After incubation, non-adherent cells were removed and adherent ce

After incubation, non-adherent cells were removed and adherent cells Enzalutamide clinical trial were harvested and counted. When the cell preparation showed ≥ 90% CD14 expression, the generation of MO and MDC

was carried out. Briefly, cells were cultured in RPMI-1640 supplemented with 10% FCS and glutamine (2 mM); granulocyte–macrophage colony-stimulating factor (GM-CSF) (50 ng/ml) (Leukomax, Schering-Plough, Dardilly, France) and interleukin (IL)-4 (40 ng/ml) (Peprotech, Rocky Hill, NJ, USA) were added for MDC generation, while G-CSF (50 ng/ml) was used for MO generation. After 5 days cells were tested for phenotype and maturation markers. Cell viability, characterization and maturation were assessed during the cell production process by light microscopy and flow cytometry using monoclonal antibodies CD1a-phycoerythrin (PE), CD14-fluorescein isothiocyanate (FITC), CD83-PE and CD86-FITC (BD, Becton Dickinson Europe, Pont-de-Claix, France). Viable cell preparations with a positivity higher than 95% for the specific markers were considered valid for subsequent analysis. MVC (Celsentri; selleck screening library Pfizer, Inc., New York, NY, USA) was dissolved in distilled water and stored

at −80°C until use. Monocytes, MO and MDCs (1 × 106/ml) were pre-incubated for different times (1–18 h) with various concentrations of MVC (0·1 µM, 1 µM, 10 µM) at 37°C under 5% CO2 atmosphere. Because, in preliminary experiments, we found no differences in incubation time, we

reported the data obtained from 18 h of MVC treatment. As controls, cells were incubated with medium alone. Drug concentrations were chosen on the basis of published data of pharmacokinetic parameters reported in MVC-treated patients [8,9]. MVC-treated cells at all concentrations used showed a viability ≥ 95%, as assessed by Trypan blue exclusion dye. The in vitro chemotactic activity was measured in an 8 µm pore size Transwell system (Becton Dickinson Europe). The following chemoattractants were used: synthetic Tolmetin peptide formyl-methionyl-leucyl-phenylalanine (fMLP) (10−5 M) (Sigma, St Louis, MO, USA), CCL5/regulated upon activation, normal T cell expressed and secreted (RANTES) (100 ng/ml), CCL4/macrophage inflammatory protein-1 (MIP-1β) (100 nM) and CCL2/monocyte chemotactic protein-1 (MCP-1) (10 ng) (R&D Systems Europe Ltd, Abingdon, UK). A bell-shaped curve described the typical migratory response of cells to increasing concentrations of chemoattractant. Thus, in preliminary experiments, we performed a full dose–response analysis and we used the optimal doses able to induce the maximum chemotactic activity in our cell systems. Cell suspensions in FCS-free RPMI-1640 were used at a concentration of 1 × 106 cells/ml.

Results: Compared with control, in the drug-naÏve group the frequ

Results: Compared with control, in the drug-naÏve group the frequency of dysfunction was significantly higher for urinary urgency (20.9% of the women, 25.9% of the men, P < 0.01), urinary incontinence (9.1%, women), retardation in initiating urination (13.1%, men); constipation (23.8%, 14.8%), diarrhea (20.3%, 21.8%); decrease in libido (42%, men), sexual intercourse (70.7%, 78.7%) orgasm

(63.6%, 65.0%), erection (92.7% of the men); and quality of life indices. No difference was found in the frequency of all three items between the drug-naÏve group and the medicated group. Conclusion: The results of the present study suggest Proteases inhibitor that major depression is a risk for all bladder, bowel and sexual dysfunction, and it significantly worsens quality of life in

patients. This finding presumably reflects that pelvic organ function is under emotional control. Amelioration of bladder, bowel, and sexual dysfunction is therefore an important target to treat patients with major depression. “
“Objectives: The present study was undertaken to investigate the association between the severity of atherosclerosis and lower urinary tract function in male patients with lower urinary tract symptoms. Methods: Male patients EPZ-6438 order with lower urinary tract symptoms were examined with routine investigation. The severity of atherosclerosis was assessed by ultrasound examination of Y-27632 2HCl carotid artery. Patients were divided into two groups: control group and atherosclerosis group. The voiding function and storage function were compared between the two groups. Results: A total of 50 men (69.9 ± 9.1 years [mean ± standard deviation]) entered the study. There was

no significant difference in age distribution (control group: 68.7 ± 7.6 years; atherosclerosis group: 72.5 ± 9.7 years) and prostate volume (control group: 26.5 ± 17.3 mL; atherosclerosis group: 22.2 ± 11.0 mL) between the two groups. In the voiding parameters, maximum flow rate in the atherosclerosis group (13.4 ± 5.5 mL/s, P < 0.05) was significantly lower than that in the control group (16.7 ± 7.7 mL/s). Postvoid residual urine volume showed no significant difference between the two groups. In the storage parameters, voided volume was significantly reduced in the atherosclerosis group (161.8 ± 46 mL, P < 0.05), as compared to control group (201.1 ± 78 mL). Moreover, daytime frequency was 7.13 ± 3.02 times in the control group, and significantly higher in the atherosclerosis group (8.75 ± 2.50 times, P < 0.05). Conclusion: Development of atherosclerosis impairs both voiding and storage function independently of age, suggesting atherosclerosis leads to lower urinary dysfunction.

In addition, LAG-3 is a negative regulator of T cell receptor (TC

In addition, LAG-3 is a negative regulator of T cell receptor (TCR)-mediated signal transduction in effector

T cells and functions in the same manner as cytotoxic T lymphocyte antigen-4 (CTLA-4) [9–12]. Finally, LAG-3 controls activated regulatory T cells (Tregs), while it is not expressed by unstimulated natural Tregs[13]. However, LAG-3 is expressed by interleukin Deforolimus supplier (IL)-10-secreting early growth response (Egr)-2+LAG-3+CD4+ Tregs associated with Peyer’s patches [14]. We have shown previously that depleting anti-LAG-3 antibodies prevented the development of alloreactive effector T cells in a heart allotransplant model in rodents and represents an effective treatment for allograft rejection [15]. In this study, we have characterized a cytotoxic anti-LAG-3 chimeric antibody (chimeric A9H12) and evaluated its potential for selective therapeutic depletion in a non-human primate model of delayed-type hypersensitivity (DTH), a low-invasive Target Selective Inhibitor Library and non-terminal model based on the induction of local T helper type 1 (Th-1)-mediated cellular immune responses [16]. Our investigation demonstrated

that LAG-3+ T lymphocytes could be depleted in vivo in primates and that this resulted in a long-lasting inhibition of immune responses in this preclinical model. C57/B6 mice were immunized three times with Chinese hamster ovary (CHO) cells transfected with human LAG-3 cDNA, followed by an intravenous (i.v.) booster injection of a recombinant hLAG-3Ig protein purified from the supernatant of transfected CHO cells. Three see more days after the boost, splenocytes were fused with the X63.AG8653 fusion partner [17] to obtain hybridoma cells, using traditional techniques. The A9H12 hybridoma was selected for its antibody-dependent cell cytotoxicity (ADCC) activity towards LAG-3 expressing cells and subcloned to yield a stable cell line. A bicistronic vector coding for the variable heavy (VH) and variable light (VL) domains of A9H12 fused to human CL kappa and CH1-hinge-CH2-CH3 immunlglobulin (Ig)G1 regions was

generated and used to transfect CHO-S cells (Invitrogen, Illkirch, France). After antibiotic selection and limiting dilutions, a stable subclone was selected to produce the chimeric A9H12 in ProCHO5 medium (Lonza, Vervier, Belgium). The product in the supernatant cell was purified by adsorption on a HiTrap recombinant Protein A FF column (GE Healthcare, Velizy, France), eluted by acid pH (Glycin HCl, 0·1 M, pH 2·8) and dialysed against phosphate-buffered saline (PBS; Invitrogen). LAG-3+ CHO cells or human peripheral blood mononuclear cells (PBMCs) stimulated with 1 µg/ml of Staphylococcal enterotoxin B (SEB; Sigma Aldrich, L’Isle D’Abeau Chesnes, France) for 48 h were used as targets. Chimeric A9H12 binding was revealed with a fluorescein isothiocyanate (FITC)-conjugated goat F(ab′)2 anti-human IgG (Southern Biotech, Birmingham, AL, USA).

To investigate the effect of IL-6

we added IL-6 neutraliz

To investigate the effect of IL-6

we added IL-6 neutralizing antibodies (MQ2-13A5, BD Biosciences) and the appropriate rat IgG1 isotype control (50 ng/mL). Basic descriptive statistics were used to describe the patient population. Data involving two time points within one population were compared using the Wilcoxon matched pair test. For differences in median between two independent groups, the Mann–Whitney U test was used to test for significance. Significance was accepted at p<0.05 indicated in the graphs by * or p<0.001 indicated by **. The authors thank W. de Jager from the Center for Molecular and Cellular Intervention for his assistance with Tigecycline clinical trial the Luminex analysis, M. Klein for technical assistance

with FACS sorting and J. Meerding for performing the CFSE assays. This study was supported by the Wilhelmina Children’s Hospital Research Fund. B. J. Prakken was supported by grants from the Dutch Organization for Scientific Research (NWO VIDI innovation grant) and the Dutch Arthritis Foundation. Conflict JAK inhibitor review of interest: The authors declare no financial or commercial conflict of interest. Detailed facts of importance to specialist readers are published as ”Supporting Information”. Such documents are peer-reviewed, but not copy-edited or typeset. They are made available as submitted by the authors. “
“Thymus colonisation and thymocyte positioning are regulated by interactions between CCR7 and CCR9, and their respective ligands, CCL19/CCL21 and CCL25. The Interleukin-2 receptor ligands of CCR7 and CCR9 also interact with the atypical receptor CCRL1 (also known as ACKR4), which is expressed in the thymus and has recently been reported to play an important role in normal αβT-cell development. Here, we show that CCRL1 is expressed within the thymic cortex, predominantly

by MHC-IIlowCD40− cortical thymic epithelial cells (TECs) and at the subcapsular zone by a population of podoplanin+ TECs in mice. Interestingly, CCRL1 is also expressed by stromal cells which surround the pericytes of vessels at the corticomedullary junction, the site for progenitor cell entry and mature thymocyte egress from the thymus. We show that CCRL1 suppresses thymocyte progenitor entry into the thymus, however, the thymus size and cellularity are the same in adult wild-type and CCRL1−/− mice. Moreover, CCRL1−/− mice have no major perturbations in T-cell populations at different stages of thymic differentiation and development, and have a similar rate of thymocyte migration into the blood. Collectively, our findings argue against a major role for CCRL1 in normal thymus development and function. This article is protected by copyright. All rights reserved “
“Epidemiological evidence on the relationship between vitamin D receptor (VDR) polymorphisms and periodontal disease is inconsistent.

If alveolar water absorption had been more important than oedema

If alveolar water absorption had been more important than oedema formation, one would have expected a clearly increased wet/dry ratio in the case of blocked ENaC [41]. Another interesting observation of the current in-vivo experiments is that co-conditioning with sevoflurane is more effective in amelioration of oxygenation than post-conditioning with the volatile anaesthetic [26]. This finding suggests that early treatment with sevoflurane could inhibit the increase of permeability and attenuate injury-induced vascular leakage. The present study has several limitations

Alisertib order that need to be addressed. Discussion from in-vitro experiments is limited, as the interaction with cells of different character is missing. Another concern lies in the experimental set-up of ALI used. Even if intratracheal application of LPS is defined as a relevant in-vitro

and in-vivo animal model for lung injury, it does Ulixertinib not fully represent ALI in patients. Therefore, conclusions cannot necessarily be translated to a clinical situation. Furthermore, due to the fact that lungs could not be utilized for both measurement of lung wet/dry ratios and lung RNA analysis, experiments had to be repeated using different animals. This, of course, may create a sample bias, which we tried to minimize by following our strict experimental protocols. Nevertheless, despite these limitations the present study provides new information regarding the protective effect of volatile anaesthetics in ALI. In conclusion, these data reveal that sevoflurane reverses the inhibitory effect of LPS on the function of ENaC and Na+/K+-ATPase in AECII in vitro. Sevoflurane almost exposure

can influence positively the course of LPS-induced lung injury with regard to oxygenation. This effect, however, seems not to be mediated by increased fluid clearance, but rather by the anti-inflammatory properties of sevoflurane leading to less oedema formation. The authors thank Irene Odermatt, art designer, Institute of Anesthesiology, University of Zurich, Switzerland, for the development of the illustrations. This work was supported by a grant of the European Society of Anaesthesiology and the Swiss National Research Foundations Grant no. 3200B0-109558. The authors have no conflicts of interest. “
“The M2 subset of macrophages has a critical role to play in host tissue repair, tissue fibrosis and modulation of adaptive immunity during helminth infection. Infection with the helminth, Fasciola hepatica, is associated with M2 macrophages in its mammalian host, and this response is mimicked by its excretory-secretory products (FhES). The tegumental coat of F.

These include upstream signalling and transcription

These include upstream signalling and transcription MK-2206 concentration factor interactions. Several members of the retinoic acid receptor (RAR) orphan receptor (ROR) family have been described as transcription factors expressed specifically in Th17 cells. These include RORα and RORγt [90–92], which are encoded by the genes RORA and RORC. RORγt is induced by TGF-β and IL-6 in naive Thp and leads to transcription of

IL-17 [90]. As expected, overexpression of RORγt promotes Th17 differentiation. However, while RORγt-deficient mice have reduced numbers of Th17 cells, the population is not depleted [90]. This is because RORα is also expressed highly in TGF-β/IL-6-induced Th17 cells [91]. This related transcription factor synergizes with RORγt to induce Th17 differentiation, and elimination of both RORα and RORγt (double-deficient animals) at the same time is required to RG7204 ic50 deplete Th17 differentiation effectively and protect against Th17-driven autoimmune diseases [91]. The Scurfy mouse (sf), an X-linked mutant strain, described in 1949 (loc. cit. [93], exhibits a series of autoimmune features including skin scaliness, diarrhoea

and death (between 2 and 4 weeks after birth) in association with CD4+ T cell hyperproliferation, multi-organ CD4+ cell infiltration [94] and over-production of several inflammatory cytokines [95]. This fatal autoimmune lymphoproliferative syndrome maps to a gene locus on the X chromosome called foxp3, which has been described as a member of the forkhead/winged-helix family of transcription factors [96]. The foxp3 gene is highly conserved between species and a mutation in the human gene, FOXP3, has been identified as the causative factor responsible for the human equivalent of Scurfy, the immunodysregulation, polyendocrinopathy

and enteropathy, X-linked syndrome (IPEX), also known as X-linked autoimmunity and allergic dysregulation syndrome (XLAAD) [19,97,98]. Both the mouse and human disease lack discrete circulating Tregs, which suggests that foxp3 and FOXP3 are essential for normal Treg development in the two species, respectively. This position is strengthened by the failure of foxp3 knock-out mice to develop circulating Tregs; these animals develop a Scurfy-like cAMP inhibitor syndrome from which they can be rescued by the adoptive transfer of Tregs from a foxp3 replete animal [99]. Furthermore, ectopic or over-expression of foxp3 in CD4+CD25- mouse cells results in development of a Treg phenotype [97,99,100]. In mice, FoxP3 expression is a good phenotypic marker of Tregs[101,102]; in humans, however, FoxP3 does not allow the unambiguous identification of Tregs[103], as FoxP3 is induced during TCR stimulation in conventional CD4+ T cells [104–106] (in much the same manner as CD25) and there is some debate as to whether the induced CD4+CD25+FoxP3+ population is suppressive or anergic [104,105].


the multiple LVAs were completed without complication


the multiple LVAs were completed without complications. The onset of postoperative cellulitis and edematous aggravation of the limb that received the minimally invasive preventive LVA procedure was not noted in any patient during 6-month follow-up period. This minimally invasive preventive LVA procedure might prevent lymphedema and improve the physical appearance of the limb with minimal scarring. Long-term follow-up will be necessary to monitor the future progression of edema in these patients. © 2013 Wiley Periodicals, Inc. Microsurgery 34:372–376, 2014. “
“Background: Several methods have been used in the management of humeral nonunions. With the advent of modern microsurgical techniques, vascularized bone grafting is becoming increasingly used to improve local biology. We report MI-503 clinical trial our experience in the use of a vascularized corticoperiosteal bone flap from the medial

femoral supracondylar region in the treatment of recalcitrant humeral nonunions. Methods: A retrospective review was performed of all patients treated with this technique over a 4-year period within our institution. Patient demographics, nonunion characteristics, complications, and long-term outcomes were analyzed. Results: Six patients underwent vascularized periosteal graft reconstruction. Prior to this, all had failed an average of three procedures with the length of nonunion ranging from 6 to 68 months. All six nonunions healed by an average of 6.8 months (range 2–12 months). Two patients required additional secondary procedures. Functional outcome improved PF-01367338 cost in all patients as

adjudged by disabilities of the arm, shoulder, and hand, Mayo elbow performance, and Constant Murley scores. Conclusions: The vascularized medial femoral condyle corticoperiosteal flap provides an additional treatment option for the management of humeral nonunions. © 2011 Wiley-Liss, Inc. Microsurgery, 2011. “
“In this study, Tacrolimus (FK506) the role of valproic acid (VPA) in protecting motoneuron after brachial plexus root avulsion was investigated in adult rats. Sixty rats were used in this study, and underwent the brachial plexus root avulsion injury, which was created by using a micro-hemostat forceps to pull out brachial plexus root from the intervertebral foramen. The animals were divided into two groups, VPA group administered with VPA dissolved in drinking water (300 mg/kg) daily, and control group had drinking water every day. The spinal cords (C5-T1) were harvested at day 1, 2, 3, 7, 14, and 28 for immunohistochemistry analysis, TUNEL staining, Nissl staining, and electron microscopy, respectively. The results showed that with VPA administration, the survival of motoneurons was promoted and the cell apoptosis was inhibited.

A number of studies comparing TCR affinities

have been re

A number of studies comparing TCR affinities

have been reported ([12-14], and references therein), and a tentative assertion made of differing affinities between click here VA- and TAPA-specific TCRs, with the virus-specific TCRs binding tighter than cancer-specific ones [12, 14-16]. However, definitive conclusions are difficult to draw for two reasons; first, only a rather limited data set is currently available, and second, small variations in affinity measurements are difficult to resolve given the inevitable methodological differences between individual laboratories. To address these issues a comprehensive panel of TCRs was investigated here (Table 1) and their affinities determined using identical methodology and equipment. The peptide antigens investigated were limited to those presented by HLA-A2, to prevent any influence from variations in CD8 coreceptor affinity between different HLA types. TCR genes were isolated from blood samples, and expressed and prepared as soluble TCRs from BMN-673 Escherichia coli as described in Materials and methods. Binding of the 24 TCRs to their specific pHLA-A2 complexes (10 VAs and 14 TAPAs)

was analyzed by surface plasmon resonance (SPR) at 25°C. The affinities, in terms of dissociation constants, (KD) and the dissociation rate constants (koff) were determined (Table 1). The half-lives (t1/2) and association rate constants (kon) were subsequently

calculated from the measured values (Table 1). Due to the limitations of SPR resolution (t1/2 = 0.5 s), dissociation rate constants could not be determined for a number of TAPA-specific TCRs that have particularly fast off-rates. Representative-binding data for high, intermediate, and low affinity TCRs are shown in Supporting Information Fig. 1. A clear pattern was observed in TCR-binding parameters correlating with the origin of the target peptide. TCRs recognizing VAs (such as those derived from HIV and influenza) exhibited relatively high affinity with KD values, between 0.18 and 25 μM (mean = 8.2 μM) while the affinity of TCRs for TAPAs ranged from 11 to 387 μM (mean = 96.6 μM). The half-lives were, in general, longer for the VA-specific TCRs (mean = 6.8 s) than for the TAPA-specific TCRs (mean = <1.8 s). These data are presented graphically in Figure 1. This represents comprehensive, single-study evidence for a variation in binding parameters between human TCRs recognizing VA and TAPA pHLAs. Where available, we find no substantial differences between the biophysical data presented here and that reported in the literature. Since each isolated TCR represents one random selection event (with the possibility of higher or lower-affinity TCRs for the same antigen being present in other donors), it was fundamental to investigate a large number of responses.

, 2008) Food poisoning caused by B cereus includes both diarrhe

, 2008). Food poisoning caused by B. cereus includes both diarrheal and emetic types, in which the involvement of enterotoxins (hemolytic and nonhemolytic enterotoxins) and an emetic toxin (cereulide) has been identified respectively

(Drobniewski, 1993; Schoeni & Wong, 2005; Arnesen et al., 2008). Enterotoxins such as cytotoxin K (CytK) or enzymes such as hemolysin II (Hly-II), phosphatidylinositol-specific Selleck Erlotinib phospholipase C (Piplc), and sphingomyelinase (Sph) are other potential virulence factors related to the pathogenicity of B. cereus (Kotitra et al., 2000; Schoeni & Wong, 2005; Arnesen et al., 2008). To date, however, there have been few reports on the virulence gene profiles of B. cereus isolates responsible for systemic infections (Kotitra et al., 2000; Dohmae et al., 2008). BSIs caused by B. cereus are usually treated with antimicrobials such as vancomycin, clindamycin, quinolones, and carbapenems. The antimicrobial susceptibility profile of clinical isolates of B. cereus has been characterized, although the Clinical and Laboratory Standards Institute (CLSI) does not define minimum inhibitory concentration (MIC) interpretative see more criteria for B. cereus (CLSI 2009). In previous studies (Kotitra et al., 2000; Luna et al., 2007; Mérens et al., 2008), most B. cereus isolates showed high MICs for β-lactams such as

penicillins and third-generation cephalosporins, and some also did so for meropenem, erythromycin, clindamycin, and sulfamethoxazole/trimethoprim. Despite recognition of B. cereus as an important causative pathogen of systemic infections, information concerning the clinical utility and the performance limitations of routine antimicrobial susceptibility Ribonucleotide reductase testings for clinical isolates of B. cereus is limited. In this study, we characterized the profiles of virulence genes and the pulsed-field gel electrophoresis (PFGE) genotypes of B. cereus isolates from blood cultures, compared antimicrobial

susceptibility results between the agar dilution, MicroScan broth microdilution, and Etest methods, and investigated the risk factors for B. cereus BSI. The strains studied were 26 clinical isolates of B. cereus recovered from blood cultures between 2006 and 2009. Each strain was isolated from different patients [female, n = 9; male, n = 17; median age: 68 years (range: 0–85 years)], who were diagnosed as having B. cereus BSIs (n = 15) or as having contaminated blood cultures (n = 11). Based on the standard of a minimum of two blood culture sets (aerobic and anaerobic cultures a set) being drawn from different sites, samples are defined as contaminated blood cultures if a single blood culture set is positive for B. cereus and the results of the positive blood culture are not compatible with signs and symptoms of blood stream infection. The clinical characteristics of the patients with BSIs or contaminated cultures are shown in Table 1.

Lysosomal storage disorders result from inherited defects in lyso

Lysosomal storage disorders result from inherited defects in lysosomal proteins [10]. These disorders can be caused either by a primary defect in a catabolic Panobinostat enzyme (e.g. Tay-Sachs and Sandhoff disease) or a defect in a transporter, channel or regulatory protein (e.g. Niemann-Pick type C (NPC1) disease). Lysosomal storage caused by a deficient lysosomal enzyme has been shown to lead to reduced iNKT cells in murine models of Sandhoff disease [11, 12], Tay-Sachs disease [11], GM1 gangliosidosis

[11-13] and Fabry disease [14, 15]. In the NPC1 mouse the numbers of iNKT cells also are greatly reduced but this is associated with impaired late-endosome/lysosome fusion in addition to the lysosomal lipid storage [11, 16]. NPC disease can be caused by mutations in one of two genes NPC1 or NPC2 [17]. Dysfunction of the NPC1 protein leads to decreased lysosomal calcium content which accounts for the failure of endocytic vesicle fusion and the complex pattern of lipid storage observed [18]. With the differential trafficking of murine and human CD1d for iNKT-cell

ligand Daporinad nmr presentation ex vivo and the requirement of normal lysosomal CD1d trafficking/function for murine iNKT-cell development in vivo, we reasoned that examining iNKT cells in NPC patients would reveal whether the findings in the murine model extends to humans. It has been reported that iNKT cells are present at normal frequencies in the peripheral blood of Fabry disease patients [19] and are slightly increased in Gaucher disease patients [20]. Here, we have studied iNKT-cell frequencies and functional responses

in NPC1 disease patients and the ability of patient-derived EBV-B-cell lines to stimulate iNKT cells. In contrast to the murine model of NPC1, we found unchanged iNKT-cell frequencies in NPC1 patients. In addition, the functional response of NPC1 iNKT cells to stimulation was normal, as was the ability of NPC1 antigen presenting cells to present a variety of iNKT cells ligands to control iNKT cells. We analysed the frequency of iNKT Staurosporine cells in the peripheral blood of controls, NPC1 patients and NPC1 heterozygote carriers by flow cytometry (gating strategy, Supporting Information Fig. 1). As previously reported [21], the frequencies of iNKT cells are very low in normal human peripheral blood, typically in the range of 0.1–1% of total T cells (Fig. 1A). In contrast to the NPC1 mouse where iNKT cells are undetectable, iNKT cells could be identified and were present at normal frequencies in the peripheral blood of NPC1 patients and heterozygotes (Fig. 1A). This indicates that fusion of late endosomes and lysosomes is not required for the generation, delivery or loading of iNKT-cell selecting ligand(s) in the thymus or for their maintenance in the periphery.