Transforming growth factor-β1 regulates chemokine and complement production by human proximal tubular epithelial cells

Transforming growth factor-γ1 regulates chemokine and complement production by human proximal tubular epithelial cells. Previously it has been demonstrated that human proximal tubular epithelial cells (PTEC) are able to produce chemokines (such as IL-8 and MCP-1) and complement components (such as C2, C3, C4 and factor H), and that production of these proteins is regulated by pro-inflammatory cytokines such as interleukin-1α (IL-1α), tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ). Since TGF-γ is also expressed in the renal interstitium during inflammation, we investigated the effect of TGF-γ on the production of chemokines and complement components by PTEC in culture. Transforming growth factor-γ1 up-regulated IL-8 production by an average of 4.11 ± 1.0-fold. macrophage chemoattractant phagocyte (MCP-1) production, on the other hand, was down-regulated by TGF-γ1 by an average of 2.2 ± 0.7-fold. The production of C3 and C4 was also down-regulated after incubation with TGF-γ1 (1.9 ± 0.3- and 3.0 ± 1.2-fold, respectively). All effects were dose- and time-dependent and were found to be specific for TGF-γ1, as assessed by inhibition of the effect with a neutralizing antibody against TGF-γ1. These data, together with the knowledge that TGF-γ, chemokines and complement components play a role in several types of renal disease, suggest that TGF-γ is involved in the regulation of local expression of chemokines and complement components by tubular cells

CsA, FK506, corticosteroids and rapamycin inhibit TNFα production by cultured PTEC

CsA, FK506, corticosteroids and rapamycin inhibit TNFα production by PTEC. In this study we investigated the effect of immunosuppressive drugs on the interleukin-1 alpha (IL-1α) enhanced tumor necrosis factor alpha (TNFα) production by proximal tubular epithelial cells (PTEC). Under basal conditions cultured PTEC produce between 0 to 390 pg/ml/105 cells of TNFα. Upon stimulation with IL-1α an enhancement of TNFα production was seen in each cell line tested, ranging from 230 to 2424 pg/ml/105 cells. The presence of cyclosporin A (CsA) during stimulation with IL-1α inhibited the enhanced TNFα production in a dose dependent fashion, with a maximal inhibition of 90% at a concentration of 250 ng/ml. Inhibition was at the level of mRNA as could be demonstrated by Northern blot analysis. FK506, corticosteroids and rapamycin also inhibited TNFa production in a dose dependent fashion, although not as effectively as CsA. Two corticosteroids were tested for their inhibitory effect on TNFa production. It was found that dexamethasone at a concentration of 10 ng/ml inhibited TNFα production for almost 40%. A 100-fold higher concentration of hydrocortisone was necessary to yield similar inhibition. The effect of rapamycin on the IL-1α enhanced TNFα production differed from the effect of CsA. While CsA induced a maximal inhibition of 90%, rapamycin only induced a maximal inhibition of 37%, and even less inhibition at higher concentrations of the drug. The presence of the various drugs was essential for their inhibitory effect, because removal of the drug from the PTEC by washing immediately resulted in loss of inhibition. Combinations of CsA and FK506 or rapamycin were not additive. However, combinations of rapamycin and FK506 were antagonistic when low concentrations of rapamycin and FK506 were used. Low concentrations of rapamycin with high concentrations of FK506 were synergistic. Since TNFα is likely to be an important mediator in renal allograft rejection, these data suggest that the beneficial effect of immunosuppressive drugs after renal transplantation may partly be due to the effect on TNFα production by renal parenchymal cells

Synergistic effect of interleukin-1 and CD40L on the activation of human renal tubular epithelial cells

Synergistic effect of interleukin-1 and CD40L on the activation of human renal tubular epithelial cells.BackgroundRenal tubular epithelial cells are a central cell type in tubulointerstitial inflammation because they can produce inflammatory mediators such as cytokines and chemokines. Several signals derived from either monocytes or activated T cells have been reported to regulate the activation of tubular epithelial cells. We studied this regulation in more detail by combined treatment with CD40 ligand and the proinflammatory cytokine interleukin-1 (IL-1) in vitro.MethodsThe regulation of cytokine and chemokine production was studied in primary cultures of human proximal tubular epithelial cells (PTECs). PTECs were activated by coculture with CD40L-transfected murine fibroblasts in combination with recombinant human cytokines. The production of IL-6, IL-8, monocyte chemoattractant protein-1 (MCP-1), and RANTES were measured by specific enzyme-linked immunosorbent assay.ResultsThe combined activation of PTECs with CD40L and IL-1 resulted in strong synergistic effects on the production of IL-6, IL-8, and RANTES, whereas only an additive stimulation of MCP-1 production was observed. The effects were specific for IL-1 and could be neutralized by the addition of the IL-1R antagonist. Both IL-1α and IL-1βbgr; showed similar effects on cytokine production by PTECs. The effects of IL-1 were dose dependent, and kinetic experiments showed that synergistic effects were observed after 24 hours of activation and remained present for at least five days. Reverse transcription-polymerase chain reaction analysis showed that human PTECs could express both IL-1α and IL-1βbgr;. The activation of PTECs with IL-1 resulted in an up-regulation of CD40 expression on these cells.ConclusionsA complex network of regulation exists for the production of cytokines and chemokines by PTECs. The combined treatment results in strong synergistic effects on IL-6, IL-8, and RANTES production. This strengthens the potential role of tubular epithelial cells in inflammatory responses within the kidney

Decreased IgA1 response after primary oral immunization with live typhoid vaccine in primary IgA nephropathy

Introduction. Patients with primary IgA nephropathy (IgAN) have an increased level of immunological memory to certain parenteral recall antigens. We recently found a deficient IgA1 immune response after intranasal challenge with a neo-antigen: cholera toxin subunit B. In the present study, we assessed the specific IgA1 and IgA2 antibody response in plasma, peripheral blood cells and mucosal secretions after primary enteral immunization. Methods. Twenty eight IgAN patients, 26 patients with non-immunological renal disease and 32 healthy subjects were immunized orally with three sequential doses of live, attenuated, Salmonella typhi Ty21a. The humoral immune response in body fluids and antibody synthesis by circulating B cells was assessed in specific ELISAs and ELIPSAs respectively. Results. Oral immunization resulted in significantly (P < 0.0001) increased IgM, IgG, IgA, IgA1 and IgA2 responses in all groups, both in plasma and in circulating B cells in vitro. The IgA1 response in plasma was significantly (P < 0.05) lower in IgAN patients, while no significant differences in IgM (P = 0.36), IgG (P = 0.79) or 1gA2 (P = 0.45) responses were found as compared with matched control groups. The amount of IgA1 synthesized by circulating B cells tended to be lower in IgAN patients. No significant IgA response after oral immunization with S. typhi Ty21a was found in saliva (P = 0.11) or tears (P = 0.10). Conclusions. These data suggest an IgA1 hyporesponsiveness in patients with IgAN that is not only apparent after primary challenge of the nasal-associated lymphoid tissue but also after presentation to the gut. Previous results after parenteral recall immunization may be explained by assuming that IgAN patients require more frequent and/or longer exposure to IgA1-inducing antigens on their mucosal surfaces before they reach protective mucosal immunity. As a consequence, overproduction of IgA1 antibodies occurs in the systemic compartment, accompanied by an increased number of IgA1 memory cells

Complement Factor C4d Is a Common Denominator in Thrombotic Microangiopathy

Complement activation has a major role in thrombotic microangiopathy (TMA), a disorder that can occur in a variety of clinical conditions. Promising results of recent trials with terminal complement-inhibiting drugs call for biomarkers identifying patients who might benefit from this treatment. The primary aim of this study was to determine the prevalence and localization of complement factor C4d in kidneys of patients with TMA. The secondary aims were to determine which complement pathways lead to C4d deposition and to determine whether complement activation results in deposition of the terminal complement complex. We examined 42 renal sections with histologically confirmed TMA obtained from a heterogeneous patient group. Deposits of C4d, mannose-binding lectin, C1q, IgM, and C5b-9 were scored in the glomeruli, peritubular capillaries, and arterioles. Notably, C4d deposits were present in 88.1% of TMA cases, and the various clinical conditions had distinct staining patterns within the various compartments of the renal vasculature. Classical pathway activation was observed in 90.5% of TMA cases. C5b-9 deposits were present in 78.6% of TMA cases and in 39.6% of controls (n=53), but the staining pattern differed between cases and controls. In conclusion, C4d is a common finding in TMA, regardless of the underlying clinical condition. Moreover, C5b-9 was present in >75% of the TMA samples, suggesting that terminal complement inhibitors may have a beneficial effect in these patients. C4d and C5b-9 should be investigated as possible diagnostic biomarkers in the clinical work-up of patients suspected of having complement-mediated TMA