Interestingly it has been reported that VCAM-1 was expressed Selleckchem Everolimus on endothelial cells according to the decreased shear stress of blood flow. Further, expression of VCAM-1 and VLA-4 was increased in active-chronic lesions of HAM/TSP. We have also reported characteristic expression of matrix metalloproteinases16 and a novel variant of CD4417 in such active-chronic lesions. Using these molecules,

HTLV-1-infected T-cells migrate into the CNS from the area where the blood flow is slow and initiate inflammatory lesions. HAM/TSP is now a well-defined clinicopahological entity in which the virus infection and the host immune responses are involved in the pathogenesis. Our series of studies mentioned here suggested that T cell-mediated chronic inflammatory processes targeting the HTLV-1 infected T-cells are the primary pathogenic mechanism of HAM/TSP (Fig. 5).18 Anatomically determined hemodynamic conditions may contribute to the localization of infected T-cells and forming of main lesions in the middle to lower thoracic spinal cord. “
“E. Zotova, C. Holmes, D. Johnston, J. W. Neal, J.

A. R. Nicoll and D. Boche (2011) Neuropathology and Applied Neurobiology37, 513–524 Microglial alterations in human Alzheimer’s disease following Aβ42 immunization GPCR Compound Library supplier Aims: In Alzheimer’s disease (AD), microglial activation prompted by the presence of amyloid has been proposed as an important contributor to the neurodegenerative process. Conversely following Aβ immunization, phagocytic microglia have been implicated in plaque removal, potentially a beneficial effect. We have investigated the effects of Aβ42 immunization on microglial activation and the relationship with Aβ42 load in human AD. Methods: Immunostaining against Aβ42 and microglia (CD68 and HLA-DR) was performed in nine immunized AD cases (iAD – AN1792, Elan Pharmaceuticals) and eight unimmunized AD (cAD) cases. Results: Although the Aβ42 load (% area stained of total area examined) was lower in the iAD than the cAD cases (P = 0.036), the CD68 load was higher (P = 0.046). In addition, in the iAD group, the CD68 level correlated with the Aβ42 load, consistent with

the immunization upregulating microglial phagocytosis when plaques are present. However, in Selleckchem Cetuximab two long-surviving iAD patients in whom plaques had been extensively cleared, the CD68 load was less than in controls. HLA-DR quantification did not show significant difference implying that the microglial activation may have related specifically to their phagocytic function. CD68 and HLA-DR loads in the pons were similar in both groups, suggesting that the differences in microglial activation in the cortex were due to the presence of AD pathology. Conclusion: Our findings suggest that Aβ42 immunization modifies the function of microglia by increasing their phagocytic activity and when plaques have been cleared, the level of phagocytosis is decreased below that seen in unimmunized AD. “
“D. Capper, M. Mittelbronn, B. Goeppert, R. Meyermann and J.

DCs stimulated directly or indirectly by PRRs from pathogens mature into a specific form and are able to activate a single specific immune response that is appropriate for the elimination of the mTOR inhibitor pathogen [32]. In this regard, DCs determine the nature of the foreign antigen and the intensity and phenotype of immune response generated. The development of different subtypes of effector

T cell differentiation, a Th1, Th2 or Th17 immune response, is dependent upon the physical interaction between the activated status of the DCs and the naive T cells [8,33] (Fig. 1). It will not be discussed in this review. It is worth mentioning that in addition to its importance in infectious diseases, TLRs also participate in inflammation and immune responses that are driven by self-, allo- or xeno-antigens [18,34,35]. TLR signalling has AZD0530 datasheet been demonstrated to be involved in the immune recognition of allo- or xenografts and the occurrence of autoimmunity [35,36]. This observation is supported strongly by the expression of TLRs on almost all immune cells and the identification of their endogenously expressing ligands by mammalian cells [9,37–39]. TLRs are expressed widely in many types of immune cells, including

DCs, T cells, neutrophils, eosinophils, mast cells, macrophages, monocytes and epithelial cells [1,40,41]. Interestingly, TLR expression is related to the functional status of different subtype T cells. TLR-3, -6, -7 and -9 have been reported to be expressed on CD4+ T cells [42]. Naive CD4+ T cells do not express significant levels of mRNA and intracellular proteins of TLR-2 and TLR-4. Only few CD3+ T cells express TLR-1, -2 or -4 on the cell surface when they have not been activated [43]. However, activated/memory T cells express appreciable levels of cell surface TLR-2 and TLR-4 [34,42]. TCR stimulation by cross-linked anti-CD3 monoclonal

antibody (mAb) induces cell surface expression of TLR-2 and TLR-4 on naive human and murine CD4+ T cells [34,44]. By contrast, TCR stimulation down-modulates significantly surface TLR-5 expression on human CD4+ T cells [45] (Table 1). TLR expression on T cells may be regulated by TCR signalling, which needs further investigation in the future. These data thus offer the possibility Fenbendazole that pathogens, via their PAMPs, may contribute directly to the perpetuation and activation of T cells. At least some TLRs may function as a co-stimulatory receptor for antigen-specific T cell responses and participate in the maintenance of T cell memory [46–48]. It has been shown that ligands for TLR-2, -3, -4, -5 and -9 enhance the proliferation and/or biological functions of conventional effector T cells [44,46,48–51]. Co-stimulation of CD4+ T effector cells with anti-CD3 mAb and TLR-5 ligand flagellin results in enhanced proliferation and production of IL-2 at levels equivalent to those achieved by co-stimulation with CD28 [52,53].

[23] Our

experimental data indicated that the globular head of C1q (gC1q) in spontaneous abortion patients showed clearly the magnitude of high intension compared with induced abortion patients (see Figure S3). Although the significance of cell surface gC1qR expression is not known, in the present set of experiments, it was observed that expression was enhanced in human placental villi tissues from patients who underwent spontaneous abortion, suggesting that its expression might play an important role during spontaneous abortion. The list of biological responses mediated by gC1qR is extensive, and gC1qR plays a major role in inflammation, infection and immune regulation.[24] When constitutively expressed in a normal murine fibroblast cell line, gC1qR induces growth perturbations, morphological abnormalities and initiates apoptosis.[25] Atezolizumab The gC1qR protein VX-809 in vivo has been extensively described in a previous study, and it is primarily an inducer of apoptosis.[14] Our study found that gC1qR was overexpressed in HTR-8/SVneo and HPT-8 cells, which in turn mediates EVCT-derived

transformed cells apoptosis (see Fig. 2 and Figure S4). Not only chemical substance can induce the expression of gC1qR gene, but also hormones such as gonadotropin can upregulate the expression of gC1qR gene. Recent cohort studies have shown that gC1qR is a conserved eukaryotic multifunctional protein that primarily localized in the mitochondrial matrix and on the cell surface. Human gC1qR is expressed as a proprotein of 282 amino acids (aa) whose first 73 amino acids, containing a mitochondrial localization signal, are required for localizing

AZD9291 mouse the protein to the mitochondria and are subsequently cleaved to generate mature gC1qR. The upregulation of mature form of gC1qR has been tied to apoptosis and autophagy via inducing mitochondrial dysfunction.[26] Increasing evidence suggests that mitochondrial dysfunction is linked to apoptosis initiated by cytotoxic factors such as ROS, which are generated in excess in defective mitochondria. These findings have focused attention on the role of the mitochondria in apoptosis. While it is not yet clear how mitochondria regulate apoptosis, it has been suggested that mitochondrial outer membrane permeabilization can occur following cellular stress, which can result in the release of apoptogenic factors (e.g. cytochrome c, Smac) into the cytosol. Data demonstrated that increased mitochondrial content at physiological levels provides protection against apoptotic cell death by decreasing caspase-dependent and caspase-independent signalling through influencing mitochondrial Ca2+-mediated apoptotic events, due to an increased sensitivity to Ca2+-induced mitochondrial membrane depolarization and mitochondrial permeability transition pore formation.[27] Our study demonstrated that gC1qR vector-treated HTR-8/SVneo and HPT-8 cells expressing gC1qR generated increased levels of ROS.

Such differences may be one of the causes of cell tropism for PrPSc accumulation, and furthermore, might result in the prion strain-specific PrPSc accumulation pattern in the brain. Species specificity in cell-free conversion has been reported

(14, 23), and the products preserve strain-specific properties (24). These data suggest that the cell-free conversion reaction mimics some aspects of in vivo conversion of PrPC into PrPSc. In this study, we demonstrated that the effect of reducing conditions and removal of Cys residues on binding and conversion differed among prion strains; indeed, these may mirror prion strain properties in vivo. In fact, classification of the five prion strains learn more by their binding and the conversion efficiencies correlated well with classification according to their biological and biochemical properties. Therefore, the

in vivo properties of each strain likely correlate with their conversion capacity. Binding and conversion assays may thus aid in the classification of prion strains. Reduction of the Z-VAD-FMK datasheet intramolecular disulfide bond did not interfere with binding of PrPSc to MoPrP and conversion of MoPrP into PrPres. However, substitution of Cys with Ser in MoPrP inhibited binding and conversion of the ME7 and Obihiro strains and conversion of the Chandler and 79A strains. Therefore, Cys residues may play a key role in the conversion and binding of Chandler and 79A, ME7, and Obihiro PrPSc. However, we cannot rule out the possibility that such a substitution alters the tertiary structure

of the prion protein. Addition of DTT significantly increased the Nintedanib (BIBF 1120) conversion efficiencies of MoPrP and the Cys-less mutant driven by mBSE PrPSc. This suggests that the effect of DTT may be mediated by a mechanism other than cleaving of the disulfide bond in MoPrP. DTT diminishes the carbohydrate binding activity of a Cys-less mutant of pigpen as well as inhibiting the intact molecule (25). Therefore, in an mBSE-seeded cell-free conversion, DTT may improve the efficiency of mBSE-seeded conversion independently of the reduction of disulfide bonds. In summary, reducing conditions did not inhibit conversion in vitro and markedly increased mBSE-seeded conversion. This suggests that cell-free conversion under reducing conditions mimics the conversion of PrPC into PrPSc within endosomes and lysosomes. In addition, classification of prion strains by their efficiency at binding and conversion of both MoPrP and its Cys-less mutant in the absence and presence of DTT correlates well with classification based on biological and biochemical properties. Therefore, the cell-free conversion assay may be useful in discriminating between prion strains. We are grateful to Dr.

Like flucytosine, Buparlisib in vivo terbinafine is usually administered in combination with other antifungal agents for the treatment of systemic infections.[21] Antifungal resistance can be intrinsic (naturally present) or acquired (developed according to environmental influences).[84, 85] Microorganisms can adapt and develop mechanisms of resistance to antifungal agents.[85] It is essential

to promote a rational use of antifungal agents in the hospital environment to decrease the occurrence of resistance but also to promote the most appropriate therapy and thereby increase survival rates in infected patients.[86] Acquired resistance of Candida spp. to azole drugs can occur by induction of the efflux pumps encoded by the MDR or CDR genes or acquisition of point mutations in the gene encoding the target enzyme (ERG11). Acquired resistance to echinocandins in Candida spp. clinical isolates comes as a result of point

mutations in the FKS1 gene or substitution of one or more amino acids in the structure of the GS enzyme.[87] Resistance to flucytosine is frequently acquired during therapy, as a result of changes in the enzyme uracil phosphoribosyltransferase (encoded by FUR1).[88] It is believed that resistance to terbinafine occurs by point mutations in the squalene epoxidase coding gene. Overexpression of CDR2 transporters FDA-approved Drug Library ic50 results in the decreased susceptibility of C. albicans to terbinafine.[21] Overuse of antifungals, especially fluconazole, promotes selection of isolates of Candida spp. resistant to azoles, which results in an increase in the incidence of infections caused by resistant Candida spp.[89, 90] Resistance to fluconazole in vitro can be

promoted by repeated exposure to the drug and it is believed that this also occurs in vivo.[91-93] The reduction of susceptibility to azole derivatives is more common among non-albicans Candida spp.[94] The frequency of invasive fungal infections and resistance to antifungal therapy has increased despite the introduction of new antifungal agents. Although antifungal susceptibility very tests are often used to select antifungals for therapy, currently, the most important function is to detect resistance.[85] A microorganism is considered resistant when it develops an infection and persists in the host, even in the presence of the maximum concentration of the drug at the site of infection.[95] In the 1990s, conventional treatment regimens for Candida spp. infections involved the use of antifungal polyenics such as amphotericin B and nystatin, and azoles such as fluconazole and itraconazole. In the following decade, voriconazole became part of the group.[96, 97] Although more effective, amphotericin B is nephrotoxic, which prevents its use in patients with chronic kidney disease. Currently, caspofungin has been used to treat infections caused by azole-resistant Candida spp. and Aspergillus spp.

: NM_017232.2); TGF-β1 forward, 5′-GACCGCAACAACGCAATCTA-3′ and TGF-β1 reverse, 5′-ACGTGTTGCTCCACAGTTGAC-3′ (GenBank accession no.: NM_021578.1); IL-6

forward, 5′-CAGCCACTGCCTTCCCTACT-3′ and IL-6 reverse, 5′-CAGTGCATCATCGCTGTTCAT-3′ (GenBank accession no.: NM_012589.1); β-actin forward, 5′-CCCGCGAGTACAACCTTCTT-3′ and β-actin reverse, 5′-CCACGATGGAGGGGAAGAC-3′ (GenBank accession no.: NM_031144.2). The significance of differences in the wound area trends between groups was evaluated using repeated-measures anova with group, time and the Decitabine group and time interaction as fixed effects. Multiple comparisons adjusted by the Bonferroni correction were performed to test the significance DNA Damage inhibitor of the differences between groups at each time point. The Wilcoxon rank sum test was used to compare the numbers of α-smooth muscle actin-positive cells between two groups. The software SAS ver. 9.1 (SAS Institute Inc., Cary, NC) was used for all statistical analyses. Values are presented as the mean and SD unless otherwise indicated.

Full-thickness wounds were created at lateroabdominal sites on both sides of each animal and kept moist until day 5. After granulation tissue had become established, 3-oxo-C12-HSL or the same concentration of DMSO was administered to the wound surface. Gross observations revealed increased wound contraction after 3-oxo-C12-HSL administration (Fig. 1a). The time course of the changes in the wound area clearly indicated accelerated wound healing at 24 h after the administration of the quorum-sensing signal (Fig. 1b). The interaction of group and time was significant (F=3.03, P=0.002), and multiple comparisons were therefore performed. The

relative areas were significantly smaller in the 3-oxo-C12-HSL group than in the vehicle group on days 6, 7, 8 and 9 (P=0.013, P<0.001, P=0.002 and P<0.001, respectively). HE staining of the granulation tissue revealed massive accumulation of fibroblasts in both groups (Fig. Thiamet G 2a). Infiltration of PMNs was also observed on the wound surface in the 3-oxo-C12-HSL group (Fig. 2b). Because wound contraction relies on the differentiation of fibroblasts to myofibroblasts, we further investigated the basis for the accelerated wound contraction by immunostaining of α-smooth muscle actin to assess myofibroblast differentiation (Ishiguro et al., 2009). The immunostaining revealed that α-smooth muscle actin-positive cells were clearly present across the granulation tissue in the 3-oxo-C12-HSL group, whereas the control DMSO group only contained α-smooth muscle actin-positive cells at the edge of the wound (Fig. 3). The number of α-smooth muscle actin-positive cells per high-power field was significantly higher in the 3-oxo-C12-HSL group than in the control group (P<0.001, Fig. 3).

Aboriginal and Torres Straight Islander (ATSI) transplant recipients have poorer allograft survival and higher rates

of acute rejection. We sought to determine whether a higher incidence of plasma cell-rich infiltrates (PCIR) could account for poorer survival. Methods:  Renal transplant biopsies performed in recipients from the Northern Territory of Australia between 1985 and 2007 were reviewed and correlated with outcome. Biopsies were designated PCIR positive when plasma cells constituted >10% of the interstitial infiltrate. Results:  Four hundred and seventy-seven biopsies from 177 recipients (108 ATSI) were performed. Median graft survival was shorter for recipients with PCIR: 4.0 years (interquartile range 2.18–6.41) versus 5.4 years (2.0–9.99) (P = 0.013).

ATSI recipients had higher rates of plasma cell-rich rejection (RR 1.76, 95% CI 1.43–2.17, Z-VAD-FMK research buy Selleckchem Ixazomib P < 0.0001), which occurred earlier (251 vs 869 days, P = 0.03) compared with non-indigenous recipients. On multivariate analysis, PCIR did not independently influence allograft survival. There was a correlation between PCIR and panel reactive antibody peak >20% (RR 1.29, 95% CI 1.03–1.56, P = 0.025), ≥5 human leukocyte antigen mismatches (RR 1.91, 1.41–2.58, p < 0.0001), increasing post-transplant infection rate (>10 infections RR 5.11, 1.69–15.5, P = 0.004), and subsequent death from septicaemia (RR 1.6, 1.17–2.18, P = 0.003). Conclusion:  PCIR is associated with infection and markers of chronic immunological stimulation but does not independently contribute to inferior renal allograft outcomes, even in ATSI recipients. “
“Aims:  Data regarding the occurrence of stroke in dialysis patients are limited and epidemiologic studies to date are controversial with respect to the stroke subtype among dialysis patients. The aim of this study was to perform a population-based study with a retrospective cohort design to investigate the risk of stroke after the initiation of haemodialysis (HD) among end-stage renal disease (ESRD) patients in Taiwan – a country with the highest incidence of ESRD in the world. Methods:  Data were retrospectively obtained from the Taiwan

National Health Insurance Research Database. In total, 644 patients who were beginning HD between 1999 and 2003 were recruited as the study cohort and 3220 patients PLEK2 matched for age and sex were included as the comparison cohort. Multivariate Cox proportional hazard regression models were used to adjust for confounding and to compare the 5 year stroke-free survival rate between these two cohorts. Results:  The incidence rate of stroke (41.76 per 1000 person-years) was significantly higher in the HD cohort than in the control cohort (24.29 per 1000 person-years). After adjusting for potential confounders, the adjusted hazard ratios of ischaemic stroke and haemorrhagic stroke were 2.16 (95% confidence interval = 1.57–2.97) and 3.78 (95% confidence interval = 1.90–7.

In contrast, L. (V.) braziliensis-infected DCs failed to up-regulate the activation markers, but exhibited an enhancement in their ability to produce TNF-α that may contribute to the local control of the parasite (23). L. (V.) braziliensis infection efficiently triggers innate immune response in DCs, helping the priming of adaptive immune response for parasite clearance, as both parasite and antigen-carrying

DCs displayed an activated phenotype despite amastigote showing higher infectivity and potential to stimulate DCs when compared with promastigote buy CAL-101 (24). Concerning the CD4+ T-cell expression, a distinct profile was noted between the two Leishmania species studied: BALB/c mice infected with L. (L.) amazonensis presented a high number of CD4+ T cells in the lesions at both 4th and 8th weeks PI (P < 0·05), just when the infection had developed a severe disease, whereas the animals infected with L. (V.) braziliensis showed a higher number (P < 0·05) of these cells only at the 8th weeks PI, just when the infection seemed to be controlled. Thus, the elevated CD4+ T-cell response in the L. (L.) amazonensis infection was preferentially characterized by a Th2 response, because higher

levels of IL-4 and IL-10 were observed in this group compared to that of the L. (V.) braziliensis infection, in which these cytokines were not detected at 8th weeks PI. In this regard, it is interesting to mention that despite Qi et al. (2001) (25) showing ACP-196 that draining lymph node cells of BALB/c mice infected with L. (L.) amazonensis may produce both Th1 selleck chemicals (IFN-γ) and Th2 (IL-4 and IL-10) cytokine profiles, the magnitude of Th2 response, linked to a higher expression of IL-4 and IL-10 cytokines, is responsible for the success of L. (L.) amazonensis infection when the levels of IFN-γ are low. In contrast,

despite the CD4+ T-cell response in the skin lesions of BALB/c mice infected with L. (V.) braziliensis showing a higher density only at the 8th weeks PI, this expression was just accompanied not only with the control of infection but also with high levels of IFN-γ, thus suggesting that the CD4+ T-cell response in L. (V.) braziliensis infection was preferentially characterized by a Th1 response. Moreover, IFN-γ is an important cytokine for the macrophage activation, leading to parasite elimination through the production of metabolites oxygen and nitrite. Thus, reduced levels of this cytokine could affect the efficiency of parasite elimination and the control of the infection (26). In this way, it should be stressed that our experiments showed that iNOS expression in the skin lesions of animals infected with L. (L.) amazonensis remained on the same level of the control group, whereas in the skin lesions of animals infected with L. (V.) braziliensis, there was a significant increase at both 4th and 8th weeks PI, suggesting an efficient T-cell immune response activation in the L. (V.

They found that in both cases the receptor/ligand interaction resulted in enhancement of mast cell activation. Moreover, it was found that Staphylococcus aureus employs CD48, together with TLR-2, to invade CBMC and to activate

the production of pro-inflammatory cytokines 12. By identifying novel receptors on mast cells, Dr. Levi-Schaffer and colleagues hope to find new “self” regulating pathways and novel functions of mast cells in different patho-physiological settings 13. Leukotrienes (LT), histamine and proteases are among the major bioactive products of mast cells. Joshua Boyce (Boston, MA) reviewed data from his laboratory on the cysteinyl (cys) leukotrienes LTC4, LTD4, and LTE4 which are known to regulate mast cell function. Cys-LT are peptide-conjugated Vismodegib mouse lipid inflammatory mediators generated by mast cells, macrophages, basophils and eosinophils when these cells are activated in both innate and adaptive immune responses. They facilitate vascular leakage, smooth muscle constriction and cell migration. Nucleotides are released with cell injury, hypoxic stress, and with activation of macrophages and mast cells, MAPK Inhibitor Library molecular weight reaching high micromolar range concentrations in extracellular fluids. Both cys-LT and nucleotides are prominent and early mediators of inflammatory responses. The G protein-coupled

receptors for cys-LTs (Cys-LT1R and Cys-LT2R) are structural homologs of the G protein-coupled receptors for nucleotides, termed purinergic (P2Y) receptors. Dr. Boyce and colleagues demonstrated that both mouse and human mast cells express Cys-LT1R and Cys-LT2R, as well as multiple P2Y receptors for both adenine (P2Y1, P2Y2, P2Y12, P2Y13) Progesterone and uracil (P2Y6)-containing nucleotides 14. They have used mast cells as a model system to demonstrate both functional and physical interactions between these receptor classes that regulate cell proliferation, survival and mediator generation 15. Complementarity between

cys-LT receptors and P2Y receptors may be part of the innate danger-sensing repertoire of mast cells. Mast cells produce histamine, which is now recognized as also being made by a variety of other types of cells. The functions of histamine production from these cells remain unknown. However, only mast cells and basophils make and store significant amounts of histamine which is recognized by four different receptors (H1R-H4R) with tissue-specific expression patterns on immune and nonimmune cells and unique signaling pathways 16. As discussed by Paul Bryce (Chicago, IL), H4R is the most recently identified member of the histamine receptor family. Three potential isoforms of H4R have been described so far, including one activating receptor and two smaller putative dominant negative receptors. The importance of mast cell-produced histamine for DC function is only just beginning to be understood.

Under these circumstances it is highly likely that presentation of autoantigen also takes place in the joint. Therefore, it could be speculated that, in RA, tolDC would ideally have the ability to act in several locations: in the rheumatoid joint to anergize autoantigen-specific effector T cells locally, and in the draining lymph node to

induce Tregs from autoantigen-specific naive T cells. However, it should be noted that T cells from RA patients can be resistant to at least some tolerogenic signals; for instance, they can resist NVP-BGJ398 research buy IL-10- and IDO-mediated suppression [90, 91]. Our tolDC operate, at least partially, via a TGF-β-dependent mechanism and inhibit proliferation and IFN-γ production of peripheral blood RA T cells in vitro (unpublished data); however, whether they can inhibit autoreactive T cells in the rheumatoid joint remains to be determined. Despite the fact that our tolDC have similar ability as mature DC to process and present exogenous antigen, tolDC have lower T cell stimulatory capacity than mature DC, in line with their lower expression of co-stimulatory molecules and low production of proinflammatory cytokines [55, 82]. Moreover, tolDC induce hyporesponsiveness (‘anergy’) in antigen-experienced memory T cells while

polarizing naive T cells towards an anti-inflammatory cytokine profile [55]. We have also shown that, in a mouse in-vivo model G protein-coupled receptor kinase of collagen-induced arthritis, murine bone marrow-derived tolDC generated with Dex, VitD3 and LPS have a therapeutic effect: treatment BI 2536 nmr of arthritic mice with tolDC (1 million cells injected intravenously three times over 8 days) reduced significantly the severity and progression of arthritis, whereas treatment with immunogenic mature DC did not reduce disease and, in fact, exacerbated arthritis [49]. Interestingly, tolDC exerted a therapeutic effect only if they had been loaded with the immunizing antigen, type

II collagen. Treatment with tolDC was associated with a reduction in Th17 cells and an enhancement of IL-10-producing T cells, and a reduction in type II collagen-specific T cell proliferation, possibly explaining their therapeutic effect. Thus, this type of tolDC is a potentially powerful tool for the treatment of RA and other autoimmune diseases. Before tolDC can be applied in a clinical trial, a protocol to generate clinical grade tolDC, compliant with current good manufacturing practice (cGMP) regulations, had to be established. For this purpose, the research-grade fetal calf serum (FCS)-containing medium was replaced with cGMP-grade medium specialized for DC (CellGro® DC medium from CellGenix, Freiburg, Germany) and LPS was replaced with MPLA, a synthetic cGMP-grade TLR-4 ligand (from Avanti Polar Lipids, Alabaster, AL, USA).