a critical regulatory factor influencing the outcome of immune responses. Recently, we demonstrated that the immunosuppressive drug rapamycin (Rapa) specifically induces apoptosis in DCs but not in other myeloid cell types. The present study unraveled the mechanism used by Rapa to induce apoptosis in human monocyte-derived DCs. Our data demonstrate that granulocyte-macrophage colony-stimulating factor (GM-CSF) preserves DC survival specifically via the phosphatidylinositol-3 lipid kinase/mammalian target of rapamycin (PI3K/mTOR) signaling pathway, which is abrogated by Rapa at the level of mTOR. Disruption of this GM-CSF signaling pathway induced loss of mitochondrial membrane potential, phosphatidyl-serine exposure, and nuclear changes. Apoptosis of these nonproliferating DCs was preceded by an up-regulation of the cell cycle inhibitor p27(KIP1). Overexpression of p27(KIP1) in DCs using adenoviral gene transduction revealed that apoptosis is directly regulated by p27(KIP1). Furthermore, both overexpression of p27(KIP1) and disruption of the GM-CSF/PI3K/mTOR signaling pathway decreased the expression of the antiapoptotic protein mcl-1. This mTOR/p27(KIP1)/mcl-1 survival seems unique for DCs and may provide novel opportunities to influence immune responses by specific interference with the life span of these cells.
Adenoviridae↗
Corticosteroids prevent generation of CD34+-derived dermal dendritic cells but do not inhibit Langerhans cell development.
Corticosteroids (CS) have been shown to exert strong inhibitory effects on dendritic cell (DC) differentiation and function. Those studies were mostly performed with monocyte-derived DC, which represents only one subpopulation from the wide variety of DC types. In the present study the effects of the CS dexamethasone and prednisolone were investigated on the differentiation of CD34(+) hemopoietic progenitor cells into 1) Langerhans cells (LC), which differentiate directly into CD1a(+) DC; and 2) dermal/interstitial DC, which differentiate via a CD14(+)CD1a(-) phenotype into CD14(-)CD1a(+) DC. CS present during the entire 11-day culture period, resulting in fully differentiated CD1a(+) DC, increased the percentage of langerin(+) DC within the CD1a(+) population. In line with these data, CS treatment during the first 6 days of differentiation reduced the development of CD14(+) dermal DC precursors and thereby seemed to support the generation of CD1a(+) LC precursors. Addition of CS from day 6 onward specifically blocked the development of CD1a(+) dermal DC by both inhibition of spontaneous and IL-4-induced differentiation of CD14(+) DC precursors into CD1a(+) DC as well as induction of apoptosis in CD14(+) DC precursors. Apoptosis was not found in CD14(+) macrophage precursors derived from the same CD34(+) progenitors. The development and function of LC were not affected by CS, as demonstrated by a normal T cell stimulatory capacity and IL-12 production. These data demonstrate that CS interfere with the normal development of DC from CD34(+) progenitors by specific induction of apoptosis in precursors of dermal/interstitial DC. In view of the different functional capacities of dermal/interstitial DC and Langerhans cells, this might affect the overall cellular immune response.
Adult↗
Fc alpha RI/CD89 circulates in human serum covalently linked to IgA in a polymeric state.
The FcR for IgA CD89/FcalphaRI, is a type I receptor glycoprotein, expressed on myeloid cells, with important immune effector functions. In vitro CD89 can be released from CD89-expressing cells upon activation. Little information is available on the existence of this soluble molecule in vivo. Using specific and sensitive ELISA techniques (detection limit 50 pg/ml), we were not able to detect circulating CD89 in human sera. However, using Western blotting, a 30-kDa soluble CD89 molecule was demonstrated in both serum and plasma. Moreover, using a specific semiquantitative dot-blot system, we found CD89 in all human sera tested (mean concentration 1900 ng/ml). Size fractionation of human serum using gel filtration chromatography showed that the CD89 molecule was predominantly present in larger molecular mass fractions. Direct complexes between IgA and CD89 were demonstrated by anti-IgA affinity purification, and when analyzed under nonreducing conditions appeared to be covalently linked. Size fractionation of affinity-purified IgA showed the presence of soluble CD89 only in the high molecular mass fractions of IgA, but not in monomeric IgA. High molecular mass complexes of CD89-IgA could be distinguished from J chain containing dimeric IgA. These data show that CD89 circulates in complex with IgA, and suggest that CD89 might contribute to the formation of polymeric serum IgA.
Antigens, CD↗
Human immature dendritic cells efficiently bind and take up secretory IgA without the induction of maturation.
Immature dendritic cells (DC) reside in peripheral tissues, where they pick up and process incoming pathogens via scavenger receptors or FcR such as FcgammaR and FcepsilonR. At mucosal surfaces, IgA is the main Ig to protect the body from incoming pathogens. In addition, DC are present in high numbers at these sites. We detected expression of FcalphaR (CD89) on the CD14+ population of CD34+ progenitor-derived DC and on monocyte-derived DC (MoDC). However, CD89 expression was strongly decreased upon differentiation from monocyte to DC. We found only minimal binding of serum IgA to MoDC but strong binding of secretory IgA (SIgA). The SIgA binding to MoDC could not be blocked by anti-CD89 blocking Abs. DC efficiently internalized SIgA, but not serum IgA, and uptake of SIgA could be blocked by specific sugars or partially by Ab reactive with mannose receptor. Importantly, binding and uptake of SIgA was not accompanied by signs of DC maturation, such as increased expression of CD86 and CD83 or induction of cytokine secretion. These data indicate that SIgA can interact with DC not via CD89, but via carbohydrate-recognizing receptors like mannose receptor and suggest that uptake of SIgA-containing immune complexes by immature DC may be a mechanism to modulate mucosal immune responses.
Antibodies↗
Antibody response against perlecan and collagen types IV and VI in chronic renal allograft rejection in the rat.
Chronic rejection is the leading cause of late renal transplant failure. Various structural lesions are observed in grafts undergoing chronic rejection including glomerular basement membrane (GBM) duplications. The well-established Fisher (F344) to Lewis (LEW) rat renal transplant model for chronic rejection was used to assess the presence and role of the humoral immune response against graft antigens during chronic rejection. LEW recipients of F344 allografts develop transplant glomerulopathy and produce IgG1 antibodies directed against F344 GBM preparations that are detectable 3 weeks after transplantation. Glomerular IgG1 deposition was observed that in vitro co-localized with a rabbit anti-rat GBM antiserum in rejecting F344 grafts; elution experiments of isolated glomeruli yielded IgG1 antibodies reactive in vitro with F344 GBM, but not LEW GBM. Prevention of acute rejection by transient treatment of the recipients with cyclosporin A completely abrogated the production of anti-GBM antibodies. Using proteomic techniques we identified the antigens recognized by the LEW posttransplant sera as being the heparan sulfate proteoglycan perlecan and the alpha1 chain of collagen type VI in association with the alpha5 chain of collagen type IV. In conclusion, LEW recipients of F344 kidney grafts produce IgG1 antibodies against donor type perlecan and alpha1(VI)/alpha5(IV) collagen and develop transplant glomerulopathy. These data implicate an important role for the humoral immune response in the development of glomerulopathy during chronic rejection.
Albuminuria↗
Production of inflammatory mediators by renal epithelial cells is insensitive to glucocorticoids.
1. In the present study we investigated the effect of glucocorticoids on the activation of renal tubular epithelial cells, which are thought to play an important role in inflammatory processes in the kidney. 2. Activation of renal epithelial cells by IL-1, TNF-alpha or CD40L resulted in increased production of cytokines and chemokines. Both in the renal epithelial cell line HK-2 and in primary cultures of human proximal tubular epithelial cells (PTEC) production of IL-6, IL-8 and monocyte chemotactic protein 1 (MCP-1) was not inhibited by glucocorticoids, independent of the stimulus. 3. In contrast, dexamethasone strongly inhibited cytokine production by immortalized renal fibroblasts and an airway epithelial cell line (A549). 4. Stimulation of renal epithelial cells resulted in activation of NF-kappaB, a pivotal transcription factor in the regulation of cytokine genes, as was shown by IkappaB-alpha degradation and increased DNA-binding activity. In contrast to dexamethasone, addition of the NF-kappaB inhibitors pyrrolidine dithiocarbamate (PDTC) and n-tosyl-l-phenylalanine chloromethyl ketone (TPCK) completely abolished cytokine and chemokine production. 5. Renal epithelial cells express abundant levels of the functional glucocorticoid receptor alpha (GRalpha) isoform and low levels of the inhibitory beta isoform (GRbeta). 6. In conclusion, cytokine production by renal epithelial cells is insensitive to the inhibitory effects of glucocorticoids. The lack of dexamethasone-mediated inhibition was specific for renal epithelial cells and could not be explained by an increased expression of the glucocorticoid receptor beta isoform.
Cells, Cultured↗
Renal tubular epithelial cell death and cyclosporin A.
BACKGROUND: The pathogenesis of chronic cyclosporin A (CsA) nephrotoxicity is largely unknown. In this study we examined whether CsA produces cell death through necrosis or apoptosis of either cultured human proximal tubular epithelial cells (PTEC) or the porcine tubular cell line LLC-PK(1). METHODS: Primary isolates of human PTEC and LLC-PK(1) cells were treated for various time periods with CsA at concentrations of 0.01-100 microg/ml. Apoptosis was studied by the assessment of annexin binding and propidium iodide uptake, the measurement of cellular DNA content and cell cycle analysis, and by the evaluation of nuclear morphology. Cell death was studied by the trypan blue exclusion method. Hypoxic conditions were simulated through chemical ATP depletion. RESULTS: In human PTEC, cell death was observed at CsA concentrations higher than 10 microg/ml; at these concentrations PTEC died as a result of necrosis and the toxicity of its vehicle Cremophore EL, and not as a result of CsA inducing apoptosis. The addition of cycloheximide to relieve a possible block in the apoptotic process had no effect on human PTEC, but did result in apoptosis of LLC-PK(1). In human PTEC, CsA did not augment cell death induced by chemical ATP depletion. CONCLUSIONS: The results of this in vitro study do not support the hypothesis that CsA directly induces cell death of proximal tubular epithelial cells.
Biopsy↗
Non-HLA humoral immunity and chronic kidney-graft loss.
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Antilymphocyte Serum↗