RESUMEN
Objective: To evaluate the cost-effectiveness of seasonal influenza vaccination, compared to no vaccination, for the elderly aged ≥60 years old in China. Methods: A static life-time Markov model is conducted to simulate the Chinese elderly population aged ≥60 years old. Taking the health care system perspective, one-year analytic cycle length is used for each influenza season. The model was assumed to be repeated until the individual reaches 100 years old. Three interventions were evaluated, including no vaccination, annual trivalent influenza vaccination, and annual quadrivalent influenza vaccination. Using the threshold of 3 times GDP per capita per Quality-adjusted life year (QALY) (193 932/QALY), the incremental cost-effectiveness ratio (ICER) was calculated to compare the cost-effectiveness of every two interventions.Model inputs like data for costs and utilities were from studies on Chinese population if they were available. QALY was used to measure health utility. One-way sensitivity analysis and probabilistic sensitivity analysis were adopted to quantify the level of confidence of the model output. Results: The total influenza associated costs of no vaccination would be 603 CNY per person, while the total costs of annual trivalent vaccination would be 1 027 CNY. Using trivalent vaccine would result in 0.007 QALY gained per person compared to no vaccination, with an increased cost of 424 CNY per person. The ICER of trivalent vaccination over no vaccination for all the elderly population in China would be 64 026 CNY per QALY gained, which was less than the threshold of 3 times GDP per capita. The total costs of annual quadrivalent vaccination would be 1 988 CNY. Using quadrivalent vaccine would result in 0.008 additional QALY gained per person compared to no vaccination, with an increased cost of 1 385 CNY per person. The ICER of quadrivalent vaccination over no vaccination would be 174 081 CNY per QALY gained, which was less than the threshold of 3 times GDP per capita. Conclusion: Vaccinating elderly population would improve health utilities at higher health care costs for the elderly. Using the threshold of 3 times GDP per capita per QALY (193 932/QALY), both trivalent and quadrivalent vaccination would be cost-effective compared to no vaccination in elderly Chinese population.
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Vacunas contra la Influenza/economía , Gripe Humana , Anciano , Anciano de 80 o más Años , China , Análisis Costo-Beneficio , Humanos , Persona de Mediana Edad , Años de Vida Ajustados por Calidad de Vida , Estaciones del AñoRESUMEN
OBJECTIVES: The oldest old population has become the fastest growing segment with excess need of care and social support, it is crucial to improve the health-related quality of life (HRQoL) of these populations. This study seeks to evaluate the health status and to investigate modifiable factors associated with health-related quality of life for oldest old adults in China. DESIGN: A cross-sectional population-based study. SETTING: Hainan Province in the south of China. PARTICIPANTS: 1,278 adults aged 80 years or older. METHODS: HRQoL was assessed by three-level EuroQol-5D scale (EQ-5D-3L) and a visual analogue scale (VAS). Demographic and health-related variables were analysed by estimating mean values and standard deviations for continuous variables, percentages and standard deviations for categorical variables. Tobit regressions, ordinary least Squared (OLS) regressions and ordered probit regressions were adopted to determine the associated factors for overall HRQoL and for each health dimension. RESULTS: Anxiety/depression was the least reported problem while mobility was the most frequently reported with problem. Female respondents had lower EQ-5D score (0.76 vs. 0.86) and VAS score (66.55 vs. 69.84) than male respondents. Better health-related quality of life was significantly associated with higher BMI, no drinking habit, more leisure activities, living with family members, good sleeping quality, closer social and family connections, fewer numbers of drugs consumed per day, without having hearing or visual impairment, and fewer chronic conditions, after controlling for potential confounders. CONCLUSION: Findings from this study suggested that quality of life was not only associated with age-related diseases, but also correlated with a range of health-related lifestyles, and factors indicating social and family support.
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Calidad de Vida/psicología , Anciano de 80 o más Años , China/epidemiología , Estudios Transversales , Femenino , Humanos , MasculinoRESUMEN
We cloned a second human beta-defensin gene, HBD-2, and determined its gene structure and expression in inflamed tissue sections. The entire gene spanned about 2 kb with two small exons and one intron. Radiation hybrid studies confirmed the location on chromosome 8p, were consistent with the order HNP-1, HBD-1 and HBD-2, and located HBD-2 as the most centromeric of the genes. By three-color fluorescence in situ hybridization on both free chromatin fiber mapping and interphase mapping, HBD-1, HBD-2 and HNP-1 were mapped to chromosome 8p23. HBD-1 was within 40-100kb of HNP-1, while HBD-2 was about 500-600 kb from HBD-1, with the most likely order HNP-1, HBD-1, HBD-2. The expression of HBD-2 was locally regulated by inflammation. HBD-2 mRNA was markedly increased in the epidermis surrounding inflamed regions, but not detectable in adjacent non-inflamed areas, a distribution that was confirmed at the peptide level by immunostaining with HBD-2 antibody. The HBD-2 gene is the first member of the human defensin family that is locally inducible by inflammation.
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Inflamación/genética , Proteínas/genética , alfa-Defensinas , beta-Defensinas , Secuencia de Bases , Clonación Molecular , ADN Complementario/genética , Defensinas , Femenino , Expresión Génica , Humanos , Inmunohistoquímica , Hibridación Fluorescente in Situ , Inflamación/metabolismo , Inflamación/patología , Masculino , Datos de Secuencia Molecular , Sondas de Oligonucleótidos/genética , Mapeo RestrictivoRESUMEN
Epithelial beta-defensins are broad-spectrum cationic antimicrobial peptides that also act as chemokines for adaptive immune cells. In the human genome, all known defensin genes cluster to a <1 Mb region of chromosome 8p22-p23. To identify new defensin genes, the DNA sequence from a contig of large-insert genomic clones from the region containing human beta-defensin-2 (HBD-2) was analyzed for the presence of defensin genes. This sequence survey identified a novel beta-defensin, termed HBD-3. The HBD-3 gene contains two exons, is located 13 kb upstream from the HBD-2 gene, and it is transcribed in the same direction. A partial HBD-3 cDNA clone was amplified from cDNA derived from IL-1beta induced fetal lung tissue. The cDNA sequence encodes for a 67 amino acid peptide that is approximately 43% identical to HBD-2 and shares the beta-defensin six cysteine motif. By PCR analysis of two commercial cDNA panels, HBD-3 expression was detected in adult heart, skeletal muscle, placenta and in fetal thymus. From RT-PCR experiments, HBD-3 expression was observed in skin, esophagus, gingival keratinocytes, placenta and trachea. Furthermore, in fetal lung explants and gingival keratinocytes, HBD-3 mRNA expression was induced by IL-1beta. Additional sequence analysis identified the HE2 (human epididymis secretory protein) gene 17 kb upstream from the HBD-3 gene. One splice variant of this gene (HE2beta1) encodes a beta-defensin consensus cysteine motif, suggesting it represents a defensin gene product. HE2beta1 mRNA expression was detected in gingival keratinocytes and bronchial epithelia using RT-PCR analysis. The discovery of these novel beta-defensin genes may allow further understanding of the role of defensins in host immunity at mucosal surfaces.
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Proteínas Portadoras , Genómica , Proteínas Recombinantes , beta-Defensinas/genética , Adulto , Secuencia de Aminoácidos , Antígenos de Superficie/genética , Secuencia de Bases , Mapeo Contig , ADN/química , ADN/genética , Exones , Femenino , Feto/metabolismo , Expresión Génica , Regulación del Desarrollo de la Expresión Génica , Genes/genética , Biblioteca Genómica , Glicopéptidos/genética , Glicoproteínas , Humanos , Intrones , Masculino , Datos de Secuencia Molecular , ARN Mensajero/genética , ARN Mensajero/metabolismo , Alineación de Secuencia , Análisis de Secuencia de ADN , Homología de Secuencia de Aminoácido , Distribución Tisular , Proteínas de Transporte VesicularRESUMEN
Twenty-six patients with non-functioning pancreatic endocrine tumor (NFPET) were operated on during a 22-year period (1968-1990) at PUMC Hospital, Beijing. Of these, 19 were female and 7 were male with a mean age of 33 years. All these tumors, including 12 malignant and 14 benign, were solitary, and most of them were well-encapsulated. Immunohistochemical staining showed 23 (88.5%) containing 1-4 kinds of peptide hormone and 18 (69.2%) being multihormonal. In 7 of the 9 tumors subjected to electron microscopic study, various amounts of neurosecretory granules were found. Tumors of this series were clinically silent, but they contained some immunoreactive peptides, although the amount of the peptides varied from tumor to tumor.
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Adenoma de Células de los Islotes Pancreáticos/química , Adenoma de Células de los Islotes Pancreáticos/patología , Adenoma de Células de los Islotes Pancreáticos/ultraestructura , Adolescente , Adulto , Anciano , Niño , Femenino , Humanos , Técnicas para Inmunoenzimas , Masculino , Microscopía Electrónica , Persona de Mediana Edad , Pronóstico , Estudios RetrospectivosRESUMEN
The effects of a partially purified antihypertensive factor (AHF) from erythrocytes of spontaneously hypertensive rats (SHR) on the blood pressure (BP) and Ca2+ influx of vascular smooth muscle (VSM) in rats were studied. The results indicated that AHF could produce a marked prolonged depressor effect and significantly inhibit the Ca2+ influx dose-dependently on both SHR and renal hypertensive rat (RHR) either in acute or in chronic experiments, but not on normotensive rats. It suggested that the inhibition of Ca2+ influx might be one of the important mechanisms for AHF as an endogenous depressor substance.
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Factores Biológicos/farmacología , Presión Sanguínea/efectos de los fármacos , Calcio/metabolismo , Músculo Liso Vascular/metabolismo , Animales , Aorta , Transporte Biológico Activo , Eritrocitos/química , Masculino , Arterias Mesentéricas , Ratas , Ratas Endogámicas SHR , Ratas Endogámicas , Ratas Endogámicas WKYRESUMEN
Cystic fibrosis mice have been generated by gene targeting but show little lung disease without repeated exposure to bacteria. We asked if murine mucosal defenses and airway surface liquid (ASL) Cl(-) were altered by the DeltaF508 cystic fibrosis transmembrane conductance regulator mutation. Naive DeltaF508 -/- and +/- mice showed no pulmonary inflammation and after inhaled Pseudomonas aeruginosa had similar inflammatory responses and bacterial clearance rates. We therefore investigated components of the innate immune system. Bronchoalveolar lavage fluid from mice killed Escherichia coli, and the microbicidal activity was inhibited by NaCl. Because beta-defensins are salt-sensitive epithelial products, we looked for pulmonary beta-defensin expression. A mouse homolog of human beta-defensin-1 (termed "MBD-1") was identified; the mRNA was expressed in the lung. Using a radiotracer technique, ASL volume and Cl(-) concentration ([Cl(-)]) were measured in cultured tracheal epithelia from normal and DeltaF508 -/- mice. The estimated ASL volume was similar for both groups. There were no differences in ASL [Cl(-)] in DeltaF508 -/- and normal mice (13.8 +/- 2.6 vs. 17.8 +/- 5.6 meq/l). Because ASL [Cl(-)] is low in normal and mutant mice, salt-sensitive antimicrobial factors, including MBD-1, may be normally active.
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Infecciones Bacterianas/prevención & control , Regulador de Conductancia de Transmembrana de Fibrosis Quística/genética , Enfermedades Pulmonares/prevención & control , Mutación/fisiología , beta-Defensinas , Administración por Inhalación , Secuencia de Aminoácidos/genética , Animales , Antibacterianos/análisis , Antibacterianos/antagonistas & inhibidores , Fenómenos Fisiológicos Bacterianos , Secuencia de Bases/genética , Líquidos Corporales/fisiología , Bronquios/metabolismo , Líquido del Lavado Bronquioalveolar/química , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos , Datos de Secuencia Molecular , Proteínas/genética , Proteínas/metabolismo , Alveolos Pulmonares/metabolismo , Cloruro de Sodio/farmacologíaRESUMEN
This study evaluated the expression and regulation of beta-defensins DEFB-104 and the recently identified DEFB-105-14 in gingival keratinocytes. Keratinocytes from healthy subjects were exposed to cytokines, Escherichia coli lipopolysaccharide or Candida species. Total RNA was extracted and defensin expression analyzed by reverse transcription-polymerase chain reaction. Three patterns of expression were seen: no expression, constitutive expression and inducible expression. Constitutive mRNA expression was evident for DEFB-104, 107, 109, 111, and 112. DEFB-108 and 114 were induced by interleukin (IL)-1beta and Candida species. For DEFB-108 expression, synergism was observed when IL-1beta was combined with tumor necrosis factor-alpha or interferon-gamma. Downregulation of DEFB-109 occurred following treatment with Candida albicans. These findings suggest a role for multiple beta-defensins in response to oral infection. Further investigation is needed to better understand their function, both in terms of antimicrobial activities and contributions to innate and acquired immunity.
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Antiinfecciosos/análisis , Encía/metabolismo , Queratinocitos/metabolismo , beta-Defensinas/análisis , Candida/fisiología , Candida albicans/fisiología , Citocinas/farmacología , Regulación hacia Abajo , Escherichia coli , Expresión Génica/genética , Regulación de la Expresión Génica/genética , Humanos , Interferón gamma/farmacología , Interleucina-1/farmacología , Lipopolisacáridos/farmacología , ARN Mensajero/análisis , Factor de Necrosis Tumoral alfa/farmacología , beta-Defensinas/genéticaRESUMEN
Human beta-defensin-1 (HBD-1) was detected in breast milk in concentrations of approximately 1 to 10 microg/mL. Breast tissue during lactation showed HBD-1 expression in mammary gland epithelia and within luminal secretions. The peptide demonstrated antimicrobial activity against Escherichia coli. HBD-1 may augment neonatal host defenses through antimicrobial effects or prime the adaptive immune system at mucosal surfaces.
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Mama/citología , Leche Humana/química , beta-Defensinas/análisis , beta-Defensinas/inmunología , Epitelio/química , Humanos , Inmunidad Materno-Adquirida/inmunología , Inmunidad Mucosa/inmunología , Lactancia/fisiología , Pruebas de Sensibilidad MicrobianaRESUMEN
SMAP29, an ovine cathelicidin, was systematically altered to create a family of 23 related peptides for MIC and minimum bactericidal concentration determinations. SMAP28, SMAP29, and a derivative of SMAP29 called ovispirin were all antimicrobial. However, many congeners of SMAP29 and ovispirin were not as active as the parent molecules. With immunoelectron microscopy, SMAP29 was seen on membranes and within the cytoplasm of Pseudomonas aeruginosa PAO1.
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Antibacterianos/farmacología , Bacterias/efectos de los fármacos , Bacterias/ultraestructura , Proteínas Sanguíneas/farmacología , Enfermedades de las Ovejas/microbiología , Ovinos/microbiología , Secuencia de Aminoácidos , Animales , Catelicidinas , Pruebas de Sensibilidad Microbiana , Microscopía Inmunoelectrónica , Datos de Secuencia MolecularRESUMEN
beta-Defensins are endogenous cationic peptides with broad-spectrum antimicrobial activity that are thought to play a role in the innate immune response. Two human beta-defensins, beta-defensin-1 (HBD-1) and beta-defensin-2 (HBD-2), have been identified. These peptides have recently been characterized in several human tissues. The presence of these peptides in the paranasal sinuses has not been investigated. We examined maxillaary sinus secretions from six patients with sinusitis and 10 patients without signs, symptoms, or radiologic evidence of sinus disease for the presence of beta-defensins. Cationic peptides were extracted from antral lavage specimens and examined for the presence of HBD-1 and HBD-2 by Western blot. Normal maxillary sinus epithelium was obtained from two patients and analyzed by RT-PCR for the presence of HBD-1 and HBD-2 mRNA. Tissue immunostaining for the two peptides was also used. Western blot analysis identified HBD-1 in two of 10 patients in the control group and in three of six patients in the sinusitis group. HBD-2 was identified in one of 10 patients in the control group and in four of six patients in the sinusitis group. RT-PCR revealed HBD-1 mRNA in one of two normal controls tested. Immunostaining localized HBD-1 and HBD-2 to the epithelial cell cytoplasm. This is the first demonstration of HBD-1 and HBD-2 production in the paranasal sinuses. In the present study, HBD-1 and HBD-2 were detected more frequently in the maxillary sinus fluid of patients with inflamed sinuses than in normal controls.
Asunto(s)
Seno Maxilar/química , beta-Defensinas/análisis , Adulto , Anciano , Femenino , Humanos , Inmunohistoquímica , Masculino , Sinusitis Maxilar/metabolismo , Persona de Mediana Edad , Membrana Mucosa/químicaRESUMEN
beta-Defensins are cationic peptides with broad-spectrum antimicrobial activity that are produced by epithelia at mucosal surfaces. Two human beta-defensins, HBD-1 and HBD-2, were discovered in 1995 and 1997, respectively. However, little is known about the expression of HBD-1 or HBD-2 in tissues of the oral cavity and whether these proteins are secreted. In this study, we characterized the expression of HBD-1 and HBD-2 mRNAs within the major salivary glands, tongue, gingiva, and buccal mucosa and detected beta-defensin peptides in salivary secretions. Defensin mRNA expression was quantitated by RNase protection assays. HBD-1 mRNA expression was detected in the gingiva, parotid gland, buccal mucosa, and tongue. Expression of HBD-2 mRNA was detected only in the gingival mucosa and was most abundant in tissues with associated inflammation. To test whether beta-defensin expression was inducible, gingival keratinocyte cell cultures were treated with interleukin-1beta (IL-1beta) or bacterial lipopolysaccharide (LPS) for 24 h. HBD-2 expression increased approximately 16-fold with IL-1beta treatment and approximately 5-fold in the presence of LPS. Western immunoblotting, liquid chromatography, and mass spectrometry were used to identify the HBD-1 and HBD-2 peptides in human saliva. Human beta-defensins are expressed in oral tissues, and the proteins are secreted in saliva; HBD-1 expression was constitutive, while HBD-2 expression was induced by IL-1beta and LPS. Human beta-defensins may play an important role in the innate defenses against oral microorganisms.
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Antiinfecciosos/metabolismo , Mucosa Bucal/metabolismo , Biosíntesis de Proteínas , Glándulas Salivales/metabolismo , beta-Defensinas , Células 3T3 , Animales , Células Cultivadas , Fraccionamiento Químico , Cromatografía Líquida de Alta Presión , Cromatografía Liquida , Defensinas , Electroforesis Capilar , Regulación de la Expresión Génica , Encía/citología , Humanos , Queratinocitos/citología , Espectrometría de Masas , Ratones , Proteínas/genéticaRESUMEN
beta-Defensins are broad spectrum antimicrobial peptides expressed at epithelial surfaces. Two human beta-defensins, HBD-1 and HBD-2, have been identified. In the lung, HBD-2 is an inducible product of airway epithelia and may play a role in innate mucosal defenses. We recently characterized rat homologs (RBD-1, RBD-2) of the human genes and used these sequences to identify novel mouse genes. Mouse beta-defensin-4 (MBD-4) was amplified from lung cDNA using polymerase chain reaction primers designed from conserved sequences of RBD-2 and HBD-2. A full-length cDNA was cloned which encodes a putative peptide with the sequence MRIHYLLFTFLLVLLSPLAAFTQIINNPITCMTNGAICWGPCPTAFRQIGNCGHFKVRCCKIR. The peptide shares approximately 40% identity with HBD-2. MBD-4 mRNA was expressed in the esophagus, tongue, and trachea but not in any of 20 other tissues surveyed. Cloning of the genomic sequence of MBD-4 revealed two nearly (>99%) identical sequences encoding MBD-4 and the presence of numerous additional highly similar genomic sequences. Radiation hybrid mapping localized this gene to a region of chromosome 8 near several other defensins, MBD-2, MBD-3, and alpha-defensins (cryptdins)-3 and -17, consistent with a gene cluster. Our genomic cloning and mapping data suggest that there is a large beta-defensin gene family in mice. Identification of murine beta-defensins provides an opportunity to understand further the role of these peptides in host defense through animal model studies and the generation of beta-defensin-deficient animals by gene targeting.
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Defensinas/genética , Esófago/metabolismo , Lengua/metabolismo , Tráquea/metabolismo , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Southern Blotting , Bovinos , Mapeo Cromosómico , Cromosomas Humanos Par 8 , Clonación Molecular , ADN Complementario/química , Defensinas/química , Femenino , Regulación de la Expresión Génica , Humanos , Ratones , Ratones Endogámicos C57BL , Datos de Secuencia Molecular , RatasRESUMEN
Endogenous peptide antibiotics are under investigation as inhaled therapeutic agents for cystic fibrosis (CF) lung disease. The bactericidal activities of five cathelicidin peptides (LL37 [human], CAP18 [rabbit], mCRAMP [mouse], rCRAMP [rat], and SMAP29 [sheep]), three novel alpha-helical peptides derived from SMAP29 and termed ovispirins (OV-1, OV-2, and OV-3), and two derivatives of CAP18 were tested by broth microdilution assays. Their MICs were determined for multiply antibiotic-resistant Pseudomonas aeruginosa (n = 24), Burkholderia cepacia (n = 5), Achromobacter xylosoxidans (n = 5), and Stenotrophomonas maltophilia (n = 5) strains isolated from CF patients. SMAP29 was most active and inhibited mucoid and nonmucoid P. aeruginosa strains (MIC, 0.06 to 8 microg/ml). OV-1, OV-2, and OV-3 were nearly as active (MIC, 0.03 to 16 microg/ml), but CAP18 (MIC, 1.0 to 32 microg/ml), CAP18-18 (MIC, 1.0 to >32 microg/ml), and CAP18-22 (MIC, 0.5 to 32 microg/ml) had variable activities. LL37, mCRAMP, and rCRAMP were least active against the clinical isolates studied (MIC, 1.0 to >32 microg/ml). Peptides had modest activities against S. maltophilia and A. xylosoxidans (MIC range, 1.0 to > 32 microg/ml), but none inhibited B. cepacia. However, CF sputum inhibited the activity of SMAP29 substantially. The effects of peptides on bacterial cell membranes and eukaryotic cells were examined by scanning electron microscopy and by measuring transepithelial cell resistance, respectively. SMAP29 caused the appearance of bacterial membrane blebs within 1 min, killed P. aeruginosa within 1 h, and caused a dose-dependent, reversible decrease in transepithelial resistance within 5 h. The tested cathelicidin-derived peptides represent a novel class of antimicrobial agents and warrant further development as prophylactic or therapeutic agents for CF lung disease.
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Péptidos Catiónicos Antimicrobianos/farmacología , Fibrosis Quística/microbiología , Resistencia a Múltiples Medicamentos/fisiología , Alcaligenes/efectos de los fármacos , Secuencia de Aminoácidos , Proteínas Sanguíneas/farmacología , Burkholderia cepacia/efectos de los fármacos , Catelicidinas , Sinergismo Farmacológico , Humanos , Pruebas de Sensibilidad Microbiana , Datos de Secuencia Molecular , Péptidos/farmacología , Pseudomonas aeruginosa/efectos de los fármacos , Homología de Secuencia de Aminoácido , Esputo/química , Stenotrophomonas maltophilia/efectos de los fármacos , Factores de TiempoRESUMEN
The presence of the complement-derived anaphylatoxin peptides, C3a and C5a, in the lung can induce respiratory distress characterized by contraction of the smooth muscle walls in bronchioles and pulmonary arteries and aggregation of platelets and leukocytes in pulmonary vessels. C3a and C5a mediate these effects by binding to their specific receptors, C3aR and C5aR, respectively. The cells that express these receptors in the lung have not been thoroughly investigated, nor has their expression been examined during inflammation. Accordingly, C3aR and C5aR expression in normal human and murine lung was determined in this study by immunohistochemistry and in situ hybridization. In addition, the expression of these receptors was delineated in mice subjected to LPS- and OVA-induced models of inflammation. Under noninflamed conditions, C3aR and C5aR protein and mRNA were expressed by bronchial epithelial and smooth muscle cells of both human and mouse lung. C3aR expression increased significantly on both bronchial epithelial and smooth muscle cells in mice treated with LPS; however, in the OVA-challenged animals only the bronchial smooth muscle cells showed increased C3aR expression. C5aR expression also increased significantly on bronchial epithelial cells in mice treated with LPS, but was not elevated in either cell type in the OVA-challenged mice. These results demonstrate the expression of C3aR and C5aR by cells endogenous to the lung, and, given the participation of bronchial epithelial and smooth muscle cells in the pathology of diseases such as sepsis and asthma, the data suggest a role for these receptors during lung inflammation.
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Antígenos CD/biosíntesis , Asma/inmunología , Bronquios/metabolismo , Complemento C3a/metabolismo , Complemento C5a/metabolismo , Endotoxemia/inmunología , Proteínas de la Membrana , Músculo Liso/metabolismo , Receptores de Complemento/biosíntesis , Aerosoles , Secuencia de Aminoácidos , Animales , Antígenos CD/genética , Asma/metabolismo , Asma/patología , Bronquios/irrigación sanguínea , Bronquios/inmunología , Bronquios/patología , Células Cultivadas , Modelos Animales de Enfermedad , Endotoxemia/metabolismo , Endotoxemia/patología , Humanos , Inyecciones Intraperitoneales , Lipopolisacáridos/administración & dosificación , Pulmón/citología , Pulmón/inmunología , Pulmón/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Datos de Secuencia Molecular , Músculo Liso/inmunología , Músculo Liso/patología , Músculo Liso Vascular/inmunología , Músculo Liso Vascular/metabolismo , Músculo Liso Vascular/patología , Ovalbúmina/administración & dosificación , Ovalbúmina/inmunología , Receptor de Anafilatoxina C5a , Receptores de Complemento/genética , Mucosa Respiratoria/irrigación sanguínea , Mucosa Respiratoria/inmunología , Mucosa Respiratoria/metabolismo , Mucosa Respiratoria/patologíaRESUMEN
Human beta-defensin-2 (HBD-2) is a member of the defensin family of antimicrobial peptides. HBD-2 was first isolated from inflamed skin where it is posited to participate in the killing of invasive bacteria and in the recruitment of cells of the adaptive immune response. Static light scattering and two-dimensional proton nuclear magnetic resonance spectroscopy have been used to assess the physical state and structure of HBD-2 in solution. At concentrations of < or = 2.4 mM, HBD-2 is monomeric. The structure is amphiphilic with a nonuniform surface distribution of positive charge and contains several key structural elements, including a triple-stranded, antiparallel beta-sheet with strands 2 and 3 in a beta-hairpin conformation. A beta-bulge in the second strand occurs at Gly28, a position conserved in the entire defensin family. In solution, HBD-2 exhibits an alpha-helical segment near the N-terminus that has not been previously ascribed to solution structures of alpha-defensins or to the beta-defensin BNBD-12. This novel structural element may be a factor contributing to the specific microbicidal or chemokine-like properties of HBD-2.
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Fragmentos de Péptidos/química , beta-Defensinas/química , Secuencia de Aminoácidos , Cristalografía por Rayos X , Humanos , Enlace de Hidrógeno , Luz , Modelos Moleculares , Datos de Secuencia Molecular , Resonancia Magnética Nuclear Biomolecular/métodos , Conformación Proteica , Estructura Secundaria de Proteína , Dispersión de Radiación , SolucionesRESUMEN
The severity of periodontal disease is dependent on a combination of host, microbial agent and environmental factors. One strong correlate related to periodontal disease pathogenesis is the immune status of the host. Here we show that human neutrophil peptide (HNP) defensins or human beta-defensins (HBD), co-administered intranasally with the antigen ovalbumin (OVA), induce unique immune responses that if used with microbial antigens may have the potential to hinder the pathogenesis of periodontal disease. C57BL/6 mice were immunized intranasally with phosphate buffered saline (PBS) containing 1 micro g HNP-1, HNP-2, HBD1 or HBD2 with and without 50 microg OVA. At 21 days, isotypes and subclasses of OVA-specific antibodies were determined in saliva, serum, nasal wash, bronchoalveolar lavage fluid, and fecal extracts. OVA-stimulated splenic lymphoid cell cultures from immunized mice were assessed for interferon (IFN)-gamma, Interleukin (IL)-4 and IL-10. In comparison with mice immunized with only OVA, HNP-1 and HBD2 induced significantly higher (P < 0.05) OVA-specific serum IgG, lower, but not significant, serum IgM and significantly lower (P < 0.05) IFN-gamma. In contrast, HNP-2 induced low OVA-specific serum IgG and higher, but not significant, serum IgM. HBD1 induced significantly higher (P < 0.05) OVA-specific serum IgG, higher, but not significant, serum IgM, and significantly higher (P < 0.05) IL-10. The elevated serum IgG subclasses contained IgG1 and IgG2b.
Asunto(s)
Adyuvantes Inmunológicos/farmacología , Defensinas/inmunología , Enfermedades Periodontales/prevención & control , Análisis de Varianza , Animales , Anticuerpos/análisis , Anticuerpos/sangre , Líquido del Lavado Bronquioalveolar/inmunología , Defensinas/farmacología , Heces/química , Femenino , Humanos , Inmunidad Activa/efectos de los fármacos , Inmunoglobulina G/análisis , Inmunoglobulina G/sangre , Inmunoglobulina M/análisis , Inmunoglobulina M/sangre , Interferón gamma/análisis , Interleucinas/análisis , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos , Ovalbúmina/inmunología , Enfermedades Periodontales/inmunología , Saliva/inmunología , alfa-Defensinas/inmunología , beta-Defensinas/inmunologíaRESUMEN
Human beta-defensins (HBDs) are antimicrobial peptides that may play a role in mucosal defense. Diminished activity of these peptides has been implicated in the pathogenesis of cystic fibrosis (CF) lung disease. We show that HBD-1 and HBD-2 mRNAs are expressed in excised surface and submucosal gland epithelia from non-CF and CF patients. The pro-inflammatory cytokine interleukin-1beta stimulated the expression of HBD-2 but not HBD-1 mRNA and peptide in primary cultures of airway epithelia. HBD-1 was found in bronchoalveolar lavage (BAL) fluid from normal volunteers, CF patients, and patients with inflammatory lung diseases, whereas HBD-2 was detected in BAL fluid from patients with CF or inflammatory lung diseases, but not in normal volunteers. Both HBD-1 and HBD-2 were found in BAL fluid in concentrations of several ng/ml, and both recombinant peptides showed salt-sensitive bactericidal activity. These data suggest that in the lung HBD-2 expression is induced by inflammation, whereas HBD-1 may serve as a defense in the absence of inflammation.
Asunto(s)
Células Epiteliales/metabolismo , Biosíntesis de Proteínas , ARN Mensajero/biosíntesis , beta-Defensinas , Células Cultivadas , Defensinas , Humanos , Hibridación in Situ , Interleucina-1/farmacología , Sistema Respiratorio/metabolismoRESUMEN
Since a negative calcium balance is present in spontaneously hypertensive rats, we searched for the gene(s) involved in this dysregulation. A cDNA library was constructed from the spontaneously hypertensive rat parathyroid gland, which is a key regulator of serum-ionized calcium. From seven overlapping DNA fragments, a 1100-base pair novel cDNA containing an open reading frame of 224 codons was reconstituted. This novel gene, named HCaRG (hypertension-related, calcium-regulated gene), was negatively regulated by extracellular calcium concentration, and its basal mRNA levels were higher in hypertensive animals. The deduced protein showed no transmembrane domain, 67% alpha-helix content, a mutated calcium-binding site (EF-hand motif), four putative "leucine zipper" motifs, and a nuclear receptor-binding domain. At the subcellular level, HCaRG had a nuclear localization. We cloned the human homolog of this gene. Sequence comparison revealed 80% homology between rats and humans at the nucleotide and amino acid sequences. Tissue distribution showed a preponderance in the heart, stomach, jejunum, kidney (tubular fraction), liver, and adrenal gland (mainly in the medulla). HCaRG mRNA was significantly more expressed in adult than in fetal organs, and its levels were decreased in tumors and cancerous cell lines. We observed that after 60-min ischemia followed by reperfusion, HCaRG mRNA declined rapidly in contrast with an increase in c-myc mRNA. Its levels then rose steadily to exceed base line at 48 h of reperfusion. HEK293 cells stably transfected with HCaRG exhibited much lower proliferation, as shown by cell count and [(3)H]thymidine incorporation. Taken together, our results suggest that HCaRG is a nuclear protein potentially involved in the control of cell proliferation.
Asunto(s)
Calcio/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Proteínas Adaptadoras Transductoras de Señales , Glándulas Suprarrenales/metabolismo , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Proteínas de Ciclo Celular , División Celular , Línea Celular , Clonación Molecular , Motivos EF Hand , Perfilación de la Expresión Génica , Humanos , Hipertensión/metabolismo , Hibridación in Situ , Riñón/metabolismo , Leucina Zippers , Microscopía Inmunoelectrónica , Datos de Secuencia Molecular , Proteínas Nucleares/química , Glándulas Paratiroides/efectos de los fármacos , Glándulas Paratiroides/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas , Ratas Endogámicas SHR , Daño por Reperfusión , Homología de Secuencia de AminoácidoRESUMEN
beta-Defensins are cationic peptides with broad-spectrum antimicrobial activity that may play a role in mucosal defenses of several organs. They have been isolated in several species, and in humans, two beta-defensins have been identified. Here, we report the identification of two genes encoding beta-defensin homologues in the rat. Partial cDNAs were found by searching the expressed-sequence-tag database, and primers were designed to generate full-length mRNA coding sequences. One gene was highly similar to the human beta-defensin-1 (HBD-1) gene and mouse beta-defensin-1 gene at both the nucleic acid and amino acid levels and was termed rat beta-defensin-1 (RBD-1). The other gene, named RBD-2, was homologous to the HBD-2 and bovine tracheal antimicrobial peptide (TAP) genes. The predicted prepropeptides were strongly cationic, were 69 and 63 residues in length for RBD-1 and RBD-2, respectively, and contained the six-cysteine motif characteristic of beta-defensins. The beta-defensin genes mapped closely on rat chromosome 16 and were closely linked to the alpha-defensins genes, suggesting that they are part of a gene cluster, similar to the organization reported for humans. Northern blot analysis showed that both RBD-1 and RBD-2 mRNA transcripts were approximately 0.5 kb in length; RBD-1 mRNA was abundantly transcribed in the rat kidney, while RBD-2 was prevalent in the lung. Reverse transcription-PCR indicated that RBD-1 and RBD-2 mRNAs were distributed in a variety of other tissues. In the lung, RBD-1 mRNA expression localized to the tracheal epithelium while RBD-2 was expressed in alveolar type II cells. In conclusion, we characterized two novel beta-defensin homologues in the rat. The rat may be a useful model to investigate the function and contribution of beta-defensins to host defense in the lung, kidney, and other tissues.