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Heparan sulfate proteoglycans (HSPGs) are well known for their proposed role in glomerular filtration. In addition, HSPGs can bind the leukocyte adhesion molecule l-selectin and chemokines, suggesting a role in inflammation. We examined a panel of biopsies representing different human primary kidney diseases for l-selectin and monocyte chemoattractant protein-1 (MCP-1) binding. In various renal diseases, l-selectin and MCP-1 binding to interstitial perivascular matrix HSPGs is increased, which is significantly associated with leukocyte influx. In proteinuric diseases, including membranous glomerulopathy, minimal change disease, but also IgA nephropathy and lupus nephritis, increased binding of l-selectin and MCP-1 to tubular epithelial cell (TEC) HSPGs is observed, which colocalizes with increased basolateral syndecan-1 and anti-heparan sulfate 10E4 staining. Short-hairpin RNA-mediated silencing demonstrates that syndecan-1 on TECs indeed mediates l-Selectin binding. Increased TEC expression of IL-8 in biopsies of proteinuric patients suggests that the increase in luminal protein may activate TECs to increase expression of l-selectin and MCP-1 binding syndecan-1. Strikingly, urinary syndecan-1 from proteinuric patients is less capable of binding l-selectin compared with urinary syndecan-1 from healthy controls, although syndecan-1 concentrations are similar in both groups. Together, our data show pronounced tubulointerstitial HSPG alterations in primary kidney disease, which may affect the inflammatory response.

Dendritic cells (DC) are the most potent antigen‐presenting cells and are therefore useful to induce immune responses against tumor cells in patients. DC can be generated in vitro from monocytes using GM‐CSF and IL‐4, the so‐called monocyte‐derived DC (MoDC). To achieve antitumor responses, MoDC must be able to migrate to the draining lymph nodes after injection to induce cytotoxic T cells. Therefore, we studied migration of MoDC in a rat model. Functional rat MoDC were generated from PVG‐RT7B rats and injected subcutaneously into PVG rats. These rat strains differ only at one epitope of the leukocyte‐common antigen, which can be recognized by the antibody His 41. The advantage is that migrated cells can be detected in the draining lymph nodes by staining sections with His 41+; thus, migration is not influenced by labeling procedures. Rat MoDC migrated to the T‐cell areas of the draining lymph nodes, just as isolated Langerhans cells or spleen DC do. In contrast, monocytes also migrated to the B‐cell areas and the medulla.

The commonly used peritoneal dialysis fluids contain glucose as the osmotic agent. Heat sterilization leads to the formation of glucose degradation products which contribute, together with glucose, to the formation of advanced glycation end-products (AGEs). AGEs have been shown to be present in the peritoneal cavity. Methods have been developed to minimize the amount of glucose degradation products in peritoneal dialysis fluids. In a rat peritoneal dialysis model, we compare the effect of a commonly used peritoneal dialysis fluid, Gambrosol, with a newly developed peritoneal dialysis fluid, PD-Bio, on the influx and functional capacity of the peritoneal cells after 2 weeks of peritoneal dialysis fluid instillation.

Abstract: Background: Immune suppression is an established consequence of surgical stress and trauma. Postoperative changes in the systemic immune system are proportional to the degree of surgical trauma and subsequent immune suppression may be implicated in the development of infectious complications and tumor metastasis formation. Laparoscopic surgery reduces the magnitude of the operative trauma and is thought to preserve postoperative immunological defenses. Methods: Relevant literature concerning postoperative immune functions and laparoscopic surgery was reviewed and clinical implications are discussed. Results: The influence of laparoscopic surgery on the postoperative systemic immune response is significantly less after laparoscopic cholecystectomy than with the conventional approach. Few immunological data are available concerning more advanced laparoscopic procedures. Various animal model studies of postoperative septic complications and tumor growth show that the postoperative preservation of the systemic immune response after laparoscopic surgery can have enormous clinical advantages. Conclusion: Laparoscopic surgery preserves the postoperative immunological defenses. In the future, this may imply a lower number of infections, less local recurrence and even fewer distant metastases. Prospective randomized studies are necessary to see whether these suspected advantages can be demonstrated in clinical practice.

In the mouse the maturation of mononuclear phagocytes was followed by comparing the ultrastructural pattern of endogenous peroxidatic activity (PA) at different time points during an acute peritonitis induced with newborn calf serum (NCS). Exudate macrophages demonstrate PA only in lysosomes, whereas resident macrophages have reaction product in the nuclear envelope (NE) and rough endoplasmic reticulum (RER). Transitional cells called "exudate-resident" macrophages have PA in the NE, RER, and some virginal lysosomes. In addition, peroxidase-negative macrophages were also present. A monoclonal antibody, F4/80, that specifically recognizes a mouse macrophage differentiation antigen (Austyn and Gordon, 1981) was used in this study. To compare the indirect immunoperoxidase labeling of this antigen and the endogenous peroxidase cytochemistry on the cellular level, a combined method was developed. Finally, the method was applied to the peritoneal cells at different time points after intraperitoneal injection of NCS in mice. The relative numbers of cells demonstrating the different patterns of endogenous PA and the proportions of each subpopulation expressing F4/80 antigen were estimated. It appeared that the expression of the antigen F4/80 coincides with the development of the resident pattern of PA. It is therefore concluded that the macrophages with the resident pattern of endogenous peroxidase are derived from monocyte-like exudate macrophages. In addition, the results indicate that both exudate-resident macrophages and at least a part of the peroxidase-negative macrophages are transitional forms.

The evidence that Kupffer cells are capable of controlling metastatic growth in the liver in vivo is largely circumstantial. The best approach when studying natural cytotoxicity activities of Kupffer cells is to investigate the effect of Kupffer cell elimination on tumour growth. Until now it has not been possible to eliminate Kupffer cells without affecting other cell populations. We have recently developed a new method to eliminate Kupffer cells selectively: intravenous injection of liposome-encapsulated (dichloromethylene)bisphosphonate (Cl2MDP-liposomes) leads to effective elimination of all Kypffer cells, without affecting non-phagocytic cells. Wag/Rij rats were injected with Cl2MDP-liposomes. After 48 h, rats were inoculated with syngeneic CC531 colon carcinoma cells by injection in the portal system. The results show a strongly enhanced tumour growth in the liver of the Cl2MDP-liposometreated rats. In these animals, livers were almost completely replaced by tumour and had increased in weight, whereas in the control groups only a few (four to eight) small (1-mm) tumour nodules were found. These data show that selective elimination of Kupffer cells results in enhanced tumour growth in the liver, implying that Kupffer cells play a crucial role in controlling tumour growth in the liver.

In the present study we examined whether mesothelial cells can ingest and digest bacteria. The results showed that all strains were ingested. Ingested staphylococci proliferated abundantly, and only a few were digested. Escherichia coli, however, was digested during the first 8 h, whereafter the mesothelial cells disintegrated and proliferation of bacteria could be observed. The clinical implications of these findings are discussed.

Summary Human long-term bone marrow cultures (HLTBMCs) were established with bone marrow samples collected from 15 patients with acute myeloid leukemia (AML) and compared with HLTBMCs from eight healthy volunteers. During 6 weeks of culture, the cellular composition of HLTBMCs was quantitatively studied. The cells of the HLTBMCs were divided into three main categories: fibroblasts, macrophages, and ‘other cells’ (endothelial cells, hematopoietic cells and undefined cells). HLTBMCs derived from healthy volunteers demonstrated a very consistent development. The number of fibroblasts increased during culture and the number of macrophages decreased, resulting in a steady state after 3 weeks of culture. In contrast, HLTBMCs derived from patients with AML showed a strikingly different pattern of irregular development and a steady state was not reached under our conditions. The APAAP technique was used to demonstrate expression of adhesion molecules. VLA2, VLA5, VLA6, LFA1, Mac1, p150/95, ß2-chain, HCAM, ICAM1, NCAM, and VCAM1 were more expressed on ‘normal’ as compared with ‘leukemic’ bone marrow stromal cells, although this reached significance only for ß2-chain and NCAM. VLA1, 3, and 4 were expressed in a higher percentage on ‘leukemic’ stroma (not significant). More expression was seen on ‘normal’ as opposed to ‘leukemic’ macrophages for the adhesion molecules tested, except for VLA5. The differences reached significance for the majority of molecules tested. It is concluded that striking differences exist in cellular composition and adhesion molecule expression between HLTBMCs from healthy individuals and those from patients with AML. This may have an impact on the pathogenesis of AML.

In this study we investigated the effect of the cytokines human granulocyte/macrophage-colony-stimulating Factor (hGM-CSF) and interferon ? (IFN?) on human Kupffer-cell-mediated cytotoxicity against the SW948 coloncarcinoma cell line. Kupffer cells were isolated from small liver wedge biopsies, taken from 14 patient who had had abdominal surgery for colon carcinoma or partial hepatectomy. The cells were incubated with hGM-CSF (100 ng/ml), or with IFN? (100 U/ml) or with their combination and the perecentage cytotoxicity was determined using a recently described modified assay. Additional experiments were performed with tumour-necrosis-factor-a(TNFa)-sensitive U937 cells as target. The TNFa secretion of Kupffer cells was measured and we evaluated the effect of TNFa on colon tumour targets. We performed human-Kupffer cell-mediated cytotoxicity blocking experiments with anti-TNFa and used paraformaldehydefixed Kupffer cells to demonstrate lysis of TNFa-sensitive WEHI-164 cells and of SW948 cells. The overall cytotoxicity against SW948 caused by unactivated Kupffer cells (n=14), and by Kupffer cells activated with hGM-CSF (n=14), IFN? (n=6) or their combination (n=6) was respectively: 19.5±2.6%, 25.3±2.9% 41±9.4% and 45.6±8% at E/T=1 and 28.2±2.9%, 35.6±3.2%, 55.6±9.7% and 62.8% at E/T=5. All differences were statistically significant (P<0.05). No growth-promoting activity by hGM-CSF on the SW948 tumour cells was observed. U937 cells were highly susceptible to Kupffer-cell-mediated cytotoxicity. The TNFa secretion by human Kupffer cells increased in parallel to their cytotoxicity after incubation with these cytokines. Soluble TNFa had only a slight anti-proliferative effect on SW948 cells, while specific anti-TNFa blocked Kupffer cell cytotoxicity by up to 80%. Finally, paraformaldehyde-fixed Kupffer cells were able to lyse WEHI-164 and SW948 cells. This indicates that expression of cell-associated TNFa is the main cytolytic mechanism of human-Kupffer-cell-mediated cytotoxicity. The implications for the use of hGM-CSF and IFN? in vivo are discussed.

Mesothelial cells (MC) are able to produce interleukin-8 (IL-8) after stimulation with IL-1 beta or tumor necrosis factor alpha. The aim of our study was to investigate whether MC are able to produce IL-8 after direct stimulation with clinically relevant bacteria. We observed a significant IL-8 response by the MC which were directly stimulated with viable staphylococci.

On light microscopical (LM) level dendritic cells (DC) isolated from lymphoid organs can be discriminated from macrophages (Mø) by the presence of acid phosphatase (APh) activity in a spot near the nucleus and constitutional expression of class II antigens. The aim of our study was to investigate whether DC and monocytes (Mo) enriched from human peripheral blood could be discriminated on the electron microscopical (EM) level. Therefore we developed a triple method by which we compared the presence of myeloperoxidase (MPO) containing vesicles, the localization of APh containing vesicles and expression of MHC class II and RFD1 (a DC-associated class II-like antigen) plasmamembrane antigens. DC, functionally characterized as potent stimulators in a MLR, are MPO-negative, whereas Mo show MPO in cytoplasmic granules. Although both DC and Mo show little APh activity at LM level, both types of cells show APh activity at the EM level but at different locations. In DC APh containing vesicles are present in a distinct juxtanuclear area, in contrast to Mo, which show APh activity in lysosomes scattered throughout the whole cytoplasm. Moreover, on both LM and EM level, DC are strongly class II positive, whereas Mo show variable labelling intensity for class II, while RFD1 was only found on DC.

Two genes involved in the enzymatic conversion of D-alanine to pyruvate are described, alnA and alnR. The alnA gene, located between ara and leu, is probably the structural gene for alaninase. The alnR gene, which can be cotransduced with thr but not with leu, could be demonstrated to be a regulatory gene with the aid of a mutation resulting in permanent repression and a thermosensitive revertant of this mutation restoring inducibility at 28°C, but not at 42°C.

It is widely accepted that the currently used dialysis solutions are not biocompatible with the peritoneal membrane. Therefore, animal studies have been performed to study different aspects of peritoneal dialysis. However, representative models mimicking the human situation are not yet available.