Fractalkine expression in human renal inflammation

Immune and inflammatory human renal disease is associated with heavy mononuclear cell infiltration. The trafficking of these cells to extravascular sites is directed by local production of chemokines. Fractalkine is the first described cell-surface anchored chemokine and has potent mononuclear cell-directed adhesion and chemotactic properties. The purpose of this study was to analyse the expression and distribution of fractalkine in human renal inflammation. In situ hybridization and immunohistochemistry were used to study renal biopsies from 15 patients with predominant glomerular inflammation (vasculitic glomerulonephritis) and 15 with predominant tubular and interstitial inflammation (acute renal allograft rejection). Controls comprised non-inflammatory glomerulonephritis and normal tissue. Fractalkine mRNA was predominantly expressed in the major compartment, glomerular or tubulointerstitial, affected by disease and with the strongest expression localized to vascular sites local to inflammation. In acute renal allograft rejection, there was increased expression of fractalkine mRNA by tubular epithelial cells. There was no expression of fractalkine by infiltrating leukocytes and there was only sparse expression in control tissue. Fractalkine mRNA expression correlated with infiltrating leukocyte subsets. Immunohistochemistry confirmed this pattern of expression, with serial section co-localization showing fractalkine expression in areas with macrophage (CD68+) and T cell (CD3+) infiltrates. These expression patterns show that fractalkine is a strong candidate for directing mononuclear cell infiltration in human renal inflammation. Copyright © 2001 John Wiley & Sons, Ltd

An in vitro model for analysing neutrophil migration into and away from the sub-endothelial space::Roles of flow and CD31

To model the later stages of neutrophil migration into tissue, we developed an assay in which human umbilical vein endothelial cells (HUVEC) were cultured on porous filters, treated with the inflammatory cytokine tumour necrosis factor-α (TNF), and then incorporated in a flow chamber. Video-microscopic observations were made of neutrophils as they were perfused over the HUVEC. When 3 μm pore filters were used (as opposed to 0.4 μm pore filters), neutrophils could be observed to migrate not only through the endothelial monolayer but also through the filter within minutes. The proportion of adherent neutrophils migrating through the endothelial monolayer and velocity of migration underneath it, were similar on the different filters, and also when neutrophils were perfused over cultures in glass capillaries, or settled on HUVEC cultured in standard plastic dishes. However, neutrophils migrated through HUVEC/filter constructs more rapidly in the flow chamber than in a standard, static, Transwell system, even though the velocities of migration under HUVEC were similar when directly observed under flow or static conditions. A function-blocking antibody against CD31 did not alter movement through the endothelial monolayer or the filter in the new flow system, but did reduce the migration velocity of neutrophils underneath the HUVEC (by 24%). Thus, we have developed a method for following each stage of neutrophil migration, including exit from the sub-endothelial space, and shown how they may be modified by applied fluid shear stress and blockade of a regulatory adhesion molecule

von Willebrand factor propeptide in malaria: evidence of acute endothelial cell activation

The pathogenicity of Plasmodium falciparum is thought to relate to the unique ability of infected erythrocytes to adhere to and subsequently activate the vascular endothelium. To study the state of endothelial activation during falciparum malaria, we measured plasma levels of both von Willebrand factor (VWF) and its propeptide, indices of chronic and acute endothelial cell perturbation, respectively. Results were correlated with clinical and biochemical markers of disease severity, including plasma lactate. Our data show that acute endothelial cell activation is a hallmark of malaria in children, indicated by a significant rise in VWF and VWF propeptide. The highest VWF and propeptide levels were seen in cerebral and non-cerebral severe malaria, and associations found between VWF propeptide level and lactate (P < 0.001). Mean VWF propeptide levels (nmol/l) were in cerebral malaria 33.4, non-cerebral severe malaria 26.3, mild malaria 22.1, non-malaria febrile illness 10.2, and controls 10.1. Differences between patient and control groups were highly significant (P < 0.005). Follow-up of 26 cerebral malaria cases showed that levels of VWF propeptide, but not VWF fell by 24 h, following the clinical course of disease and recovery. These novel findings potentially implicate acute, regulated exocytosis of endothelial cell Weibel-Palade bodies in the pathogenesis of Plasmodium falciparum malari

Host response to cytoadherence in Plasmodium falciparum

Cytoadherence of PRBCs (Plasmodium falciparum-infected red blood cells) to host endothelium has been associated with pathology in severe malaria, but, despite extensive information on the primary processes involved in the adhesive interactions, the mechanisms underlying the disease are poorly understood. Endothelial cells have the ability to mobilize immune and pro-adhesive responses when exposed to both PRBCs and TNF (tumour necrosis factor). in addition, there is also an up-regulation by PRBCs and TNF and a concurrent down-regulation of a range of genes involved in inflammation and cell death, by PRBCs and TNF. We propose that the balance between positive and negative regulation will contribute to endothelial pathology during malarial infection. Apposition of PRBCs has been shown by a number of groups to activate signalling pathways. This is dependent, at least in part, on the cytoadherence characteristics of the invading isolate, such that the avidity of the PRBC for the receptor on host endothelium is proportional to the level of activation of the signalling pathways. An understanding of the post-adhesive processes produced by cytoadherence may help us to understand the variable pathology seen in malaria and to design appropriate therapies to alleviate severe disease