1 AR is due to host immune responses towards antigens on the transplanted kidney that are foreign to the host, most importantly the human leucocyte antigens (HLA).2 Incompatible HLA can be recognized by alloreactive T cells through antigen-presenting cells (APC) either of donor organ origin (direct allorecognition) or in recipient host (indirect allorecognition).2,3 Effector host CD4+ and CD8+ T cells then home to the graft where they produce inflammatory cytokines and mediate direct destruction

of graft tissue.4 A number of products of cellular infiltration of the kidney have been studied as potential urinary biomarkers of rejection, including urinary Granzyme B and CD103.5 Other cell types in the kidney are also involved in the rejection process CAL-101 in vivo and may be useful potential markers for rejection. In particular, tubular epithelial cells (TEC) are able to https://www.selleckchem.com/products/Y-27632.html respond to inflammation and provide a rich source of potentially useful biomarkers into the urine for monitoring kidney function following transplant. A biomarker is defined as ‘a cellular, biochemical, molecule or genetic alteration by which a biological process can be recognized and/or monitor and has diagnostic and prognostic

utility’.6 Biomarkers may be membrane molecules (or fragments) shed following cleavage by proteolytic enzymes (either expressed by TEC or by infiltrating leucocytes at the local injury site) or secreted molecules such as cytokines. Such biomarkers may either be constitutively expressed or released by enhanced proteolytic activity present during inflammation, or alternatively, biomarkers may be absent in steady state, but

selectively upregulated during inflammation.7 In addition, oxidative stress, bacterial infection or inflammation, may induce alternate protein synthesis pathways, or induce alternate mRNA splicing, resulting in the secretion of ‘cell-associated’ molecules and peptides into biological fluids.7 Proteins associated with exosomes (100 nm lipid-bound particles) have also been discovered in urine8 and may provide an additional source selleck chemical of biomarkers.9,10 Detection of protein biomarkers generally involves a colorimetric or fluorescent system such as ELISA, Luminex® Beads and flow cytometry. Recently, proteomics have provided a comprehensive protein profile for analysing graft status. The proteomic approach used electrophoresis or chromatography techniques and mass spectrometry of graft biopsies, plasma and urine. Sigdel et al., in a comparative analysis of AR patients’ urine proteomic profiles with those of healthy controls and stable graft function established by Adachi et al.11 and Gonzalez et al.

This is illustrated in infection with Chlamydia trachomatis, Salmonella Typhimurium, Francisella Doramapimod cost tularensis, Mycobacterium tuberculosis, and Leishmania major. In the case of Chlamydia infection, B-cell-deficient and FcγR-deficient mice not only have a significantly higher mortality rate than wild-type mice in lethal challenges but also show reduced Th1 responses and fail to mount an efficient DTH response 37–39. Analogous observations have been made when protection against S. Typhimurium was studied. As seen in Chlamydia infection, B-cell-deficient mice are not protected against lethal challenge with Salmonella and develop reduced Th-cell responses marked by lower levels

of IFN-γ and IL-2 40, 41. In addition, when Ab-opsonized Salmonella are added to DCs in vitro, the DCs process and present Ag more efficiently than in the absence of Abs and

are able to stimulate enhanced T-cell activation 42, 43. The interaction of Abs with activating LY2157299 in vitro FcγRs is also required for optimal protection in M. tuberculosis-infected mice, as aerogenically infected μMT mice, as well as FcRγ−/− mice, show exacerbated immunopathology corresponding with elevated pulmonary recruitment of neutrophils and increased levels of IL-10 in the lung 44, 45. On the contrary, mice lacking inhibitory FcγRIIB manifest enhanced mycobacterial control upon infection and have increased Montelukast Sodium levels of Th1-promoting IL-12 45. Similarly, the protective effect of passively transferred Abs against Francisella has been attributed to limited sequelae associated with infection and an increased T-cell response in the presence of Abs 46, 47. Protective

effects of Ab–FcR interactions on T-cell responses have also been described for infections with intracellular parasites as described in the next section. In the absence of specific Abs, L. major amastigotes are phagocytosed by macrophages, which present Ag on MHC class II and activate memory, but not naïve, T cells; however, when L. major-specific IgG is present, amastigotes are taken up by DCs via FcγRI and II, which results in DC activation and production of IL-12 by the DCs. In contrast to macrophages, DCs are able to effectively prime naïve T cells and the resulting Th1 response leads to smaller lesions and reduced parasite burden 48. On the contrary, the presence of Abs during Leishmania mexicana infection drives a Th2 response and leads to the development of chronic lesions, whereas FcRγ−/− mice are resistant and able to resolve lesions by mounting a Th1 response. Resistance to L. mexicana is also observed in FcγRIII−/− mice, indicating that this receptor is responsible for shifting T-cell responses toward a Th2 phenotype via IL-10 production by macrophages 49, 50.

We therefore hypothesized that low levels of NKG2D ligands in vancomycin-treated mice could be explained by a less proinflammatory milieu

in the gut further regulated by the gut microbiota. To test if a less immune-suppressed intestinal environment could play a role in the potential gut microbiota-mediated suppression of NKG2D ligands on IECs, IL-10 B6 KO mice were compared with wild-type B6 mice as IL-10 is a key immunoregulatory cytokine counteracting the production of several proinflammatory cytokines and which PKC412 concentration thereby acts as an essential immunosuppressant in the gastrointestinal tract [37]. NKG2D ligand expression on epithelial cells isolated from the entire small intestine was significantly higher (p < 0.001) in IL-10 KO mice compared with B6 mice which indicate an, at least indirect, suppressive role of IL-10 in NKG2D ligand expression (Fig. 6). In order to alter the gut microbiota in a less-extreme way, male B6 mice were fed with a diet supplemented with XOS. XOS are a prebiotic candidate that stimulates microbes in the gut, such as bifidobacteria that may have beneficial effects on the host including anti-inflammatory effects on the immune system

to proliferate [38]. Thus, XOS feeding induces changes in the gut microbiota without compromising the physiologically normal functions of the gut, as opposed to antibiotic treatment, and may therefore in future treatment Lapatinib purchase strategies be considered as a better opportunity to correct dysbiosis. The NKG2D expression on duodenal IECs in B6 mice fed with XOS diet was found to be significantly lower compared than that in mice fed with standard diet (Fig. 7). In addition, click here the MFI was also

significantly lower (Table 1). It is therefore likely that the gut microbiota profile obtained after XOS feeding suppresses NKG2D ligand expression. Next, we analyzed the proportions of A. muciniphila in the XOS-fed mice, as we had seen an inverse correlation between this bacteria and the NKG2D ligand expression in the vancomycin-treated mice. Interestingly, this inverse correlation was clearly observed in the XOS-fed mice which also had significantly higher proportions of A. muciniphila in the gut compared with that in the control group (Fig. 7C). Our observations suggest that the gut microbiota strongly influences the expression of NKG2D ligands on small IECs. Germ-free mice lacking a commensal microbiota had an increased surface expression of NKG2D ligands, and a similar result was seen during ampicillin treatment which depleted most of the murine commensal bacteria. The NKG2D ligand expression returned to lower levels seen in the untreated mice after ampicillin treatment ended.

All participants were recruited between May 2008 and March 2009 at the Ottawa Hospital, Ontario, Canada. The participants were classified into three groups, namely healthy controls, latent TB and active TB. The demographic data including age, gender and ethnicity are listed in Table 1. Eleven participants who were tested negative to tuberculin skin test (TST), which was defined as having

induration of ≤5 mm, were considered to be healthy controls. Twenty-four participants with latent TB infection were diagnosed Dorsomorphin by positive TST (induration ≥10 mm) without any clinical and radiological evidence of active disease. Nine active TB patients were diagnosed on the basis of positive acid-fast bacilli staining and culture from sputum, bronchoalveolar

lavage or lymph nodes. Two patients had extrapulmonary Romidepsin supplier tuberculosis (TB lymphadenitis and cystitis). None of the latent TB individuals had any active infections at the time of blood acquisitions. All participants with latent and active TB infection were enrolled prior to receiving medication for tuberculosis. All participants were HIV seronegative. Informed consent was given by all participants based on the study protocol, which was approved by the Research Ethics Boards of the Ottawa Hospital Research Institute. The peripheral heparinized blood (20–30 ml) was collected and used for whole blood cytokine assay and for PBMC intracellular cytokine assay. M. bovis culture filtrate (CF) was a gift from Dr Bryan D. Rennie (Health

Canada, Ottawa, Ontario, Canada). This culture Protirelin filtrate is 99% identical to M. tuberculosis. Phorbol 12-myristate-13 acetate (PMA) (Sigma-Aldrich, St Louis, MO, USA) and ionomycin (Invitrogen, Burlington, Ontario, Canada) were used to stimulate cells as a positive control in a whole blood assay. The following antibodies were used for surface and intracellular staining: anti-human-CD3-fluorescein isothiocyanate (FITC), IFN-γ-FITC, CD8-phycoerythrin (PE)-Texas Red (ECD), CD14-ECD (Beckman Coulter, Mississauga, Ontario, Canada); CD4-allophycocyanin and cyanin (APC Cy7), CD8-PE, CD25-PE, IL-22-PE, IL-17-APC Cy7 (R&D Systems, Minneapolis, MN, USA) and CD15-FITC (Sigma-Aldrich). Anti-CD4-APC Cy7 antibody listed above was used in all experiments gating for CD4 T cells. Up to five antibodies were used in each experiment.

7 The pathological findings in the central nervous system of affected humans and animals, characterized by atrophy and the absence of inflammatory changes, were such that intoxication was strongly implicated. A number of possibilities EPZ-6438 mouse including Mn, CS2, Cu, Zn, Tl, Se, As, and V were considered. In 1959, Takeuchi read the previous description

of human alkylmercury poisoning made by Hunter and Russell.8 This led him to the notion that the neurological disorder seen around Minamata Bay must have been caused by alkylmercury compounds. In the meantime, he and his colleagues were able to demonstrate the feeding animals with fish or shellfish from Minamata Bay could produce a similar neurological disorder. This finding, which was consistent with the possibility of foodborne intoxication, was soon confirmed by Hosokawa and his collaborators. Investigation revealed that the chemical plant had been utilizing mercuric sulfate as the catalyst for acetaldehyde synthesis in sharply increasing amounts and discarding the waste catalyst into the effluent outlet directly connected to the sea. It was strongly suggested that the inorganic mercury discharged from the plant was somehow responsible for the disease. However, there was a missing link between the

organic and inorganic forms of mercury. Soon afterwards, BMN 673 datasheet a second outbreak of Minamata disease took place between 1964 and 1965, in Niigata approximately 250 km north of Tokyo. This outbreak was the subject of detailed studies by Tsubaki and other researchers from Niigata University School of Medicine.9–11 Mercuric catalyst for acetaldehyde synthesis was again identified as the culprit. A difference from the Minamata outbreak was that a river (the Agano River) rather than the sea was polluted. Two important discoveries soon followed. In 1961, Uchida and his associate at the Department of Biochemistry, Kumamoto University School of Medicine, succeeded in detecting a methylmercury

compound (methylmercury sulfide) in shellfish samples taken from Minamata Bay. In 1962, Irukayama and his colleagues at the Department of Hygiene, Kumamoto University School of Medicine, identified methylmercuric chloride in sludge from the acetaldehyde plant and the bottom sediment of the effluent channel. He postulated that it was formed from mercuric sulfate Protein kinase N1 as a by-product in the reaction for acetaldehyde synthesis. The causal links between the source and the disease thus became evident. It should be added that Hosokawa independently succeeded in detecting a methylmercuric compound in the effluent of the plant at about the same time. This achievement was published by Eto et al. in 2001.12 After 1995, the political problems related to MD were resolved in Japan and new facts have been gradually revealed. For example, Nishimura2 and Nishimura and Okamoto3 reported that large amounts of Me-Hg were generated by the chemical processes of the Chisso Co.

In addition to higher basal proliferation, draining LN cells from B10.S mice immunized with 3B3/PLP139–151/CFA showed much higher proliferation upon antigen restimulation (Fig. 5A). The treatment dramatically enhanced both IFN-γ- and IL-17-producing CD4+ T cells, while the treatment did not increase IL-4/IL10-producing T cells (Fig. 5B). Consistently, the 3B3-treated mice became susceptible to the development of EAE, with over 70% of B10.S mice developing this website EAE (Fig. 5C and Table 2). To further examine the effect of high-avidity anti-Tim-1 as a co-adjuvant on DCs and effector and regulatory T cells, we generated B10.S Foxp3/GFP ‘knock-in’ mice. The ‘knock-in’ mice were immunized with

3B3 or control rIgG in immunogenic emulsion. DCs, Foxp3−CD4+ effector T cells (Teffs), and Foxp3+CD4+ Tregs were Vemurafenib molecular weight isolated from spleen and lymph nodes of the mice and analyzed in criss-cross proliferation assays (Fig. 6A). Teffs from 3B3-treated

mice showed stronger proliferation and produced higher levels of IFN-γ and IL-17 upon antigen restimulation than Teffs from rIgG-treated mice. More interestingly, DCs from 3B3-treated mice induced higher Teffs proliferation and IFN-γ and IL-17 production than DCs from rIgG-treated mice (Fig. 6A). The frequency of Foxp3+ Tregs in spleens, lymph nodes, or the CNS was not significantly affected by 3B3 treatment (Fig. 6D and data not shown). However, Foxp3+ Tregs from 3B3-treated mice was less efficient in suppressing Teff proliferation in the cultures where Foxp3− Teffs and DCs were obtained from

rIgG-treated B10.S mice (Fig. 6B). Phenotypically, 3B3 in PLP139–151/CFA emulsion promoted DC activation as the treatment significantly upregulated the intensity of costimulatory molecules CD80, CD86, and MHC class II (Fig. 6C). In the CNS, treatment with the high-avidity anti-Tim-1 resulted in more mononuclear cell infiltration, containing high frequencies/numbers of CD11c+ DCs and CD4+ T cells (Fig. 6D and data not shown). Although the frequency of CD4+Foxp3+ Tregs in 3B3-treated mice was not dramatically decreased, significantly more Foxp3+ Tregs in the CNS of 3B3-treated MRIP mice produced proinflammatory cytokine IL-17 (7.85±2.36% from 3B3-treated mice versus 1.85±0.96% from rIgG-treated mice, n=3; p<0.05). In addition, the frequency of CNS-infiltrating CD4+Foxp3− Teffs producing IFN-γ and/or IL-17 was also increased in 3B3-treated mice (Fig. 6D). Moreover, similar to the observation in Fig. 5B, control rIgG-treated B10.S mice showed a very low percentage of IL-17-producing Teffs in the CNS, which was dramatically increased by the high-avidity anti-Tim-1 treatment (Fig. 6D). DCs are professional APCs with a remarkable capacity to activate naïve T cells and prime T-cell responses, therefore providing a link between innate and adaptive immunity.

The role of gut bacteria in immunological responses to C. parvum infection in mice has not been investigated directly, but studies suggest that bacteria are not so important in establishing the inflammatory response. Following infection of gnotobiotic Rucaparib molecular weight and conventionally reared lambs no differences between the groups in intestinal pathology or clinical signs were observed [68]. With piglets, intestinal inflammation and patent infection lasted longer in gnotobiotic animals than in control animals, suggesting the presence of intestinal bacteria provided a partial barrier to infection and also reduced immunopathology [69]. As Cryptosporidium

is a minimally invasive parasite and infects only epithelial cells whereas T. gondii infects most nucleated cell types, the role of bacteria in the immune response might be expected to differ. The induction of IL-12 expression by murine dendritic cells by T. gondii antigen in the absence of intestinal bacteria has been shown to be dependent largely on TLR11 recognition of the parasite protein profilin

[67]. A recent report described production of exceptionally high levels of IL-12 by cultured mouse spleens after addition of C. parvum profilin but the cell types producing IL-12 and TLR involvement in activation were not identified [70]. However, it has been reported recently that human dendritic RNA Synthesis inhibitor cells that do not have functional TLR11 produced significant amounts of IL-12 when exposed to C. parvum sporozoite antigen [45]. Results of murine investigations have confirmed a protective role for TLRs against C. parvum infection. Juvenile MyD88−/− mice had heavier infection burdens than control mice [71] while, compared with control animals, TLR4−/− mice took longer to clear infection why from the intestine and bile ducts and had an altered and

enhanced hepatic inflammatory response [72]. Weaned malnourished mice had increased susceptibility to infection compared with control animals that correlated with depleted intestinal expression of TLR2 and TLR4, but not TLR9 [73]. In a study with neonatal mice, administration of the TLR9 ligand CpG reduced the parasite load at the peak of infection by up to 95% and these mice had significantly increased expression of IFN-γ and IL-12 compared with controls [74]. In similar experiments with adult malnourished mice, only a modest reduction in the parasite load was obtained after CpG treatment [66]. The variation between degrees of resistance to infection induced by CpG in these two studies could be related to the different infection models employed or might imply that controlling infection by TLR9 stimulation is more readily achieved in the neonatal mouse. Figure 2 summarizes some of the major points regarding innate immune responses during C. parvum infection, combining in vitro and in vivo observations (predominantly with mice).

7B). TdTom-transduced cells expressed red tdTom protein spread throughout the cytoplasm (Fig. 1B-iv) and similarly to untransduced CTLs (Supporting Information Fig. 7A) relocalized GZMB-containing granules expressing Lamp-1 to the CTL/target contact zone (Fig. 1B-iv). Mathematical analyses showed that GZMB-tdTom colocalized with Lamp-1 and GZMB (Pearson’s Rr coefficient around 0.55) whereas tdTom did not show any colocalization (Rr 0.1) (Supporting Information Fig. 7C). Following TCR/antigen

engagement, calcium flux and PKC activation are important signals for gene activation and granule migration to the CTL/target contact zone preceding degranulation 4, 8. CTLs preloaded with Fluo-4 were used to monitor by video microscopy the Ca++ fluxes and the redistribution of GZMB-tdTom-containing granules. When GZMB-tdTom-transduced MI-503 manufacturer P14-TCR CTLs faced a specific target, an attachment signal preceded a rapid Ca++ flux (10–20 s) and granule translocation to the contact zone occurring at various times (20–480 s) (Fig. 1C-i and ii, Supporting Information Fig. 7D, Video 1). No significant signal was observed when the CTLs were facing control targets (Fig. 1C-iii and iv, Video 2). These kinetics are in agreement with published studies using CTL clones 6, 9. We used the Lamp-1 exposure method to assess CTL degranulation in response to antigenic stimulation and

to observe the fate of GZMB-tdTom during that process. GZMB-tdTom-transduced P14-TCR CTLs exposed Lamp-1

in response to gp33-loaded RMA-S, the extent of buy Staurosporine degranulation being dependent on peptide concentration (Fig. 2A). The percent of GZMB-tdTom fluorescent Urocanase CTLs markedly decreased (from 20% for non-stimulated or control-peptide stimulated CTLs to 13% for CTLs activated with 10−6 M gp33-loaded RMA-S), with a level of GZMB-tdTom fluorescence much lower in Lamp-1–positive (MRFI 422 (MRFI, mean relative fluorescence intensity)) as compared to Lamp-1–negative (607) CTLs. GZMB expression as measured on fixed and permeabilized cells were also reduced (about 50%) in the antigen-activated CTLs (data not shown). These results suggest that the whole GZMB-tdTom fusion protein was released during degranulation. Similarly, analysis of GZMB-tdTom-transduced OT1-TCR-Gzmb-KO (Gzmb, GZMB-encoding gene) CTLs, in which the only source of GZMB is GZMB-tdTom, showed that expression of GZMB-tdTom as well as GZMB was markedly decreased upon CTL activation with OVA-expressing cells (Supporting Information Fig. 8). We also found that the capacity of GZMB-tdTom-transducted P14-TCR CTLs to kill specific targets was not affected as compared to that of untransduced CTLs (Fig. 2B). To our knowledge, two attempts at expressing fluorescent GZMB fusion proteins have been reported, but they were not expressed in CTLs 10, 11.

The cytoplasmic expression strongly correlated with IL-1α expression (ρ = 0.9583). The cytoplasmic colocalization of HMGB1 and IL-1α was histologically confirmed in cells with collapsing nuclei by the double-staining method. The IgG4/IgG

indexes varied case by case. IL-6 and TLR4 expressions may influence IgG4/IgG index. The nuclei of cells with both IL-1α and HMGB1 expressions in the cytoplasm collapse in the cell death stage. The cooperative high expression of TLR4, IL-6, IL-18, MyD88 and HMGB1 suggest their AZD2014 solubility dmso critical roles in the inflammation circuit. “
“R. D. Jolly, N. R. Marshall, M. R. Perrott, K. E. Dittmer, K. M. Hemsley and H. Beard (2011) Neuropathology and Applied Neurobiology37, 414–422 Intracisternal enzyme replacement therapy in lysosomal storage diseases: routes of absorption into brain Aims: The research concerns enzyme replacement therapy in lysosomal storage diseases with central nervous system involvement. The principle aim was to understand the routes of entry of enzyme into the brain when delivered directly into the cerebrospinal fluid (CSF) via the cerebellomedullary cistern. Methods: Pathways for absorption of replacement enzyme were investigated in dogs with mucopolysaccharidosis IIIA (MPSIIIA) following intracisternal HSP inhibitor injections of human recombinant N-sulphoglucosamine

sulphohydrolase (rhSGSH, EC3.10.1.1) by light and confocal microscopy using chromogenic and fluorescent immune probes. Results: Enzyme entered the brain superficially by penetration of the pia/glia limitans interface, but the main route was perivascular along large veins, arteries and arterioles extending onto capillaries. It further dispersed into surrounding neuropil to be taken up by neurones, macrophages, astrocytes and oligodendroglia. Enzyme also entered the lateral ventricles adjacent to the choroid plexus, probably also by the tela choroidea and medullary velum, with further spread throughout Beta adrenergic receptor kinase the ventricular system

and spinal canal. There was secondary spread back across the ependyma into nervous tissue of brain and spinal cord. Conclusions: Enzyme mainly enters the brain by a perivascular route involving both arteries and veins with subsequent spread within the neuropil from where it is taken up by a proportion of neurones and other cells. Penetration of enzyme through the pia/glia limitans is minor and superficial. “
“I. El Ayachi, N. Baeza, C. Fernandez, C. Colin, D. Scavarda, P. Pesheva and D. Figarella-Branger (2010) Neuropathology and Applied Neurobiology36, 399–410 KIAA0510, the 3′-untranslated region of the tenascin-R gene, and tenascin-R are overexpressed in pilocytic astrocytomas Aims: Studying the molecules and signalling pathways regulating glioma invasiveness is a major challenge because these processes determine malignancy, progression, relapse and prognosis.

Comparison of the CD11b activation epitope on peripheral and transmigrated neutrophils was analysed by Wilcoxon matched pairs test. Correlations between IL-8, both endogenous and recombinant, and the expression of CD11b were analysed by Spearman’s rank order analysis. Significant correlations with a regression coefficient (R) ≥0.7

were further analysed. A P < 0.05 was considered significant. The median number of transmigrated cells per skin chamber was 2.14 (1.59–3.96) 106 cells with 82.1 (77.6–84.8) % granulocytes, 13.2 (10.3–16.4) % monocytes and 2.82 (2.37–4.37) % lymphocytes. In the peripheral circulation, the number of leucocytes was 4.85 (3.6–6.1)*109 leucocytes/l with 54.1 (50.5–58.9) % granulocytes, 8.51 (7.61–10.5) % monocytes and 33.3 (30.8–37.9) % lymphocytes. From the original skin blister, analysed for CD11b activation after 14 h of incubation, Opaganib price the median number of LY2109761 solubility dmso extravasated leucocytes was 0.11 (0.04–0.14) million cells per skin blister. The expression of CD11b activation epitope on extravasated neutrophils from the original 14-h blister was 73.2 (18.9–83.4) % and corresponding expression on circulating neutrophils was 1.96 (1.29–2.14) %, P = 0.04. The concentration of IL-8 in serum was 8.5 (1.9–11) pg/ml and in the 14-h blister fluid 338 (194–10,627) pg/ml, P = 0.04. The concentration of soluble mediators in serum and

in the skin chamber fluid was assessed by Milliplex multi-analysis and ELISA and

is presented in Fig. 1. Significantly higher concentrations of soluble markers were detected in the skin chamber fluid compared to that in serum for all markers Liothyronine Sodium except eotaxin (P < 0.01 for IL-4 and IL-10 and P < 0.001 for the remaining markers). TCC was analysed in chamber fluid, and median concentration was 28 (17–40) AU/ml. Figure 2 demonstrates the correlation between the number of in vivo extravasated neutrophils and the concentration of IL-8 in the chamber fluid at P < 0.05 and R = 0.79. In addition, the number of in vivo extravasated neutrophils also correlated with IL-1β, R = 0.83; IL-6, R = 0.73; IL-7 R = 0.71; and TNF-α, R = 0.71, all at P < 0.05. When the total number of extravasated leucocytes were analysed, the corresponding numbers were IL-8, R = 0.83; IL-1β, R = 0.81; IL-6, R = 0.72; IL-7 R = 0.71 and TNF-α, R = 0.70, also at P < 0.05. Following in vitro transmigration, the mean percentage of neutrophils that migrated towards chamber fluid was 34.2 ± 5.4% and towards cell culturing medium was 1.16 ± 0.55%. The percentage of transmigrated neutrophils correlated with the concentration of IL-8 (R = 0.79), IL-1β (R = 0.77) and TNFα (R = 0.79) at P < 0.05. The expression of CD11b activation epitope was measured following in vitro incubation with serum and skin chamber fluid. Figure 3 displays the expression of CD11b activation epitope following incubation with 50% serum, 50% skin camber fluid or IL-8 at 100 ng/ml.