For MIF stimulation, 1 × 107 spleen cells were incubated for 24 hr in RPMI-1640 medium containing 100 ng/ml recombinant MIF as described previously.29 Splenocytes (1 × 106 cells) were incubated with anti-CD74 (Santa Cruz Biotechnologies, Santa Cruz, CA), anti-CD44 (Southern Biotechnology Associates, Birmingham, AL), or anti-B220 (eBioscience, San Diego, CA) specific antibodies and analysed by flow cytometry. For Annexin-V and propidium iodide staining, cells were analysed using the Phosphatidyl Serine Detection Kit (IQ Products, Groningen, the Netherlands), according to the manufacturer’s instructions, and were analysed by FACS. Lysates extracted

from either B cells, brain hippocampi or kidneys were separated on SDS–PAGE as described previously.8 The membranes were Cilomilast incubated with the antibodies anti-CD74, Pexidartinib supplier anti-Bcl-2, anti-Bcl-xL (Santa Cruz Biotechnologies) and anti-β-actin (Sigma-Aldrich, Poole, UK) antibodies. Membranes were incubated with the appropriate second antibody coupled to horseradish peroxidase. Detection was performed using the enhanced chemiluminescence method. Densitometric units were determined using the NIH Image program (National Institutes of Health, Bethesda, MD). Total RNA was prepared from isolated B cells, brain hippocampi or kidneys using TRI Reagent (Molecular Research Center, Cincinnati,

OH). Complementary DNA was prepared and real-time reverse transcription-PCR was performed using the LightCycler system (Roche,

Mannheim, Germany), according to the manufacturer’s instructions. The following primer sequences were used (forward and reverse, respectively): CD74 (5′-CAACGCGACCTCATCT-3′, 5′-TGTTGCCGTACTTGGTAA-3′), CD44 (5′-GCTATCCTGGCCTACC-3′, 5′-TGTCCTACCACAACCCAACT-3′), MIF (5′-CGCTTTGTACCGTCCT-3′, 5′-CGTGCCGCT-AAAATCA-3′), Bcl-xL (5′-GGACCGCGTATCAGAG-3′, 5′-GCATTGTTCCCGTAGAG-3′), Bcl-2 (5′-CCATGTGGCTATGCG-3′, 5′-ATCAGCCACGCCTAA-3′), β-actin (5′-GTGACGTTGACATCCG-3′, 5′-CAGTAACAGTCCGCCT-3′). The levels of β-actin were used for normalizing the expression levels of selleck screening library the studied genes. Results are presented relative to the vehicle-treated group (considered as 100%). Statistical analysis was performed using Mann–Whitney U-test and Student’s t-test. Values of P < 0·05 were considered significant. Eight-month-old BWF1 mice with established disease were divided into three groups (n = 8 to n = 12) and injected subcutaneously with hCDR1, the scrambled peptide (both 50 μg per mouse) or vehicle alone, once a week for 10 weeks. The clinical data of three treatment experiments are summarized in Table 1. It can be seen in the table that mice treated with the vehicle or with the control peptide exhibited high levels of anti-dsDNA autoantibodies. In mice treated with hCDR1, however, these levels were significantly reduced.

Finally, the IL-10 (Th2) reduction could be suggestive of its therapeutic use in TAO. Based on the unclear TAO pathogenesis, particularly the involvement of immune trigger mechanisms of vascular disease, further studies should be carried out to reveal the role of the immune disorder in TAO progression. Finally, the discovery of IL-17 and its association with inflammation and autoimmune pathology

has reshaped our viewpoint regarding the pathogenesis of TAO, which was based previously on the Th1–Th2 paradigm. The inflammatory profile status is not exclusive to TAO; in other diseases, where damage to the vascular wall is recorded, the scenario of increased proinflammatory and detrimental anti-inflammatory Vorinostat cytokines has also been described this website for other types of vasculitis. In order to analyse the behaviour/onset of inflammatory status, it would be useful to have evidence of those cytokine profiles in healthy smokers along the time course of consumption and in non-diseased TAO patients. Unfortunately, we could not obtain this information from consulting the patients’ charts. In addition

to the Th17 profile, the development of autoimmunity could be defined clearly by monitoring autoantibodies and autoreactive T cells along the time course of TAO, which was not performed in the present study. None. This work was supported in part by FAEPA (HCFMRP/USP), CNPq and FAPESP 09/50508-5. “
“Precise identification of NK-cell populations in humans and nonhuman primates has been confounded by imprecise phenotypic definitions. A common definition used in nonhuman primates, including chimpanzees, is CD3−CD8α+CD16+, and this is the dominant NK-cell phenotype in peripheral

blood. However, recent data suggest that in chimpanzees a rare CD8α−CD16+ population also exists. Herein, we present evidence validating the existence of this rare subset in chimpanzee peripheral blood, but also demonstrating that gating on CD3−CD8α−CD16+ cells can inadvertently include a large number of CD16+ myeloid DCs (mDCs). We confirmed the inclusion of mDCs in CD3−CD8α−CD16+ gated cells by demonstrating high expression of CD11c, BDCA-1 and HLA-DR, and by SB-3CT the lack of expression of NKp46 and intracellular perforin. We also functionally validated the CD8α− NK-cell and mDC populations by mutually exclusive responsiveness to a classical NK-cell stimulus, MHC class I-deficient cells, and a prototypic mDC stimulus, poly I:C, respectively. Overall, these data demonstrate common problems with gating of NK cells that can lead to erroneous conclusions and highlight a critical need for consensus protocols for NK-cell phenotyping. Because of their potent ability to kill virus-infected or neoplastic cells without prior sensitization, NK cells are often characterized as the major effector cells of the innate immune system.

0 mg/day. However, urine protein further increased beyond 1 g/g Cr, and serum creatinine increased and C-reactive protein also increased, accompanied by skin rash Selleckchem NVP-AUY922 and dyspnoea. Allograft biopsy was conducted (2.5 years post transplant). The biopsy showed diffuse glomerular endocapillary proliferation and swelling of glomerular endothelial cells (Fig. 1). The presence of glomerular basement

membrane injury was present. Immunofluorescence showed no significant immune deposit including C4d. Intraluminal proliferating cells were mostly CD34+, indicating that the majority of them were endothelial cells. On the contrary, CD4+ or CD8+, i.e. T cells or CD68+ macrophages were few by immunohistochemistry. These findings suggested that the injury was mediated by direct insult to the endothelium, such as drug-induced injury, and was not mediated by alloantigen-directed immunological insult. No endarteritis or tubulointerstitial lesion was observed. No donor-specific antibody was detected with flow-bead analysis. EVR was discontinued and TACER was returned to the previous dose. https://www.selleckchem.com/products/byl719.html Proteinuria only decreased to 0.5 g/g Cr and methylprednisolone mini-pulse therapy was given (125 mg ×3), resulting in improvement of proteinuria to 0.1 g/g Cr. Other symptoms have also disappeared. Allograft biopsy taken 6 months later still showed mild and focal endocapillary

proliferation but those lesions had significantly improved compared with the previous biopsy (Fig. 2). Glomerular injury accompanied by de novo proteinuria in this case is assumed to be caused by everolimus. The presence of glomerulitis supports antibody-mediated rejection (AMR) as a possible cause. However, glomerular endothelial injury was not associated with either lymphocyte margination or C4d deposition and the proliferating cells were mostly endothelial cells, indicating the injury was mediated by drug rather than alloimmune response. Furthermore, the lack of donor-specific antibody and the reversal of clinical and pathological findings only by low-dose steroid therapy are unsupportive of

AMR. The presence of basement membrane injury could be mediated by Fossariinae CNI toxicity. In this case, the deterioration of the clinical data occurred after adding EVR. The presence of underlying CNI-mediated glomerular injury could not be excluded but the injury, severe enough to induce significant clinical presentation was likely to be triggered by EVR. Typically, proteinuria induced by mTORi is believed to be mainly mediated by podocyte injury.[5] Reports of glomerulonephritis induced by EVR, as in our case, are scarce.[6] Reluctance to biopsy would have resulted in attributing the cause of proteinuria to typical adverse effect of EVR in general. We could fortunately reverse the graft injury after recognizing the presence of glomerulonephritis and this case suggests the importance of clarifying the pathology by allograft biopsy.

Thus, in this study we investigated the effects of sMD-2 and sCD14 on the growth of both Gram-negative and Gram-positive bacteria. E. coli O111:B4 LPS (Sigma-Aldrich, St Louis, MO, USA) was re-purified according to Hirschfeld et al. (20). PG from Bacillus subtilis (Sigma-Aldrich) was confirmed to possess no TLR4-stimulatory activity up to 10 μg/ml. Unless otherwise noted, all other chemicals were from Wako Pure Chemical Industries (Osaka, Japan). The coding region of human MD-2 lacking its signal

sequence was amplified by PCR from pEIAV-hMD-2 as described previously (21) and subcloned into the yeast expression vector pGAPZα (Invitrogen, Carlsbad, CA, USA) with an N-terminal 6× histidine Everolimus tag sequence, resulting in plasmid pGAPZα-hMD-2. The coding region of human CD14 lacking its signal sequence and the sequence encoding the eight C-terminal amino acids (22) was subcloned into pGAPZα GPCR Compound Library order with an N-terminal 6× histidine tag sequence, resulting in plasmid pGAPZα-hCD14. A plasmid encoding a CD14 mutant lacking amino acids 57 to 64 was generated by PCR from pGAPZα-hCD14 using primers 5′-GACACGGTCAAGGCTCTC-3′ and 5′-CGCATCGACGCGCTTTAG-3′. The deletion was confirmed by automated DNA sequencing. Human MD-2 and CD14 in yeast were purified as previously described (7). pGAPZα-hMD-2

and pGAPZα-hCD14 were expressed in a Pichia expression system (Invitrogen) and purified with a Ni2+-column (Novagen, Madison, WI, USA) under denaturing conditions according to the manufacturer’s recommendations. E. coli DH5α (Invitrogen) and B. subtilis NBRC3134 were inoculated in LB broth and bacillus broth (10 g/l polypeptone, 2 g/l yeast extract, 1 g/l MgSO4·7H2O,

http://www.selleck.co.jp/products/cetuximab.html pH 7.0), respectively and incubated at 37°C for 18 hr. After incubation, each culture was diluted to 2 × 105 CFU/ml for E. coli and 4 × 104 CFU/ml for B. subtilis with phenol red-free DMEM (Gibco, Eggenstein, Germany). Either sMD-2 or sCD14 (0.25–1 μg/ml each) was added to the culture, and myosin (Sigma-Aldrich; 1 μg/ml), which had been confirmed to have no effects on bacterial growth, was added as a control. These were cultured at 37°C for up to 18 hr. The number of viable cells was measured by plating cultures on either LB agar for E. coli or bacillus broth agar for B. subtilis and counting the number of colonies (CFU/ml). The viability of bacteria was also measured using the MTS assay in the CellTiter 96 AQueous One Solution Cell Proliferation Assay kit (Promega, Madison, WI, USA) according to the manufacturer’s recommendations. Wells in 96-well plates were coated with PG (250 pg/ml) in PBS at 37°C for 3 hr. After washing five times with PBST, the wells were blocked by incubating with 0.2% BSA (Sigma-Aldrich) in PBS at 4°C overnight. After five washes with PBST, either His-tagged sMD-2 or sCD14 was added at the indicated concentration, and the plates incubated at 37°C for 1 hr.

To assess the phosphorylated status of the tau protein recognized by PHF-1, we performed double labelling, combining PHF-1 in green colour and thiazin red (TR) in red colour (Figure 3a, in colour scale yellow colour represents equal contribution of both markers). TR is an analogue of naphthol-based azo structures whose functional

characteristic is to bind β-pleated sheet structures [34]. None surprisingly, well defined NFTs detected by PHF-1 were found in a fibrillar state as revealed by the yellow tonality (Figure 3c). Similar results were observed in additional pathology like NFTs in earlier state were coexisting events were found, named phosphorylation and TR labelling (Figure 3b). Interestingly, some early aggregates labelled by PHF-1 were found with little fibrillar structure, as revealed

by the intense green tonality (Figure 3a, white arrows). Note Acalabrutinib that the presence of nuclei reveals the early state of the NFT-like structure (Figure 3a, blue). Overall, PHF-1 marker is able to detect the classical NFT structure in fibrillar conformation, but more importantly is also capable of detecting early phospho aggregates that are not yet in fibrillar conformation. Our group previously BMN 673 ic50 reported that cleavage of tau protein is sequential starting by the carboxyl terminus, with cleavage at D421 being an early event and cleavage at E391 occurring latter in AD [8, 24, 32]. Taking advantage of this finding, we wanted to evaluate if phosphorylation Fludarabine molecular weight of tau protein was present in coexistence with both cleavage events (Figure 4Ai,Aii), and more importantly, if phosphorylation suffers any changes during the tau abnormal processing. In this regard we found coexistence of phosphorylation at site Ser396 with either, D421 or E391 truncated tau (Figure 4c blue arrow and stars, and 4f arrows and stars). Some NFT pathology was found with nothing but phosphorylation at site Ser396 (Figure 4a,c white arrows). Interestingly,

some NFT pathology (population A) showed an elevated level of phosphorylation at site Ser396 with lowest levels of E391 truncated tau (Figure 4d–f white arrow), while, some others (population B) showed an increased level of E391 truncated tau with lowest levels of phosphorylation at site Ser396 (Figure 4d–f blue arrows). Relative expression analysis confirmed the two populations; one with significantly elevated presence of phosphorylation (Figure 4B,D, population A) and the other with significantly elevated presence of cleavage at E391 (Figure 4C,D, population B). Further analysis of both cleavage events (D421 and E391) revealed that almost all the structures containing truncated tau also comprised phosphorylated tau (Figure 4E). In summary, phosphorylation of tau protein appears as single event and remains during early and advanced proteolytic events.

This result is largely driven by lower staff

costs, and better health outcomes for survival and quality of life. Expanding the proportion of haemodialysis patients managed at home is likely to produce cost savings. “
“Aim:  To summarize the clinical and pathological features of renal amyloidosis in order to achieve early diagnosis. Methods:  EX-527 The clinical and pathological data of 32 patients with renal amyloidosis, diagnosed by renal biopsy in one renal centre, were retrospectively analyzed. Immunohistochemistry of amyloid A protein and immunoglobulin light chains was further performed on the renal specimens for further classification. Results:  Twenty-four out of the 32 patients (75%) were not considered to have renal amyloidosis by local physicians; 91.7% (22/24) of them had at least one of the following signs: bodyweight loss, organ enlargement and decreased blood pressure. Twenty-nine

out of the 32 selleck screening library patients (90.6%) were over 40 years, 30 patients (93.8%) had nephrotic syndrome, and 21 patients (65.6%) were found to have monoclonal light chain in serum or urine by immunofixation. Six patients (18.8%) were negative by Congo red stain and were diagnosed as having early renal amyloidosis by electron microscopy. Twenty-eight patients were diagnosed as having AL amyloidosis, two were suspected of having AL amyloidosis, one had AA amyloidosis and the status of the remaining patient was undetermined. Conclusion:  Renal amyloidosis is frequently neglected by local physicians in China. Middle-aged nephrotic patients with weight loss, organ enlargement and monoclonal light chains in serum

or urine should be highly suspected of the disease. Renal biopsies, especially electron microscopy, play a crucial ID-8 role in the early diagnosis of renal amyloidosis. “
“We present a case of an unsensitized patient with end-stage kidney disease secondary to atypical haemolytic uremic syndrome (aHUS) with mutations in CD46/MCP and CFH who developed severe, intractable antibody-mediated rejection (ABMR) unresponsive to therapy post kidney transplantation. There were no haematological features of thrombotic microangiopathy. The patient received standard induction therapy and after an initial fall in serum creatinine, severe ABMR developed in the setting of urosepsis. Despite maximal therapy with thymoglobulin, plasma exchange and methylprednisolone, rapid graft loss resulted and transplant nephrectomy was performed. Luminex at 4 weeks showed a new DSA and when repeated after nephrectomy showed antibodies to each of the 5 mismatched antigens with high MFI. The rate of recurrence of disease in patients with aHUS referred for transplantation is 50% and is associated with a high rate of graft loss. It is dependent in part on the nature of the mutation with circulating factors CFH and CFI more likely to cause recurrent disease than MCP which is highly expressed in the kidney.

[44] Treatment of NZB/W F1 mice with soluble TACI-Ig fusion protein prevented the development

of proteinuria and prolonged the survival of the animals.[44] These findings underscored the involvement of TSA HDAC mw BLys and its receptors in the development of SLE and hence the TACI-Ig was proposed as a promising treatment for human autoimmune disease. Furthermore, mice treated with exogenous BLys showed increased numbers of anti-chromatin B cells and augmented anti-dsDNA production.[45] Deletion of either BLys or BR3 critically impaired B cell maturation beyond the transitional developmental stages.[37, 40, 44, 46] T cell-deficient BAFF transgenic (Tg) mice developed SLE similar to T cell-sufficient BAFF Tg mice, and such features were associated with innate B lymphocyte ABT-263 mouse activation and pro-inflammatory autoantibodies release. These data suggest that a dysregulated innate activation of B cells alone can drive disease independently of the T cells.[47] In human lupus patients, the circulating BLys level was raised in human lupus and is correlated with the anti-dsDNA level.[48] In a survey which measured the serum BLys level and disease activities, healthy subjects universally exhibits a normal longitudinal serum BLys profile, whereas escalated BLys level was observed in SLE patients (persistent rise in 25% and intermittent increase in another 25% of patients).

Increased cerebrospinal fluid levels of a proliferation-inducing ligand (APRIL) are also observed SLE patients with neuropsychiatric manifestations. The antagonism of BLys has been one of the important progresses in the treatment of SLE. Recently, belimumab Phloretin was approved by the Food and Drug Administration (FDA) for the treatment of SLE. The efficacy and safety of belimumab in active SLE had been evaluated by two large multicentre randomized control trials. Both studies have demonstrated that the use of belimumab is associated with significant improvement in the SLE Responder Index (defined as ≥4 points improvement in SLEDAI) at 52 weeks, reduced SLE activity

and severe flares, as well as a comparable tolerability profile to placebo.[33, 34] Analysis of the pooled data from these two large trials showed that belimumab treatment improved overall SLE disease activity mostly in the musculoskeletal and mucocutaneous organ domains and less deterioration occurred in the haematological, immunological and renal domains.[49] In a post-hoc analysis of the BLISS study, the rates of renal flare, renal remission, renal organ disease improvement, proteinuria reduction and serologic activity all favoured belimumab, although the between-group differences in most renal outcomes were not significant. Among the 267 patients with renal involvement at baseline, belimumab resulted in greater renal improvement among patients receiving mycophenolate mofetil or those with active serology at baseline when compared with placebo.

We previously reported that an increased visceral fat area (VFA) determined using computed tomography scans was associated with atherosclerosis in hemodialysis patients. However, whether a high VFA is associated with increased cardiovascular mortality in hemodialysis patients remains unknown. Therefore, we investigated the relationship between VFA and prognosis in hemodialysis patients. Methods: VFA

check details was estimated in 126 patients on maintenance hemodialysis using computed tomography scans. These patients were followed for 60 months. Results: Kaplan-Meier analysis revealed that the cardiovascular survival rate was significantly lower in the high VFA group, with a VFA of 71.5 cm2 or greater, than in the low VFA group, with a VFA of less than 71.5 cm2. Hazards ratio of clinical characteristics of subjects for cardiovascular deaths were see more calculated in the univariate cox analyses. A high VFA, but not high BMI or WC was an independent predictor of cardiovascular deaths. In the multivariable analyses, we adjusted for significant factors such as age, LDL, CTR and High

VFA in univariate analyses. High VFA was an independent predictor of cardiovascular deaths. Conclusion: These results suggest that an increased VFA is a stronger risk factor than body mass index or waist circumference for cardiovascular deaths in hemodialysis patients. Measuring VFA may be recommended for predicting the risk of cardiovascular diseases in hemodialysis patients. In addition, interventions to reduce an increased VFA may be effective in preventing cardiovascular deaths in these patients. PEI-LIN CHUNG1, TSAI JEN-PI2, CHANG CHIEN-HWA3 1Department of Nursing, Buddhist Dalin Tzu Chi General Hospital; 2Department

of Nephrology, Buddhist Dalin Tzu Chi General Hospital; 3Department of Cardiac Surgery, Buddhist Dalin Tzu Chi General Hospital Introduction: Arteriovenous shunt infection is a major morbidity of chronic maintenance hemodialysis (HD) patients. This study was conducted RVX-208 to determine the risk factors at the development of shunt infection. Methods: From 2007 April to 2013 August, there were 1048 patients received shunt creation, which included arteriovenous fistula (AVF), arteriovenous graft (AVG) and arteriovenous fistula transposition (AVFT), and had regular follow up at our hospital. Shunt infection was defined by clinical impressions and wound/blood culture reports. Results: During this period, 54 HD patients (5.13%) were diagnosed to have shunt infection (2 AVF, 49 AVG, 3 AVFT). The pathogens were gram positive 68% (39/57), gram negative 12.3% (7/57), no growth 14% (8/57) and not known 5.3% (3/57). Patients who had shunt infection were older (69.21 ± 10.5 vs. 65.47 ± 12.98, p = 0.015) and used more AVG (90.

e. age and sex. Receiver operating characteristics (ROC) of the logistic models included age and sex; age, sex, and MMP-8; age, sex, and MPO; and age, sex, MMP-8,

and MPO. Multiple linear regression analyses were performed for the patients with arterial disease and the relation between covariates, and the dependent variable was evaluated with regression coefficients (β values) and their 95% CIs. Analyses were performed using the spss 15.0 statistical package (SPSS Inc., Chicago, IL, USA). Characteristics of the patients with arterial disease and their sub-groups are presented in Table 1. When compared to the healthy reference subjects (n = 100), the patients (n = 126) were older [59.0 (56.0–61.0) versus 70.1 (60.1–75.6) years,

P < 0.001], and more frequently males (53.0% versus 77.8%, P < 0.001). In the univariate analyses, the patients with arterial disease had higher serum MMP-8 (P < 0.001) this website and TIMP-1 (P = 0.04) concentrations, as well as MMP-8/TIMP-1 ratios (P < 0.001) than check details in the reference sera (Table 2). On the contrary, the patients had lower serum MPO concentrations than the healthy subjects (P < 0.001, Table 2). In the scatter plot of the values measured from the samples obtained from the patients (Fig. 1A,C,E,G,H) and healthy subjects (Fig. 1B,D,F), MMP-8 had a strong positive correlation with MPO and HNE (Fig. 1A–D), and HNE had a positive correlation with MPO (Fig. 1E,F), whereas only weak correlations were found between MMP-8 and MMP-1 and MMP-13 as indicated by r values (Fig. 1G,H). In the forward stepwise multiple logistic regression analysis, where the serum concentrations of patients were compared to those of the reference values adjusted for age and gender, the male

gender (OR = 2.51, 95% CI = 1.29-4.90, P < 0.01), age (OR = 1.18/year, Tangeritin 95% CI = 1.1–1.25, P < 0.001), elevated MMP-8 (OR= 1.30/ng/ml, 95% CI = 1.2–1.4, P < 0.001), and decreased MPO concentrations (OR = 0.97/ng/ml, 95% CI = 0.96–0.98, P < 0.001) were found to be associated with arterial disease. As seen in the ROC-curve of the logistic models, the advancing age, male gender, elevated serum MMP-8, and decreased MPO levels were cumulatively associated with arterial disease when compared to age and sex only (Fig. 2). In the multiple linear regressions analyses (Table 3), MMP-8 concentration had a positive correlation with HNE and hsCRP concentrations (Model I, Table 3). Similarly, MPO concentration had a positive correlation with HNE concentration (Model II, Table 3), while HNE correlated with serum MMP-8 and MPO concentrations (Model III, Table 3). On the other hand, chlamydial LPS in serum (serum cLPS) had a positive correlation with LBP, LDL cholesterol, MMP-13, and interleukin-6 (IL-6) concentrations (Model IV, Table 3).

20 It is more likely that some alleles exhibit a less restrictive peptide binding while other MHC class I alleles show a more restrictive peptide binding pattern. Most MHC class I molecules have been studied in detail and peptide anchor positions have been identified. HLA-A*0201 preferentially recognizes peptides with the amino acids leucine

or methionine at position Pirfenidone 2 and valine or leucine at the C-terminus.24 Only two out of 17 epitopes showed ‘correct’ anchor residues; others had either one ‘correct’ anchor residue or residues with similar hydrophobic groups at these positions. Other peptides, such as SQIMYNYPA (TB10.42–10), shared almost none of the previously reported preferred residues, but they strongly stabilized the HLA-A*0201 monomer, sufficient for tetramer production. For many of the TB10.4 peptides, we could identify Everolimus solubility dmso extensive ‘cross-binding’ to different MHC class I molecules. MHC molecules have been divided into supertypes based on similar binding preferences for peptides.25 Some of the cross-binding could be a result of the fact that the alleles belong to the same supertype, or to supertypes with similar binding preferences. However, the majority of the peptides identified in the current study bound to alleles that exhibited very different binding preferences; for example, they bound both to HLA-A and HLA-B alleles.

This observation has previously been postulated to represent an exception rather than a rule.26,27 Only recently, more systematic studies of HIV

epitopes and human papillomavirus (HPV) epitopes showed that this phenomenon may be more common.28,29 In the context of nonviral pathogens, ‘cross-binding’ has previously been reported for the Mtb protein Ag85B19 and the tumour-associated protein 5T4.20 The extensive promiscuity of peptides in their binding to different MHC class I molecules may certainly be of clinical importance considering the vast number of alleles that exist. A peptide capable of binding to Pregnenolone many different MHC class I molecules is more likely to be presented by a majority of people, a situation that could facilitate vaccine design, as fewer epitopes are needed to cover large population cohorts. While this might be positive from the perspective of vaccine design, it may also mean that a narrow focus on a few epitopes, presented by a high number of alleles to CD8+ T cells, could lead to ‘immune exhaustion’ or escape mutations. Escape mutations are common in viral epitopes but have not yet been reported for Mtb epitopes. This may be a result of the fact that the mutation rate for immunogenic Mtb proteins has been shown to be quite low;30 in addition, data comparing a comprehensive panel of Mtb isolates using genome-wide analysis are lacking at this time. In the case of TB10.4, similar epitopes from the closely related proteins TB10.3 and TB12.