PBMC (4 × 105 cells/well in 100 µl) were incubated in duplicate with 5 µM of each peptide in complete medium with 50 UI/ml interleukin (IL)-2 (Boehringer, Mannheim, Germany) for 48 h. Plates were washed and 100 µl of polyclonal rabbit anti-human IFN-γ antibodies (Genzyme) diluted 1:250 were added. After overnight incubation at +4°C, plates were washed and 100 µl of polyclonal biotin-conjugated goat anti-rabbit IgG antibodies (Boehringer) diluted 1:500 were added for 2 h at 37°C. The plates were washed and incubated

with alkaline phosphatase-labelled extravidin (diluted 1/5000; Sigma-Aldrich Chimie SARL, Lyon, France) for 1 h. Chromogenic alkaline phosphatase substrate (Bio-Rad Laboratories, Hercules, CA, USA) was added to the wells

to develop spots. Blue spots Hydroxychloroquine order were counted with an automatic Copanlisib microscope (Zeiss Apparatus; Carl Zeiss, Göttingen, Germany). Negative controls were PBMC incubated in complete medium alone. Positive controls were obtained by activating PBMC with 50 ng/ml phorbol myristate acetate and 500 ng/ml ionomycin (Sigma-Aldrich Chimie SARL) (2000 cells/well). Only large spots with fuzzy borders were scored as IFN-γ-spot-forming cells (SFC). Responses were considered significant when the mean number of SFC by 106 cells in two experimental wells was superior to the highest either mean number of SFC in the negative control (PBMC alone) plus 3 standard deviations or number of SFC in the negative control (PBMC alone) plus 25 SFC/106 cells. HLA molecules were purified from human Epstein–Barr virus (EBV)-transformed cell lines by using affinity columns coupled to various immunoglobulins (Igs), as described previously [27,28]. After denaturation in urea plus NaOH, HLA heavy chains and β2m were separated from endogenous peptides then incubated with different concentrations of exogenous peptides (10−4–10−10 M)

and β2m. Reassembled HLA/peptide complexes were trapped in microtitration plate wells coated with anti-HLA monoclonal Igs, as described in Bourgault et al.[27]. Correctly folded HLA complexes were revealed only with alkaline phosphatase-coupled antiβ2m Igs (M28) with 4-methyl-umbelliferyl phosphate as a substrate (M-8883; Sigma-Aldrich Chimie SARL). Fluorescence was measured at 360/460 nm in a Microfluor reader (Victor 1420; Wallac, Turku, Finland). Results were expressed as the lowest peptide concentrations yielding a significant binding (20% of maximal fluorescence). Purification of HLA-DR molecules and peptide binding assays were performed as described previously [29,30]. These assays are specific for the HLA-DR molecules predominant in the European and North American populations, which are also frequent globally.

High-throughput analysis of fungal cells walls as well as in-depth sequencing of the meta-transcriptome of eukaryotes during the interaction with the host immune system will soon offer a novel window into the integrated functioning of the mycobiota and microbiota. We would like to thank Luigina Romani for the histological images, Francesco Strati and Tobias Weil for the helpful discussion, and Andrea Mancini for helping in figure editing. This work was supported by funding from the European Community’s Integrative Project FP7, SYBARIS (Grant Agreement 242220,

www.sybaris.eu). The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The authors declare no financial or commercial conflict of interest. “
“Nematode infections are generally followed by high rates Selleck AP24534 PD0332991 of reinfection, leading to elevated prevalence in endemic areas. Therefore, the effective control of nematode infections depends on understanding the induction and regulation of protective mechanisms. However, most experimental models for protective immune response against nematodes use high parasite exposure, not always reflecting

what occurs naturally in human populations. In this study, we tested whether infecting mice with different Strongyloides venezuelensis larvae loads would affect protective responses against reinfection. Interestingly, we found that a previous infection with 10–500 larvae conferred high rate of protection against reinfection with S. venezuelensis in mice, by destroying large numbers of migrating larvae. However, low-dose priming did not abolish adult worm maturation, as detected in high-dose primed group. Results also indicated that a previous low-dose infection delayed the development

of cellular infiltrate, while a high inoculum rapidly induced these inflammatory features. Cytokine production by splenocyte cultures of challenge infected mice demonstrated that low-dose priming had increased production of IL-4 and IFN-γ, while high-dose induced IL-4 production but not IFN-γ. Our data support the hypothesis Tryptophan synthase that low-dose nematode infection does not induce a polarized type-2 immune response, allowing adult worm survival. Gastrointestinal (GI) nematode parasites represent a very important cause of disorders in humans and animals. Geohelminth species belonging to the genus Ascaris, Ancylostoma, Necator, Strongyloides and Trichuris infect more than 1 billion people worldwide, causing 1 million deaths annually and are most frequent in children in developing countries, located mainly in tropical and subtropical regions (1–3). Apart from the relatively elevated mortality, children severely infected by these nematodes can show delayed growth, affected cognitive function and reduced educational achievement (4–6).

Results: 139 of 204 patients completed the survey (68%). The most prevalent symptom was pruritus (71.2%), with over 90% of peritoneal dialysis patients reporting this symptom. Next most common was “weakness or a lack of energy” (70.5%), and restless legs (69.7%). Of concern, moderate or severe pain was present in 66.7% of the conservatively

managed find protocol patients, and in 25% overall. “Feeling depressed” was also self-reported in 43.9% of patients, with 60% of home haemodialysis and 58% of PD patients feeling depressed. Conclusions: Our results are similar to previous published reports, but this survey is one of the few which reports across a “whole of service” renal population. Renal patients suffer a significant symptom burden, and an important challenge is how to better support and reliably identify renal patients with high Ponatinib clinical trial symptom burdens. Shared care clinics with nephrologists, palliative specialists, nurses and other allied health staff are an increasingly common model of care. 217 A RETROSPECTIVE STUDY OF ANTI-GBM DISEASE AT WAIKATO HOSPITAL, NEW ZEALAND B MAHBUB, P SIZELAND Waikato Hospital, Hamilton, New Zealand Aim: To evaluate the efficacy of plasmapheresis in patients with anti GBM disease who presented with severe renal impairment in conjunction with extensive crescent formation on renal biopsy. Background: Anti GBM disease is a rare entity. It classically

presents with the syndrome of glomerulonephritis and/or pulmonary haemorrhage. The majority of patients have a combination of immunosuppression and plasmapheresis at the time of their diagnosis. Recent KDIGO guidelines do not support disease specific treatment in patients with severe AKI, 100% crescents on biopsy and no pulmonary haemorrhage. Morin Hydrate Methods: We reviewed patients (2001–2013) who had serologic and/or biopsy proven

anti GBM disease at Waikato Hospital using the hospital database. Results: 17 patients were identified. Age range was 16 to 81 (median of 47), 10 male and 7 female, 10 Maori and 7 European. One was noncompliant. 14 (82%) were ANCA negative. Anti GBM antibody titre ranged from 1:40 to 1:1280 with a median of 1:320. Serum creatinine level at presentation ranged from 70 to 2,080 with an average of 760 μmol/L. 12 (71%) had severe renal impairment (creatinine level of >500). 13 patients had 100% crescents, 3 did not have biopsy (normal renal function) and one biopsy was not available. 10 (59%) had pulmonary haemorrhage. All of our patients received immunosuppression and plasmapheresis. All patients with renal impairment at presentation required and remained on dialysis long term. 2 (12%) patients died, one within a month and the other one 8 years after diagnosis. Conclusions: Plasmapheresis has no effect on renal recovery in patients with anti GBM disease who have severe renal impairment.

3B). GF109203X, an inhibitor of both classical and novel PKC isoforms, could prevent Nur77 and Nor-1 nuclear/cytoplasmic shuttling in PMA/or HK434/ionomycin stimulated thymocytes (Fig. 3B and data not shown). We have previously shown that PMA/ionomycin signals target Nur77 to

the mitochondria, where the protein binds to Bcl-2 in thymocytes 20. To determine if specific activation of PKC could induce Nur77/Bcl-2 association, we treated thymocytes with ionomycin in the absence and presence of PKC ligand, HK434 or PMA. Figure 4A shows that treatment of thymocytes with ionomycin alone cannot induce Nur77/Bcl-2 or Nor-1/Bcl-2 association. Yet, when thymocytes were stimulated with HK434/ionomycin, anti-Nur77 and anti-Nor-1 but not control selleck chemicals llc antibodies could pull down Bcl-2. The HK434-induced association of Nur77 and Bcl-2 could be interrupted when cells were stimulated in the presence of PKC inhibitor, Gö6976 (Fig. 4A). It should be noted that the Nur77 and Nor-1 being pulled down in the presence of the PKC inhibitors PF-01367338 research buy represents the nuclear localized form of these proteins, as Nur77 and Nor-1 are unable to target the mitochondria when PKC proteins are inhibited. The PMA/ionomycin induced Nur77/Bcl-2 association could only be disrupted with GF109203X pre-treatment. Thymocytes stimulated with PMA/ionomycin in the presence of classical PKC

inhibitor, Gö6976 show similar levels of Bcl-2 association with Nur77 as compared to thymocytes stimulated in the absence of inhibitor (Fig. 4B). Similarly, the association between Nor-1 and Bcl-2 induced by PMA/ionomycin is disrupted only when nPKC in addition to cPKC isoforms are inhibited by GF 109203X (Fig. 4B). Nur77′s targeting of Bcl-2 induces a conformational change in which the buried BH3 domain of Bcl-2 is exposed 20–22, 47. Similar to anti-CD3/CD28 and PMA/ionomycin

treatment, stimulation with HK434/ionomycin induces a Bcl-2 conformational change in stimulated thymocytes (Fig. 5A). This Bcl-2 conformational change Cyclooxygenase (COX) was blocked in thymocytes pre-incubated with Gö6976 and GF109203X. The cPKC inhibitor was also effective in blocking the conversion of Bcl-2 induced by anti-CD3/CD28 antibody treatment. In contrast, only the inhibitor of both classical and novel PKC could block the Bcl-2/BH3 exposure in PMA/ionomycin stimulated thymocytes. The exposure of Bcl-2 is restricted to DP thymocytes. There was no conversion of Bcl-2 observed in DN, CD4+ SP or CD8+ SP cells (Fig. 5B). Ionomycin treatment alone is unable to induce the BH3 conformational change within Bcl-2 (Fig. 5B). These data combined suggest that cPKC isoenzymes are responsible for Nur77/Nor-1 mitochondrial targeting and the subsequent conversion of Bcl-2 into a killer molecule in HK434/ionomycin- and anti-CD3/CD28-treated thymocytes. Yet, nPKC proteins regulate Nur77 and Nor-1 subcellular localization following PMA/ionomycin stimulation.

The mechanism of andrographolide induced AKI is unclear. Some authors have postulated that andrographolide induced AKI may be caused by its intrinsic nephrotoxicity, its high distribution in kidney tubular, and its unstable water solubility originating from diterpene lactone structure with conjugated

double bond in it.[37] However, none of these hypotheses have been approved. In our idea, since the manifestation of andrographolide induced AKI is similar to suprofen induced AFPS, their mechanism might share some similarities also. It is believed that the inhibitory effects of NSAIDs on prostaglandin Epigenetics inhibitor synthesis play important roles in AFPS.[33-36] A recent study shows that andrographolide also has inhibitory effects on prostaglandin E2 (PGE2) production,[38] which hints that andrographolide induced AKI and suprofen induced AFPS may share this similar mechanism. The limitations

of our study are obvious. Because this is a study using spontaneously reported cases, some important data are missing or inadequate. For example, creatine kinase levels were absent in nearly all the cases; however, the possibility of rhabdomyolysis was scarce according to the authors’ judgment. Second, the number of 26 cases is not large; however, andrographolide induced AKI may be underreported, ERK inhibitor as it is an adverse event. An efficient pharmacovigilance system may be lacking, as it is common for traditional medicine. It is hard to know the true country-wide incidence of this situation. However, the frequent occurrence of this adverse event does result in a strong reaction from the official authorities like CFDA,[13, 14] and

causes much academic concern in China. Furthermore, some cases exist but were not included in this analysis. For instance, 80 cases of Telomerase AKI had been reported to CFDA to 2007,[37] but detailed data were not available. There are also some cases of andrographolide induced AKI reported as case series, however, they were not included in this analysis due to the lack of sufficient individual patient information.39,40 Third, our review was limited to Chinese-language literature. Although we also searched English-language literature and retrieved zero results, it should be noted that there may be published, non-Chinese and non-English reports available, especially in Asian areas other than China, where andrographolide was also widely used, such as India, Thailand, and Malaysia etc.[9] Overall, our work represents the first summary of spontaneously reported cases of andrographolide induced AKI in English literature. Although the number of 26 cases is not large, the results are sufficient to raise the concern on the safety of andrographolide, particularly AKI induced by andrographolide. The high incidence of flank pain and subsequently reversible renal failure makes it similar to suprofen induced AFPS.

[4-9] Hessell et al.[10] showed that an HIV-specific neutralizing Ku 0059436 antibody mutated in the Fc position was no longer able to elicit Fc-mediated functions, such as ADCC, and that the efficacy in preventing simian/human immunodeficiency virus (SHIV) infection of macaques was significantly decreased, suggesting that the ADCC function is important for the protection afforded by neutralizing antibodies. There is a more limited understanding of the role of ADCC in the small subset of HIV-infected subjects who naturally control chronic infection, although the role

of cytotoxic T lymphocytes and neutralizing antibodies has been extensively studied.[11-24] We previously detected ADCC-mediated NK-cell activation in a small cohort of six subjects with slow HIV progression, but found no clear correlation with the magnitude of the ADCC response and control of Luminespib price HIV. A recent study of 22 subjects indicated that elite controllers of HIV infection (subjects with consistent plasma HIV levels of < 50 copies/ml) have higher levels of ADCC antibodies than viraemic subjects, with an absence of correlation between cytotoxic T lymphocytes and neutralizing antibodies.[6] Whether these results are generalized across larger numbers of long-term slow-progressors

(LTSP) subjects is not clear. In addition, the HIV epitopes targeted by efficient ADCC are unknown but would logically be interesting vaccine targets. We analysed ADCC responses using an assay studying antibody-mediated interferon- γ (IFN-γ) and CD107a expression of NK cells. We

studied serum samples from 139 HIV-infected subjects not on anti-retroviral therapy; 65 subjects were LTSP who maintained a CD4 T-cell count of > 500/μl for at least 8 years after infection and the remaining 74 subjects were non-LTSP. We found that ADCC responses in LTSP subjects were broadly reactive against multiple HIV proteins and that LTSP subjects disproportionally targeted three specific ADCC epitopes within Vpu (viral protein U). The characteristics of the 139 subjects are shown in Table 1. All subjects were HIV-infected Isotretinoin and not on anti-retroviral therapy at the time of sampling. Subjects enrolled in both cohorts provided written informed consent and the relevant human research ethics committees approved all studies. Subjects were recruited both through the Long-term non-progressor network co-ordinated by the Kirby Institute, Sydney, Australia and through the Melbourne Sexual Health Centre, Australia. Sixty-five of the subjects met the pre-defined criteria as LTSPs, being HIV-positive for more than 8 years without anti-retroviral therapy and maintaining a peripheral CD4+ T-cell count above 500 cells/μl. There were no viral load entry criteria. The remaining 74 subjects did not meet the criteria for LTSP (i.e. had not maintained CD4 T-cell counts > 500 cells/μl for 8 years). For both cohorts, serum for ADCC testing was derived from the earliest time-point available.

Thus, IgG-mediated protective immunity Dabrafenib appears to act predominately against the larval stages of the parasite, which are also the major stimulus for acquired immunity and the target of acquired responses [36]. The next challenge will be to determine the mechanisms by which IgG antibodies target H. p. bakeri larvae. Numerous possibilities exist, perhaps acting in parallel or even synergistically, including neutralization of larval products required for tissue migration/feeding and for evasion of the

immune response or antibody-dependent cellular activation and the consequent destruction or trapping of larvae by immune cells. Of note, macrophages are also required for protective immunity against H. p. bakeri [73], and both antibodies and macrophages are abundant in the Th2-type granuloma surrounding the larvae [55, 73]. These findings raise the possibility that antibodies may activate macrophages to kill or trap parasitic larvae. Whether this occurs still needs to be determined, but it is known that larvae can survive in the granuloma for a long time, as they can be re-activated to continue their growth and maturation into fecund adults by treatment with immunosuppressive corticosteroids as

long as 3 weeks after challenge infection [74]. The entrapment of larvae in granuloma and their eventual destruction could involve binding of IgG to the high- or low-affinity receptors, FcγRI and FcγRIII, known to be expressed by macrophages [75]. Alternatively, antibodies may act in an indirect manner https://www.selleckchem.com/products/PD-0332991.html by promoting the recruitment of immune cells into

the granuloma or by activating complement. In this Selleck Pembrolizumab regard, a recent publication indicated that antibodies play an important role in mediating the production of basophils within the bone marrow following H. p. bakeri infection [72]. However, specific depletion of basophils had a minor impact on larval killing, indicating that this is not the major pathway of antibody-mediated protective immunity [72]. As discussed, H. p. bakeri forms a chronic infection in most mouse strains following primary infection. In the poor responder strain, C57BL/6, B-cell deficiency had little impact on the development of adult worms 14 days following infection [55]. However, fecundity was strikingly increased and remained high for several weeks following primary infection of B cell–deficient mice [55]. Primary infection with H. p. bakeri infection elicits a striking, but largely polyclonal, IgG and IgE response, and the observed impact on worm fecundity could be ascribed to low-affinity IgG antibodies, [55]. These low-affinity IgG antibodies were present even in naïve animals presumably in response to environmental antigens or intestinal bacteria and were amplified by infection [55]. This contrasts with the ability of antibodies to provide protective immunity against challenge infections, where high-affinity parasite-specific antibodies are necessary. Thus, early production of polyclonal antibodies following primary infection with H.

We conclude that modification of antigen with either 3-sulfo-LewisA or tri-GlcNAc enhances cross-presentation and permits Th1 skewing, through specific targeting of the MR, which may be beneficial for DC-based vaccination strategies to treat cancer. Activation of antigen-specific cytotoxic T cells is crucial for the induction of adequate

anti-tumor immunity. Since most tumor cells are poorly immunogenic, optimal presentation VEGFR inhibitor of tumor-derived antigens in MHC class I molecules on the surface of antigen presenting cells is required. An important mechanism that allows DCs to present exogenous antigens, such as tumor-derived antigens, in MHC class I molecules is cross-presentation 1. At tumor lesions, multiple factors and cells are present that prevent the proper activation of DCs that enter the lesion to sample for antigens 2, 3. Consequently, these DCs will not be able to properly activate antigen-specific CD8+ T cells in the tumor-draining LN. To obtain therapeutic anti-tumor immunity powerful vaccination protocols are required. Current strategies focus

either on ex vivo loading of autologous DCs as well as specifically targeting of antigens to DCs in vivo. These new therapies may be combined with a Treg depletion regimen, as these cells have been shown to block anti-tumor immune responses 3–6. As a classical C-type lectin Sirolimus mw receptor (CLR), the mannose receptor (MR) binds carbohydrate structures such as mannose, fucose and N-acetylglucosamine (GlcNAc) in a calcium-dependent manner 7, 8. Besides these carbohydrate structures, the MR has recently also been reported to bind sulfated sugars, such as sulfated oligosaccharides of the blood group antigens LewisA (LeA) and LewisX (LeX) 8–10. Binding of these types of ligands occurs via the cysteine-rich (CR) region of the MR and in a calcium-independent manner 8. The MR has been proposed to mediate antigen uptake and presentation by DCs based on the finding that mannosylated proteins are presented DOK2 more efficiently than non-mannosylated proteins 11, 12. Fusion of an MR-specific monoclonal antibody to tumor antigens enhanced MHC class I-restricted T-cell

responses 13. Additionally, the glycoprotein ovalbumin (OVA), which contains mannose residues, was reported to be endocytosed through the MR, upon which the antigen was transferred to early endosomes, resulting in strong cross-presentation 14, 15. By contrast MR-mediated uptake of OVA did not induce CD4+T-cell responses. Processing of native glycosylated OVA in the early endosomes occurs in a TAP-dependent manner. Transport of TAP from ER to the endosomes is mostly, but not entirely, dependent on toll-like receptor-4 (TLR4)/MyD88 signaling 15. Although these studies report that the MR is an endocytic receptor for mannosylated OVA, in the human setting mannose may simultaneously target other CLR such as DC-SIGN, which shares specificity for mannose 16.

We measured participants’ own QOL and that of two hypothetical colorectal cancer health states using a rating scale, and a utility-based QOL measure, the time trade-off, with extremes of 0 (death) and 1 (full health). Results:  Recipients of kidney transplants (n = 79) had the highest mean QOL weights of 0.79 (standard deviation (SD) = 0.34) compared with participants with CKD 3–5 (n = 53) with mean QOL weights of 0.70 (SD = 0.39), and those on dialysis (n = 89), who had the lowest mean QOL weights of 0.62 (SD = 0.41) (P = 0.02). Having early and advanced stage colorectal cancers were valued at mean QOL weights of 0.44 (SD = 0.41) selleck inhibitor and 0.12 (SD = 0.25) among people with moderate stage

CKD; 0.45 (SD = 0.39) and 0.11 (SD = 0.24) among dialysis patients; 0.62 (SD = 0.36) and 0.18 (SD = 0.29) among kidney transplant recipients. Conclusions:  People with CKD have poor

QOL. Having coexistent illnesses such as cancer further reduces the overall well-being of individuals with kidney disease. In addition to the development of effective screening and treatment programs to improve cancer outcomes in people with CKD, our study also highlights the need for effective interventions to improve the QOL in people with learn more CKD, particularly those with major comorbidities like cancer. “
“Background:  Haemodialysis (HD) circuits are known to produce microemboli. Patent foramen ovale (PFO) may be important in HD patients by allowing right to left intracardiac shunting of microemboli (blood clots or microbubbles), which may pass into the cerebral circulation. Methods:  We undertook bubble contrast transthoracic echocardiography to identify PFO in HD patients and in a control population of peritoneal dialysis patients. We interrogated draining arteriovenous fistulae to confirm that microemboli are created during HD. We then

undertook transcranial Doppler scanning of the middle cerebral artery before Baricitinib and during dialysis, with and without Valsalva augmentation, to detect cerebral microemboli in HD patients and in the control group. Results:  Eighty patients (age 60.4 ± 15.0 years) were recruited to the study. In 12 of 51 HD patients and five of 29 peritoneal dialysis patients a PFO was found (21.3%). Ultrasound scanning of draining arteriovenous fistulae showed a significant difference in the number of microemboli before (1.63 ± 3.47 hits per 5 min) and during (31.6 ± 28.9 hits per 5 min) HD (P = 0.012). However, there was no evidence of microembolization to the middle cerebral artery before or during HD in the study or control groups. Conclusions:  Although microemboli are detectable in the draining arteriovenous fistulae of patients undergoing HD, there was no evidence of cerebral microembolization in the middle cerebral artery during HD in those with or without a PFO. The results contrast with previous reports demonstrating microemboli in the carotid circulation during HD.

Importantly, these in silico investigations could be used to design experiments distinguishing between the two explanations above. In summary, the virtual NOD mouse was built to reproduce untreated

pathogenesis and responses to interventions (internal validation). The virtual NOD mouse also predicted most responses accurately to interventions not used in model construction (external validation). In the few instances where the virtual NOD mouse did not match the reported therapeutic response, a closer examination highlighted potential conflicts within the published data, in some cases providing a basis for clarifying laboratory experiments. The model as described Sirolimus is ready for in silico research. It can be updated to accommodate new data or to address additional biology not currently within the model scope. Model updates may include new validation tests to ensure MK-8669 that the modifications are consistent with the reported biology. The Type 1 Diabetes PhysioLab platform is a physiologically based mathematical model of type 1 diabetes pathogenesis in a NOD mouse, designed to facilitate type 1 diabetes research and accelerate development of human therapies. The model has a graphical user interface and incorporates much of the known biology in the PLN and islets, which sets the stage for its use as an educational and research tool to illustrate complex biological relationships at

these important sites. Because data are used to define qualitatively or quantitatively the biological relationships that are embedded in the model, the model can also be used as a data archive or continuing repository.

Beyond these applications, the model simulates the represented biology, providing a mathematically integrated description which is consistent with published experimental data. Generating this description was an intensive and iterative process, which refined our understanding and interpretation of the published literature. Montelukast Sodium For example, the initial modelling exercise did not include the representation of a distinct tolerogenic DC phenotype. With the initial representation, late and transient LipCl2MDP-mediated depletion of macrophages and DCs reduced the cellular infiltrate and delayed disease onset but did not provide sustained protection despite the presence of Treg cells. Briefly, when LipCl2MDP was cleared from the system, phagocyte populations recovered and re-established a diabetogenic environment and a corresponding destructive cellular infiltrate. With no data to suggest a direct effect of LipCl2MDP on Treg cell populations, the next plausible scenario was an effect mediated through phagocytes. The representation of tolerogenic DCs was based largely on data from outside the NOD mouse literature (e.g. [99–101]), and included regulation by cytokines and cell contact.