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1.
Mucosal Immunol ; 11(2): 486-495, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-28853442

RESUMO

Protective efficacy of Bacillus Calmette-Guérin (BCG) may be affected by the methods and routes of vaccine administration. We have studied the safety and immunogenicity of oral (PO) and/or intradermal (ID) administration of BCG in healthy human subjects. No major safety concerns were detected in the 68 healthy adults vaccinated with PO and/or ID BCG. Although both PO and ID BCG could induce systemic Th1 responses capable of IFN-γ production, ID BCG more strongly induced systemic Th1 responses. In contrast, stronger mucosal responses (TB-specific secretory IgA and bronchoalveolar lavage T cells) were induced by PO BCG vaccination. To generate preliminary data comparing the early gene signatures induced by mucosal and systemic BCG vaccination, CD4+ memory T cells were isolated from subsets of BCG vaccinated subjects pre- (Day 0) and post-vaccination (Days 7 and 56), rested or stimulated with BCG infected dendritic cells, and then studied by Illumina BeadArray transcriptomal analysis. Notably, distinct gene expression profiles were identified both on Day 7 and Day 56 comparing the PO and ID BCG vaccinated groups by GSEA analysis. Future correlation analyses between specific gene expression patterns and distinct mucosal and systemic immune responses induced will be highly informative for TB vaccine development.


Assuntos
Vacina BCG/imunologia , Pulmão/imunologia , Células Th1/fisiologia , Tuberculose/imunologia , Vacinação/métodos , Administração Oral , Adolescente , Adulto , Anticorpos Antibacterianos/metabolismo , Antígenos CD4/metabolismo , Dinamarca , Feminino , Seguimentos , Perfilação da Expressão Gênica , Humanos , Imunidade nas Mucosas , Imunoglobulina A Secretora/metabolismo , Injeções Intradérmicas , Interferon gama/metabolismo , Pulmão/microbiologia , Ativação Linfocitária , Masculino , Pessoa de Meia-Idade , Transcriptoma , Adulto Jovem
2.
Am J Respir Crit Care Med ; 163(4): 1002-9, 2001 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-11282780

RESUMO

We hypothesized that ischemia/reperfusion (I/R) injury of the liver during normotensive gram-negative bacteremic sepsis alters the kinetics of circulating endotoxin, tumor necrosis factor-alpha (TNF-alpha), and coinduced mediators, thereby exacerbating sepsis-induced lung inflammation. Liver and lung dysfunction were studied after hematogenous infection of Sprague-Dawley rats with 10(9) Escherichia coli serotype O55:B5 (EC) and 90 min of secondary hepatic ischemia in EC + I/R and saline-infused (normal saline NS) x I/R rats, followed by brief (1 h) or longer reperfusion (24 h). TNF- alpha:leukotriene interactions in this model were examined using the 5-lipoxygenase-activating protein inhibitor MK-886. Compared with sham-operated EC + Sham animals, peak serum endotoxin, TNF-alpha, alanine aminotransferase, interleukin-6 (IL-6), and hepatic neutrophil (PMN) influx were higher in EC + I/R rats through 24 h (p < 0.05) despite comparable arterial pressure. Lung PMN influx and wet/dry weight ratios were likewise enhanced in EC + I/R versus EC + Sham or NS + I/R rats. MK-886 attenuated TNF-alpha concentrations and ischemic liver injury but not mortality. Thus, focal hepatic I/R augments circulating endotoxin, TNF-alpha, and postbacteremic lung inflammation early after normotensive E. coli bacteremic sepsis.


Assuntos
Bacteriemia/fisiopatologia , Infecções por Escherichia coli/fisiopatologia , Hepatopatias/fisiopatologia , Pneumonia/fisiopatologia , Traumatismo por Reperfusão/fisiopatologia , Animais , Bacteriemia/complicações , Modelos Animais de Doenças , Endotoxinas/análise , Infecções por Escherichia coli/complicações , Mediadores da Inflamação/análise , Isquemia/complicações , Isquemia/fisiopatologia , Inibidores de Lipoxigenase/farmacologia , Fígado/irrigação sanguínea , Fígado/efeitos dos fármacos , Hepatopatias/complicações , Masculino , Neutrófilos/fisiologia , Pneumonia/complicações , Ratos , Ratos Sprague-Dawley , Traumatismo por Reperfusão/complicações , Sensibilidade e Especificidade , Análise de Sobrevida , Fator de Necrose Tumoral alfa/análise
3.
Am J Physiol Regul Integr Comp Physiol ; 279(1): R99-R108, 2000 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-10896870

RESUMO

Reductions in hepatic O(2) delivery are common early after gram-negative bacteremic sepsis owing to cardiopulmonary dysfunction and derangements in sinusoidal perfusion. Although gram-negative endotoxin and cellular hypoxia independently enhance activation of nuclear factor-kappaB (NF-kappaB) via generation of reactive O(2) species (ROS), the combination of these stimuli downregulates hepatic TNF-alpha gene expression. Here we tested the hypothesis that hypoxic suppression of postbacteremic TNF-alpha gene expression is transcriptionally mediated by reduced activation of NF-kappaB. Buffer-perfused rat livers (n = 52) were studied over 180 min after intraportal infection at t = 0 with 10(9) live Escherichia coli (EC), serotype O55:B5, or 0.9% NaCl controls under normoxic conditions, compared with 0.5 h of constant-flow hypoxia (PO(2) approximately 41 +/- 7 Torr) beginning at t = 30 min, followed by 120 min of reoxygenation. In parallel studies, tissue was obtained at peak hypoxia (t = 60 min). To determine the role of xanthine oxidase (XO)-induced ROS in modulating NF-kappaB activity after hypoxia/reoxygenation (H/R), livers were pretreated with the XO inhibitor allopurinol, with results confirmed in organs of tungstate-fed animals. Electrophoretic mobility shift assays were performed on nuclear extracts of whole liver lysates using (32)P-labeled oligonucleotides specific for NF-kappaB. Compared with normoxic EC controls, hypoxia reduced postbacteremic NF-kappaB nuclear translocation and TNF-alpha bioactivity, independent of reoxygenation, tissue levels of reduced glutathione, or posthypoxic O(2) consumption. XO inhibition reversed the hypoxic suppression of NF-kappaB nuclear translocation and ameliorated decreases in cell-associated TNF-alpha. Thus decreases in hepatic O(2) delivery reduce postbacteremic nuclear translocation of NF-kappaB and hepatic TNF-alpha biosynthesis by signaling mechanisms involving low-level generation of XO-mediated ROS.


Assuntos
Bacteriemia/metabolismo , Hipóxia/metabolismo , Fígado/metabolismo , NF-kappa B/metabolismo , Fator de Necrose Tumoral alfa/biossíntese , Alopurinol/farmacologia , Animais , Eletroforese em Gel de Poliacrilamida , Inibidores Enzimáticos/farmacologia , Infecções por Escherichia coli/metabolismo , Expressão Gênica , Técnicas In Vitro , Testes de Função Hepática , Masculino , NF-kappa B/análise , Perfusão , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Estresse Fisiológico/metabolismo , Fator de Necrose Tumoral alfa/genética , Xantina Oxidase/antagonistas & inibidores , Xantina Oxidase/metabolismo
4.
Am J Physiol Lung Cell Mol Physiol ; 278(6): L1289-96, 2000 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-10835336

RESUMO

Episodes of tissue hypoxia and reoxygenation frequently occur during gram-negative bacteremia that progresses to septic shock. However, few studies have evaluated modulation by hypoxia and reoxygenation of the proinflammatory cytokine gene expression that is normally induced by gram-negative bacteremia or endotoxemia. In buffer-perfused organs, hypoxia downregulates Escherichia coli-induced expression of tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta in the liver but upregulates these cytokines in the lungs. To identify molecular mechanisms underlying these events, we investigated the effects of brief (1.5-h) hypoxia on TNF-alpha and IL-1beta expression in cultured RAW 264.7 cells during their continuous exposure to lipopolysaccharide (LPS) endotoxin derived from E. coli (serotype 055:B5) for up to 24 h. IL-1beta and TNF-alpha concentrations in cell lysates and culture supernatants were measured by ELISA, and steady-state mRNA was measured by Northern analysis. LPS-induced IL-1beta synthesis was downregulated by hypoxia at both the protein and mRNA levels despite no change in cellular redox status as measured by levels of GSH. In contrast, LPS-induced TNF-alpha production was unaffected by hypoxia as assessed by cell lysate mRNA and lysate and supernatant protein levels. Nuclear runoff analysis showed that downregulation of IL-1beta gene expression by hypoxia occurred transcriptionally. Allopurinol or catalase treatment did not alter modulation of LPS-induced IL-1beta expression by hypoxia, suggesting that this suppression was not caused by reactive oxygen species. Cycloheximide pretreatment suggested that hypoxia-induced downregulation of IL-1beta expression did not require de novo protein synthesis.


Assuntos
Hipóxia/genética , Interleucina-1/genética , Lipopolissacarídeos/farmacologia , Fator de Necrose Tumoral alfa/genética , Animais , Carbocianinas/farmacologia , Catalase/farmacologia , Linhagem Celular , Homeostase , Interleucina-1/antagonistas & inibidores , Camundongos , RNA Mensageiro/antagonistas & inibidores , RNA Mensageiro/metabolismo , Transcrição Gênica , Fator de Necrose Tumoral alfa/metabolismo
5.
Crit Care Med ; 27(7): 1230-51, 1999 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-10446814

RESUMO

OBJECTIVES: The purpose of this study was to determine whether apoptosis is a major mechanism of cell death in patients with sepsis. The activities of caspase-3 and the antiapoptotic protein, BCL-2, were investigated also. DESIGN: A prospective study of 20 patients who died of sepsis and multiple organ dysfunction was performed. The control group of 16 patients consisted of critically ill, nonseptic patients who were evaluated either prospectively (7) or retrospectively (9). In addition, normal colon sections from seven patients who had bowel resections were included. Apoptosis was evaluated in hematoxylin and eosin-stained specimens by deoxyuridine triphosphate nick end-labeling (TUNEL) and by DNA gel electrophoresis. SETTING: Two academic medical centers. PATIENTS: Critically ill patients. MEASUREMENTS AND MAIN RESULTS: In septic patients, apoptosis was detected in diverse organs by all three methods with a predominance in lymphocytes and intestinal epithelial cells. Hematoxylin and eosin-stained specimens from septic patients demonstrated at least focal apoptosis in 56.3% of spleens, 47.1% of colons, and 27.7% of ileums. Indirect evidence of lymphocyte apoptosis in septic patients included extensive depletion of lymphocytes in white pulp and a marked lymphocytopenia in 15 of 19 patients. Hematoxylin and eosin from nonseptic patients' tissues revealed a low level of apoptosis in one patient only. The TUNEL method increased in positivity with a delay in tissue fixation and was highly positive in many tissues from both septic and nonseptic patients. Immunohistochemical staining for active caspase-3 showed a marked increase in septic vs. nonseptic patients (p < .01), with >25% to 50% of cells being positive focally in the splenic white pulp of six septic but in no nonseptic patients. CONCLUSIONS: We conclude that caspase-3-mediated apoptosis causes extensive lymphocyte apoptosis in sepsis and may contribute to the impaired immune response that characterizes the disorder.


Assuntos
Apoptose/fisiologia , Insuficiência de Múltiplos Órgãos/patologia , Sepse/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Autopsia , Biomarcadores , Estudos de Casos e Controles , Caspase 3 , Caspases/metabolismo , Precursores Enzimáticos/metabolismo , Feminino , Humanos , Imuno-Histoquímica , Intestinos/patologia , Linfócitos/fisiologia , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Choque Séptico/patologia , Baço/patologia
7.
J Surg Res ; 76(2): 179-84, 1998 May.
Artigo em Inglês | MEDLINE | ID: mdl-9698520

RESUMO

INTRODUCTION: Muramlytripeptide phosphatidylethanolamine (MTP) stimulates synthesis of cytokines by hepatic Kupffer cells. We have shown in a perfused rat liver model that secondary ischemia/reperfusion (I/R) downregulates tumor necrosis factor alpha (TNF-alpha) expression after Escherichia coli (EC) bacteremia. Here, we tested the hypothesis that pretreatment with MTP restores cytokine response after sequential bacteremia and I/R. METHODS: Thirty-eight livers were studied in eight groups after intraportal injection of 10(9) live EC or normal saline (NS): (1) normoxic EC; (2) EC + I/R (ischemia began 30 min after EC followed by 2 h of reperfusion); (3) normoxic NS controls; and (4) NS + I/R. Four groups of rats received 300 micrograms of MTP i.v. 24 h prior to liver harvesting; (5) MTP + EC; (6) MTP + EC + I/R; (7) MTP + NS; and (8) MTP + NS + I/R. Bioactive and antigenic TNF-alpha, PGE2 and bacterial clearance were assessed. RESULTS: MTP increased bioactive TNF-alpha response to EC (MTP + EC vs EC controls: 685 +/- 255 U/ml vs 250 +/- 180 U/ml, P < 0.02). I/R did not downregulate TNF-alpha in animals treated with MTP (MTP + NS vs MTP + NS +I/R, P = 0.83). Pretreatment with MTP restored TNF-alpha after I/R MTP + EC + I/R vs EC + I/R: 671 +/- 215 U/ml vs 27 +/- 14 U/ml, P < 0.001) to levels similar to those found in the MTP + EC group (MTP + EC + I/R vs MTP + EC: 671 +/- 215 U/ml vs 685 +/- 255 U/ml, P = 0.75). Finally, bacterial clearance was increased in groups which received MTP. CONCLUSION: In vivo administration of MTP increases hepatic TNF-alpha response to intraportal EC bacteremia by a PGE2 independent mechanism. This response is maintained even after subsequent ischemia and reperfusion.


Assuntos
Acetilmuramil-Alanil-Isoglutamina/análogos & derivados , Bacteriemia/metabolismo , Fatores Imunológicos/farmacologia , Fígado/metabolismo , Fosfatidiletanolaminas/farmacologia , Traumatismo por Reperfusão/metabolismo , Fator de Necrose Tumoral alfa/biossíntese , Acetilmuramil-Alanil-Isoglutamina/farmacologia , Animais , Dinoprostona/metabolismo , Infecções por Escherichia coli , Cinética , Fígado/irrigação sanguínea , Fígado/efeitos dos fármacos , Masculino , Ratos , Ratos Sprague-Dawley
8.
Am J Respir Crit Care Med ; 157(5 Pt 1): 1550-8, 1998 May.
Artigo em Inglês | MEDLINE | ID: mdl-9603137

RESUMO

We employed a bile duct ligation (BDL) model of cholestatic liver injury to test the hypothesis that this form of preexisting hepatic dysfunction alters the kinetics of circulating TNF-alpha and IL-6 after Escherichia coli endotoxemia, thereby augmenting mortality and lung injury by a TNF-alpha:leukotriene (LT) axis of inflammation. Male rats were catheterized 13 d after BDL or sham surgery and studied while awake 18 to 24 h later. Cholestasis after BDL was confirmed by baseline serum bilirubin (BDL = 7.34 +/- 0.72 mg/dl, mean +/- SEM, n = 17 versus Sham = 0.25 +/- 0.07, n = 20; p < 0.005) and histopathology. Sham and BDL animals received E. coli lipopolysaccharide serotype O55:B5 (LPS, 5 mg/kg i.v.) or 0.9% NaCl (NS) ending at t = 0 and were monitored over 24 h for vital signs and hemodynamics. In parallel studies, lipoxygenase inhibition was performed using diethylcarbamazine or the 5-lipoxygenase activating-protein inhibitor MK-886. Blood was collected at baseline and at t = 1.5, 3.5, and 24 h for formed elements and for serum endotoxin, TNF-alpha, IL-6, bilirubin, and alanine aminotransferase (ALT). Organs were evaluated at 24 h for histopathology, including neutrophil (PMN) densities and wet/dry weight (W/D) ratios. Cholestasis reduced survival after otherwise nonlethal endotoxemia, with seven of 11 BDL + LPS rats dying within 24 h versus no deaths in BDL + NS (n = 6), Sham + LPS (n = 14), or Sham + NS (n = 6) animals (p < 0.01). Despite equivalent serum endotoxin between groups, circulating TNF-alpha was 8-fold higher in BDL + LPS than in Sham + LPS rats at 1.5 and 3.5 h (p < 0.001), whereas serum TNF-alpha did not differ between BDL + NS and Sham + NS rats. IL-6 likewise was increased differentially by 1.5 h in BDL + LPS animals (11.98 +/- 2.42 ng/ml) versus Sham + LPS rats (3.05 +/- 0.58 ng/ml, p < 0.05). Hypothermia, bradycardic hypotension, and leukopenia were most severe and prolonged in BDL + LPS rats, which also had significantly higher ALT values, W/D ratios, and organ PMN counts. LT inhibition failed to reduce BDL-related differences in serum cytokines or survival after endotoxemia. Thus, cholestasis augments inflammatory responses to gram-negative endotoxemia, sensitizing the host to enhanced fluid flux in multiple organs and to mortality by a LT-independent mechanism.


Assuntos
Colestase/complicações , Endotoxemia/sangue , Infecções por Escherichia coli/sangue , Interleucina-6/sangue , Hepatopatias/complicações , Fator de Necrose Tumoral alfa/análise , Alanina Transaminase/sangue , Animais , Bilirrubina/sangue , Ducto Colédoco , Endotoxemia/complicações , Endotoxemia/mortalidade , Endotoxemia/patologia , Infecções por Escherichia coli/complicações , Infecções por Escherichia coli/mortalidade , Infecções por Escherichia coli/patologia , Ligadura , Inibidores de Lipoxigenase/farmacologia , Fígado/patologia , Hepatopatias/etiologia , Hepatopatias/patologia , Pulmão/patologia , Masculino , Neutrófilos/patologia , Ratos , Ratos Sprague-Dawley
9.
Am J Respir Crit Care Med ; 157(2): 629-37, 1998 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-9476882

RESUMO

Decreases in the alveolar O2 tension commonly follow gram-negative bacteremic shock that progresses to the acute respiratory distress syndrome (ARDS). To examine the effects of alveolar hypoxia and reoxygenation (H/R) on postbacteremic pulmonary cytokine expression, lungs from Sprague-Dawley rats (n = 43) were perfused over 180 min after hematogenous infection with 10(9) live Escherichia coli serotype O55:B5 (EC) or infusion of 0.9% NaCl (NS). Compared with normoxic EC and NS controls, EC + H/R and NS + H/R lungs received 90 min of constant-flow hypoxia followed by 60 min of reoxygenation. Perfusates were cultured and analyzed for TNF-alpha, IL-1alpha, IL-1beta, and PGE2 while monitoring pulmonary artery pressure (Ppa). Changes in the filtration coefficient (Kf) were evaluated at 180 min when cytokine mRNA levels were assessed in lung homogenates. Transcripts of the anti-inflammatory cytokine TGF-beta1 and of inducible cyclooxygenase (COX-2) were similarly analyzed. For equivalent EC clearance, Ppa, and Kf as in normoxic EC, postbacteremic H/R increased TNF-alpha gene expression and doubled the export of TNF-alpha from the lungs, an effect not blocked by allopurinol. IL-1alpha transcripts were also increased in EC + H/R versus EC lungs, in contrast to the lack of change in IL-1beta, TGF-beta, or COX-2 mRNA levels, or in cell-associated or circulating IL-1beta and PGE2. Thus, gram-negative bacteremic lung infection and secondary alveolar H/R upregulate the expression of specific inflammatory cytokines compared with pulmonary infection under normoxic conditions, independently of xanthine oxidase-induced O2 radicals. These findings identify the alveolar PO2 as a potent immunomodulatory signal whose reductions early after gram-negative sepsis may enhance lung inflammation in ARDS.


Assuntos
Bacteriemia/genética , Citocinas/genética , Infecções por Escherichia coli/genética , Expressão Gênica , Hipóxia/genética , Alvéolos Pulmonares , Animais , Expressão Gênica/fisiologia , Técnicas In Vitro , Interleucina-1/genética , Pulmão/efeitos dos fármacos , Pulmão/fisiopatologia , Pneumopatias/genética , Masculino , Oxigênio/farmacologia , Perfusão , Ratos , Ratos Sprague-Dawley , Fator de Necrose Tumoral alfa/genética
12.
New Horiz ; 5(3): 233-8, 1997 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-9259336

RESUMO

OBJECTIVE: To review the literature addressing use of the pulmonary artery catheter (PAC) to augment oxygen delivery to supranormal levels in critical illness. DATA SOURCE: All pertinent English language articles dealing with use of the PAC to augment oxygen delivery to supranormal levels in critical illness were retrieved from 1988 through 1996. STUDY SELECTION: Articles were chosen if material relevant to supranormal oxygen delivery in critically ill patients was studied or reviewed. DATA EXTRACTION: Methodology of the available studies was analyzed. From the articles selected, information was obtained regarding changes in outcome associated with prospective goal-oriented hemodynamic intervention using a PAC to augment systemic oxygen delivery to supranormal levels prior to high-risk surgery. Patient outcome information was also obtained regarding use of the PAC to augment oxygen delivery to supranormal values begun after the onset of the systemic inflammatory response syndrome (SIRS) following sepsis or trauma. DATA SYNTHESIS: Inadequate evidence exists to accurately determine if PAC use to augment systemic oxygen delivery improves outcome in either of these circumstances. CONCLUSION: Further research must be performed before a recommendation can be made about goal-oriented hemodynamic intervention utilizing the PAC to augment oxygen delivery to supranormal levels prior to high-risk surgery. PAC-guided hemodynamic intervention to augment oxygen delivery to supranormal values in patients with SIRS-related organ dysfunction from sepsis, trauma, or postoperative complications is not recommended at this time. Carefully designed, multicenter, randomized, controlled trials are needed to establish whether augmenting oxygen delivery improves organ-specific outcomes and survival under each of these circumstances.


Assuntos
Cateterismo de Swan-Ganz , Estado Terminal , Consumo de Oxigênio , Oxigênio/sangue , Humanos , Síndrome de Resposta Inflamatória Sistêmica/sangue , Resultado do Tratamento
13.
J Crit Care ; 12(1): 28-38, 1997 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-9075062

RESUMO

PURPOSE: During gram-negative bacteremia (GNB), tumor necrosis factor-alpha (TNF-alpha) is a critical early mediator of host defense, whose overexpression can initiate acute lung injury, multiple organ failure, and death. In this study we evaluated the ability of a chimeric fusion protein containing two extracellular domains of the human p80 TNF-alpha receptor and the Fc region of human IgG1 (TNFR:Fc) to reduce circulating TNF-alpha, and to ameliorate organ injury and improve survival in a rodent model of GNB during immunosuppression-related neutropenia. MATERIALS AND METHODS: Conscious catheterized male rats (n = 37) with stable cyclophosphamide-induced neutropenia were infected intravenously (i.v.) with 5 x 10(9) live Escherichia coli (EC, serotype O55:B5) ending at t = 0. All animals received antibiotics (penicillin/ amikacin sulfate) at t = 0.5 and t = 8 hours, and 0.9% sodium chloride (normal saline solution (NS), 1 mL/h) from t = 0 to 8 hours. Subgroups were post-treated at t = 0.5 hours with a 1.0 mL i.v. dose of TNFR:Fc (60, 600, or 1,200 micrograms; Immunex), 600 micrograms of human IgG1-kappa or IgG1-lambda (Sigma), or NS alone (controls). A separate TNFR:Fc pretreatment subgroup received 600 micrograms/rat of the fusion protein 5 minutes before starting EC infusion. Hemodynamics were monitored continuously through t = 24 hours, and arterial samples were collected at baseline and at t = 1.5, 4.5, 8, and 24 hours after EC were analyzed for blood gases, quantitative culture, serum endotoxin, bioactive and antigenic TNF-alpha, and formed elements. Postmortem tissues were examined for histopathologic changes. RESULTS: Compared with antibiotic-treated and fluid-supported controls, TNFR:Fc dose-dependently reduced bioactive but not antigenic TNF-alpha without altering bacterial clearance, serum endotoxin, or 24-hour survival. Of note, 600 micrograms of IgG1-kappa or IgG1-lambda attenuated peak bioactive TNF-alpha to a similar degree as 1,200 micrograms TNFR:Fc, and also significantly reduced serum endotoxin levels. Nevertheless, by t = 8 hours all bacteremic rats were hypothermic with tachypnea-related hypocarbia and hyperoxemia and were thrombocytopenic. At death, all subgroups showed similar hepatic glycogen depletion and pulmonary congestion with perivascular edema and alveolar hemorrhage. CONCLUSIONS: Although TNFR:Fc and its idiotypic control IgG1 reduced circulating bioactive TNF-alpha, neither treatment prevented progression of lethal shock with attendant organ injury in this conscious, antibiotic-treated and fluid-resuscitated model of immunosuppression-related GNB.


Assuntos
Antígenos CD/imunologia , Bacteriemia/terapia , Infecções por Escherichia coli/terapia , Imunoglobulina G/imunologia , Receptores de IgG/imunologia , Receptores do Fator de Necrose Tumoral/imunologia , Proteínas Recombinantes de Fusão/uso terapêutico , Fator de Necrose Tumoral alfa/efeitos dos fármacos , Animais , Bacteriemia/imunologia , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Infecções por Escherichia coli/imunologia , Masculino , Neutropenia/complicações , Ratos , Ratos Sprague-Dawley , Receptores Tipo II do Fator de Necrose Tumoral , Síndrome do Desconforto Respiratório/microbiologia , Análise de Sobrevida
14.
JAMA ; 277(6): 482-7, 1997 Feb 12.
Artigo em Inglês | MEDLINE | ID: mdl-9020273

RESUMO

OBJECTIVE: To test the effect of a novel bradykinin antagonist, deltibant (CP-0127), on survival, organ dysfunction, and other outcomes in patients with the systemic inflammatory response syndrome (SIRS) and presumed sepsis. DESIGN: Multicenter, randomized, placebo-controlled, double-blind, parallel, dose-ranging trial. Follow-up for 28 days or until death. SETTING: A total of 47 US referral hospitals. PATIENTS: A total of 504 patients with SIRS and documented evidence of infection plus either hypotension or dysfunction of 2 organ systems. INTERVENTIONS: Three-day continuous intravenous infusion of either placebo or 1 of 3 doses (0.3, 1.0, or 3.0 microg x kg(-1) x min(-1)) of deltibant. Concurrent therapy at the discretion of the treating physician. MAIN OUTCOME MEASURE: Risk-adjusted, 28-day, log-normal intent-to-treat survival analysis. Risk adjustment was performed using a study-specific risk model derived from the APACHE III database. RESULTS: Deltibant had no significant effect on risk-adjusted 28-day survival. In a posthoc analysis, risk-adjusted 7-day survival showed a nonsignificant trend toward improvement (P=.09). The 28-day risk-adjusted survival in the prospectively defined subset of patients with gram-negative infections showed a statistically significant improvement (P=.005). CONCLUSIONS: Deltibant may have some effect on survival in patients with SIRS and gram-negative sepsis; however, additional studies would be required to prove this.


Assuntos
Antagonistas dos Receptores da Bradicinina , Peptídeos/uso terapêutico , Síndrome de Resposta Inflamatória Sistêmica/tratamento farmacológico , APACHE , Método Duplo-Cego , Esquema de Medicação , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Infecções por Bactérias Gram-Negativas/fisiopatologia , Humanos , Hipotensão/etiologia , Pessoa de Meia-Idade , Insuficiência de Múltiplos Órgãos/etiologia , Peptídeos/administração & dosagem , Sepse/tratamento farmacológico , Sepse/fisiopatologia , Análise de Sobrevida , Síndrome de Resposta Inflamatória Sistêmica/fisiopatologia
15.
Crit Care Med ; 25(1): 111-20, 1997 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-8989186

RESUMO

OBJECTIVES: Dimorphic Candida albicans spp. increasingly cause lethal septic shock and disseminated infection in the critically ill. Following candidemia, production of specific fungal exotoxins coincident with the yeast to hyphal phenotypic transition is believed to be important in the pathogenesis of Candida septic shock. However, overexpression of the pleiotropic cytokine tumor necrosis factor (TNF)-alpha by the host following hyphal germination is also thought to be a mechanism of Candida-related cardiopulmonary dysfunction, as well as of bacteremic shock. In this study, we hypothesized that increases in circulating TNF-alpha coinciding with the yeast to hyphal transition modulate the onset and progression of shock with multiple organ injury early after hematogenous candidiasis. DESIGN: Prospective, controlled laboratory animal study. SETTING: University hospital animal research facility. SUBJECTS: Pathogen-free, male Sprague-Dawley rats (n = 26). INTERVENTIONS: Conscious, antibiotic-treated animals with chronic indwelling carotid arterial and jugular venous catheters were intravenously infected with 10(9) viable blastoconidia of the C. albicans clinical pathogen, CA-MEN (n = 10), over 30 mins and ending at t = 0 hr, compared with an equivalent inoculum of its viable agerminative mutant, CA-MM2002 n = 11), or an intravenous infusion of 0.9% sodium chloride (n = 5). MEASUREMENTS AND MAIN RESULTS: Mean arterial pressure (MAP), pulse rate, respiratory frequency, rectal temperature, acid-base status, quantitative blood cultures, circulating alanine aminotransferase (ALT), and bioactive TNF-alpha were serially measured in all three groups over 24 hrs or until death. Organ cultures, wet/dry weight ratios, and histopathologic changes in the lungs, heart, liver, and kidneys were determined in Candida-infected and 0.9% sodium chloride (normal saline)-infused subgroups at 6 and 24 hrs. Animals hematogenously infected with the C. albicans clinical isolate developed lethal nonendotoxemic shock in < or = 6 hrs (MAP 49 +/- 7 mm Hg [SEM]; p < .05 vs. t = 0 hr), and at death (7.0 +/- 0.3 hrs) were acidotic, hypocapnic, and hypothermic (rectal temperature 33.2 +/- 0.7 degrees C). Despite similar peak concentrations of circulating fungal colony-forming units (cfu) and kinetics of vascular clearance in both Candida-infected groups, survival and MAP in rats challenged with the agerminative C. albicans mutant were unchanged for > 8 hrs, as were pH, Pco2, and rectal temperature. No germination of the agerminative fungal strain occurred in vivo over 6 hrs. Serum TNF was nearly undetectable at t = 0 hr in all three groups. Although shock developed soon after fungemia with the C. albicans clinical isolate, TNF-alpha concentrations did not increase above normal saline values in either candidemic group at t = 1.5, 4.5, or 6 hrs (17 +/- 7 vs. 14 +/- 1 U/mL in the parent C. albicans organism vs. its agerminative mutant at t = 6 hrs). Greater numbers of agerminative C. albicans than its dimorphic parent strain were recovered from the lungs (5.41 +/- 1.0 vs. 2.02 +/- 0.38 x 10(7) cfu/g, respectively; p < .05) and kidneys (p < .01). By 24 hrs, modest germination of the mutant Candida strain was observed in the tissues. However, lung wet/dry ratios, intrapulmonary hyphal proliferation, and alveolar hemorrhage were all greater after infection with the parent fungal isolate. Likewise, myocardial necrosis and hepatic glycogen depletion with vacuolization were more severe after infection with the C. albicans clinical isolate vs. candidemia with its agerminative mutant, although serum ALT values did not differ between these groups. CONCLUSIONS: Lethal C. albicans sepsis with lung injury and multiple organ damage are temporally associated with the in vivo yeast to hyphal transition in this model. However, this candidemic septic shock syndrome is modulated by circulating fungal virulence factors or host mediators other than TNF-alpha, a cytokine considered essen


Assuntos
Candida albicans/crescimento & desenvolvimento , Candida albicans/patogenicidade , Candidíase/complicações , Candidíase/microbiologia , Modelos Animais de Doenças , Fungemia/complicações , Fungemia/microbiologia , Choque Séptico/microbiologia , Fator de Necrose Tumoral alfa/análise , Animais , Sangue/microbiologia , Candida albicans/metabolismo , Candidíase/sangue , Doenças Cardiovasculares/etiologia , Fungemia/sangue , Fígado/microbiologia , Pulmão/microbiologia , Masculino , Ratos , Ratos Sprague-Dawley , Choque Séptico/sangue , Choque Séptico/mortalidade , Fatores de Tempo
17.
Am J Physiol ; 271(5 Pt 2): R1311-8, 1996 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-8945969

RESUMO

Hepatic cytokine gene expression is independently stimulated by circulating microbial products and reductions in the cellular O2 supply. Although these stimuli occur sequentially after gram-negative bacteremia, it is unknown whether their interplay augments production of interleukin (IL)-1 by the liver. We studied the effects of intraportal Escherichia coli (EC) bacteremia and secondary constant-flow hypoxia (Po2, approximately 46 Torr for 30 min) on IL-1 alpha and IL-1 beta gene expression in ex situ buffer-perfused rat livers over 180 min (n = 67). At t = 0, normoxic EC and normal saline (NS) controls received 10(9) live EC serotype 055:B5 and 0.9% NaCl, respectively; in livers subjected to EC+hypoxia-reoxygenation (H/R) and NS+H/R, hypoxia began 0.5 h after EC or NS and was followed by 120 min of reoxygenation. Portal and hepatic venous perfusates were serially analyzed for bacterial colony-forming units, O2 uptake, and aspartate aminotransferase. At 60 min (peak hypoxia) and 180 min, cDNAs for IL-1 alpha and IL-1 beta were hybridized to whole liver RNA, and IL-1 beta protein levels in venous perfusates were assessed. Intrahepatic levels of reduced glutathione (GSH) were measured as an index of oxidative stress. Compared with normoxic EC, IL-1 alpha transcripts decreased at 180 min in EC+H/R livers (P < 0.0001) as did IL-1 beta mRNA (P < 0.05), despite similar EC clearance, GSH levels, posthypoxic O2 uptake, and aspartate aminotransferase release. Hepatic secretion of IL-1 beta likewise fell in EC+H/R vs. EC controls (P < 0.005). Prostaglandin H synthase-2 (COX-2) message accumulation was not enhanced by H/R, and indomethacin did not reverse H/R-mediated suppression of IL-1 production. In contrast, H/R-related falls in EC-induced IL-1S expression were reversed by allopurinol or catalase. Thus brief hypoxic stress of the liver causing neither GSH depletion nor functional impairment downregulates postbacteremic IL-1 expression by a mechanism involving O2 radicals but not cyclooxygenase metabolites.


Assuntos
Infecções por Escherichia coli/genética , Expressão Gênica , Hipóxia/genética , Interleucina-1/genética , Fígado/fisiopatologia , Estresse Fisiológico/genética , Animais , Bacteriemia/genética , Bacteriemia/metabolismo , Bacteriemia/fisiopatologia , Infecções por Escherichia coli/metabolismo , Infecções por Escherichia coli/fisiopatologia , Glutationa/metabolismo , Hipóxia/metabolismo , Hipóxia/fisiopatologia , Interleucina-1/metabolismo , Fígado/metabolismo , Masculino , Oxigênio/fisiologia , Perfusão , Prostaglandina-Endoperóxido Sintases/metabolismo , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Estresse Fisiológico/metabolismo , Estresse Fisiológico/fisiopatologia
20.
Am J Physiol ; 270(1 Pt 2): R289-97, 1996 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-8769813

RESUMO

We tested the hypothesis that gram-negative bacteremia (GNB) and brief (30 min) reductions in the hepatic O2 supply by low-flow ischemia differentially modulate tumor necrosis factor-alpha (TNF-alpha) gene expression owing to sequence-specific activation of cyclooxygenase vs. complement (C) pathways. Buffer-perfused Sprague-Dawley rat livers (n = 82) were studied over 180 min after intraportal 10(9) live E. coli serotype 055:B5 (EC) or 0.9% NaCl (NS) at t = 0. Compared with EC and NS controls receiving constant-flow perfusion, sequential GNB and ischemia/reperfusion (I/R) were studied in EC + 30 I/R and NS + 30 I/R livers, in which 30 min of ischemia (I) beginning 0.5 h after EC or NS was followed by 120 min of reperfusion (R). This sequence was reversed in 30 I/R + EC and 30 I/R + NS groups. Bacterial clearance, bioactive and antigenic TNF-alpha, prostaglandin E2 (PGE2), and hepatic O2 uptake and performance were serially assessed. Venous TNF-alpha increased in EC controls to peak at 155 +/- 29 U/ml after 180 min (P < 0.001 vs. NS controls) as did hepatic TNF-alpha mRNA. Both TNF-alpha transcripts and protein levels were markedly attenuated in EC + 30 I/R (P < 0.001 vs. EC) despite equivalent EC clearance by Kupffer cells. Indomethacin (10(-5) M) decreased I/R-induced PGE2 secretion and restored TNF-alpha to control levels. In contrast, TNF-alpha levels in 30 I/R + EC perfusates exceeded those of EC + 30 I/R livers (P < 0.05) and were indistinguishable from EC controls. Allopurinol pretreatment but not heat inactivation of C or infusion of soluble human complement receptor type 1 inhibited TNF-alpha production in 30 I/R + EC organs. These results identify a novel sequence-dependent interaction whereby hepatic O2 deprivation after GNB downregulates TNF-alpha via generation of cyclooxygenase metabolites, whereas ischemia preceding GNB increases cytokine expression via reactive O2 species but not C activation.


Assuntos
Escherichia coli/fisiologia , Expressão Gênica , Isquemia/genética , Circulação Hepática , Traumatismo por Reperfusão/genética , Fator de Necrose Tumoral alfa/genética , Animais , Inibidores de Ciclo-Oxigenase/farmacologia , Citocinas/genética , Técnicas In Vitro , Isquemia/patologia , Fígado/patologia , Masculino , Oxigênio/fisiologia , Ratos , Ratos Sprague-Dawley , Traumatismo por Reperfusão/patologia
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