The multifaceted nature of aminopeptidases ERAP1, ERAP2, and LNPEP: from evolution to disease

In the human genome, the aminopeptidases ERAP1, ERAP2 and LNPEP lie contiguously on chromosome 5. They share sequence homology, functions and associations with immune-mediated diseases. By analyzing their multifaceted activities as well as their expression in the zoological scale, we suggest here that the progenitor of the three aminopeptidases might be LNPEP from which the other two aminopeptidases could have derived by gene duplications. We also propose that their functions are partially redundant. More precisely, the evolutionary story of the three aminopeptidases might have been dictated by their role in regulating the renin–angiotensin system, which requires their controlled and coordinated expression. This hypothesis is supported by the many species that lack one or the other gene as well as by the lack of ERAP2 in rodents and a null expression in 25% of humans. Finally, we speculate that their role in antigen presentation has been acquired later on during evolution. They have therefore been diversified between those residing in the ER, ERAP1 and ERAP2, whose role is to refine the MHC-I peptidomes, and LNPEP, mostly present in the endosomal vesicles where it can contribute to antigen cross-presentation or move to the cell membrane as receptor for angiotensin IV. Their association with autoinflammatory/autoimmune diseases can therefore be two-fold: as “contributors” to the shaping of the immune-peptidomes as well as to the regulation of the vascular response

Fibroblasts to keratinocytes redox signaling: The possible role of ROS in psoriatic plaque formation

Although the role of reactive oxygen species-mediated (ROS-mediated) signalling in physiologic and pathologic skin conditions has been proven, no data exist on the skin cells ROS-mediated communication. Primary fibroblasts were obtained from lesional and non-lesional skin of psoriatic patients. ROS, superoxide anion, calcium and nitric oxide levels and lipoperoxidation markers and total antioxidant content were measured in fibroblasts. NADPH oxidase activity and NOX1, 2 and 4 expressions were assayed and NOX4 silencing was performed. Fibroblasts and healthy keratinocytes co-culture was performed. MAPK pathways activation was studied in fibroblasts and in co-cultured healthy keratinocytes. Increased intracellular calcium, •NO and ROS levels as well as an enhanced NADPH oxidase 4 (NOX4)–mediated extracellular ROS release was shown in lesional psoriatic vs. control fibroblasts. Upon co-culture with lesional fibroblasts, keratinocytes showed p38 and ERK MAPKs pathways activation, ROS, Ca2+ and •NO increase and cell cycle acceleration. Notably, NOX4 knockdown significantly reduced the observed effects of lesional fibroblasts on keratinocyte cell cycle progression. Co-culture with non-lesional psoriatic and control fibroblasts induced slight cell cycle acceleration, but notable intracellular ROS accumulation and ERK MAPK activation in keratinocytes. Collectively, our data demonstrate that NOX4 expressed in dermal fibroblasts is essential for the redox paracrine regulation of epidermal keratinocytes proliferation

Coerced Hospital Admission and Symptom Change-A Prospective Observational Multi-Centre Study

PMCID: PMC3227658 Freely available online via CCplosone.org

Clinical decision making and outcome in the routine care of people with severe mental illness across Europe (CEDAR)

Aims. There is a lack of knowledge on clinical decision making and its relation to outcome in the routine treatment of people with severe mental illness. This study examined preferred and experienced clinical decision making from the perspectives of patients and staff, and how these affect treatment outcome. Methods. CEDAR (ISRCTN75841675) is a naturalistic prospective observational study with bimonthly assessments during a 12-month observation period. 588 adults with severe mental illness were consecutively recruited from caseloads of community mental health services at the six study sites (Germany, UK, Italy, Hungary, Denmark, and Switzerland). Clinical decision making was measured using two instruments (Clinical Decision Making Style Scale. CDMS;Clinical Decision Making Involvement and Satisfaction Scale, CDIS) from patient and staff perspectives. Outcomes assessed were unmet needs (Camberwell Assessment of Need Short Appraisal Schedule, CANSAS). Mixed-effects multinomial regression was used to examine differences in involvement in and satisfaction with actual decision making. The effect of clinical decision making on outcome was examined using hierarchical linear modelling controlling for covariates. Results. Shared decision making was preferred by patients (2=135.08; p<0.001) and staff (2=368.17; p<0.001). Decision making style of staff significantly affected unmet needs over time, with unmet needs decreasing more in patients whose clinicians preferred active to passive (-0.406 unmet needs per two months, p=0.007) or shared (-0.303 unmet needs per two months, p=0.015) decision making. Conclusions. A shift from shared to active involvement of patients is indicated, including the development and rigorous test of targeted interventions

ERAP1 and ERAP2 Haplotypes Influence Suboptimal HLA-B*27:05-Restricted Anti-Viral CD8+ T Cell Responses Cross-Reactive to Self-Epitopes

The human leukocyte antigen (HLA)-B*27 family of alleles is strongly associated with ankylosing spondylitis (AS), a chronic inflammatory disorder affecting the axial and peripheral joints, yet some HLA-B*27 variants not associated with AS have been shown. Since no major differences in the ligandome of associated compared to not-associated alleles have emerged, a plausible hypothesis is that the quantity rather than the quality of the presented epitopes makes the difference. In addition, the Endoplasmic Reticulum AminoPeptidases (ERAPs) 1 and 2, playing a crucial role in shaping the HLA class I epitopes, act as strong AS susceptibility factors, suggesting that an altered peptidome might be responsible for the activation of pathogenic CD8+ T cells. In this context, we have previously singled out a B*27:05-restricted CD8+ T cell response against pEBNA3A (RPPIFIRRL), an EBV peptide lacking the B*27 classic binding motif. Here, we show that a specific ERAP1/2 haplotype negatively correlates with such response in B*27:05 subjects. Moreover, we prove that the B*27:05 allele successfully presents peptides with the same suboptimal N-terminal RP motif, including the self-peptide, pDYNEIN (RPPIFGDFL). Overall, this study underscores the cooperation between the HLA-B*27 and ERAP1/2 allelic variants in defining CD8+ T cell reactivity to suboptimal viral and self-B*27 peptides and prompts further investigation of the B*27:05 peptidome composition

Demonstration and Comparison of Operation of Photomultiplier Tubes at Liquid Argon Temperature

Liquified noble gases are widely used as a target in direct Dark Matter searches. Signals from scintillation in the liquid, following energy deposition from the recoil nuclei scattered by Dark Matter particles (e.g. WIMPs), should be recorded down to very low energies by photosensors suitably designed to operate at cryogenic temperatures. Liquid Argon based detectors for Dark Matter searches currently implement photo multiplier tubes for signal read-out. In the last few years PMTs with photocathodes operating down to liquid Argon temperatures (87 K) have been specially developed with increasing Quantum Efficiency characteristics. The most recent of these, Hamamatsu Photonics Mod. R11065 with peak QE up to about 35%, has been extensively tested within the R&D program of the WArP Collaboration. During these testes the Hamamatsu PMTs showed superb performance and allowed obtaining a light yield around 7 phel/keVee in a Liquid Argon detector with a photocathodic coverage in the 12% range, sufficient for detection of events down to few keVee of energy deposition. This shows that this new type of PMT is suited for experimental applications, in particular for new direct Dark Matter searches with LAr-based experiments

Interaction Pattern of Arg 62 in the A-Pocket of Differentially Disease-Associated HLA-B27 Subtypes Suggests Distinct TCR Binding Modes

The single amino acid replacement Asp116His distinguishes the two subtypes HLA-B*2705 and HLA-B*2709 which are, respectively, associated and non-associated with Ankylosing Spondylitis, an autoimmune chronic inflammatory disease. The reason for this differential association is so far poorly understood and might be related to subtype-specific HLA:peptide conformations as well as to subtype/peptide-dependent dynamical properties on the nanoscale. Here, we combine functional experiments with extensive molecular dynamics simulations to investigate the molecular dynamics and function of the conserved Arg62 of the α1-helix for both B27 subtypes in complex with the self-peptides pVIPR (RRKWRRWHL) and TIS (RRLPIFSRL), and the viral peptides pLMP2 (RRRWRRLTV) and NPflu (SRYWAIRTR). Simulations of HLA:peptide systems suggest that peptide-stabilizing interactions of the Arg62 residue observed in crystal structures are metastable for both B27 subtypes under physiological conditions, rendering this arginine solvent-exposed and, probably, a key residue for TCR interaction more than peptide-binding. This view is supported by functional experiments with conservative (R62K) and non-conservative (R62A) B*2705 and B*2709 mutants that showed an overall reduction in their capability to present peptides to CD8+ T cells. Moreover, major subtype-dependent differences in the peptide recognition suggest distinct TCR binding modes for the B*2705 versus the B*2709 subtype