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M B van Werkhoven,
J Damman,
M C R F van Dijk,
M R Daha,
I J de Jong,
A Leliveld,
C Krikke,
H G Leuvenink,
H van Goor,
W J van Son,
P Olinga, J-L Hillebrands,
M A J Seelen
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ABSTRACT: Kidneys retrieved from brain-dead donors have impaired allograft function after transplantation compared to kidneys from living donors. Donor brain death (BD) triggers inflammatory responses, including both systemic and local complement activation. The mechanism by which systemic activated complement contributes to allograft injury remains to be elucidated. The aim of this study was to investigate systemic C5a release after BD in human donors and direct effects of C5a on human renal tissue. C5a levels were measured in plasma from living and brain-dead donors. Renal C5aR gene and protein expression in living and brain-dead donors was investigated in renal pretransplantation biopsies. The direct effect of C5a on human renal tissue was investigated by stimulating human kidney slices with C5a using a newly developed precision-cut method. Elevated C5a levels were found in plasma from brain-dead donors in concert with induced C5aR expression in donor kidney biopsies. Exposure of precision-cut human kidney slices to C5a induced gene expression of pro-inflammatory cytokines IL-1 beta, IL-6 and IL-8. In conclusion, these findings suggest that systemic generation of C5a mediates renal inflammation in brain-dead donor grafts via tubular C5a-C5aR interaction. This study also introduces a novel in vitro technique to analyze renal cells in their biological environment.
American Journal of Transplantation 02/2013; · 6.39 Impact Factor
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ABSTRACT: Smooth muscle cells (SMCs) play a key role in the pathogenesis of occlusive vascular diseases, including transplant vasculopathy. Neointimal SMCs in experimental renal transplant vasculopathy are graft-derived. We propose that neointimal SMCs in renal allografts are derived from the vascular media resulting from a transplantation-induced phenotypic switch. We examined the molecular changes in the medial microenvironment that lead to phenotypic modulation of SMCs in rat renal allograft arteries with neointimal lesions. Dark Agouti donor kidneys were transplanted into Wistar Furth recipients and recovered at day 56. Neointimal and medial layers were isolated using laser microdissection. Gene expression was analyzed using low-density arrays and confirmed by immunostaining. In allografts, neointimal SMCs expressed increased levels of Tgf β1 and Pdgfb. In medial allograft SMCs, gene expression of Ctgf, Tgf β1 and Pdgfrb was upregulated. Gene expression of Klf4 was upregulated as well, while expression of Sm22α was downregulated. Finally, PDGF-BB-stimulated phenotypically modulated SMCs, as evidenced by reduced contractile function in vitro which was accompanied by increased Klf4 and Col1α1, and reduced α-Sma and Sm22α expression. In transplant vasculopathy, neointimal PDGF-BB induces phenotypic modulation of medial SMCs, through upregulation of KLF4 in the media to contribute to (further) expansion of the neointima.
American Journal of Transplantation 03/2012; 12(6):1429-40. · 6.39 Impact Factor
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J Damman,
M R Daha,
H G Leuvenink,
H van Goor, J L Hillebrands,
M C van Dijk,
B G Hepkema,
H Snieder,
J van den Born,
M H de Borst,
S J Bakker,
G J Navis,
R J Ploeg,
M A Seelen
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ABSTRACT: Local renal complement activation by the donor kidney plays an important role in the pathogenesis of renal injury inherent to kidney transplantation. Contradictory results were reported about the protective effects of the donor C3F allotype on renal allograft outcome. We investigated the influence of the donor C3F allotype on renal transplant outcome, taking all different donor types into account. C3 allotypes of 1265 donor-recipient pairs were determined and divided into four genotypic groups according to the C3F allotype of the donor and the recipient. The four genotypic groups were analyzed for association with primary nonfunction (PNF), delayed graft function, acute rejection, death-censored graft survival and patient survival. Considering all donor types, multivariable analysis found no association of the donor C3F allotype with renal allograft outcome. Also, for living and deceased brain-dead donors, no association with allograft outcome was found. Post hoc subgroup analysis within deceased cardiac dead (DCD) donors revealed an independent protective association of donor C3F allotype with PNF. This study shows that the donor C3F allotype is not associated with renal allograft outcome after kidney transplantation. Subgroup analysis within DCD donors revealed an independent protective association of the donor C3F allotype with PNF, which is preliminary and warrants further validation.
American Journal of Transplantation 12/2011; 12(3):660-8. · 6.39 Impact Factor
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H Rienstra,
M Boersema,
G Onuta,
M W Boer,
A Zandvoort,
M van Riezen,
J Rozing,
H van Goor,
G J Navis,
E R Popa, J L Hillebrands
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ABSTRACT: Chronic transplant dysfunction (CTD) is the leading cause for limited kidney graft survival. Renal CTD is characterized by interstitial and vascular remodeling leading to interstitial fibrosis, tubular atrophy and transplant vasculopathy (TV). The origin of cells and pathogenesis of interstitial and vascular remodeling are still unknown. To study graft-versus-recipient origin of interstitial myofibroblasts, vascular smooth muscle cells (SMCs) and endothelial cells (ECs), we here describe a new rat model for renal CTD using Dark Agouti kidney donors and R26 human placental alkaline phosphatase transgenic Fischer344 recipients. This model showed the development of CTD within 12 weeks after transplantation. In interstitial remodeling, both graft- and recipient-derived cells contributed to a similar extent to the accumulation of myofibroblasts. In arteries with TV, we observed graft origin of neointimal SMCs and ECs, whereas in peritubular and glomerular capillaries, we detected recipient EC chimerism. These data indicate that, within the interstitial and vascular compartments of the transplanted kidney, myofibroblasts, SMCs and ECs involved in chronic remodeling are derived from different sources and suggest distinct pathogenetic mechanisms within the renal compartments.
American Journal of Transplantation 04/2009; 9(3):463-72. · 6.39 Impact Factor
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ABSTRACT: Previously, we reported that exclusive breastfeeding delayed and partially protected bio-breeding diabetes-prone (BBDP) rats from spontaneous autoimmune diabetes development. To investigate whether this protection results from modulation of the (mucosal) immune system, the present study was designed to analyse the effect of nutrition early in life on the immune status of BBDP rats.
The breastfeeding period of BBDP pups was extended or not, while allowing half of the pups to eat during that period whereas the other half received only breast milk. Cytokine profiles as well as naturally occurring regulatory T-cell frequencies were measured over time in the mesenteric lymph nodes (MLNs) and spleen.
Prolonged exclusive breastfeeding partially protects against autoimmune diabetes development and resulted in elevated levels of natural regulatory T cells (CD4(+) CD25(+) FoxP3(+)) in MLNs and spleen directly after weaning and throughout life. Stimulation of MLN cells from rats that ingested solid food during the nursing period showed massive secretion of interferon gamma (IFN-gamma), interleukin (IL)-4 and IL-10, whereas MLN cells from exclusive breastfed rats did not. In contrast, transforming growth factor beta (TGF-ss) was secreted equally by all groups.
Prolonged exclusive breastfeeding partially protects BBDP rats from autoimmune diabetes development. Interestingly, ingestion of solid food during the weaning period completely abolishes this protective effect. The protective effect of exclusive breastfeeding correlates with higher levels of naturally occurring regulatory T cells throughout life and low cytokine secretion at weaning.
Diabetes/Metabolism Research and Reviews 04/2009; 25(4):380-7. · 3.37 Impact Factor
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ABSTRACT: Atherosclerosis is the main contributor to cardiovascular disease and leads to intimal plaque formation, which may progress to plaque rupture with subsequent thromboembolic events and/or occlusion of the arterial lumen. There is increasing evidence that the development or progression of atherosclerosis is associated with advanced glycation endproducts (AGEs). AGEs are a heterogeneous group of compounds formed by the non-enzymatic reaction of reducing sugars with proteins, lipids, and nucleic acids. An increased understanding of the mechanisms of formation and interaction of AGEs has allowed the development of several potential anti-AGE strategies. This review summarizes AGE formation and biochemistry, the pathogeneic role of AGEs in cardiovascular disease, anti-AGE therapies and clinical relevance to vascular surgery.
European journal of vascular and endovascular surgery: the official journal of the European Society for Vascular Surgery 09/2008; 36(2):125-31. · 2.92 Impact Factor
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ABSTRACT: Although advances in graft procurement, preservation, matching and immunosuppression have all contributed to today's outstanding short-term graft survival rates after solid organ transplantation, similar success has not been achieved in preventing chronic transplant dysfunction (CTD) and extending long-term graft survival. CTD is being recognized as one of the major causes of long-term (>5 years) graft loss. CTD is featured by obliteration of the vascular lumen as a result of occlusive neointima formation referred to as transplant arteriopathy (TA). Uncontrolled proliferation (hyperplasia) of smooth muscle cells (SMCs) is in major part responsible for neointima formation in TA. Endothelial cells (ECs) cover the neointima and form the barrier between the circulating blood and the vascular wall. The etiology of TA is largely unknown and development of TA is refractory to most anti-rejection therapies. Recent data attribute an important role to host-derived vascular progenitor cells in the development of TA, and these cells may be recruited from various sources including the bone marrow. Vascular progenitor cells are potential targets for therapeutic intervention to attenuate TA development. Therefore, understanding the molecular pathways that promote recruitment of vascular progenitor cells, that determine their differentiation fate, and that determine the proliferative capacity of their progeny, is warranted to dissect their detrimental and possible beneficial effects in the development of TA.
Current Genomics 09/2005; 6(6):431-437. · 2.41 Impact Factor
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B W A van der Strate, J L Hillebrands,
S S Lycklama à Nijeholt,
L Beljaars,
C A Bruggeman,
M J A Van Luyn,
J Rozing,
T H The,
D K F Meijer,
G Molema,
M C Harmsen
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ABSTRACT: The role of leukocytes in the in vivo dissemination of cytomegalovirus was studied in this experiment. Rat cytomegalovirus (RCMV) could be transferred to rat granulocytes and monocytes by cocultivation with RCMV-infected fibroblasts in vitro. Intravenous injection of purified infected granulocytes or monocytes resulted in a systemic infection in rats, indicating that our model is a powerful tool to gain further insight into CMV dissemination and the development of new antivirals.
Journal of Virology 11/2003; 77(20):11274-8. · 5.40 Impact Factor
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Journal of Antimicrobial Chemotherapy 07/2001; 47(6):894-5. · 5.07 Impact Factor
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ABSTRACT: The development of transplant arteriosclerosis (TA) is today's most important problem in clinical organ transplantation. Histologically, TA is characterized by perivascular inflammation and progressive intimal thickening. Current thought on this process of vascular remodeling assumes that neointimal vascular smooth muscle (VSM) cells and endothelium in TA are graft-derived, holding that medial VSM cells proliferate and migrate into the subendothelial space in response to signals from inflammatory cells and damaged graft endothelium. Using MHC class I haplotype-specific immunohistochemical staining and single-cell PCR analyses, we show that the neointimal alpha-actin-positive VSM cells in rat aortic or cardiac allografts are of recipient and not of donor origin. In aortic but not in cardiac allografts, recipient-derived endothelial cells (ECs) replaced donor endothelium. Cyclosporine treatment prevents neointima formation and preserves the vascular media in aortic allografts. Recipient-derived ECs do not replace graft endothelium after cyclosporine treatment. We propose that, although it progresses beyond the needs of functional repair, TA reflects the activity of a normal healing process that restores vascular wall function following allograft-induced immunological injury.
Journal of Clinical Investigation 07/2001; 107(11):1411-22. · 15.39 Impact Factor
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ABSTRACT: We showed previously that our intrathymic immune modulation protocol induces virtually permanent graft survival of simultaneously transplanted cardiac allografts in MHC-incompatible rat strain combinations. It is, however, unknown whether this procedure prevents the development of graft arterial disease (GAD).
Male AO recipient rats were intrathymically inoculated with 2.5x10(7) PVG splenocytes immediately followed by heterotopic transplantation of a PVG cardiac allograft (day 0). Immunosuppression consisted of 1 ml of antilymphocyte serum i.p. (day 0) and cyclosporine i.m. (15 mg/kg body weight) on days 1, 2, and 3 posttransplantation. Histological analysis, mixed lymphocyte reactions, and intragraft cytokine mRNA expression were performed at several time points after engraftment.
Histological analysis revealed that GAD was already present 14 days after transplantation. At 200 days, virtually all vessels were affected and over 80% of the vessels showed severe intimal lesions. Infiltrate analysis displayed massive parenchymatous infiltrates (CD8+ cells and ED1+ macrophages) 2 weeks after transplantation. At later time points, infiltrates became epicardial and/or blood vessel associated and mainly consisted of CD4+, CD8+, and B cells. Mixed lymphocyte reactions showed nonspecifically decreased responses at 60 days but complete restoration of these responses at later time points (120 to 280 days). Intragraft cytokine mRNA expression showed decreased interleukin-2/interferon-gamma and sustained interleukin-10 expression 2 weeks after transplantation. Transforming growth factor-beta mRNA expression was increased >200 days after transplantation.
Intrathymic immune modulation does not abolish alloreactivity, and despite induction of long-lasting graft survival, this procedure does not prevent and may even facilitate the development of GAD.
Transplantation 04/2001; 71(7):914-24. · 4.00 Impact Factor
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ABSTRACT: The salivary gland is the preferred organ for cytomegalovirus (CMV) replication and viral persistence. In order to identify the nature of infected cells and to study viral replication in more detail, several experiments were conducted. Using the rat CMV (RCMV) model, acutely infected young adult rats (6 weeks of age) and new-born rats (3 days of age) were infected, and submandibular, parotid and sublingual glands were harvested at different time points after infection. For identification of the nature of infected cells, immunohistochemistry, in situ hybridisation and electron microscopic techniques were used. In young adult animals, the submandibular gland was the preferred organ for RCMV replication. The parotid and sublingual glands contained fewer viruses than the submandibular gland. In contrast, in new-born rats, the main site of RCMV replication was the sublingual gland, while the submandibular and parotid glands contained low amounts of virus. No virus could be detected in the parotid glands. In all glands of RCMV-infected animals, the infection was exclusively confined to striated duct cells. Infection resulted in a cellular inflammatory response which was mostly located in the interlobular duct region, whereas only few inflammatory cells were found in the neighbourhood of infected striated duct cells. This phenomenon may contribute to the long persistence of the virus in this organ.
Archiv für Pathologische Anatomie und Physiologie und für Klinische Medicin 11/2000; 437(4):413-21. · 2.49 Impact Factor
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ABSTRACT: In chronic allografts, graft vessels eventually develop so-called "transplant vascular sclerosis" or "intimal hyperplasia". A major question is whether the cells in the neointima are donor or recipient derived. The process of transplant vascular sclerosis closely resembles the remodeling of the vascular wall as seen when synthetic biodegradable small caliber vascular grafts are implanted. In this model, the cells in the newly developing neointima as well as neomedia are, by definition, recipient derived. By using cardiac allografts as well as aortic allografts exchanged between a female donor and a male recipient (rats), the origin of the neointimal vascular smooth muscle cells could be traced by looking for the Y-chromosome in isolated (alpha-actin positive) intimal cells using PCR. In both models these intimal cells were found to be of recipient-origin. It is proposed, that, basically, this remodeling process is part of a normal healing process. Whereas in biodegradable grafts this "healing process" appears to be self limiting, in allografts the process goes on beyond the needs of functional repair, eventually, in some cases, leading to total vascular occlusion. Future therapeutic protocols might try and aim at controlling this essentially normal repair process.
Critical Reviews in Immunology 02/2000; 20(1):85-8. · 3.32 Impact Factor
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Transplantation Proceedings 06/1999; 31(3):1563-6. · 1.00 Impact Factor
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ABSTRACT: The role of the liver in the regulation of systemic insulin levels is not well understood. The reported extraction rates vary between 0 to 85%, and extraction of a constant fraction of 50% of the portally delivered insulin is generally assumed. In the present study, we have investigated the role of the liver in the regulation of systemic insulin levels in the normal rat. Insulin was infused into the portal vein of conscious and freely moving rats in doses of 20, 40, 80 pmol/min during 15 min to mimic the gradual release of insulin by the native endocrine rat pancreas. The profiles of plasma insulin and glucose levels in the systemic circulation were compared to those obtained after direct infusion into the systemic circulation. The effect of intraportal and direct systemic infusion on plasma insulin and blood glucose levels were virtually similar where 20 pmol/min was applied. But, these effects were different if the dose was 40 pmol/min, and this difference increased when the dose was increased to 80 pmol/min, since hypoglycemia was less severe and normoglycemia was restored more rapidly with portal than with systemic infusion. Thus, our results show that the fraction of intraportally infused insulin reaching the systemic circulation decreases with higher doses of insulin. This suggests that the liver contains adaptable mechanisms to reduce the systemic insulin levels.
Hormone and Metabolic Research 01/1999; 30(12):721-5. · 2.19 Impact Factor
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Transplantation Proceedings 03/1998; 30(2):484. · 1.00 Impact Factor
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ABSTRACT: An intraperitoneally located and prevascularized expanded polytetrafluoroethylene solid support is potentially a suitable transplantation site for encapsulated pancreatic islets, because it allows for both the implantation of a large volume islet graft in the immediate vicinity of blood vessels, and its complete removal. The present study investigates the efficacy of such solid supports for the implantation of nonencapsulated islet isografts in streptozotocin diabetic rat recipients. These solid supports were always coated with acidic fibroblast growth factor, because we found that this growth factor enhances the neovascularization. The success rates of 5-microl (group A) and 10-microl (group B) islet isografts in solid supports were compared with the success rates of 5-microl (group C) and 10-microl (group D) islet isografts implanted in the unmodified peritoneal cavity. Four of seven rats in group A and all seven rats in group B became normoglycemic for at least 6 months. Only two of eight rats in group C and four of eleven rats in group D showed normoglycemia. The normoglycemia lasted for at least 6 months in zero of two animals in group C and in three of four animals in group D. Because of the low success rates in groups C and D, intravenous and oral glucose testing were restricted to the successful recipients in groups A and B. Glucose tolerance was found to be proportional to the grafted islet volume but, expectedly, in both groups the glucose tolerance and the insulin responses were somewhat lower than in controls. Thus, the prevascularized expanded polytetrafluoroethylene solid support, rather than the unmodified peritoneal cavity, is an efficacious transplantation site, potentially suitable for encapsulated islets.
Transplantation 04/1997; 63(6):824-30. · 4.00 Impact Factor
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Transplantation Proceedings 33(1-2):324-5. · 1.00 Impact Factor
