Adaptations of Natural Killer Cells to Self-MHC Class I.

Natural Killer (NK) cells use germ line encoded receptors to detect diseased host cells. Despite the invariant recognition structures, NK cells have a significant ability to adapt to their surroundings, such as the presence or absence of MHC class I molecules. It has been assumed that this adaptation occurs during NK cell development, but recent findings show that mature NK cells can also adapt to the presence or absence of MHC class I molecules. Here, we summarize how NK cells adjust to changes in the expression of MHC class I molecules. We propose an extension of existing models, in which MHC class I recognition during NK cell development sequentially instructs and maintains NK cell function. The elucidation of the molecular basis of the two effects may identify ways to improve the fitness of NK cells and to prevent the loss of NK cell function due to persistent alterations in their environment.

Lower incisor dentoalveolar compensation and symphysis dimensions among Class I and III malocclusion patients with different facial vertical skeletal patterns

Objective: To compare lower incisor dentoalveolar compensation and mandible symphysis morphology among Class I and Class III malocclusion patients with different facial vertical skeletal patterns. Materials and Methods: Lower incisor extrusion and inclination, as well as buccal (LA) and lingual (LP) cortex depth, and mandibular symphysis height (LH) were measured in 107 lateral cephalometric x-rays of adult patients without prior orthodontic treatment. In addition, malocclusion type (Class I or III) and facial vertical skeletal pattern were considered. Through a principal component analysis (PCA) related variables were reduced. Simple regression equation and multivariate analyses of variance were also used. Results: Incisor mandibular plane angle (P < .001) and extrusion (P  =  .03) values showed significant differences between the sagittal malocclusion groups. Variations in the mandibular plane have a negative correlation with LA (Class I P  =  .03 and Class III P  =  .01) and a positive correlation with LH (Class I P  =  .01 and Class III P  =  .02) in both groups. Within the Class III group, there was a negative correlation between the mandibular plane and LP (P  =  .02). PCA showed that the tendency toward a long face causes the symphysis to elongate and narrow. In Class III, alveolar narrowing is also found in normal faces. Conclusions: Vertical facial pattern is a significant factor in mandibular symphysis alveolar morphology and lower incisor positioning, both for Class I and Class III patients. Short-faced Class III patients have a widened alveolar bone. However, for long-faced and normal-faced Class III, natural compensation elongates the symphysis and influences lower incisor position.

T Cell Priming by Activated Nlrc5-Deficient Dendritic Cells Is Unaffected despite Partially Reduced MHC Class I Levels.

NLRC5, a member of the NOD-like receptor (NLR) protein family, has recently been characterized as the master transcriptional regulator of MHCI molecules in lymphocytes, in which it is highly expressed. However, its role in activated dendritic cells (DCs), which are instrumental to initiate T cell responses, remained elusive. We show in this study that, following stimulation of DCs with inflammatory stimuli, not only did NLRC5 level increase, but also its importance in directing MHCI transcription. Despite markedly reduced mRNA and intracellular H2-K levels, we unexpectedly observed nearly normal H2-K surface display in Nlrc5(-/-) DCs. Importantly, this discrepancy between a strong intracellular and a mild surface defect in H2-K levels was observed also in DCs with H2-K transcription defects independent of Nlrc5. Hence, alongside with demonstrating the importance of NLRC5 in MHCI transcription in activated DCs, we uncover a general mechanism counteracting low MHCI surface expression. In agreement with the decreased amount of neosynthesized MHCI, Nlrc5(-/-) DCs exhibited a defective capacity to display endogenous Ags. However, neither T cell priming by endogenous Ags nor cross-priming ability was substantially affected in activated Nlrc5(-/-) DCs. Altogether, these data show that Nlrc5 deficiency, despite significantly affecting MHCI transcription and Ag display, is not sufficient to hinder T cell activation, underlining the robustness of the T cell priming process by activated DCs.

Direct Interrogation of Viral Peptides Presented by the Class I HLA of HIV-Infected T Cells

Identification of CD8+ cytotoxic T lymphocyte (CTL) epitopes has traditionally relied upon testing of overlapping peptide libraries for their reactivity with T cells in vitro. Here, we pursued deep ligand sequencing (DLS) as an alternative method of directly identifying those ligands that are epitopes presented to CTLs by the class I human leukocyte antigens (HLA) of infected cells. Soluble class I HLA-A*11:01 (sHLA) was gathered from HIV-1 NL4-3-infected human CD4+ SUP-T1 cells. HLA-A*11:01 harvested from infected cells was immunoaffinity purified and acid boiled to release heavy and light chains from peptide ligands that were then recovered by size-exclusion filtration. The ligands were first fractionated by high-pH high-pressure liquid chromatography and then subjected to separation by nano-liquid chromatography (nano-LC)–mass spectrometry (MS) at low pH. Approximately 10 million ions were selected for sequencing by tandem mass spectrometry (MS/MS). HLA-A*11:01 ligand sequences were determined with PEAKS software and confirmed by comparison to spectra generated from synthetic peptides. DLS identified 42 viral ligands presented by HLA-A*11:01, and 37 of these were previously undetected. These data demonstrate that (i) HIV-1 Gag and Nef are extensively sampled, (ii) ligand length variants are prevalent, particularly within Gag and Nef hot spots where ligand sequences overlap, (iii) noncanonical ligands are T cell reactive, and (iv) HIV-1 ligands are derived from de novo synthesis rather than endocytic sampling. Next-generation immunotherapies must factor these nascent HIV-1 ligand length variants and the finding that CTL-reactive epitopes may be absent during infection of CD4+ T cells into strategies designed to enhance T cell immunity.

Genèse de l’immunopeptidome du CMH de classe I

La différentiation entre le « soi » et le « non-soi » est un processus biologique essentiel à la vie. Les peptides endogènes présentés par les complexes majeurs d’histocompatibilité de classe I (CMH I) représentent le fondement du « soi » pour les lymphocytes T CD8+. On donne le nom d’immunopeptidome à l’ensemble des peptides présentés à la surface cellulaire par les molécules du CMH I. Nos connaissances concernant l’origine, la composition et la plasticité de l’immunopeptidome restent très limitées. Dans le cadre de cette thèse, nous avons développé une nouvelle approche par spectrométrie de masse permettant de définir avec précision : la nature et l’abondance relative de l’ensemble des peptides composant l’immunopeptidome. Nous avons trouvé que l’immunopeptidome, et par conséquent la nature du « soi » immun, est surreprésenté en peptides provenant de transcrits fortement abondants en plus de dissimuler une signature tissu-spécifique. Nous avons par la suite démontré que l’immunopeptidome est plastique et modulé par l’activité métabolique de la cellule. Nous avons en effet constaté que les modifications du métabolisme cellulaire par l’inhibition de mTOR (de l’anglais mammalian Target Of Rapamycin) provoquent des changements dynamiques dans la composition de l’immunopeptidome. Nous fournissons également la première preuve dans l’étude des systèmes que l’immunopeptidome communique à la surface cellulaire l’activité de certains réseaux biochimiques ainsi que de multiples événements métaboliques régulés à plusieurs niveaux à l’intérieur de la cellule. Nos découvertes ouvrent de nouveaux horizons dans les domaines de la biologie des systèmes et de l’immunologie. En effet, notre travail de recherche suggère que la composition de l’immunopeptidome est modulée dans l’espace et le temps. Il est par conséquent très important de poursuivre le développement de méthodes quantitatives au niveau des systèmes qui nous permettront de modéliser la plasticité de l’immunopeptidome. La simulation et la prédiction des variations dans l’immunopeptidome en réponse à différents facteurs cellulaires intrinsèques et extrinsèques seraient hautement pertinentes pour la conception de traitements immunothérapeutiques.

Effect of the unfolded protein response on MHC class I antigen presentation

Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal.

Rapid acidification and alkylation: Redox analysis of the MHC class I pathway

The technique of rapid acidification and alkylation can be used to characterise the redox status of oxidoreductases, and to determine numbers of free cysteine residues within substrate proteins. We have previously used this method to analyse interacting components of the MHC class I pathway, namely ERp57 and tapasin. Here, we have applied rapid acidification alkylation as a novel approach to analysing the redox status of MHC class I molecules. This analysis of the redox status of the MHC class I molecules HLA-A2 and HLA-B27, which is strongly associated with a group of inflammatory arthritic disorders referred to as Spondyloarthropathies, revealed structural and conformational information. We propose that this assay provides a useful tool in the study of in vivo MHC class I structure. (c) 2008 Elsevier B.V. All rights reserved.

Comparative Histopathological Analysis of Human Pulps after Class I Cavity Preparation with a High-Speed Air-Turbine Handpiece or Er:YAG Laser

The purpose of this study was to comparatively evaluate the response of human pulps after cavity preparation with different devices. Deep class I cavities were prepared in sound mandibular premolars using either a high-speed air-turbine handpiece (Group 1) or an Er: YAG laser (Group 2). Following total acid etching and the application of an adhesive system, all cavities were restored with composite resin. Fifteen days after the clinical procedure, the teeth were extracted and processed for analysis under optical microscopy. In Group 1 in which the average for the remaining dentin thickness (RDT) between the cavity floor and the coronal pulp was 909.5 mu m, a discrete inflammatory response occurred in only one specimen with an RDT of 214 mu m. However, tissue disorganization occurred in most specimens. In Group 2 (average RDT = 935.2 mu m), the discrete inflammatory pulp response was observed in only one specimen (average RDT = 413 mu m). It may be concluded that the high-speed air-turbine handpiece caused greater structural alterations in the pulp, although without inducing inflammatory processes.

Frequency of class I anti-HLA alloantibodies in patients infected by HIV-1

O objetivo deste estudo foi avaliar a presença de aloanticorpos anti-HLA classe I em pacientes infectados pelo HIV-1 e relacioná-la aos diferentes cursos clínicos da doença. Amostras de sangue de 145 indivíduos HIV positivo foram coletadas em tubos com EDTA. A infecção pelo HIV-1 foi confirmada por teste ELISA e a presença de aloanticorpos anti-HLA classe I determinada em seguida. A evolução clínica foi definida como rápida (<1 ano entre diagnóstico e morte), moderada (1-3 anos) ou lenta (>3 anos). A presença de aloanticorpos anti-HLA classe I foi menor em indivíduos saudáveis em relação aos infectados pelo HIV-1 (4,2% contra 32,4%). Porém, a distribuição destes aloanticorpos entre os indivíduos infectados foi igual, independente da evolução clínica. Deste modo, a presença de aloanticorpos anti-HLA classe I não é um fator determinante na piora clínica do paciente.

Influence of Ridge Inclination and Implant Localization on the Association of Mandibular Kennedy Class I Removable Partial Denture

The aim of this study was to evaluate the tendency of displacement of the supporting structures of the distal extension removable partial denture (DERPD) associated to the implant with different inclinations of alveolar ridge and implant localizations through a two-dimensional finite-element method. Sixteen mandibular models were fabricated, presenting horizontal, distally descending, distally ascending, or descending-ascending ridges. All models presented the left canine and were rehabilitated with conventional DERPD or implant-retained prosthesis with the ERA system. The models were obtained by the AutoCAD software and transferred to the finite-element software ANSYS 9.0 for analysis. A force of 50 N was applied on the cusp tips of the teeth, with 5 points of loading of 10 N. The results were visualized by displacement maps. For all ridge inclinations, the assembly of the DERPD with distal plate retained by an anterior implant exhibited the lowest requisition of the supporting structures. The highest tendency of displacement occurred in the model with distally ascending ridge with incisal rest. It was concluded that the association of the implant decreased the displacement of the DERPD, and the anterior positioning of the implant associated to the DERPD with the distal plate preserved the supporting structures for all ridges.

Influence of Implant Inclination Associated With Mandibular Class I Removable Partial Denture

The aim of this study was to use two-dimensional finite element method to evaluate the displacement and stress distribution transmitted by a distal extension removable partial denture (DERPD) associated with an implant placed at different inclinations (0, 5, 15, and 30 degrees) in the second molar region of the edentulous mandible ridge. Six hemimandibular models were created: model A, only with the presence of the natural tooth 33; model B, similar to model A, with the presence of a conventional DERPD replacing the missing teeth; model C, similar to the previous model, with a straight implant (0 degrees) in the distal region of the ridge, under the denture base; model D, similar to model C, with the implant angled at 5 degrees in the mesial direction; model E, similar to model C, with the implant angled at 15 degrees in the mesial direction; and model F, similar to ME, with the implant angled at 30 degrees in the mesial direction. The models were created with the use of the AutoCAD 2000 program (Autodesk, Inc, San Rafael, CA) and processed for finite element analysis by the ANSYS 8.0 program (Swanson Analysis Systems, Houston, PA). The force applied was vertical of 50 N on each cusp tip. The results showed that the introduction of the RPD overloaded the supporting structures of the RPD and that the introduction of the implant helped to relieve the stresses of the mucosa alveolar, cortical bone, and trabecular bone. The best stress distribution occurred in model D with the implant angled at 5 degrees. The use of an implant as a support decreased the displacement of alveolar mucosa for all inclinations simulated. The stress distribution transmitted by the DERPD to the supporting structures was improved by the use of straight or slightly inclined implants. According to the displacement analysis and von Mises stress, it could be expected that straight or slightly inclined implants do not represent biomechanical risks to use.

Evaluation of Bone Insertion Level of Support Teeth in Class I Mandibular Removable Partial Denture Associated With an Osseointegrated Implant: A Study Using Finite Element Analysis

Purpose: This study evaluated the influence of distal extension removable partial denture associated with implant in cases of different bone level of abutment tooth, using 2D finite element analysis.Materials and Methods: Eight hemiarch models were simulated: model A-presenting tooth 33 and distal extension removable partial denture replacing others teeth, using distal rest connection and no bone lost; model B-similar to model A but presenting distal guide plate connection; model C-similar to model A but presenting osseointegrated implant with ERA retention system associated under prosthetic base; model D-similar to model B but presenting osseointegrated implant as described in model C; models E, F, G, and H were similar to models A, B, C, and D but presenting reduced periodontal support around tooth 33. Using ANSYS 9.0 software, the models were loaded vertically with 50 N on each cusp tip. For results, von Mises Stress Maps were plotted.Results: Maximum stress value was encountered in model G (201.023 MPa). Stress distribution was concentrated on implant and retention system. The implant/removable partial denture association decreases stress levels on alveolar mucosa for all models.Conclusions: Use of implant and ERA system decreased stress concentrations on supporting structures in all models. Use of distal guide plate decreased stress levels on abutment tooth and cortical and trabecular bone. Tooth apex of models with reduced periodontal support presented increased stress when using distal rest. (Implant Dent 2011;20:192-201)

Comparative Histopathological Analysis of Human Pulps after Class I Cavity Preparation with a High-Speed Air-Turbine Handpiece or Er:YAG Laser

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effect of Conventional and Resin-modified Glass-Ionomer Liner on Dentin Adhesive Interface of Class I Cavity Walls After Thermocycling

Objective: The aim of this in vitro study was to analyze the effect of glass-ionomer cement as a liner on the dentin/resin adhesive interface of lateral walls of occlusal restorations after thermocycling.Materials and Methods: Occlusal cavities were prepared in 60 human molars, divided into six groups: no liner (1 and 4); glass-ionomer cement (GIC, Ketac Molar Easymix, 3M ESPE) (2 and 5); and resin-modified glass-ionomer cement (RMGIC, Vitrebond, 3M ESPE) (3 and 6). Resin composite (Filtek Z250, 3M ESPE) was placed after application of an adhesive system (Adper Single Bond 2, 3M ESPE) that was mixed with a fluorescent reagent (Rhodamine B) to allow confocal microscopy analysis. Specimens of groups 4, 5 and 6 were thermocycled (5 degrees C-55 degrees C) with a dwell time of 30 seconds for 5000 cycles. After this period, teeth were sectioned in approximately 0.8-mm slices. One slice of each tooth was randomly selected for confocal microscopy analysis. The other slices were sectioned into 0.8 nun x 0.8 mm beams, which were submitted to microtensile testing (MPa). Data were analyzed using two-way ANOVA and Tukey test (p < 0.05).Results: There was no detectedstatistical difference on bond strength among groups (alpha < 0.05). Confocal microscopy analysis showed a higher mean gap size in group 4(12.5 mu m) and a higher percentage of marginal gaps in the thermocycled groups. The RNIGIC liner groups showed the lowest percentage of marginal gaps.Conclusions: Lining with RMGIC resulted in less gap formation at the dentin/resin adhesive interface after artificial aging. RMGIC or GIC liners did not alter the microtensile bond strength of adhesive system/resin composite to dentin on the lateral walls of Class I restorations.

Longitudinal study of mandibular behavior in Class I subjects with vertical and horizontal growth

OBJETIVO: avaliar as possíveis diferenças no comportamento mandibular em indivíduos Classe I com crescimento vertical e horizontal. MÉTODOS: a amostra desse estudo consistiu de 20 indivíduos Classe I não tratados, sendo o grupo 1 composto por 10 indivíduos com padrão de crescimento vertical e o grupo 2 por 10 indivíduos com padrão de crescimento horizontal, pertencentes aos arquivos do Burlington Growth Center, University of Toronto, no Canadá, acompanhados radiograficamente nas idade de 9, 12 e 21 anos. Determinou-se, por meio de telerradiografias cefalométricas, em norma lateral, os valores médios para a avaliação longitudinal do comportamento da mandíbula utilizando as medidas SNB, Co-Gn, SN.GoMe, altura facial anterior e altura facial posterior. RESULTADOS: o valor de SNB e Co-Gn foram maiores no grupo com crescimento horizontal em todas as idades. A medida Sn.GoMe foi significativamente menor no grupo com crescimento horizontal, a altura facial anterior (AFH) apresentou valores menores nos indivíduos com padrão de crescimento horizontal, e a altura facial posterior (PFH) apresentou valores menores nos indivíduos com crescimento vertical. CONCLUSÃO: as comparações longitudinais das tendências de crescimento de indivíduos Classe I indicam que existe diferenças significativas entre os dois grupos. A mandíbula apresentou tendência à rotação horária no grupo 1. O grupo 2 exibiu tendência à característica de indivíduos braquicefálicos, na forma facial, devido ao déficit no desenvolvimento vertical na altura facial anterior.