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Rationale: The biological mechanisms of long-term cognitive impairment and disability after critical illness are unclear.Objectives: To test the hypothesis that markers of acute inflammation and coagulation are associated with subsequent long-term cognitive impairment and disability.Methods: We obtained plasma samples from adults with respiratory failure or shock on Study Days 1, 3, and 5 and measured concentrations of CRP (C-reactive protein), IFN-γ, IL-1ß, IL-6, IL-8, IL-10, IL-12, MMP-9 (matrix metalloproteinase-9), TNF-α (tumor necrosis factor-α), soluble TNF receptor 1, and protein C. At 3 and 12 months after discharge, we assessed global cognition, executive function, and activities of daily living. We analyzed associations between markers and outcomes using multivariable regression, adjusting for age, sex, education, comorbidities, baseline cognition, doses of sedatives and opioids, stroke risk (in cognitive models), and baseline disability scores (in disability models).Measurements and Main Results: We included 548 participants who were a median (interquartile range) of 62 (53-72) years old, 88% of whom were mechanically ventilated, and who had an enrollment Sequential Organ Failure Assessment score of 9 (7-11). After adjusting for covariates, no markers were associated with long-term cognitive function. Two markers, CRP and MMP-9, were associated with greater disability in basic and instrumental activities of daily living at 3 and 12 months. No other markers were consistently associated with disability outcomes.Conclusions: Markers of systemic inflammation and coagulation measured early during critical illness are not associated with long-term cognitive outcomes and demonstrate inconsistent associations with disability outcomes. Future studies that pair longitudinal measurement of inflammation and related pathways throughout the course of critical illness and during recovery with long-term outcomes are needed.
Assuntos
Biomarcadores/sangue , Transtornos da Coagulação Sanguínea/sangue , Proteína C-Reativa/análise , Disfunção Cognitiva/sangue , Inflamação/sangue , Fatores Reguladores de Interferon/sangue , Metaloproteinases da Matriz/sangue , Fatores de Necrose Tumoral/sangue , Idoso , Estado Terminal , Pessoas com Deficiência , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Estudos ProspectivosRESUMO
Previous studies have suggested that pathogenic variants in interferon regulatoryse factor 6 (IRF6) can account for almost 70% of familial Van der Woude Syndrome (VWS) cases. However, gene modifiers that account for the phenotypic variability of IRF6 in the context of VWS remain poorly characterized. The aim of this study was to report a family with VWS with variable expressivity and to identify the genetic cause. A 4monthold boy initially presented with cleft palate and bilateral lower lip pits. Examination of his family history identified similar, albeit milder, clinical features in another four family members, including bilateral lower lip pits and/or hypodontia. Peripheral blood samples of eight members in this threegeneration family were subsequently collected, and wholeexome sequencing was performed to detect pathogenic variants. A heterozygous missense IRF6 variant with a c.1198C>T change in exon 9 (resulting in an R400W change at the amino acid level) was detected in five affected subjects, but not in the other three unaffected subjects. Moreover, subsequent structural analysis was indicative of damaged stability to the structure in the mutant IRF protein. Wholetranscriptome sequencing, expression analysis and Gene Ontology enrichment analysis were conducted on two groups of patients with phenotypic diversity from the same family. These analyses identified significant differentially expressed genes and enriched pathways in these two groups. Altogether, these findings provide insight into the mechanism underlying the variable expressivity of VWS.
Assuntos
Anormalidades Múltiplas/genética , Fenda Labial/genética , Fissura Palatina/genética , Cistos/genética , Características da Família , Fatores Reguladores de Interferon/genética , Lábio/anormalidades , Mutação de Sentido Incorreto , Polimorfismo Genético , Anormalidades Múltiplas/sangue , Adulto , Anodontia/sangue , Anodontia/complicações , Anodontia/genética , Criança , Pré-Escolar , China , Fenda Labial/sangue , Fenda Labial/complicações , Fissura Palatina/sangue , Fissura Palatina/complicações , Cistos/sangue , Cistos/complicações , Éxons , Feminino , Humanos , Lactente , Fatores Reguladores de Interferon/sangue , Masculino , Anamnese , Pessoa de Meia-Idade , Linhagem , Fenótipo , TranscriptomaRESUMO
Systemic Lupus Erythematosus (SLE) is a heterogeneous autoimmune disease, which currently lacks specific diagnostic biomarkers. The diversity within the patients obstructs clinical trials but may also reflect differences in underlying pathogenesis. Our objective was to obtain protein profiles to identify potential general biomarkers of SLE and to determine molecular subgroups within SLE for patient stratification. Plasma samples from a cross-sectional study of well-characterized SLE patients (n = 379) and matched population controls (n = 316) were analyzed by antibody suspension bead array targeting 281 proteins. To investigate the differences between SLE and controls, Mann-Whitney U-test with Bonferroni correction, generalized linear modeling and receiver operating characteristics (ROC) analysis were performed. K-means clustering was used to identify molecular SLE subgroups. We identified Interferon regulating factor 5 (IRF5), solute carrier family 22 member 2 (SLC22A2) and S100 calcium binding protein A12 (S100A12) as the three proteins with the largest fold change between SLE patients and controls (SLE/Control = 1.4, 1.4, and 1.2 respectively). The lowest p-values comparing SLE patients and controls were obtained for S100A12, Matrix metalloproteinase-1 (MMP1) and SLC22A2 (padjusted = 3 × 10-9, 3 × 10-6, and 5 × 10-6 respectively). In a set of 15 potential biomarkers differentiating SLE patients and controls, two of the proteins were transcription factors, i.e., IRF5 and SAM pointed domain containing ETS transcription factor (SPDEF). IRF5 was up-regulated while SPDEF was found to be down-regulated in SLE patients. Unsupervised clustering of all investigated proteins identified three molecular subgroups among SLE patients, characterized by (1) high levels of rheumatoid factor-IgM, (2) low IRF5, and (3) high IRF5. IRF5 expressing microparticles were analyzed by flow cytometry in a subset of patients to confirm the presence of IRF5 in plasma and detection of extracellular IRF5 was further confirmed by immunoprecipitation-mass spectrometry (IP-MS). Interestingly IRF5, a known genetic risk factor for SLE, was detected extracellularly and suggested by unsupervised clustering analysis to differentiate between SLE subgroups. Our results imply a set of circulating molecules as markers of possible pathogenic importance in SLE. We believe that these findings could be of relevance for understanding the pathogenesis and diversity of SLE, as well as for selection of patients in clinical trials.
Assuntos
Biomarcadores/sangue , Fatores Reguladores de Interferon/sangue , Lúpus Eritematoso Sistêmico/imunologia , Transportador 2 de Cátion Orgânico/sangue , Proteína S100A12/sangue , Adulto , Análise por Conglomerados , Estudos Transversais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fenótipo , Proteômica , Proteínas Proto-Oncogênicas c-ets/metabolismoRESUMO
BACKGROUND: Tumor-infiltrating lymphocytes influence the prognosis of solid tumors, including ovarian cancer (OC). The immunoregulatory transcription factor (IRF4) is mainly expressed in plasma cells and regulates immunoglobulin class switch recombination as well as plasma cell differentiation. Therefore, we analyzed the impact of IRF4 expression in a consecutive cohort of OC patients. METHODS: IRF4 expression was evaluated by immunostaining. Differences in IRF4 expression among the subgroups of the established clinical-pathological features like age, histological subtype, tumor stage, histological grading, postoperative tumor burden, and completeness of chemotherapy were determined by χ2 test. The impact of IRF4 expression on progression-free survival (PFS) and overall survival (OS) was examined by univariate and multivariate Cox analysis adjusted for established clinical-pathological factors and Kaplan-Meier survival analysis. RESULTS: 114 patients entered this study. IRF4 was expressed in 51.7% of the entire cohort. 72.3% patients with high-grade serous OC showed IRF4 expression compared to 37.3% patients with a non-high-grade serous OC (p < 0.001). Univariate Cox-regression analysis revealed no prognostic impact of IRF4 expression in terms of PFS (p = 0.35) and OS (p = 0.98). Kaplan-Meier plots failed to show any prognostic impact for PFS (p = 0.35) and OS (p = 0.98), too. Established clinical-pathological factors retained their prognostic impact as tumor stage in terms of PFS (< 0.001) and as postoperative residual tumor burden (p = 0.04), tumor stage (< 0.001), histological grade (p = 0.02), and completeness of chemotherapy (p < 0.001) in terms of OS, respectively. CONCLUSION: Immunohistochemically determined IRF4 expression correlated with high-grade serous OC. However, it failed to show any prognostic impact in this cohort of 114 patients.
Assuntos
Biomarcadores Tumorais/análise , Fatores Reguladores de Interferon/sangue , Neoplasias Ovarianas/patologia , Adulto , Idoso , Carcinoma Epitelial do Ovário , Estudos de Coortes , Cistadenocarcinoma Seroso/sangue , Cistadenocarcinoma Seroso/patologia , Intervalo Livre de Doença , Feminino , Humanos , Imuno-Histoquímica , Estimativa de Kaplan-Meier , Pessoa de Meia-Idade , Neoplasias Ovarianas/metabolismo , Neoplasias Ovarianas/mortalidade , Prognóstico , Estudos RetrospectivosRESUMO
BACKGROUND Community-acquired pneumonia (CAP) is a common disease with significant morbidity and mortality. Interferon regulatory factor 5 (IRF5), which induces type I interferons (IFNs) and cytokines such as interleukin (IL)-6, tumor necrosis factor (TNF)-α, IL-10, and interferon gamma-induced protein (IP)10, is a key transcription factor involved in controlling the expression of proinflammatory cytokines and responses to infection. Here, we carefully investigated the role of IRF5 in regulating immune responses to CAP. MATERIAL AND METHODS QRT-PCR was used to detect the mRNA levels of IRF5, IL-6, IL-10, IP10, TNF-α, and IFN-α in the peripheral blood of 71 CAP patients and 31 healthy controls, as well as in the bronchoalveolar lavage cells of 20 patients with CAP and 23 patients with lung cancer (using samples from the unaffected lung). Flow cytometry was performed to detect the protein level of IRF5, and a CBA flex set was used to detect the levels of these cytokines in the volunteers. RESULTS The expression levels of IRF5 and its related cytokines were significantly increased in CAP patients compared with the controls. Additionally, IRF5, IL-6, IL-10, and IP10 levels were found to be related with the severity of CAP. Furthermore, the levels of IRF5 and IFN-a increased significantly in the early phase of pneumonia caused by influenza virus infection. CONCLUSIONS IRF5 and its related inflammatory cytokines are associated with the severity, prognosis, and causative pathogen of CAP patients. This finding may provide new drug targets for the prevention and treatment of severe pneumonia caused by influenza virus.
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Infecções Comunitárias Adquiridas/imunologia , Citocinas/biossíntese , Fatores Reguladores de Interferon/biossíntese , Pneumonia/imunologia , Adulto , Líquido da Lavagem Broncoalveolar , Estudos de Casos e Controles , Quimiocina CXCL10/sangue , Quimiocina CXCL10/genética , Infecções Comunitárias Adquiridas/sangue , Infecções Comunitárias Adquiridas/genética , Infecções Comunitárias Adquiridas/microbiologia , Citocinas/sangue , Citocinas/genética , Feminino , Humanos , Fatores Reguladores de Interferon/sangue , Fatores Reguladores de Interferon/genética , Interferon-alfa/sangue , Interferon-alfa/genética , Interleucina-10/sangue , Interleucina-10/genética , Interleucina-6/sangue , Interleucina-6/genética , Masculino , Pessoa de Meia-Idade , Pneumonia/sangue , Pneumonia/genética , Pneumonia/microbiologia , Prognóstico , RNA Mensageiro/biossíntese , RNA Mensageiro/sangue , RNA Mensageiro/genética , Fator de Necrose Tumoral alfa/sangue , Fator de Necrose Tumoral alfa/genéticaRESUMO
Johne's disease is a chronic wasting disease of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP), resulting in inflammation of intestines and persistent diarrhea. The initial host response against MAP infections is mainly regulated by the Th1 response, which is characterized by the production of IFN-γ. With the progression of disease, MAP can survive in the host through the evasion of the host's immune response by manipulating the host immune response. However, the host response during subclinical phases has not been fully understood. Immune regulatory genes, including Th17-derived cytokines, interferon regulatory factors, and calcium signaling-associated genes, are hypothesized to play an important role during subclinical phases of Johne's disease. Therefore, the present study was conducted to analyze the expression profiles of immune regulatory genes during MAP infection in whole blood. Different expression patterns of genes were identified depending on the infection stages. Downregulation of IL-17A, IL-17F, IL-22, IL-26, HMGB1, and IRF4 and upregulation of PIP5K1C indicate suppression of the Th1 response due to MAP infection and loss of granuloma integrity. In addition, increased expression of IRF5 and IRF7 suggest activation of IFN-α/ß signaling during subclinical stages, which induced indoleamine 2,3-dioxygenase mediated depletion of tryptophan metabolism. Increased expression of CORO1A indicate modulation of calcium signaling, which enhanced the survival of MAP. Taken together, distinct host gene expression induced by MAP infection indicates enhanced survival of MAP during subclinical stages.
Assuntos
Doenças dos Bovinos/patologia , Mycobacterium avium subsp. paratuberculosis/patogenicidade , Paratuberculose/patologia , Transcriptoma , Animais , Bovinos , Doenças dos Bovinos/imunologia , Doenças dos Bovinos/microbiologia , Citocinas/sangue , Citocinas/genética , Citocinas/metabolismo , Regulação para Baixo , Proteína HMGB1/sangue , Proteína HMGB1/genética , Proteína HMGB1/metabolismo , Fatores Reguladores de Interferon/sangue , Fatores Reguladores de Interferon/genética , Fatores Reguladores de Interferon/metabolismo , Proteínas dos Microfilamentos/sangue , Proteínas dos Microfilamentos/genética , Proteínas dos Microfilamentos/metabolismo , Paratuberculose/imunologia , Paratuberculose/microbiologia , Fosfotransferases (Aceptor do Grupo Álcool)/sangue , Fosfotransferases (Aceptor do Grupo Álcool)/genética , Fosfotransferases (Aceptor do Grupo Álcool)/metabolismo , Transdução de Sinais/genética , Células Th1/citologia , Células Th1/imunologia , Células Th1/metabolismo , Regulação para CimaRESUMO
Introduction: Overactivation of the type I interferon (IFN) signature has been observed in several systemic autoimmune conditions, such as Systemic Lupus Erythematosus (SLE) or Rheumatoid Arthritis (RA). Impaired control of Interferon-Responding Genes (IRGs) expression by their regulatory mechanisms, including Interferon Regulatory Factors (IRFs), may underlie these findings and it may explain the heterogeneity observed among these conditions. In the present study we aimed to evaluate the associations between IRF4 gene expression and those of IRGs in SLE and RA patients to gain insight about its links with the IFN signature as well as to explore the potential clinical relevance of these associations. Methods: The gene expression of IRF4 and IRGs (IFI44, IFI44L, IFI6, and MX1) in peripheral blood was analyzed in 75 SLE patients, 98 RA patients, and 28 healthy controls. A group of 13 biological-naïve RA patients was prospectively followed upon TNFα-blockade. The associations among IRF4 and IRGs were evaluated by principal component analyses (PCA), correlations and network analyses. Publicly available datasets were used for replication. Results: A broad activation of IRGs was observed in autoimmune patients, although certain heterogeneity can be distinguished, whereas IRF4 was only upregulated in RA. The differential expression of IRF4 in RA was then confirmed in publicly available gene expression datasets. PCA revealed different associations among IRF4 and IRGs in each condition, which was later confirmed by correlation and network analyses. Cluster analysis identified 3 gene expression signatures on the basis of IRF4 and IRGs expression which were differentially used by SLE and RA patients. Cluster III was associated with markers of disease severity in SLE patients. Cluster II, hallmarked by IRF4 upregulation, was linked to clinical stage and mild disease course in RA. TNFα-blockade led to changes in the association between IRF4 and IRGs, whereas increasing IRF4 expression was associated with a good clinical outcome in RA. Conclusions: The differential expression of IRF4 and IRGs observed in SLE and RA can delineate gene expression signatures associated with clinical features and treatment outcomes. These results support a clinically-relevant phenomenon of shaping of the IFN signature by IRF4 in autoimmune patients.
Assuntos
Antígenos/genética , Artrite Reumatoide/genética , Proteínas do Citoesqueleto/genética , Fatores Reguladores de Interferon/genética , Lúpus Eritematoso Sistêmico/genética , Proteínas Mitocondriais/genética , Proteínas de Resistência a Myxovirus/genética , Transcriptoma , Proteínas Supressoras de Tumor/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígenos/sangue , Artrite Reumatoide/sangue , Distribuição de Qui-Quadrado , Análise por Conglomerados , Proteínas do Citoesqueleto/sangue , Feminino , Seguimentos , Humanos , Fatores Reguladores de Interferon/sangue , Interferon Tipo I/genética , Lúpus Eritematoso Sistêmico/sangue , Masculino , Pessoa de Meia-Idade , Proteínas Mitocondriais/sangue , Proteínas de Resistência a Myxovirus/sangue , Estudos Prospectivos , Estatísticas não Paramétricas , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Proteínas Supressoras de Tumor/sangue , Adulto JovemRESUMO
OBJECTIVE: To investigate the value of cysteine-rich secretory protein LCCL domain-containing 2 (CRISPLD2) in diagnosis and prognosis in patients with sepsis. METHODS: Clinical data of patients admitted to intensive care unit (ICU) of the First Hospital of Hebei Medical University from December 2014 to December 2016 were retrospectively analyzed. According to the severity of sepsis, the patients were divided into three groups: sepsis patients, severe sepsis patients and septic shock patients, and 100 healthy persons were enrolled as control group. Levels of serum CRISPLD2, procalcitonin (PCT) and C-reactive protein (CRP), acute physiology and chronic health evaluation II (APACHE II) score, sequential organ failure assessment (SOFA) score, and 28-day prognosis were recorded. Analysis of the correlation between CRISPLD2 and PCT, CRP, APACHE II score, SOFA score was done. The receiver operating characteristic (ROC) curve was plotted for the CRISPLD2 value for the diagnosis and prognosis in patients with sepsis. RESULTS: A total of 115 patients with sepsis were enrolled in this study, including 52 sepsis, 48 severe sepsis, and 15 septic shock; 29 patients died after 28 days, 28-days mortality rate was 25.2%. There was no significant difference in CRISPLD2 between sepsis and healthy control group (mg/L: 204.1±74.5 vs. 211.3±12.0, P > 0.05); the level of CRISPLD2 in septic shock group was significantly lower than that in sepsis group and severe sepsis group (mg/L: 139.0±55.0 vs. 240.2±89.6, 233.0±8.9, both P < 0.05). The level of PCT, CRP and APACHE II score, SOFA score in sepsis patients were significantly higher than those in healthy control group, and increased with the severity of sepsis. There was no statistically significant difference in CRISPLD2 level between the dead and the survival of sepsis, and the levels of PCT and CRP in death group were significantly higher. The levels of CRISPLD2 were significantly negative correlated with the levels of PCT, CRP, APACHE II score and SOFA score (r values were -0.089, -0.431, -0.115, -0.201, respectively, all P < 0.05). It was shown by ROC curve analysis that the area under ROC curve (AUC) and 95% confidence interval (95%CI) of CRISPLD2, PCT, CRP for diagnosis of sepsis were 0.907 (0.871-0.944), 0.922 (0.886-0.958), 0.916 (0.878-0.954) respectively, all P = 0.000; when the cut-off value of CRISPLD2 > 216.0 mg/L, the sensitivity was 96.7%, and the specificity was 92.6%, which power lied between PCT and CRP. The AUC of CRISPLD2 for prognosis was significantly lower than that of PCT [0.617 (0.507-0.727) vs. 0.786 (0.668-0.903), P < 0.01]; when the cut-off value of CRISPLD2 was 103.5 mg/L, the sensitivity was 100%, and the specificity was 25.6%. CONCLUSIONS: CRISPLD2 is a potential biomarker in sepsis, but cannot predict the prognosis of patients with sepsis.
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Moléculas de Adesão Celular/sangue , Fatores Reguladores de Interferon/sangue , Sepse/diagnóstico , Sepse/terapia , Estudos de Casos e Controles , Humanos , Prognóstico , Estudos Retrospectivos , Sensibilidade e Especificidade , Sepse/sangueRESUMO
Using an immunohistochemistry (IHC) based method, diffuse large B-cell lymphoma (DLBCL) can be classified into germinal center B-cell (GCB) and non-GCB subtypes. However, the prognostic value of Hans algorithm was contradictory in the literature. Using IHC and fluorescence in situ hybridization, we analyzed the antibodies applied in Hans algorithm and other genetic factors in 601 DLBCL patients and prognostic value of Hans algorithm in 306 cases who were treated with chemoimmunotherapy. The results showed that patients with GCB subtype have better overall survival (OS) and progression-free survival (PFS) than non-GCB cases. However, to some extent, double positive (CD10(+)MUM1(+), DP) and triple negative (CD10(-)Bcl6(-)MUM(-), TN) showed different clinical characteristics and prognosis to others that were assigned to the same cell-of-origin group. The DP group showed similar OS (median OS: both not reached, P = 0.3650) and PFS (median PFS: 47.0 vs. 32.7 months, P = 0.0878) with the non-GCB group while the TN group showed similar OS (median OS: both not reached, P = 0.9278) and PFS (median PFS: both not reached, P = 0.9420) with the GCB group. In conclusion, Recognition of specific entities in Hans algorithm could help us to accurately predict outcome of the patients and choose the best clinical management for them.
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Algoritmos , Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Biomarcadores/sangue , Fatores Reguladores de Interferon/sangue , Linfoma Difuso de Grandes Células B , Neprilisina/sangue , Proteínas Proto-Oncogênicas c-bcl-6/sangue , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Intervalo Livre de Doença , Feminino , Humanos , Linfoma Difuso de Grandes Células B/sangue , Linfoma Difuso de Grandes Células B/tratamento farmacológico , Linfoma Difuso de Grandes Células B/mortalidade , Masculino , Pessoa de Meia-Idade , Taxa de SobrevidaRESUMO
INTRODUCTION: Previous studies have shown that cysteine-rich secretory protein containing LCCL domain 2 (CRISPLD2) is a novel lipopolysaccharide (LPS)-binding protein, and the upregulation of CRISPLD2 expression protects mice against LPS-induced lethality. The aim of this study was to examine the expression of CRISPLD2 in patients with sepsis and characterize the association of this protein with procalcitonin. METHODS: The expression of CRISPLD2 was determined in 100 healthy volunteers and 119 septic patients. According to the definition of sepsis, patients were divided into three groups sepsis, severe sepsis, and septic shock. The relationship between CRISPLD2 levels and procalcitonin was also examined and statistically analyzed. RESULTS: The CRISPLD2 levels in healthy individuals were 219.3±69.1 µg/ml. Patients with sepsis exhibited higher CRISPLD2 levels than observed in healthy individuals (pâ=â0.001), but CRISPLD2 expression was not upregulated in patients with septic shock. No significant differences were observed between the levels of CRISPLD2 in surviving and non-surviving spesis patients. CRISPLD2 levels were negatively correlated with procalcitonin levels (râ=â-0.334, p<0.001). CONCLUSIONS: The present study is the first to demonstrate the decreased expression of CRISPLD2 in septic shock and its association with PCT in sepsis. Further studies are needed to clarify the potential association between CRISPLD2 expression and clinical outcomes to determine if it could be used as a novel sepsis biomarker.
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Calcitonina/sangue , Moléculas de Adesão Celular/sangue , Fatores Reguladores de Interferon/sangue , Precursores de Proteínas/sangue , Choque Séptico/sangue , Adulto , Idoso , Peptídeo Relacionado com Gene de Calcitonina , Estudos de Casos e Controles , Moléculas de Adesão Celular/genética , Feminino , Expressão Gênica , Humanos , Fatores Reguladores de Interferon/genética , Masculino , Pessoa de Meia-Idade , Sepse/sangue , Sepse/mortalidade , Choque Séptico/mortalidade , Resultado do TratamentoRESUMO
LPS is an immunostimulatory component of Gram-negative bacteria. Acting on the immune system in a systemic fashion, LPS exposes the body to the hazard of septic shock. In this study we report that cysteine-rich secretory protein LCCL domain containing 2 (CRISPLD2/Crispld2; human and mouse/rat versions, respectively), expressed by multitissues and leukocytes, is a novel LPS-binding protein. As a serum protein, median CRISPLD2 concentrations in health volunteers and umbilical cord blood samples are 607 microg/ml and 290 microg/ml, respectively. Human peripheral blood granulocytes and mononuclear cells including monocytes, NK cells, and T cells spontaneously release CRISPLD2 (range, 0.2-0.9 microg/ml) and enhance CRISPLD2 secretion (range, 1.5-4.2 microg/ml) in response to stimulation of both LPS and humanized anti-human TLR4-IgA Ab in vitro. CRISPLD2 exhibits significant LPS binding affinity similar to that of soluble CD14, prevents LPS binding to target cells, reduces LPS-induced TNF-alpha and IL-6 production, and protects mice against endotoxin shock. In in vivo experiments, serum Crispld2 concentrations increased in response to a nontoxic dose of LPS and correlated negatively with LPS lethality, suggesting that CRISPLD2 serum concentrations not only are indicators of the degree of a body's exposure to LPS but also reflect an individual's LPS sensitivity.
Assuntos
Moléculas de Adesão Celular/imunologia , Fatores Reguladores de Interferon/imunologia , Lipopolissacarídeos/imunologia , Proteínas Recombinantes/imunologia , Choque Séptico/imunologia , Animais , Anticorpos Monoclonais/farmacologia , Moléculas de Adesão Celular/sangue , Moléculas de Adesão Celular/efeitos dos fármacos , Moléculas de Adesão Celular/metabolismo , Feminino , Granulócitos/imunologia , Granulócitos/metabolismo , Humanos , Fatores Imunológicos/farmacologia , Fatores Reguladores de Interferon/sangue , Fatores Reguladores de Interferon/efeitos dos fármacos , Fatores Reguladores de Interferon/metabolismo , Interleucina-6/imunologia , Interleucina-6/metabolismo , Estimativa de Kaplan-Meier , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Lipopolissacarídeos/metabolismo , Lipopolissacarídeos/farmacologia , Camundongos , Camundongos Endogâmicos BALB C , Monócitos/imunologia , Monócitos/metabolismo , Proteínas Recombinantes/farmacologia , Choque Séptico/prevenção & controle , Linfócitos T/imunologia , Linfócitos T/metabolismo , Receptor 4 Toll-Like/imunologia , Receptor 4 Toll-Like/metabolismo , Fator de Necrose Tumoral alfa/imunologia , Fator de Necrose Tumoral alfa/metabolismoRESUMO
OBJECTIVE: The aim was to investigate the level of interferon regulatory factor (IRF) 1, 3, and 7 in peripheral blood cells from patients with primary Sjogren syndrome (pSS) and to determine whether and where IRF1 exists in the parotid glands of pSS. METHODS: Peripheral blood cells and parotid gland biopsy specimens from patients with pSS were studied. The IRF1, IRF3, and IRF7 gene mRNA levels in peripheral blood cells were calculated by using real-time PCR. The IRF1-positive cells in the parotid glands with pSS were observed by using immunohistochemistry and immunofluorescence. Statistical analysis was performed by Student t test. RESULTS: Compared with 24 control samples, the IRF1 mRNA levels in peripheral blood cells of 37 cases with pSS were up-regulated (P < .05), but the IRF3 and IRF7 mRNA levels of pSS were not up-regulated (P > .05). Relative quantitative levels of IRF1 mRNA were 2.17-fold higher in pSS patients than control subjects. The IRF1-positive cells of the pSS group were localized in the epithelial islands, lymphocytes, and ductal epithelial cells of the parotid glands. In all control subjects, the IRF1-positive cells were localized only to the ductal epithelial cells of parotid glands as determined by immunohistochemical staining or immunofluorescence. The scores of IRF1-positive cells of pSS were significantly higher than that of control samples (P < .05). CONCLUSION: These findings indicate that IRF1 mRNA levels are up-regulated in the peripheral blood cells of pSS patients. Also, IRF1-positive cells exist in the epithelial islands, lymphocytes, and ductal epithelial cells of the parotid glands of individuals affected by pSS, but are limited to the ductal epithelial cells of healthy control subjects.
Assuntos
Fatores Reguladores de Interferon/biossíntese , Síndrome de Sjogren/imunologia , Síndrome de Sjogren/metabolismo , Adulto , Idoso , Estudos de Casos e Controles , Células Epiteliais/metabolismo , Feminino , Imunofluorescência , Expressão Gênica , Humanos , Fator Regulador 1 de Interferon/biossíntese , Fator Regulador 1 de Interferon/genética , Fator Regulador 3 de Interferon/biossíntese , Fator Regulador 3 de Interferon/sangue , Fator Regulador 3 de Interferon/genética , Fator Regulador 7 de Interferon/biossíntese , Fator Regulador 7 de Interferon/sangue , Fator Regulador 7 de Interferon/genética , Fatores Reguladores de Interferon/sangue , Fatores Reguladores de Interferon/genética , Linfócitos/metabolismo , Masculino , Pessoa de Meia-Idade , Glândula Parótida/metabolismo , RNA Mensageiro/sangue , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Síndrome de Sjogren/sangue , Síndrome de Sjogren/genética , Regulação para Cima , Adulto JovemRESUMO
In ruminants, pregnancy results in up-regulation of a large number of IFN-stimulated genes (ISG) in the uterus. Recently, one of these genes was also shown to increase in peripheral blood leukocytes (PBL) during early pregnancy in sheep. Our working hypothesis is that conceptus signaling activates maternal gene expression in PBL in dairy cattle. The objectives of this study were to characterize ISG expression in PBL from pregnant (n = 20) and bred, nonpregnant (n = 30) dairy cows. Steady-state levels of mRNA for Mx1, Mx2, beta2-microglobulin, ISG-15, IFN regulatory factor-1, and IFN regulatory factor-2 were quantified. Holstein cows were synchronized to estrus and artificially inseminated (d 0). Blood samples were collected (coccygeal venipuncture) on d 0 and 16, 18, and 20 d after insemination for progesterone analysis and PBL isolation. Pregnancy was confirmed by transrectal ultrasonography at approximately 40 d after breeding. A status x day interaction was detected for Mx1, Mx2, and ISG-15 gene expression. When analyzed within day, levels of mRNA for ISG-15 and Mx1 were greater in pregnant compared with bred, nonpregnant cows on d 18 and 20, respectively. Expression of the Mx2 gene increased in the pregnant group compared with bred, nonpregnant cows on d 16, 18, and 20 after insemination. beta2-Microglobulin, IFN regulatory factor-1, and IFN regulatory factor-2 were not different between groups. The results clearly indicated that components of the innate immune response are activated in PBL during the period of pregnancy recognition and early embryo signaling. The physiological implications of these changes on maternal immune function are as yet unknown; however, they do provide a unique opportunity to identify bred, nonpregnant, cows 18 d after insemination in dairy cattle.