Análise da eficácia do instrumento RECIPROC #25 em atingir o forame apical sem glide path

O presente estudo avaliou a eficáciado instrumento Reciproc#25 em atingir o forame de canais de molares inferiores sem um glide path manual prévio. Para isso, uma amostra geral de 300 molares inferiores foi radiografada e previamente selecionada quanto ao grau de curvatura segundo critério de Schneider sendo divididos em classes I e II. Após a aplicação dos critérios de inclusão e exclusão, uma amostragem total de 502 canais radiculares foi incluída para formar os grupos experimentais: 253 canais no grupo de molares inferiores classe I e 249 no grupo de molares classe II. Todos os canais foram instrumentados diretamente com a lima 25, sem nenhum glide path prévio, seguindo criteriosamente as diretrizes do fabricante. Os dados foram descritos como a frequência da distribuição do número de canais (%) nos quais foi possível chegar ao forame apical sem a necessidade de glide path, assim como o número de fraturas em cada grupo. Os resultados compilados dos 2 grupos experimentais mostraram que em 93,4% do total dos canais instrumentados, o instrumento R25 foi capaz de ir até o forame apical sem a necessidade de glide path. Em 6,4% do total dos canais, o instrumento R25 não chegou até o forame apical e em somente 0,2% dos casos ocorreu fratura da lima (um caso no grupo classe I, enquanto no grupo classe II não houve nenhuma ocorrência de fratura). O teste Qui-quadrado foi realizado para verificar se uma determinada classe de canais encontra-se mais associada ou não a necessidade de glide path quando o sistema Reciproc é usado. No grupo de molares classe II houve maior número de canais (23) que o instrumento R25 não foi capaz de ir até o forame apical do que no grupo de molares classe I (9), sendo essa diferença estatisticamente significante (Qui-quadrado, p=0,020, X2=5,452). Dentro das condições experimentais do presente estudo, pode-se concluir então quea lima R25 mostrou uma alta eficácia em instrumentar toda a extensão dos canais de molares inferiores classe I e II sem a necessidade de glide path prévio. Além disso, o sistema de instrumentação proposto mostrou-se altamente seguro quanto ao índice de fratura.

Reprodução e parasitismo em Melanoides tuberculata (Müller, 1774) (Gastropoda, Thiaridae) da Vila do Abraão, Ilha Grande, Angra dos Reis, RJ

Melanoides tuberculata (Müller, 1774), molusco exótico límnico de origem afroasiática, foi registrado pela primeira vez na Vila do Abraão, Ilha Grande, Angra dos Reis, Rio de Janeiro, em 2005. Atua como primeiro hospedeiro intermediário de diversos trematódeos de importância médica. As populações são compostas majoritariamente por fêmeas partenogenéticas que possuem um marsúpio onde se desenvolvem os juvenis. Os objetivos foram verificar: a presença de machos; se existem flutuações na produção de ovos e juvenis ao longo do tempo; quando a maturidade sexual é atingida e, a existência de parasitos relacionando sua presença com o número de ovos e juvenis. Para a contagem de ovos e juvenis, foram utilizadas fêmeas coletadas bimestralmente de setembro/07 a outubro/10, separadas em quatro classes de tamanho, segundo o diâmetro da concha: Classe I: < 3mm; Classe II: 3 a 5,99mm; Classe III: 6 a 8,99mm; Classe IV: > 9mm. Separamos cinco fêmeas/classe, totalizando 20 fêmeas/coleta, exceto nas amostras de setembro/07 (16 exemplares), abril/09 (19 exemplares) e de abril/10 (13 exemplares), totalizando 348 fêmeas. Para observação dos parasitos, ovos e juvenis o teto da cavidade palial foi retirado e o marsúpio dissecado. Classificamos os juvenis em classes de acordo com o número de voltas: com menos de duas; com duas a quatro e com mais de quatro. Os dados foram analisados utilizando Excel e SYSTAT 12. Para a histologia da gônada e do marsúpio, utilizamos cinco exemplares coletados mensalmente de setembro/12 a fevereiro/13, de cada classe de tamanho, totalizando 20 espécimes/coleta, exceto em outubro/12, quando 16 espécimes foram usados, pois somente um espécime da Classe IV foi encontrado. Os espécimes foram fixados em formalina Millonig de Carson, descalcificados em EDTA, incluídos em parafina e corados em hematoxilina e eosina. Foram encontrados 16 machos. Quantificamos 24.694 ovos e 31.474 juvenis, com as seguintes médias: Classe I: 0 ovos e 0,31 juvenis; Classe II: 24,19 ovos e 47,56 juvenis; Classe III: 114,84 ovos e 155,78 juvenis; Classe IV: 146,76 ovos e 158,41 juvenis. O teste de Kruskal-Wallis mostrou que existe variação significativa no número de ovos e juvenis ao longo do ano (p < 0,01 para ambos) e entre o número de ovos e juvenis entre as diferentes classes de tamanho de concha (p <0,01 para ambos). Foram contabilizados 27.877 juvenis com menos de duas voltas, 3.251 juvenis com duas a quatro voltas e 346 juvenis com mais de quatro voltas. Dentre 348 fêmeas, 111 estavam parasitadas (32% do total) por Centrocestus formosanus (Nishigori, 1924). O Teste de Mann-Whitney, levando em consideração todas as fêmeas, demonstrou que fêmeas parasitadas apresentaram menor número de ovos e juvenis que as não parasitadas ( p<0,01). Excluindo as fêmeas da Classe 1, o resultado do teste foi o mesmo (p< 0,01). Concluímos que: a população de M. tuberculata da Vila do Abraão não é formada somente por fêmeas; as fêmeas se reproduzem o ano todo, pois foram encontrados ovos e juvenis em todas as coletas; que quanto maior a fêmea há mais ovos e, em média, há mais juvenis no marsúpio; que a maturidade sexual é alcançada com aproximadamente 3 mm de diâmetro de concha, devido a presença de ovócitos vitelogênicos tardios e, que o parasitismo afeta negativamente M. tuberculata, reduzindo o número de ovos e juvenis formados

Qualidade de vida em pacientes com más oclusões de Classe III tratados com o protocolo do benefício antecipado

O protocolo do benefício antecipado é uma modalidade de tratamento ortocirúrgico que não envolve o preparo ortodôntico prévio. De acordo com os preceitos da Odontologia baseada em evidências, é essencial que se conheça o impacto dessa modalidade de tratamento na vida diária dos pacientes, uma vez que, para ser considerada viável, deve-se comprovar que ela oferece benefícios significativos para a qualidade de vida. Esse estudo objetivou conhecer os efeitos do tratamento ortocirúrgico com o protocolo do benefício antecipado na qualidade de vida e na autopercepção estética dos pacientes, durante dois anos de acompanhamento, e compará-los com os percebidos pelos pacientes tratados pela técnica tradicional. A amostra foi constituída por dezesseis pacientes, sendo oito no grupo tratado com o benefício antecipado (GBA) e oito no grupo tratado com a técnica ortocirúrgica tradicional. A qualidade de vida dos pacientes foi avaliada com três questionários: o OQLQ (Orthognathic Quality of Life Questionnaire), o OHIP-14 (Oral Health Impact Profile Short Version) e o SF-36 (Medical Outcomes Study 36 Item Short-Form Health Survey), em suas versões traduzidas e validadas para o português, e a autopercepção estética e a gravidade da má oclusão foram avaliadas com o Índice de Necessidade de Tratamento Ortodôntico (IOTN). Os exames foram repetidos em sete momentos de avaliação dos participantes: no exame inicial (T0), um mês depois do início do tratamento (T1), três meses depois do início do tratamento (T2), seis meses depois do início do tratamento (T3), um ano depois do início do tratamento e dois anos após o início do tratamento ou no término do tratamento ortocirúrgico (T5). Para ambos os grupos, houve um tempo pós-operatório (TPO) que foi realizado entre duas e três semanas após a cirurgia ortognática. A análise dos dados foi realizada com os testes de Mann-Whitney e de Friedman. Os pacientes do grupo GBA tiveram uma redução significativa no OQLQ (p<0,001) e no OHIP-14 (p<0,001) após dois anos de avaliação. Essa melhora foi progressiva e iniciada após a realização da cirurgia ortognática. O SF-36 apresentou melhoras significativas nas dimensões de capacidade funcional, limitação por aspectos físicos e aspectos sociais (p<0,001). A autopercepção estética comportou-se de maneira similar, com uma melhora progressiva e significativa (p<0,001), acompanhada de uma melhora significativa na gravidade da má oclusão (p<0,001). Porém os indivíduos que removeram o aparelho tiveram OHRQoL e autopercepção estética melhores em relação aos pacientes que não finalizaram o tratamento no período de dois anos no grupo GBA (N=4). No grupo GTT nenhum paciente foi operado após os dois anos de acompanhamento, e pioras significativas foram observadas no OQLQ (p<0,001) e no OHIP-14 (p<0,001) e na autopercepção estética (p<0,001). O CPO-D não teve alteração significativa para nenhum dos dois grupos. Concluiu-se que o tratamento ortocirúrgico com o protocolo do benefício antecipado gerou efeitos mais positivos na qualidade de vida, na autopercepção estética e na gravidade da má oclusão do que o tratamento ortocirúrgico tradicional após dois anos.

Alorreatividade dos enxertos ósseos homólogos na reconstrução alveolar em humanos

Enxerto ósseo homólogo é utilizado independentemente da compatibilidade HLA entre doador e receptor ou uso de drogas imunossupressoras. Considerando o volume de transplantes ósseos realizados no Brasil e o possível efeito deletério da sensibilização HLA para o transplante de órgãos sólidos, este estudo tem como objetivo avaliar a alorreatividade do enxerto ósseo homólogo fresco-congelado utilizado na reconstrução alveolar com finalidade de reabilitação oral com prótese sobre implantes. Anticorpos anti-HLA e anti-MICA foram monitorados através do teste Labscreen Mixed, nos intervalos 0, 7, 30, 90 e 180 pós transplante ósseo em 15 pacientes (6 homens e 9 mulheres, idade média 58,1, DP=10,1) que estavam em tratamento no Instituto de Odontologia da Pontifícia Universidade Católica do Rio de Janeiro. Caso resultado do teste Mixed fosse positivo (Razão de fundo normatizado, NBG>4,5) o teste Labscreen Single (tecnologia antígeno único por pérola, SABA) era realizado para verificar se os anticorpos anti-HLA eram específicos ao doador. Nenhum paciente relatou transplante prévio, 4 relataram transfusão prévia e todas as mulheres relataram gravidez. Dez pacientes não apresentaram reação positiva no dia 0 sendo considerados não sensibilizados previamente (NSP); destes, 6 pacientes permaneceram sem nenhuma evidência de sensibilização, 2 pacientes apresentaram reação positiva para Classe I e II; 2 para Classe I apenas; e 2 para MICA, sendo considerados sensibilizados pelo enxerto ósseo oral. Dois pacientes apresentaram aumento de Intensidade Média de Fluorescência (ΔMFI>1000) de anticorpos específicos ao doador para Classe I e Classe II, e 2 somente para Classe II, demonstrando uma reação específica ao doador. Os resultados sugerem uma alorreatividade HLA oscilatória ao enxerto ósseo homólogo em reconstruções alveolares, confirmada pela formação de anticorpos anti-HLA específicos ao doador em 4 pacientes (27%) da amostra.

HLA-DRB1 allele polymorphisms in genetic susceptibility to esophageal carcinoma

AIM: To probe into the genetic susceptibility of HLA-DRB1 alleles to esophageal carcinoma in Han Chinese in Hubei Province. METHODS: HLA-DRB1 allele polymorphisms were typed by polymerase chain reaction with sequence-specific primers (PCR-SSP) in 42 unrelated patients with esophageal cancer and 136 unrelated normal control subjects and the associated HLA-DRB1 allele was measured by nucleotide sequence analysis with PCR.SAS software was used in statistics. RESULTS: Allele frequency (AF) of HLA-DRB1*0901 was significantly higher in esophageal carcinoma patients than that in the normal controls (0.2500 vs0.1397, P=0.028, the odds ratio 2.053, etiologic fraction 0.1282). After analyzed the allele nucleotide sequence of HLA-DRB1*0901 which approachs to the corresponded exon 2 sequence of the allele in genebank. There was no association between patients and controls in the rested HLA-DRB1 alleles. CONCLUSION: HLA-DRB1*0901 allele is more common in the patients with esophageal carcinoma than in the healthy controls, which is positively associated with the patients of Hubei Han Chinese. Individuals carrying HLA-DRB1*0901 may be susceptible to esophageal carcinoma.

Identification and analysis of a CpG motif that protects turbot (Scophthalmus maximus) against bacterial challenge and enhances vaccine-induced specific immunity

Oligodeoxynucleotides (ODNs) containing unmethylated CpG motifs in certain contexts are known to be immunostimulatory in vertebrate systems. CpG ODNs with immune effects have been identified for many fish species but, to our knowledge, not for turbot. In this study, a turbot-effective CpG ODN, ODN 205, was identified and a plasmid, pCN5, was constructed which contains the CpG motif of ODN 205. When administered into turbot via intraperitoneal (i.p.) injection, both ODN 205 and pCN5 could (i) inhibit bacterial dissemination in blood in dose and time dependent manners, and (ii) protect against lethal bacterial challenge. Immunological analyses showed that in vitro treatment with ODN 205 stimulated peripheral blood leukocyte proliferation, while i.p. injection with ODN 205 enhanced the respiratory burst activity, chemiluminescence response, and acid phosphatase activity of turbot head kidney macrophages. pCN5 treatment-induced immune responses similar to those induced by ODN 205 treatment except that pCN5 could also enhance serum bactericidal activity in a calcium-independent manner. To examine whether ODN 205 and pCN5 had any effect on specific immunity, ODN 205 and pCN5 were co-administered into turbot with a Vibrio harveyi subunit vaccine, DegQ. The results showed that pCN5, but not ODN 205, significantly increased the immunoprotective efficacy of DegQ and enhanced the production of specific serum antibodies in the vaccinated fish. Further analysis indicated that vaccination with DegQ in the presence of pCN5 upregulated the expression of the genes encoding MHC class II alpha, IgM, Mx, and IL-8 receptor. Taken together, these results demonstrate that ODN 205 and pCN5 can stimulate the immune system of turbot and induce protection against bacterial challenge. In addition, pCN5 also possesses adjuvant property and can potentiate vaccine-induced specific immunity. (C) 2010 Elsevier Ltd. All rights reserved.

Investigation of polyhydroxyalkanoate production by an activated sludge microbial consortium treating artificial dairy wastewater

Petrochemical plastics/polymers are a common feature of day to day living as they occur in packaging, furniture, mobile phones, computers, construction equipment etc. However, these materials are produced from non-renewable materials and are resistant to microbial degradation in the environment. Considerable research has therefore been carried out into the production of sustainable, biodegradable polymers, amenable to microbial catabolism to CO2 and H2O. A key group of microbial polyesters, widely considered as optimal replacement polymers, are the Polyhydroxyalkaonates (PHAs). Primary research in this area has focused on using recombinant pure cultures to optimise PHA yields, however, despite considerable success, the high costs of pure culture fermentation have thus far hindered the commercial viability of PHAs thus produced. In more recent years work has begun to focus on mixed cultures for the optimisation of PHA production, with waste incorporations offering optimal production cost reductions. The scale of dairy processing in Ireland, and the high organic load wastewaters generated, represent an excellent potential substrate for bioconversion to PHAs in a mixed culture system. The current study sought to investigate the potential for such bioconversion in a laboratory scale biological system and to establish key operational and microbial characteristics of same. Two sequencing batch reactors were set up and operated along the lines of an enhanced biological phosphate removal (EBPR) system, which has PHA accumulation as a key step within repeated rounds of anaerobic/aerobic cycling. Influents to the reactors varied only in the carbon sources provided. Reactor 1 received artificial wastewater with acetate alone, which is known to be readily converted to PHA in the anaerobic step of EBPR. Reactor 2 wastewater influent contained acetate and skim milk to imitate a dairy processing effluent. Chemical monitoring of nutrient remediation within the reactors as continuously applied and EBPR consistent performances observed. Qualitative analysis of the sludge was carried out using fluorescence microscopy with Nile Blue A lipophillic stain and PHA production was confirmed in both reactors. Quantitative analysis via HPLC detection of crotonic acid derivatives revealed the fluorescence to be short chain length Polyhydroxybutyrate, with biomass dry weight accumulations of 11% and 13% being observed in reactors 1 and 2, respectively. Gas Chromatography-Mass Spectrometry for medium chain length methyl ester derivatives revealed the presence of hydroxyoctanoic, -decanoic and -dodecanoic acids in reactor 1. Similar analyses in reactor 2 revealed monomers of 3-hydroxydodecenoic and 3-hydroxytetradecanoic acids. Investigation of the microbial ecology of both reactors as conducted in an attempt to identify key species potentially contributing to reactor performance. Culture dependent investigations indicated that quite different communities were present in both reactors. Reactor 1 isolates demonstrated the following species distributions Pseudomonas (82%), Delftia acidovorans (3%), Acinetobacter sp. (5%) Aminobacter sp., (3%) Bacillus sp. (3%), Thauera sp., (3%) and Cytophaga sp. (3%). Relative species distributions among reactor 2 profiled isolates were more evenly distributed between Pseudoxanthomonas (32%), Thauera sp (24%), Acinetobacter (24%), Citrobacter sp (8%), Lactococcus lactis (5%), Lysinibacillus (5%) and Elizabethkingia (2%). In both reactors Gammaproteobacteria dominated the cultured isolates. Culture independent 16S rRNA gene analyses revealed differing profiles for both reactors. Reactor 1 clone distribution was as follows; Zooglea resiniphila (83%), Zooglea oryzae (2%), Pedobacter composti (5%), Neissericeae sp. (2%) Rhodobacter sp. (2%), Runella defluvii (3%) and Streptococcus sp. (3%). RFLP based species distribution among the reactor 2 clones was as follows; Runella defluvii (50%), Zoogloea oryzae (20%), Flavobacterium sp. (9%), Simplicispira sp. (6%), Uncultured Sphingobacteria sp. (6%), Arcicella (6%) and Leadbetterella bysophila (3%). Betaproteobacteria dominated the 16S rRNA gene clones identified in both reactors. FISH analysis with Nile Blue dual staining resolved these divergent findings, identifying the Betaproteobacteria as dominant PHA accumulators within the reactor sludges, although species/strain specific allocations could not be made. GC analysis of the sludge had indicated the presence of both medium chain length as well short chain length PHAs accumulating in both reactors. In addition the cultured isolates from the reactors had been identified previously as mcl and scl PHA producers, respectively. Characterisations of the PHA monomer profiles of the individual isolates were therefore performed to screen for potential novel scl-mcl PHAs. Nitrogen limitation driven PHA accumulation in E2 minimal media revealed a greater propensity among isoates for mcl-pHA production. HPLC analysis indicated that PHB production was not a major feature of the reactor isolates and this was supported by the low presence of scl phaC1 genes among PCR screened isolates. A high percentage distribution of phaC2 mcl-PHA synthase genes was recorded, with the majority sharing high percentage homology with class II synthases from Pseudomonas sp. The common presence of a phaC2 homologue was not reflected in the production of a common polymer. Considerable variation was noted in both the monomer composition and ratios following GC analysis. While co-polymer production could not be demonstrated, potentially novel synthase substrate specificities were noted which could be exploited further in the future.

Assessment of the immunomodulatory effects of raw/farm milk and probiotic bacteria

The overall aims of this study were to investigate the differences between raw/farm milk and pasteurised milk with respect to potential immune modifying effects following consumption and investigate the bacterial composition of raw milk compared to pasteurised milk. Furthermore, in this thesis, panels of potential probiotic bacteria from the Bifidobacterium and Lactobacillus genera were investigated. The overall bacterial composition of raw milk was compared with pasteurised milk using samples obtained from commercial milk producers around Ireland using next generation sequencing technology (454 pyrosequencing). Here the presence of previously unrecognised and diverse bacterial populations in unpasteurised cow’s milk was identified. Futhermore the bacterial content of pasteurised milk was found to be more diverse than previously thought. The global response of the adenocarcinoma cell line HT-29 to raw milk and pasteurised milk exposures were also characterised using whole genome microarray technology. Over one thousand differentially expressed genes were identified which were found to be involved in a plethora of cellular functions. Interestingly a reduction in immune related activity (e.g. Major histocompatability complex class II signalling and T and B cell proliferation) was identified in cells exposed to pasteurised milk compared with raw milk exposures. Further studies comparing human cell response to raw versus pasteurised milk was performed using peripheral blood mononuclear cells (PBMC) from healthy donors. A reduction in CD14 was identified following raw milk exposures compared with pasteurised milk and the pattern of cytokine production may indicate that gram positive bacteria in the raw milk were contributing to the differences in the cellular response to raw versus pasteurised milk. Panels of potentially probiotic bacteria (comprising of lactobacilli and bifidobacteria) were further assessed for immunomodulatory capabilities using cell culture based models. Gene expression and cytokine production were used to evaluate stimulated and unstimulated (LPS) cellular responses as well as interaction mechanisms

Differential T and B cell responses against Mycobacterium tuberculosis heparin-binding hemagglutinin adhesin in infected healthy individuals and patients with tuberculosis.

Because only 10% of individuals infected with Mycobacterium tuberculosis will eventually develop disease, antigens that are recognized differently by the immune systems of infected healthy and diseased subjects may constitute potential vaccine candidates. Here, the heparin-binding hemagglutinin adhesin (HBHA) is identified as such an antigen. Lymphocytes from 60% of healthy infected individuals (n=25) produced interferon (IFN)-gamma after stimulation with HBHA, compared with only 4% of patients with active tuberculosis (n=24). In the responders, both CD4(+) and CD8(+) cells secreted HBHA-specific IFN-gamma, and the antigen was presented by both major histocompatibility complex class I and II molecules. In contrast to the reduced ability of patients with tuberculosis to produce HBHA-specific IFN-gamma, most of them (82%) produced anti-HBHA antibodies, compared with 36% of the infected healthy subjects. These observations indicate that HBHA is recognized differently by the immune systems of patients with tuberculosis and infected healthy individuals and might provide a marker for protection against tuberculosis.

From clonal selection to immune networks: induction of silent idiotypes.

Journal Article

Polyacid-modified composite resins ("compomers") and their use in clinical dentistry

OBJECTIVES: This paper describes the chemistry and properties of polyacid-modified composite resins ("compomers") designed for use in clinical dentistry, and reviews the literature in this area. METHODS: Information has been obtained from over 50 published articles appearing in the dental and biomaterials literature, with studies being principally identified through MedLine. RESULTS: Published work shows that polyacid-modified composite resins constitute a discrete class of polymeric repair material for use in dentistry. Their distinction is that they contain hydrophilic components, and these cause water to be drawn into the material following cure. This triggers an acid-base reaction, and gives the materials certain clinically-desirable properties (fluoride release, buffering capability) that are also associated with glass-ionomer cements. The water uptake leads to a decline in certain, though not all, physical properties. However, clinical studies have shown these materials to perform acceptably in a variety of applications (Class I, Class II and Class V cavities, as fissure sealants and as orthodontic band cements), especially in children's teeth. CONCLUSIONS/SIGNIFICANCE: Polyacid-modified composite resins constitute a versatile class of dental repair material, whose bioactivity confers clinical advantages, and which are particularly useful in children's dentistry.

Botany origin and protein content of homemade honeys from Galicia hives (NW Spain)

The botanic origin and the protein content of 15 honeys from small bee farms exploitations of Galicia, for family consume, were studied; the aim is to check if the protein wealth and the pollen wealth are dependent parameters. Seven honeys resulted to be Rhamnus frangula unifloral (pollen patterns with low diversity), two Castanea sativa Miller unifloral, other one heather unifloral, and five was multifloral honeys of various pollen patterns (four Castanea predominant and one Rhamnus frangula predominant). Their pollen wealth was low; eight honeys classified in the Maurizio Class I, 3 in Class II, 2 in Class III, and one in Maurizio Class IV. There has been a wide variability in its protein content (0.09- 4.83 mg prot./g honey). The relative amount of pollen from different taxa has a direct or inverse proportionality to wealth protein.

The pathophysiology of eosinophilic esophagitis: Altered mechanisms of antigen detection in the esophagus

Recently, a chronic idiopathic disease of the esophagus has emerged, which is now known as eosinophilic esophagitis (EoE). Incomplete knowledge regarding the pathogenesis of EoE has limited treatment options. EoE is known to be a Th2-type immune-mediated disorder. Based on previous studies in both patients and experimental models, it is possible that an abnormal reaction to antigen mediates the pathophysiology of EoE. In this thesis, symptoms and signs unique to EoE were identified by an age-matched, case-controlled study of 326 patients with EoE and gastroesophageal reflux disease. The molecular mechanisms involved in antigen detection in the esophagus, in relation to EoE were then investigated. Esophageal epithelial cells were found, for the first time, to be capable of acting as non-professional antigen presenting cells, with the ability to engulf, process and present antigen on MHC class II to T helper lymphocytes. Antigen presentation by esophageal epithelial cells was induced by interferon-γ, which is increased in biopsies from patients with EoE. Next, it was discovered that esophageal epithelial cell lines expressed functional toll-like receptor (TLR) 2 and TLR3, but in esophageal mucosal biopsies only infiltrating immune cells (including eosinophils) expressed TLR2 and TLR3. Finally, the potential involvement of IgE in the pathogenesis of esophageal inflammation was investigated. IgE in the esophagus was found to be present on mast cells, which are increased in density in the esophageal mucosae of patients with EoE and especially those with a history of atopy. Mechanisms of antigen detection may mediate the pathophysiology of EoE in the esophagus through antigen presentation by epithelial cells, detection by TLRs on immune cells and detection through IgE on mucosal mast cells. Together, these findings demonstrate that mechanisms of antigen detection may actually contribute to the pathophysiology of EoE. Through increased understanding of the mechanisms of EoE, the results of this thesis may contribute to future therapy.

Cloning and characterization of the mouse homologue of the human dendritic cell maturation marker CD208/DC-LAMP

DC-LAMP, a member of the lysosomal-associated membrane protein (LAMP) family, is specifically expressed by human dendritic cells (DC) upon activation and therefore serves as marker of human DC maturation. DC-LAMP is detected first in activated human DC within MHC class II molecules-containing compartments just before the translocation of MHC class II-peptide complexes to the cell surface, suggesting a possible involvement in this process. The present study describes the cloning and characterization of mouse DC-LAMP, whose predicted protein sequence is over 50% identical to the human counterpart. The mouse DC-LAMP gene spans over 25 kb and shares syntenic chromosomal localization (16B2-B4 and 3q26) and conserved organization with the human DC-LAMP gene. Analysis of mouse DC-LAMP mRNA and protein revealed the expression in lung peripheral cells, but also its unexpected absence from mouse lymphoid organs and from mouse DC activated either in vitro or in vivo. In conclusion, mouse DC-LAMP is not a marker of mature mouse DC and this observation raises new questions regarding the role of human DC-LAMP in human DC.

Th2 lymphoproliferative disorder of LatY136F mutant mice unfolds independently of TCR-MHC engagement and is insensitive to the action of Foxp3+ regulatory T cells.

Mutant mice where tyrosine 136 of linker for activation of T cells (LAT) was replaced with a phenylalanine (Lat(Y136F) mice) develop a fast-onset lymphoproliferative disorder involving polyclonal CD4 T cells that produce massive amounts of Th2 cytokines and trigger severe inflammation and autoantibodies. We analyzed whether the Lat(Y136F) pathology constitutes a bona fide autoimmune disorder dependent on TCR specificity. Using adoptive transfer experiments, we demonstrated that the expansion and uncontrolled Th2-effector function of Lat(Y136F) CD4 cells are not triggered by an MHC class II-driven, autoreactive process. Using Foxp3EGFP reporter mice, we further showed that nonfunctional Foxp3(+) regulatory T cells are present in Lat(Y136F) mice and that pathogenic Lat(Y136F) CD4 T cells were capable of escaping the control of infused wild-type Foxp3(+) regulatory T cells. These results argue against a scenario where the Lat(Y136F) pathology is primarily due to a lack of functional Foxp3(+) regulatory T cells and suggest that a defect intrinsic to Lat(Y136F) CD4 T cells leads to a state of TCR-independent hyperactivity. This abnormal status confers Lat(Y136F) CD4 T cells with the ability to trigger the production of Abs and of autoantibodies in a TCR-independent, quasi-mitogenic fashion. Therefore, despite the presence of autoantibodies causative of severe systemic disease, the pathological conditions observed in Lat(Y136F) mice unfold in an Ag-independent manner and thus do not qualify as a genuine autoimmune disorder.