Transplantation is the golden standard for the treatment of end-stage renal disease. During this process, the transplanted organ is often damaged. In this thesis, we investigated whether activation... Show moreTransplantation is the golden standard for the treatment of end-stage renal disease. During this process, the transplanted organ is often damaged. In this thesis, we investigated whether activation of the complement system, part of our innate immune system, plays a local role. We reviewed that properdin, the only known positive regulator of the complement system, was detected in serum, plasma and urine from patients with various complement-mediated renal diseases. In protocol biopsies obtained 10 days after transplantation, properdin was found deposited in addition to complement activation markers. Next, we showed that dendritic cells secrete properdin and a decrease in properdin levels during dendritic cell- T-cell interaction resulted in reduced T-cell proliferation and activation. We also showed that properdin is able to bind to surfaces of both viable and dead cells, contributing to complement activation. Macrophages can also produce properdin and negative regulators factor H and its splice variant FHL-1. Increasing knowledge on complement factor production by other cells than hepatocytes, including immune cells, hints towards a local role of the complement system in various processes. These findings contribute to a better understanding of the local role of the complement system and are important for the applications of (new) complement-inhibiting drugs. Show less
Essen, M.F. van; Schlagwein, N.; Gijlswijk-Janssen, D.J. van; Ruben, J.M.; Kooten, C. van; COMBAT Consortium 2022
The complement system does not only play an important role in the defence against microorganism and pathogens, but also contributes to the regulation of innate and adaptive immunity. Especially... Show moreThe complement system does not only play an important role in the defence against microorganism and pathogens, but also contributes to the regulation of innate and adaptive immunity. Especially activation fragments C3a and C5a and complement activation at the interface of antigen presenting cell (APC) and T cell, were shown to have a role in T cell activation and proliferation. Whereas most complement factors are produced by the liver, properdin, a positive regulator of the C3 convertase, is mainly produced by myeloid cells. Here we show that properdin can be detected in myeloid cell infiltrate during human renal allograft rejection. In vitro, properdin is produced and secreted by human immature dendritic cells (iDCs), which is further increased by CD40-L-matured DCs (mDCs). Transfection with a specific properdin siRNA reduced properdin secretion by iDCs and mDCs, without affecting the expression of co-stimulatory markers CD80 and CD86. Co-culture of properdin siRNA-transfected iDCs and mDCs with human allogeneic T cells resulted in reduced T cell proliferation, especially under lower DC-T cell ratio's (1:30 and 1:90 ratio). In addition, T cell cytokines were altered, including a reduced TNF-alpha and IL-17 secretion by T cells co-cultured with properdin siRNA-transfected iDCs. Taken together, these results indicate a local role for properdin during the interaction of DCs and allogeneic T cells, contributing to the shaping of T cell proliferation and activation. Show less
This thesis focuses on describing novel methodology for studying the complement system in experimental rodent models, on fundamental aspects of mouse complement, and the activation and... Show moreThis thesis focuses on describing novel methodology for studying the complement system in experimental rodent models, on fundamental aspects of mouse complement, and the activation and contribution of complement system in experimental renal ischemia/reperfusion (I/RI) and renal autoimmune disease animal models. Our findings show that transient and acute complement activation is observed following renal I/RI and that the developed and validated methodology for analysis of systemic complement can be a valuable addition to complement analysis in preclinical studies. Our novel findings included the characterisation of C5a receptor C5L2 and its contribution to renal I/RI. Our results revealed injurous contribution to the pathology of renal I/RI, in similar degree as with C5aR. Using anti-GBM renal autoimmune disease as an experimental model, we showed for the first time in vivo that properdin, the only positive regulator of complement, may direct complement activation independent of classical ligands such as C3. Other findings included the characterisation of drastic gender difference in mouse complement, where female mice have near-deficient terminal pathway serum activity, and the impact of therapeutic dietary restriction on serum complement, which was shown to downregulate the hepatic expression of the terminal pathway components in serum. Show less
Dixon, K.O.; O'Flynn, J.; Klar-Mohamad, N.; Daha, M.R.; Kooten, C. van 2017
The complement system has been shown to have a role in various systemic autoimmune (AI) diseases which have a renal component. This includes systemic lupus erythematosus (SLE), goodpastures... Show moreThe complement system has been shown to have a role in various systemic autoimmune (AI) diseases which have a renal component. This includes systemic lupus erythematosus (SLE), goodpastures syndrome and anti-neutrophil cytoplasmic antibody (ANCA) associated vasculitides. In particular the classical pathway (CP) of complement is involved in SLE, with the exact mechanism of how these SLE autoantigens along with their autoantibodies interact with complement in the glomerulus remaining ambiguous. Furthermore, recent studies have demonstrated the expanding role of the alternative pathway (AP) of complement activation in steady state or in aspects of ANCA associated vasculitides or anti-GBM disease. The precise means of activation of the AP in these immune complex mediated disease settings remains unclear. Regulation is imperative in the AP due to its inherent ability to autoactivate. Factor H and properdin are both key opposing regulators in the AP with novel emerging roles in initiation and control of the AP. Deciphering the specific modes of AP activation and involvement is an ever expanding field requiring further elucidation, with previously unexpected roles for AP components revealing the diversity and complexity of this ancient pathway. Show less
The innate immune system plays an important role in solid organ transplantation. This thesis focuses on the role of the lectin pathway of complement activation in kidney and simultaneous pancreas... Show moreThe innate immune system plays an important role in solid organ transplantation. This thesis focuses on the role of the lectin pathway of complement activation in kidney and simultaneous pancreas-kidney transplantation (SPKT) and describes the role of properdin in tubular complement activation and clearance of apoptotic cells. Mannose-binding lectin (MBL) is the initiating molecule of the lectin pathway of complement activation and its levels are largely determined by frequently occurring polymorphisms of the MBL gene. Our studies show that high MBL levels in the transplant recipient are associated with poorer graft survival and in the case of SPKT also with poorer survival of the recipient. While low MBL levels are associated with better survival we also show that low MBL does increase the risk for infectious complications after SPKT. Properdin is classically described as a stabilizer of the alternative pathway of complement activation. This thesis points towards a new role for properdin by describing a direct interaction with apoptotic cells and with the tubular epithelium of the kidney. By directly binding to these cellular surfaces properdin may contribute to the clearance of apoptotic cells and participate in complement activation and renal damage in proteinuric states. Show less
