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OBJECTIVES: To assess tooth discoloration induced by different hydraulic calcium silicate-based cements (HCSCs), including effects of blood and placement method. MATERIALS AND METHODS: Eighty bovine teeth cut to a length of 18 mm (crown 8 mm, root 10 mm) were randomly assigned to 10 groups (n = 8), receiving orthograde apical plug treatment (APT). Apical plugs were 4 mm in length and made of ProRoot MTA (Dentsply), Medcem MTA (Medcem), TotalFill BC RRM Fast Set Putty (Brasseler), or Medcem Medical Portland Cement (Medcem) plus bismuth oxide (Bi2O3) with and without bovine blood. Further, orthograde (with or without preoperative adhesive coronal dentin sealing) and retrograde APT were compared. Teeth were obturated with gutta-percha and sealer, sealed with composite and stored in distilled water. Tooth color was measured on apical plug, gutta-percha/sealer, and crown surface before treatment versus 24 h, 1, 3, 6, 12, and 24 months after treatment by spectrophotometry. Color difference (ΔE) values were calculated and analyzed by Shapiro-Wilk test, ANOVA with post hoc tests, Friedman test, t test, and post hoc tests with Bonferroni correction (α = .05). RESULTS: Tooth discoloration occurred in all groups with no significant differences between HCSCs (p > .05). After 24 months, color changes were prominent on roots but insignificant on crowns. Blood contamination induced a significantly decreased luminescence (p < .05). Blood had a stronger impact on tooth color than Bi2O3. No relevant effects of retrograde placement (p > .05) or preoperative dentin sealing (p > .05) were detected. CONCLUSIONS: Apical plugs of the tested HCSCs cause discoloration of bovine roots, but not discoloration of bovine tooth crowns within a 24-month period. CLINICAL RELEVANCE: APT should be performed carefully while avoiding direct contact with the coronal dentin, and in that case no aesthetic impairments occur.
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Materiais Restauradores do Canal Radicular , Descoloração de Dente , Animais , Compostos de Cálcio/efeitos adversos , Bovinos , Combinação de Medicamentos , Materiais Restauradores do Canal Radicular/efeitos adversos , Silicatos/efeitos adversos , Descoloração de Dente/induzido quimicamenteRESUMO
Vector-borne diseases often originate from wildlife and can spill over into the human population. One of the most important determinants of vector-borne disease transmission is the host preference of mosquitoes. Mosquitoes with a specialised host preference are guided by body odours to find their hosts in addition to carbon dioxide. Little is known about the role of mosquito host preference in the spillover of pathogenic agents from humans towards animals and vice versa. In the Republic of Congo, the attraction of mosquitoes to primate host odours was determined, as well as their possible role as malaria vectors, using odour-baited traps mimicking the potential hosts of mosquitoes. Most of the mosquito species caught showed a generalistic host preference. Anopheles obscurus was the most abundant Anopheles mosquito, with a generalistic host preference observed from the olfactory response and the detection of various Plasmodium parasites. Interestingly, Culex decens showed a much higher attraction towards chimpanzee odours than to human or cow odours. Human Plasmodium parasites were observed in both human and chimpanzee blood, although not in the Anopheles mosquitoes that were collected. Understanding the role of mosquito host preference for cross-species parasite transmission provides information that will help to determine the risk of spillover of vector-borne diseases.
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Anopheles/fisiologia , Quimiotaxia , Culex/fisiologia , Odorantes , Pan troglodytes , Plasmodium/isolamento & purificação , Zoonoses/transmissão , Animais , Anopheles/parasitologia , Congo , Culex/parasitologia , Comportamento Alimentar , Malária/transmissão , Malária/veterinária , Masculino , Mosquitos Vetores/parasitologia , Mosquitos Vetores/fisiologiaRESUMO
This corrects the article DOI: 10.1103/PhysRevLett.119.162501.
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The charge and magnetic form factors, F_{C} and F_{M}, respectively, of ^{3}He are extracted in the kinematic range 25 fm^{-2}≤Q^{2}≤61 fm^{-2} from elastic electron scattering by detecting ^{3}He recoil nuclei and scattered electrons in coincidence with the two High Resolution Spectrometers of the Hall A Facility at Jefferson Lab. The measurements find evidence for the existence of a second diffraction minimum for the magnetic form factor at Q^{2}=49.3 fm^{-2} and for the charge form factor at Q^{2}=62.0 fm^{-2}. Both minima are predicted to exist in the Q^{2} range accessible by this Jefferson Lab experiment. The data are in qualitative agreement with theoretical calculations based on realistic interactions and accurate methods to solve the three-body nuclear problem.
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Aim: 20 years after establishment of the National Breastfeeding Committee, the present work, based on published data on breastfeeding, is aimed at providing insight into the development of breastfeeding behaviour in Germany. Methods: To identify relevant publications, a comprehensive literature search was conducted in PubMed and Web of Science using the search terms "breast feeding" or "breastfeeding" in combination with "Germany". The publication period was limited to the period 1995-2014. Results: A total of 35 studies with data on breastfeeding for the birth cohorts of 1990-2012 were identified. Most of the data had been collected in regional or local surveys, often retrospectively. About 60% of the studies had been conducted with the primary aim of collecting data on breastfeeding or infant nutrition. Over the past 2 decades, breastfeeding rates were always relatively high at the beginning (72-97%). However, they declined significantly within the first 2 months, and by the age of 6 months, only about 50% of infants were still breastfed. Conclusion: Breastfeeding support and early assistance should be offered to a greater extent in order to achieve sustainable improvement of breastfeeding frequency and duration in Germany. Regarding the quality of data collected on breastfeeding, it seems crucial to implement standardised approaches to monitor breastfeeding in Germany.
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Aleitamento Materno/estatística & dados numéricos , Aleitamento Materno/tendências , Comportamento Materno , Adolescente , Adulto , Distribuição por Idade , Feminino , Alemanha/epidemiologia , Humanos , Recém-Nascido , Pessoa de Meia-Idade , Adulto JovemRESUMO
UNLABELLED: This study describes bone mass changes during pregnancy and lactation measured by a special ultrasound method. Pregnant women showed a decrease of bone mass followed by a stable bone mass while breast-feeding afterwards. Later in life, there is a recovery of bone mass loss. INTRODUCTION: The aim of this study was to evaluate bone changes during pregnancy using the radiation-free method of quantitative ultrasonometry (QUS). METHODS: One hundred twenty-five pregnant women who underwent prenatal care were included in this study. Ultrasound measurement of the calcaneus was performed in each trimester and then 6 weeks, 3 months, and 1 year postpartum. The calcaneal QUS measurements were carried out using the Achilles plus device (GE/Lunar Corporation, Madison, WI). Three ultrasound variables were measured: speed of sound (SOS, m/s), broadband ultrasound attenuation (BUA, dB/MHz), and the "stiffness index" (expressed as the percentage of the mean value in young adults). SOS and BUA raw data result in the t-score and z-score. RESULTS: A complete panel of six measurements was acquired over the time period in 101 patients (80.8%). Forty-two percent of the included patients were primipara, while 58% had given birth to at least one child (47%) previously. There was a statistically significant change of the t-score (tv = 2.14, p = 0.035) and the stiffness index (tv = 2.46, p = 0.016) from the second to the third trimester, followed by a plateau during lactation. Interestingly, the t-score remained stable during lactation, regardless of the duration of lactation (<3 months, 3-6 months, and >6 months). CONCLUSIONS: Young primiparas who had a sedentary adolescence were at the highest risk of bone loss during pregnancy. Bone loss that occurred during pregnancy was typically recovered later on, based on unknown molecular and biochemical mechanisms that must be elucidated with further studies.
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Densidade Óssea/fisiologia , Reabsorção Óssea/diagnóstico por imagem , Calcâneo/diagnóstico por imagem , Lactação/fisiologia , Gravidez/fisiologia , Adulto , Feminino , Humanos , Estudos Longitudinais , UltrassonografiaRESUMO
New results are reported from a measurement of π^{0} electroproduction near threshold using the p(e,e^{'}p)π^{0} reaction. The experiment was designed to determine precisely the energy dependence of s- and p-wave electromagnetic multipoles as a stringent test of the predictions of chiral perturbation theory (ChPT). The data were taken with an electron beam energy of 1192 MeV using a two-spectrometer setup in Hall A at Jefferson Lab. For the first time, complete coverage of the Ï_{π}^{*} and θ_{π}^{*} angles in the pπ^{0} center of mass was obtained for invariant energies above threshold from 0.5 up to 15 MeV. The 4-momentum transfer Q^{2} coverage ranges from 0.05 to 0.155 (GeV/c)^{2} in fine steps. A simple phenomenological analysis of our data shows strong disagreement with p-wave predictions from ChPT for Q^{2}>0.07 (GeV/c)^{2}, while the s-wave predictions are in reasonable agreement.
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The charge form factor of 4He has been extracted in the range 29 fm(-2) ≤ Q2 ≤ 77 fm(-2) from elastic electron scattering, detecting 4He recoil nuclei and electrons in coincidence with the high resolution spectrometers of the Hall A Facility of Jefferson Lab. The measurements have uncovered a second diffraction minimum for the form factor, which was predicted in the Q2 range of this experiment. The data are in qualitative agreement with theoretical calculations based on realistic interactions and accurate methods to solve the few-body problem.
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OBJECTIVES: Modern adhesives and composites allow the restoration of deep defects. In such cases, the matrix technique is particularly challenging, and excess composite is a common problem. Removing such overhangs with a scalpel has already been described as a substance preserving or selective finishing technique. Clinically, restoration margins may appear as a white line after scalpel finishing, and it is unclear whether this line represents a marginal gap and/or whether scalpel finishing promotes marginal gap formation. Therefore, the aim of this study was to investigate the influence of scalpel finishing of deep Class II composite restorations on marginal gap formation. METHODS AND MATERIALS: Standardized mesioocclusal-distal (MOD) cavities were prepared and restored in 60 human molars randomly divided into six finishing protocol groups: G1, scalpels (SC); G2, oscillating files (OF); G3, finishing strips (FS); G4, scalpels and finishing strips (SC+FS); G5, scalpels and polishing discs (SC+PD); G6, polishing discs alone (PD, controls). The groups were additionally assigned to finishing and polishing in a phantom head (groups 1-4) or hand-held setting (groups 5-6) to simulate clinical and in-vitro research conditions, respectively. After restoration, artificial aging was performed by thermocycling (5-55°C, 2500 cycles) and mechanical loading (50 newtons (N) with 500,000 cycles) prior to scanning electron microscopy analysis of proximal restoration margin quality on the mesial and distal surfaces (n=120) of each tooth. Outcomes (perfect margin, marginal gap, overhang, marginal fracture) were statistically analyzed by t-test, Mann-Whitney U test, single-factor analysis of variance, post-hoc t-test, Kruskal-Wallis test and Dunn-Bonferroni correction for multiple group comparisons. Cohen's effect size d(Cohen) was calculated to show the strength of the relationship between variables. RESULTS: Overall, marginal quality was significantly better in the hand-held setting (SC+PD and PD) than the phantom head setting (SC, OF, FS, SC+FS). The best marginal quality was achieved with oscillating files in the phantom head setting and with scalpels plus polishing discs in the hand-held setting. Marginal gaps occurred significantly more often with scalpels, but the proportion of gaps was very low and clinically insignificant. Finishing strips were the least effective instruments for removing overhangs but performed better in combination with scalpels. CONCLUSIONS: Scalpel finishing can effectively and gently remove overhangs from enamel. However, blades should be used with caution as they can cut the dentin and cementum. Scalpel finishing does not lead to a clinically relevant increase in marginal gaps, but should be followed by polishing, whenever possible, to eliminate any marginal fractures that might be present.
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Adaptação Marginal Dentária , Restauração Dentária Permanente , Humanos , Restauração Dentária Permanente/métodos , Resinas Compostas , Esmalte Dentário , Dente Molar/cirurgia , Cimentos de Resina , Preparo da Cavidade Dentária , Teste de MateriaisRESUMO
The endoscopic laser balloon ablation system affords a unique view of the beating heart for visual guidance in pulmonary vein (PV) isolation. A 66-year-old patient was admitted for catheter ablation of atrial fibrillation (AF). While encircling the left superior PV, AF terminated into sinus rhythm, which was diagnosed by observing sudden regularization of previously rapidly fibrillating atrial tissue demonstrating the unique endoscopic video function.
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Fibrilação Atrial/patologia , Fibrilação Atrial/cirurgia , Endoscópios , Sistema de Condução Cardíaco/cirurgia , Terapia a Laser/instrumentação , Veias Pulmonares/cirurgia , Cirurgia Assistida por Computador/instrumentação , Idoso , Desenho de Equipamento , Análise de Falha de Equipamento , Feminino , Humanos , Veias Pulmonares/patologia , Resultado do TratamentoRESUMO
Administration of an artificial peptide (pConsensus) based on anti-DNA IgG sequences that contain major histocompatibility complex class I and class II T-cell determinants, induces immune tolerance in NZB/NZW F1 female (BWF1) mice. To understand the molecular basis of CD8(+) Ti-mediated suppression, we previously performed microarray analysis to identify genes that were differentially expressed following tolerance induction with pCons. CD8(+) T cells from mice tolerized with pCons showed more than two-fold increase in Ifi202b mRNA, an interferon inducible gene, versus cells from untolerized mice. Ifi202b expression increased through weeks 1-4 after tolerization and then decreased, reapproaching baseline levels at 6 weeks. In vitro polyclonal activation of tolerized CD8(+) T cells significantly increased Ifi202b mRNA expression. Importantly, silencing of Ifi202b abrogated the suppressive capacity of CD8(+) Ti cells. This was associated with decreased expression of Foxp3, and decreased gene and protein expression of transforming growth factor (TGF)ß and interleukin-2 (IL-2), but not of interferon (IFN)-γ, IL-10, or IL-17. Silencing of another IFN-induced gene upregulated in tolerized CD8(+) T cells, IFNAR1, had no effect on the ability of CD8(+) T cells to suppress autoantibody production. Our findings indicate a potential role for Ifi202b in the suppressive capacity of peptide-induced regulatory CD8(+) Ti cells through effects on the expression of Foxp3 and the synthesis of TGFß.
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Anticorpos Antinucleares/biossíntese , Linfócitos T CD8-Positivos/imunologia , DNA/imunologia , Tolerância Imunológica , Peptídeos e Proteínas de Sinalização Intracelular/fisiologia , Animais , Anticorpos Antinucleares/química , Linfócitos B/metabolismo , Linfócitos T CD8-Positivos/metabolismo , Citocinas/metabolismo , Modelos Animais de Doenças , Feminino , Fatores de Transcrição Forkhead/genética , Fatores de Transcrição Forkhead/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/imunologia , Inativação Gênica , Tolerância Imunológica/efeitos dos fármacos , Tolerância Imunológica/genética , Região Variável de Imunoglobulina/imunologia , Região Variável de Imunoglobulina/farmacologia , Fatores Imunológicos/imunologia , Fatores Imunológicos/farmacologia , Peptídeos e Proteínas de Sinalização Intracelular/genética , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Lúpus Eritematoso Sistêmico/genética , Lúpus Eritematoso Sistêmico/imunologia , Lúpus Eritematoso Sistêmico/metabolismo , Camundongos , Camundongos Endogâmicos NZB , Peptídeos , RNA Mensageiro/genética , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismoRESUMO
Therapeutic agents currently in use to treat systemic lupus erythematosus (SLE) are predominantly immunosuppressive agents with limited specificities. Multiple groups, including ours, have illustrated that inducing tolerance in SLE animal models ameliorates disease symptoms and increases survival. We examined if oral administration of a tolerogenic peptide could affect SLE disease progression. The pConsensus (pCons) peptide, based on protein sequences of anti-double stranded (anti-ds)DNA antibodies, induces tolerance through upregulation of regulatory T cells when administered intravenously. Six different forms of pCons, including multiple antigenic peptides (MAP) and cyclic peptides made up of L- and D-amino acids, at three different concentrations, were fed to BWF1 SLE-susceptible mice for 30 weeks. Mice fed 100 µg of L-MAP or D-MAP had less cumulative proteinuria and serum anti-dsDNA antibody levels than controls. In addition, animals in these groups also survived significantly longer than controls with a corresponding increase in serum transforming growth factor beta (TGFß, implying a protective role for pCons-induced regulatory T cells. Oral administration of a tolerogenic peptide is a safe, effective method for ameliorating SLE disease manifestations and prolonging survival in SLE-prone mice. Induction of oral tolerance using modified pCons peptides could lead to a novel targeted therapy for human SLE.
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Anticorpos Antinucleares/biossíntese , Tolerância Imunológica/imunologia , Imunossupressores , Lúpus Eritematoso Sistêmico , Nefrite/tratamento farmacológico , Peptídeos , Administração Oral , Animais , Anticorpos Antinucleares/sangue , Humanos , Imunossupressores/administração & dosagem , Imunossupressores/imunologia , Imunossupressores/uso terapêutico , Lúpus Eritematoso Sistêmico/tratamento farmacológico , Lúpus Eritematoso Sistêmico/imunologia , Lúpus Eritematoso Sistêmico/patologia , Camundongos , Nefrite/patologia , Nefrite/fisiopatologia , Compostos Orgânicos , Peptídeos/administração & dosagem , Peptídeos/imunologia , Peptídeos/uso terapêutico , Taxa de Sobrevida , Linfócitos T Reguladores/imunologia , Fator de Crescimento Transformador beta/sangue , Fator de Crescimento Transformador beta/imunologiaRESUMO
Systemic lupus erythematosus (SLE) is a complex autoimmune disease characterized by a hyperactive immune system, including activation of autoreactive T and B cells. These studies demonstrate that administration of recombinant galectin-1, a ß-galactose binding protein, to SLE-prone (NZB × NZW) F1 mice reduced lymphocyte activation, inhibited serum anti-double-stranded DNA(dsDNA) IgG antibody production, decreased the incidence of proteinuria, and increased survival rate. In addition, recombinant galectin-1'-treated mice had a higher frequency of Foxp3 expression, which suggested an increase in the percentage of peripheral regulatory T cells. Consistent with the finding that there were fewer activated T lymphocytes, ex vivo T cells from mice treated with recombinant galectin-1 exhibited less proliferation in response to TCR stimulation. Furthermore, these cells were less efficient at lipid raft clustering in response to TCR/CD28 engagement, consistent with published reports that galectin-1 can reorganize the synaptic contact to interfere with TCR signaling and activation to prevent T cell activation. Aged galectin-1-deficient mice had higher serum levels of antibodies against dsDNA, elucidating a role for endogenous galectin-1 in decreasing susceptibility to autoimmunity. Together, the findings highlight galectin-1 as a novel potential therapeutic immune modulator for treatment of lupus-like disease.
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Autoanticorpos/sangue , Galectina 1/uso terapêutico , Lúpus Eritematoso Sistêmico/tratamento farmacológico , Ativação Linfocitária/efeitos dos fármacos , Linfócitos T/efeitos dos fármacos , Animais , DNA/imunologia , Regulação para Baixo , Avaliação Pré-Clínica de Medicamentos , Feminino , Fatores de Transcrição Forkhead/metabolismo , Galectina 1/farmacologia , Humanos , Imunoglobulina G/imunologia , Lúpus Eritematoso Sistêmico/imunologia , Lúpus Eritematoso Sistêmico/metabolismo , Microdomínios da Membrana/efeitos dos fármacos , Camundongos , Camundongos da Linhagem 129 , Camundongos Endogâmicos NZB , Camundongos Knockout , Proteinúria/etiologia , Proteinúria/prevenção & controle , Receptores de Antígenos de Linfócitos T/metabolismo , Proteínas Recombinantes/uso terapêutico , Baço/metabolismoRESUMO
The Lupus Foundation of America (LFA) convened an international working group to obtain a consensus definition of disease flare in lupus. With help from the Paediatric Rheumatology International Trials Organization (PRINTO), two web-based Delphi surveys of physicians were conducted. Subsequently, the LFA held a second consensus conference followed by a third Delphi survey to reach a community-wide agreement for flare definition. Sixty-nine of the 120 (57.5%) polled physicians responded to the first survey. Fifty-nine of the responses were available to draft 12 preliminary statements, which were circulated in the second survey. Eighty-seven of 118 (74%) physicians completed the second survey, with an agreement of 70% for 9/12 (75%) statements. During the second conference, three alternative flare definitions were consolidated and sent back to the international community. One hundred and sixteen of 146 (79.5%) responded, with agreement by 71/116 (61%) for the following definition: "A flare is a measurable increase in disease activity in one or more organ systems involving new or worse clinical signs and symptoms and/or laboratory measurements. It must be considered clinically significant by the assessor and usually there would be at least consideration of a change or an increase in treatment." The LFA proposes this definition for lupus flare on the basis of its high face validity.
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Lúpus Eritematoso Sistêmico/diagnóstico , Terminologia como Assunto , Doença Aguda , Técnica Delphi , Humanos , InternacionalidadeRESUMO
The HIV-1-specific cytotoxic T lymphocyte (CTL) response is temporally associated with the decline in viremia during primary HIV-1 infection, but definitive evidence that it is of importance in virus containment has been lacking. Here we show that in a patient whose early CTL response was focused on a highly immunodominant epitope in gp 160, there was rapid elimination of the transmitted virus strain and selection for a virus population bearing amino acid changes at a single residue within this epitope, which conferred escape from recognition by epitope-specific CTL. The magnitude (> 100-fold), kinetics (30-72 days from onset of symptoms) and genetic pathways of virus escape from CTL pressure were comparable to virus escape from antiretroviral therapy, indicating the biological significance of the CTL response in vivo. One aim of HIV-1 vaccines should thus be to elicit strong CTL responses against multiple codominant viral epitopes.
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Síndrome da Imunodeficiência Adquirida/virologia , HIV-1/imunologia , Linfócitos T Citotóxicos/imunologia , Viremia/imunologia , Síndrome da Imunodeficiência Adquirida/imunologia , Proteína gp160 do Envelope de HIV/imunologia , Humanos , Epitopos Imunodominantes/imunologia , Sondas de OligonucleotídeosRESUMO
The human immunodeficiency virus type 1 (HIV-1) encodes a protein, called Vpr, that prevents proliferation of infected cells by arresting them in G2 of the cell cycle. This Vpr-mediated cell-cycle arrest is also conserved among highly divergent simian immunodeficiency viruses, suggesting an important role in the virus life cycle. However, it has been unclear how this could be a selective advantage for the virus. Here we provide evidence that expression of the viral genome is optimal in the G2 phase of the cell cycle, and that Vpr increases virus production by delaying cells at the point of the cell cycle where the long terminal repeat (LTR) is most active. Although Vpr is selected against when virus is adapted to tissue culture, we show that selection for Vpr function in vivo occurs in both humans and chimpanzees infected with HIV-1. These results suggest a novel mechanism for maximizing virus production in the face of rapid killing of infected target cells.
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Ciclo Celular/fisiologia , Produtos do Gene vpr/biossíntese , HIV-1/fisiologia , Animais , Divisão Celular , Linhagem Celular , Fase G2 , Produtos do Gene vpr/fisiologia , Infecções por HIV/virologia , Humanos , Células Jurkat , Cinética , Modelos Biológicos , Pan troglodytes , Reação em Cadeia da Polimerase , Provírus/fisiologia , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/metabolismo , Vírus da Imunodeficiência Símia/fisiologia , Linfócitos T , Transfecção , Produtos do Gene vpr do Vírus da Imunodeficiência HumanaRESUMO
The viral accessory protein Vpx is required for productive in vitro infection of macrophages by simian immunodeficiency virus from sooty mangabey monkeys (SIV(SM)). To evaluate the roles of Vpx and macrophage infection in vivo, we inoculated pigtailed macaques intravenously or intrarectally with the molecularly cloned, macrophage tropic, acutely pathogenic virus SIV(SM) PBj 6.6, or accessory gene deletion mutants (deltaVpr or deltaVpx) of this virus. Both wild-type and SIV(SM) PBj deltaVpx viruses were readily transmitted across the rectal mucosa. A subsequent 'stepwise' process of local amplification of infection and dissemination was observed for wild-type virus, but not for SIV(SM) PBj deltaVpx, which also showed considerable impairment of the overall kinetics and extent of its replication. In animals co-inoculated with equivalent amounts of wild-type and SIV(SM) Pbj deltaVpx intravenously or intrarectally, the deltaVpx mutant was at a strong competitive disadvantage. Vpx-dependent viral amplification at local sites of initial infection, perhaps through a macrophage-dependent mechanism, may be a prerequisite for efficient dissemination of infection and pathogenic consequences after exposure through either mucosal or intravenous routes.
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Macrófagos/imunologia , Macrófagos/virologia , Vírus da Imunodeficiência Símia/patogenicidade , Proteínas Virais Reguladoras e Acessórias/fisiologia , Animais , Cercocebus atys , Genótipo , Imuno-Histoquímica , Hibridização In Situ , Mucosa Intestinal/virologia , Macaca nemestrina , Síndrome de Imunodeficiência Adquirida dos Símios/imunologia , Síndrome de Imunodeficiência Adquirida dos Símios/virologia , Vírus da Imunodeficiência Símia/genética , Vírus da Imunodeficiência Símia/fisiologia , Carga Viral , Replicação ViralRESUMO
Tolerizing mice polygenically predisposed to lupus-like disease (NZB/NZW F1 females) with a peptide mimicking anti-DNA IgG sequences containing MHC class I and class II T cell determinants (pConsensus, pCons) results in protection from full-blown disease attributable in part to the induction of CD4(+)CD25(+)Foxp3+ and CD8(+)Foxp3+ regulatory T cells. We compared 45 000 murine genes in total white blood cells (WBC), CD4(+) T cells, and CD8(+) T cells from splenocytes of (NZBxNZW) F1 lupus-prone mice tolerized with pCons vs untreated naïve mice and found two-fold or greater differential expression for 448 WBC, 174 CD4, and 60 CD8 genes. We identified differentially expressed genes that played roles in the immune response and apoptosis. Using real-time PCR, we validated differential expression of selected genes (IFI202B, Bcl2, Foxp3, Trp-53, CCR7 and IFNar1) in the CD8(+)T cell microarray and determined expression of selected highly upregulated genes in different immune cell subsets. We also determined Smads expression in different immune cell subsets, including CD4(+) T cells and CD8(+) T cells, to detect the effects of TGF-beta, known to be the major cytokine that accounts for the suppressive capacity of CD8(+) Treg in this system. Silencing of anti-apoptotic gene Bcl2 or interferon genes (IFI202b and IFNar1 in combination) in CD8(+) T cells from tolerized mice did not affect the expression of the other selected genes. However, silencing of Foxp3 reduced expression of Foxp3, Ifi202b and PD1-all of which are involved in the suppressive capacity of CD8(+) Treg in this model.
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Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD8-Positivos/metabolismo , DNA/imunologia , Imunoglobulinas/imunologia , Lúpus Eritematoso Sistêmico/imunologia , Animais , Apoptose/genética , Linfócitos T CD4-Positivos/citologia , Linfócitos T CD8-Positivos/citologia , Feminino , Perfilação da Expressão Gênica , Lúpus Eritematoso Sistêmico/genética , Camundongos , Reação em Cadeia da Polimerase , Regulação para CimaRESUMO
Neonatal exposure to antigen is believed to result in T cell clonal inactivation or deletion. Here we report that, contrary to this notion, neonatal injection of BALB/c mice with a hen egg lysozyme peptide 106-116 in putative "tolergenic" doses induced a T cell proliferative and an immunoglobulin G (IgG) antibody (Ab) response of both T helper cell 1 (Th1)- (IgG2a, IgG2b, and IgG 3) and Th2-dependent (IgG1) isotopes. Upon subsequent challenge with the peptide in complete Freund's adjuvant in adult life, although this neonatal regimen suppressed proliferation and the production of Th1 cytokines (interleukin[IL]-2 and interferon gamma), Th2 cytokine (IL-5, IL-4, and IL-10) secretion was increased, and the serum levels of Th1- and Th2-dependent isotypes of peptide-specific Ab remained elevated. The in vitro proliferative unresponsiveness in Th1 cells could be reversed by Abs to Th2 cytokines (IL-4 and IL-10). Thus, neonatal treatment with a peptide antigen induces T cell priming including production of IgG Abs of both Th1- and Th2-dependent isotypes. Upon subsequent peptide exposure, the peptide-specific T cell responses undergo an effective class switch in the direction of Th2, resulting in T cell proliferative unresponsiveness. Accordingly, this shift towards increased Ab production to autoantigen could be deleterious in individuals prone to antibody-mediated diseases. Indeed, neonatal treatment with a self-autoantigenic peptide from an anti-DNA monoclonal Ab (A6H 58-69) significantly increased the IgG anti-double-stranded DNA Ab levels in lupus-prone NZB/NZW F1 mice, despite suppressing peptide-specific T cell proliferation. This adverse clinical response is in sharp contrast to the beneficial outcome of neonatal treatment with autoantigens in Th1-mediated autoimmune diseases, such as autoimmune encephalomyelitis, as reported by others. A Th1 to Th2 immune deviation can explain the discordant biological responses after the presumed induction of neonatal tolerance in autoantibody- vs. Th-1 mediated autoimmune diseases.
Assuntos
Animais Recém-Nascidos/imunologia , Autoimunidade , Ativação Linfocitária , Muramidase/imunologia , Fragmentos de Peptídeos/imunologia , Sequência de Aminoácidos , Animais , Anticorpos/imunologia , Tolerância Imunológica , Imunização , Imunoglobulina G/biossíntese , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos NZB , Dados de Sequência Molecular , Linfócitos T/imunologia , Linfócitos T Auxiliares-Indutores/imunologiaRESUMO
(NZB x NZW) F1 (BWF1) mice develop spontaneous T cell autoimmunity to VH region determinants of syngeneic anti-DNA before the onset of clinical disease. In this study, we characterized the immunogenicity, MHC binding, and lymphokine secretion patterns induced by T cell determinants from the VH region of one such anti-DNA mAb (A6.1) and examined their role in the regulation of autoimmunity. Determinants were identified by proliferation of syngeneic splenic T cells from young, unprimed BWF1 mice in response to overlapping 12-mer peptides representing the entire VH region sequence. Immunization of young BWF1 mice with any of three determinants (A6H 34-45 [p34], A6H 58-69 [p58], and A6H 84-95 [p84]) elicited proliferative responses upon in vitro recall. Upon immunization with the whole A6.1 molecule, however, proliferative responses could be recalled only to the p58 peptide, defining this as immunodominant. The other two peptides (p34 and p84) elicited minimal or no proliferation and could be termed cryptic. Proliferative responses elicited by the cryptic determinants were restricted by a single class II (I-Ed for p34 and I-Au for p84), whereas the immunodominant p58 determinant was restricted by both I-Ed and I-Eu. The cryptic p34 and p84 bound strongly to I-Ed and I-Au, respectively, whereas the immunodominant p58 peptide bound poorly to I-Ed. A6H p84 elicited T cells that secreted lymphokines in a pattern consistent with a Th1-like phenotype, whereas p58 induced a Th2-like cytokine pattern. Immunization with p34 or p84, or adoptive transfer of a p84-reactive T cell line to young BWF1 mice significantly increased IgG anti-DNA levels, accelerated nephritis, and decreased survival. In conclusion, in BWF1 mice, autoreactive T cells recognizing both cryptic and dominant self-determinants on anti-DNA autoantibodies escape deletion or anergy induction. Furthermore, since these cells are spontaneously activated before the onset of clinical disease, they may be involved in the development of the autoimmune process.