RESUMO
OBJECTIVES: Evaluate efficacy and safety of tabalumab, a human IgG4 monoclonal antibody that binds and neutralises membrane and soluble B-cell activating factor (BAFF) versus placebo plus standard of care (SoC) in patients with systemic lupus erythematosus (SLE). METHODS: This phase III, 52-week study randomised 1164 patients with moderate-to-severe SLE (Safety of Estrogens in Lupus Erythematosus National Assessment-SLE Disease Activity Index ≥6 at baseline). Patients received SoC plus subcutaneous injections of tabalumab or placebo, starting with a loading dose (240â mg) at week 0 and followed by 120â mg every twoâ weeks (120 Q2W, n=387), 120â mg every fourâ weeks (120 Q4W, n=389) or placebo Q2W (n=388). PRIMARY ENDPOINT: proportion of patients achieving SLE Responder Index 5 (SRI-5) response at week 52. RESULTS: Similar proportions of patients in each group achieved SRI-5 response at week 52 (120 Q2W: 31.8%; 120 Q4W: 35.2% and placebo: 29.3%). Key secondary endpoints were not met. In a sensitivity analysis not excluding patients who decreased antimalarials or immunosuppressants, SRI-5 response was achieved with 120 Q4W (37.0% vs 29.8% placebo; p=0.021), but not 120 Q2W (34.1%; p=0.171). Significant reductions in anti-dsDNA antibodies, increases in C3 and C4, and reductions in total B cells and immunoglobulins were observed with tabalumab. No differences were observed between treatment groups in percentage of deaths (120 Q2W: 0.8%; 120 Q4W: 0.5%; placebo: 0.5%), serious adverse events (AEs) (range 11.1-14.4%) or treatment-emergent AEs (range 81.1-82.3%). CONCLUSIONS: Tabalumab had biological activity-changes in anti-dsDNA, complement, B cells and immunoglobulins-consistent with BAFF pathway inhibition. Key clinical efficacy endpoints did not achieve statistical significance. Safety profiles were similar with tabalumab and placebo. TRIAL REGISTRATION NUMBER: NCT01196091.
Assuntos
Anticorpos Monoclonais/administração & dosagem , Fator Ativador de Células B/antagonistas & inibidores , Lúpus Eritematoso Sistêmico/tratamento farmacológico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos Antinucleares/sangue , Anticorpos Monoclonais Humanizados , Autoanticorpos/sangue , Fator Ativador de Células B/administração & dosagem , Linfócitos B/metabolismo , Biomarcadores/sangue , População Negra , Complemento C3/metabolismo , Complemento C4/metabolismo , Relação Dose-Resposta a Droga , Método Duplo-Cego , Esquema de Medicação , Feminino , Humanos , Injeções Subcutâneas , Lúpus Eritematoso Sistêmico/etnologia , Masculino , Pessoa de Meia-Idade , Índice de Gravidade de Doença , Resultado do Tratamento , Adulto JovemRESUMO
INTRODUCTION: Anti-RNP autoantibodies occur either in mixed connective tissue disease (MCTD) (with a frequently favorable prognosis), or in systemic lupus erythematosus (SLE) cases with aggressive major organ disease. It is uncertain how to assess for the risk of severe disease in anti-RNP + patients. METHODS: Following institutional review board-approved protocols, clinical data and blood were collected from patients with known or suspected anti-RNP autoimmunity and normal controls in a cohort study. Samples were screened for parameters of immune activation. Groups were compared based on clinical diagnoses, disease classification criteria, disease activity and specific end-organ clinical manifestations. RESULTS: Ninety-seven per cent of patients satisfying Alarcon-Segovia MCTD criteria also met Systemic Lupus International Collaborating Clinic (SLICC) SLE criteria, while 47% of the anti-RNP + SLE patients also met MCTD criteria. Among SLICC SLE patients, MCTD criteria were associated with reduced rates of renal disease (odds ratio (OR) 4.3, 95% confidence interval (CI) 1.3-14.0), increased rates of Raynaud's phenomenon (OR 3.5, 95% CI 1.3-9.5) and increased serum B-cell maturation antigen, transmembrane activator and CAML interactor and TNFα levels. Circulating immune markers and markers of type I interferon activation were not effective at distinguishing clinical subgroups. CONCLUSIONS: Among anti-RNP patients, the question of MCTD versus SLE is not either/or: most MCTD patients also have lupus. MCTD classification criteria (but not a broad set of immune markers) distinguish a subset of SLE patients at reduced risk for renal disease.
Assuntos
Autoanticorpos/sangue , Doença Mista do Tecido Conjuntivo/diagnóstico , Ribonucleoproteínas/antagonistas & inibidores , Adulto , Anticorpos Antinucleares/imunologia , Autoanticorpos/metabolismo , Biomarcadores/sangue , Estudos de Casos e Controles , Estudos de Coortes , Feminino , Expressão Gênica , Humanos , Lúpus Eritematoso Sistêmico/sangue , Lúpus Eritematoso Sistêmico/diagnóstico , Lúpus Eritematoso Sistêmico/imunologia , Masculino , Pessoa de Meia-Idade , Doença Mista do Tecido Conjuntivo/sangue , Doença Mista do Tecido Conjuntivo/imunologia , RNA/sangue , RNA/genética , Doença de Raynaud/imunologia , Ribonucleoproteínas/imunologia , Medição de Risco/métodosRESUMO
Antibody heteroaggregates have been used to render human peripheral blood T cells lytic for specified targets. The heteroaggregates contain anti-T3 covalently linked to antibodies against nominal target cell antigens. Such heteroaggregates bind target cells directly to T3 molecules on effector cells and trigger target cell lysis. Freshly prepared human PBL, when coated with anti-T3-containing heteroaggregates, are lytic without further stimulation, although brief exposure to crude lymphokine-containing supernatants or recombinant IL-2, but not recombinant IFN-gamma, enhances the activity. The effector cells are T8+, and when fully stimulated, their lytic activity approaches that of some cloned CTL. When T cells are treated with heteroaggregate, washed, and incubated at 37 degrees C in medium not containing heteroaggregate, they retain activity for at least 24 h. The results of this study suggest a strategy in which heteroaggregate-coated T cells could be used in vivo to mount a lytic response against pathogenic cells such as tumor cells or virus-infected cells.
Assuntos
Anticorpos Monoclonais/imunologia , Anticorpos/imunologia , Citotoxicidade Imunológica , Linfócitos T/imunologia , Citotoxicidade Celular Dependente de Anticorpos , Humanos , Interferon gama/farmacologia , Interleucina-2/farmacologia , Cinética , Ativação Linfocitária , Neoplasias/imunologiaRESUMO
A patient had von Hippel-Lindau disease, a functional intrathoracic paraganglioma (pheochromocytoma), bilateral adrenal pheochromocytomas, and a para-adrenal pheochromocytoma. Seven other members of the patient's family had features of von Hippel-Lindau disease and one, a cousin, had medullary carcinoma of the thyroid. This is the first report of a pheochromocytoma arising outside the abdomen in von Hippel-Lindau disease and the 25th report of intrathoracic pheochromocytoma in the literature. The association between von Hippel-Lindau disease and pheochromocytoma is reviewed.
Assuntos
Neoplasias das Glândulas Suprarrenais/genética , Angiomatose/genética , Neoplasias Primárias Múltiplas , Feocromocitoma/genética , Neoplasias Torácicas/genética , Doença de von Hippel-Lindau/genética , Adulto , Diagnóstico Diferencial , Feminino , Humanos , LinhagemRESUMO
The role of T cells in the pathogenesis of systemic lupus erythematosus (SLE) is reviewed with a focus on autoantigen-specific T cells in SLE. The initial clue to a role for T cells in SLE was histopathologic studies demonstrating extensive infiltration of T cells at the sites of inflammation. Later studies, showing association between HLA polymorphisms and specific autoantibodies, directly implicated a role for T cells in autoantibody production. More recently, we and others have identified and characterized autoantigen-specific T cells in SLE. We review these studies on the role of autoantigen-specific T cells in SLE and present new findings on the molecular characterization of T cell immunity to Sm-B, Sm-D and U1-70kD small nuclear ribonucleoprotein (snRNP) autoantigens.
Assuntos
Lúpus Eritematoso Sistêmico/patologia , Linfócitos T/imunologia , Autoantígenos/imunologia , Humanos , Lúpus Eritematoso Sistêmico/imunologia , Ribonucleoproteínas Nucleares Pequenas/imunologia , Proteínas Centrais de snRNPRESUMO
We describe in this report a new strategy to directly sequence polymerase chain reaction-amplified human leucocyte antigen DRB genes using biotinylated allele-specific synthetic oligonucleotide primers coupled to streptavidin-coated magnetic beads. The use of allele-specific primers in the polymerase chain reaction allows for selective amplification of DNA from one haplotype, which when combined with the direct sequencing technique circumvents the need for DNA cloning prior to sequencing. We demonstrate here that this method can be used to characterize human leucocyte antigen DRB genes among heterozygous individuals. This method can be used for the rapid analysis of highly polymorphic genes among individuals heterozygous at the gene of interest.
Assuntos
Antígenos HLA-DR/genética , Reação em Cadeia da Polimerase/métodos , Alelos , Sequência de Bases , Clonagem Molecular , Genes , Humanos , Dados de Sequência Molecular , Oligodesoxirribonucleotídeos , Polimorfismo GenéticoRESUMO
The U1-70kD autoantigen is a major target of B cell responses in patients with connective tissue diseases (CTD). T cell responses are important in the pathogenesis of CTD, however little is known about autoantigen-specific T cells in these diseases. We have recently proven that U1-70kD-reactive human T cells exist. To further characterize these autoreactive T cells, U1-70kD-reactive T cell clones have been generated from patients with CTD using either a recombinant fusion protein or synthetic peptides spanning the U1-70kD polypeptide. T cell receptors (TCR) isolated from the U1-70kD-reactive T cell clones were sequenced and the third complementarity-determining region (CDR3) compared to determine if a common motif was present. mAb blocking of antigen-induced proliferation was done to determine the HLA restriction element used in recognition of the U1-70kD autoantigen by T cells. The results presented here indicate that TCRAV CDR3 usage is highly restricted among U1-70kD autoantigen-specific human T cells clones derived from CTD patients with distinctive structural features. Furthermore, the recognition of the U1-70kD autoantigen occurs in the context of HLA-DR.
Assuntos
Autoantígenos/imunologia , Doenças do Tecido Conjuntivo/imunologia , Receptores de Antígenos de Linfócitos T alfa-beta , Ribonucleoproteína Nuclear Pequena U1/imunologia , Linfócitos T/imunologia , Sequência de Aminoácidos , Células Clonais , Simulação por Computador , Doenças do Tecido Conjuntivo/etiologia , Sequência Conservada , Rearranjo Gênico do Linfócito T , Antígenos HLA-DR/imunologia , Humanos , Região Variável de Imunoglobulina , Lúpus Eritematoso Sistêmico/etiologia , Lúpus Eritematoso Sistêmico/imunologia , Doença Mista do Tecido Conjuntivo/etiologia , Doença Mista do Tecido Conjuntivo/imunologia , Modelos Moleculares , Dados de Sequência Molecular , Fragmentos de Peptídeos/imunologia , Análise de Sequência de DNARESUMO
Myxobolus cerebralis, the myxosporean parasite-causing salmonid whirling disease, was first reported among rainbow trout (Oncorhynchus mykiss) in Germany in 1903. The parasite was reported for the first time in North America in 1958 among hatchery-reared trout in the eastern USA, presumably arriving with frozen trout shipments from Europe. A comparison of 18S and ITS-1 ribosomal DNA sequences was conducted to identify potential strain differences between selected geographic isolates of this parasite from Europe and North America. Only fourteen of 1700 base pairs were different in the 18S rRNA gene from isolates obtained from California and West Virginia in the USA, and the Federal German Republic. No evidence for strain differences was obtained from ITS-1 sequences that were found to be identical among all parasite isolates. This finding is consistent with the hypothesis that the parasite was recently introduced to the USA from Europe.
Assuntos
DNA de Protozoário/genética , DNA Ribossômico/genética , Eucariotos/genética , Doenças dos Peixes/parasitologia , Infecções Protozoárias em Animais/parasitologia , Animais , Composição de Bases , Sequência de Bases , Eucariotos/classificação , Alemanha , Dados de Sequência Molecular , Oncorhynchus mykiss/parasitologia , Estados UnidosRESUMO
We studied a large North American Caucasian population of patients with biopsy proven IgA nephropathy for polymorphisms of HLA-A,B,C, HLA-DR, HLA-DQ and HLA-DP using a combination of serologic phenotyping and polymerase chain reaction sequence specific oligonucleotide probe (PCR-SSOP) hybridization genotyping. We also examined patients for polymorphisms of immunoglobulin by determining Gm and Km allotypes. When compared to healthy local controls there was an apparent decrease in HLA-DR5 (DRw11, DRw12) suggesting that this allele had a protective effect on disease susceptibility. None of the previously reported HLA-DQ associations found among Japanese or british Caucasian patients were found among this large North American Caucasian population. The HLA-DPB1*0601 genotype was increased among patients, but this was not significant when corrected for multiple comparisons. There were no differences in the distribution of Gm or Km allotypes among patients versus controls, regardless of whether they were stratified into those with progressive or non-progressive renal disease. Taken together, these findings suggest that there is substantial genetics heterogeneity in susceptibility to IgA nephropathy among different ethnic and/or geographically distinct populations.
Assuntos
Genes de Imunoglobulinas , Genes MHC da Classe II , Genes MHC Classe I , Glomerulonefrite por IGA/genética , Antígenos HLA/genética , Imunoglobulina A/genética , Alótipos de Imunoglobulina/genética , Alelos , Estudos de Casos e Controles , Frequência do Gene , Predisposição Genética para Doença , Glomerulonefrite por IGA/etnologia , Glomerulonefrite por IGA/imunologia , Antígeno HLA-DR5/genética , Teste de Histocompatibilidade , Humanos , Alótipos Gm de Imunoglobulina/genética , Cadeias Leves de Imunoglobulina , Reação em Cadeia da Polimerase , População Branca/genéticaRESUMO
OBJECTIVE: To examine relationships among changes in self-efficacy and changes in other clinically relevant outcome measures. METHOD: Subjects (n = 44) were participants in a prospective, randomized stress-management study followed over 15 months. Outcome measures included self-efficacy, depression, pain, health status, and disease activity. RESULTS: Correlational analyses revealed significant associations between changes in self-efficacy (particularly total self-efficacy) and changes in selected measures of depression, pain, health status, and disease activity. The observed associations were not due to changes in medication regimen or to nonadherence to the stress-management program. CONCLUSIONS: Evidence is provided that induced changes in self-efficacy following a stress-management program were significantly related to other clinically important outcome measures.
Assuntos
Atividades Cotidianas , Artrite Reumatoide/psicologia , Terapia de Relaxamento/normas , Autocuidado , Autoimagem , Estresse Psicológico/etiologia , Estresse Psicológico/prevenção & controle , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos ProspectivosRESUMO
One hundred and nineteen Merriam's wild turkeys (Meleagris gallopavo merriami) and 31 domestic chickens coexisting on a ranch in west-central Colorado (USA) were surveyed for mycoplasmosis by serologic and cultural methods. Although no clinical signs were apparent in any wild turkeys tested, 51 (43%) had positive rapid plate agglutination (RPA) reactions for M. gallisepticum (MG) and/or M. synoviae (MS); 37% of 56 adults and 48% of 63 subadults were classified as positive reactors to MG and/or MS. No turkeys tested in 1992 (n = 61) and 17 (29%) of 58 turkeys tested in 1993 were RPA-positive for M. meleagridis (MM). Hemagglutination inhibition (HI) test results were negative for MG, MS and MM as were most enzyme-linked immunosorbent assay (ELISA) test reactions (MG = 99%, MS = 93%, MM = 87%). Immunoblotting showed mild to moderate reactivity to MG proteins in 49% of 41 samples tested. Most chickens were strongly positive for MS by RPA (81%), HI (58%) and ELISA (87%); 48% also were positive for MG by RPA but all were MG-negative by HI and ELISA. No pathogenic mycoplasmas were isolated from either group of birds. Mycoplasma gallopavonis was commonly identified from the wild turkeys, and M. gallinaceum was isolated from both the chickens and wild turkeys. In a transmission study conducted in 1994, disease-free domestic turkeys failed to seroconvert when co-housed with wild turkeys from this population that were RPA-positive for MG. Collectively, the results of this study were inconclusive regarding the status of pathogenic mycoplasmas within this wild turkey population.
Assuntos
Doenças das Aves/epidemiologia , Galinhas , Infecções por Mycoplasma/veterinária , Perus , Testes de Aglutinação/veterinária , Animais , Animais Domésticos , Animais Selvagens , Anticorpos Antibacterianos/sangue , Colorado/epidemiologia , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Testes de Inibição da Hemaglutinação/veterinária , Immunoblotting , Masculino , Mycoplasma/imunologia , Infecções por Mycoplasma/epidemiologia , Prevalência , Estudos SoroepidemiológicosRESUMO
A process was developed for seed culture expansion (3.6 million-fold) using 5% of the hemicellulose hydrolysate from dilute acid pretreatment as the sole organic nutrient and source of sugar. Hydrolysate used for seed growth was neutralized with ammonia and combined with 1.0mM sodium metabisulfite immediately before inoculation. This seed protocol was tested with phosphoric acid pretreated sugarcane and sweet sorghum bagasse using a simplified process with co-fermentation of fiber, pentoses, and hexoses in a single vessel (SScF). A 6h liquefaction (L) step improved mixing prior to inoculation. Fermentations (L+SScF process) were completed in 72 h with high yields (>80 gal/US ton). Ethanol titers for this L+SScF process ranged from 24 g/L to 32 g/L, and were limited by the bagasse concentration (10% dry matter).
Assuntos
Celulose/metabolismo , Escherichia coli/metabolismo , Etanol/metabolismo , Lignina/metabolismo , Saccharum/microbiologia , Sementes/química , Sorghum/microbiologia , Fermentação/fisiologia , Lignina/química , VaporRESUMO
Microaeration (injecting air into the headspace) improved the fermentation of hemicellulose hydrolysates obtained from the phosphoric acid pretreatment of sugarcane bagasse at 170°C for 10 min. In addition, with 10% slurries of phosphoric acid pretreated bagasse (180°C, 10 min), air injection into the headspace promoted xylose utilization and increased ethanol yields from 0.16 to 0.20 g ethanol/g bagasse dry weight using a liquefaction plus simultaneous saccharification and co-fermentation process (L+SScF). This process was scaled up to 80 L using slurries of acid pretreated bagasse (96 h incubation; 0.6L of air/min into the headspace) with ethanol yields of 312-347 L (82-92 gal) per tone (dry matter), corresponding to 0.25 and 0.27 g/g bagasse (dry weight). Injection of small amounts of air into the headspace may provide a convenient alternative to subsurface sparging that avoids problems of foaming, sparger hygiene, flotation of particulates, and phase separation.
Assuntos
Ar , Biotecnologia/métodos , Celulose/química , Escherichia coli/metabolismo , Fermentação/efeitos dos fármacos , Ácidos Fosfóricos/farmacologia , Saccharum/química , Reatores Biológicos/microbiologia , Biotecnologia/instrumentação , Carboidratos/química , Escherichia coli/efeitos dos fármacos , Hidrólise , Vapor , SulfitosRESUMO
The addition of reduced sulfur compounds (thiosulfate, cysteine, sodium hydrosulfite, and sodium metabisulfite) increased growth and fermentation of dilute acid hydrolysate of sugarcane bagasse by ethanologenic Escherichia coli (strains LY180, EMFR9, and MM160). With sodium metabisulfite (0.5mM), toxicity was sufficiently reduced that slurries of pretreated biomass (10% dry weight including fiber and solubles) could be fermented by E. coli strain MM160 without solid-liquid separation or cleanup of sugars. A 6-h liquefaction step was added to improve mixing. Sodium metabisulfite also caused spectral changes at wavelengths corresponding to furfural and soluble products from lignin. Glucose and cellobiose were rapidly metabolized. Xylose utilization was improved by sodium metabisulfite but remained incomplete after 144 h. The overall ethanol yield for this liquefaction plus simultaneous saccharification and co-fermentation process was 0.20 g ethanol/g bagasse dry weight, 250 L/tonne (61 gal/US ton).
Assuntos
Escherichia coli/metabolismo , Etanol/metabolismo , Fermentação , Ácidos Fosfóricos/metabolismo , Saccharum/metabolismo , Compostos de Enxofre/metabolismo , BiomassaRESUMO
Hexose and pentose sugars from phosphoric acid pretreated sugarcane bagasse were co-fermented to ethanol in a single vessel (SScF), eliminating process steps for solid-liquid separation and sugar cleanup. An initial liquefaction step (L) with cellulase was included to improve mixing and saccharification (L+SScF), analogous to a corn ethanol process. Fermentation was enabled by the development of a hydrolysate-resistant mutant of Escherichia coli LY180, designated MM160. Strain MM160 was more resistant than the parent to inhibitors (furfural, 5-hydroxymethylfurfural, and acetate) formed during pretreatment. Bagasse slurries containing 10% and 14% dry weight (fiber plus solubles) were tested using pretreatment temperatures of 160-190°C (1% phosphoric acid, 10 min). Enzymatic saccharification and inhibitor production both increased with pretreatment temperature. The highest titer (30 g/L ethanol) and yield (0.21 g ethanol/g bagasse dry weight) were obtained after incubation for 122 h using 14% dry weight slurries of pretreated bagasse (180°C).
Assuntos
Celulose/metabolismo , Escherichia coli/classificação , Escherichia coli/metabolismo , Etanol/metabolismo , Hidrolisados de Proteína/metabolismo , Saccharum/metabolismo , Saccharum/microbiologia , Celulase/química , Celulose/química , Escherichia coli/genética , Mutação , Especificidade da EspécieAssuntos
Ferro/análise , Fígado/análise , Magnetismo/instrumentação , Animais , Engenharia Biomédica , Métodos , RatosRESUMO
OBJECTIVE: To observe the order of development of anti-U1 RNP peptide antibodies in humans. METHODS: Immunoblots against Jurkat cell lysates were performed on 5,882 serum samples from 3,668 patients referred on clinical grounds for RNP antibody testing to a reference laboratory between 1989 and 1999. In patients from whom multiple samples were drawn, we determined the order in which IgG antibodies to the U1 RNP peptides A, B'/B, C, D, and 70 kd appeared. RESULTS: One hundred sixty-three patients with serial samples were identified in whom antibodies to at least one U1 RNP peptide initially were not present but later appeared. The first RNP antibodies to appear were most often directed against the 70 kd and B'/B peptides (P < 0.01). Antibodies to the A and C peptides usually developed after other RNP peptide antibodies, and antibodies to D often emerged only after immunity to multiple other U1 RNP proteins had appeared. B'/B, but not 70 kd, was a frequent early target of spreading after initial immunity to other RNP peptides. CONCLUSION: Orderly patterns of emergence of U1 RNP peptide antibodies appear to exist in humans. Two peptides, 70 kd and B'/B, show characteristics of early immunogens in the development of human RNP immunity.
Assuntos
Doenças Autoimunes/sangue , Soros Imunes/imunologia , Ribonucleoproteína Nuclear Pequena U1/imunologia , Anticorpos/sangue , Especificidade de Anticorpos , Humanos , Imunidade , Epitopos Imunodominantes , Peptídeos/sangue , Peptídeos/imunologiaRESUMO
A defining feature of mixed connective tissue disease (MCTD) is the presence of antibodies against the U1-ribonucleoprotein (RNP) complex, but other autoantibodies in MCTD have recently been described. Research has also further elucidated the immune responses directed against U1-RNP in humans and in murine models of disease. Hypotheses implicating modified self-antigens and/or infectious agents in the pathogenesis of MCTD have been advanced. Links between the immunologic and clinical phenomena in MCTD are emerging. Longitudinal study of patients with MCTD highlights the impact of pulmonary hypertension on disease outcome.