RESUMO
Patients with infection, sepsis, severe sepsis, or septic shock were compared to each other and to healthy controls with regard to serum levels of biomarkers and clinical symptoms. Of the 15 biomarkers assayed, 9 were detectable in patients, and 4, in controls. Both proinflammatory and anti-inflammatory cytokines were detected in the patients. No single biomarker could differentiate the 3 septic levels of severity from each other; however, interleukin (IL) 1 receptor antagonist (IL-1ra) had the best sensitivity and specificity for differentiating sepsis and severe sepsis from septic shock. IL-6 was the only cytokine able to differentiate infected patients without signs of sepsis from those with sepsis. Although IL-1ra, IL-6, IL-8, and monocyte chemoattractant protein 1 could differentiate infection, sepsis, and severe sepsis from septic shock, the biomarkers could not differentiate sepsis from severe sepsis. The top scoring pair algorithm with clinical and biomarker analyses was able to correctly diagnose those with sepsis who will progress to a more severe state.
Assuntos
Biomarcadores/sangue , Sepse/diagnóstico , Sepse/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: Candida glabrata causes infections associated with severe sepsis, production of high concentrations of cytokines/chemokines, and high mortality. This study describes the effects of anidulafungin (ANF) and voriconazole (VRC), singly and in combination, on the production of eight cytokines/chemokines by human monocyte-derived macrophages (MDM) infected with C. glabrata or activated by lipopolysaccharide (LPS). METHODS: MDM monolayers were established, infected with C. glabrata or activated with LPS, and then treated with high or low concentrations of ANF, VRC, or both. Cytokine/chemokine levels in MDM supernatants were determined. RESULTS: Levels of cytokines/chemokines were significantly elevated in supernatants of infected or LPS-activated MDM. Except for interleukin-10, all significant decreases in cytokine/chemokine concentrations (p < 0.01) occurred in supernatants of infected MDM treated with high concentrations of ANF or ANF + VRC. CONCLUSIONS: Decreases in cytokine/chemokine levels in supernatants of infected MDM treated with high concentrations of ANF or ANF + VRC suggest that similar treatment could improve survival in patients with severe, invasive C. glabrata infections and markedly elevated levels of serum cytokines/chemokines.
Assuntos
Antifúngicos/farmacologia , Candida glabrata/patogenicidade , Quimiocinas/metabolismo , Citocinas/metabolismo , Equinocandinas/farmacologia , Macrófagos/metabolismo , Pirimidinas/farmacologia , Triazóis/farmacologia , Anidulafungina , Humanos , Lipopolissacarídeos/toxicidade , Macrófagos/efeitos dos fármacos , VoriconazolRESUMO
Serious Candida glabrata infections, which can be difficult to treat, are often treated with echinocandins. We compared in vitro the effects of high and low concentrations of 3 echinocandins (micafungin [MCF], caspofungin [CAS], and anidulafungin [ANF]), voriconazole (VRC), and amphotericin B (AmB), singly and VRC in combination with MCF, CAS, and ANF, on the production of cytokines/chemokines by human monocyte-derived macrophages (MDM). MDM were activated by infection with C. glabrata or lipopolysaccharide (LPS). Luminex multi-analyte microsphere technology was used for cytokine/chemokine analysis. Concentrations of cytokines/chemokines were significantly elevated following activation by infection or LPS. Treatment with high concentrations of echinocandins, singly or in combination with VRC, was most effective in lowering the elevated cytokine/chemokine levels. This effect occurred only with MDM activated by infection with C. glabrata and not with LPS. Treatment with VRC or AmB alone had little or no effect on cytokine/chemokine levels. In severe C. glabrata infection associated with very high concentrations of dysregulated cytokines/chemokines, echinocandins, singly or in combination with VRC, may decrease cytokine/chemokine concentrations and thus may improve host survival.
Assuntos
Antifúngicos/farmacologia , Candida glabrata/imunologia , Citocinas/biossíntese , Equinocandinas/farmacologia , Lipopolissacarídeos/imunologia , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Candidíase/imunologia , Candidíase/metabolismo , Quimiocinas/biossíntese , Humanos , Ativação de Macrófagos/imunologiaRESUMO
Infections caused by fluconazole-resistant Candida glabrata and Candida krusei are increasingly common causes of morbidity and mortality. We investigated the intracellular killing of fluconazole-resistant C. glabrata and C. krusei by cytokine-activated human monocyte-derived macrophages (MDM) in the presence and absence of voriconazole. For C. glabrata, MDM were activated with either granulocyte-macrophage colony-stimulating factor (GM-CSF) or interferon gamma (IFN-gamma) before infection, after infection, or both before and after infection, whereas for C. krusei MDM were activated with cytokines both before and after infection. Activated MDM were infected, treated with voriconazole, and then lysed, and viable yeast in the lysates enumerated at 0, 24, or 48 h after infection. In the presence of voriconazole (2.5 x MIC), the best activity against C. glabrata occurred when MDM were activated with GM-CSF for 24 h before infection as well as after infection or when they were activated for 24 h before infection alone. A lesser effect was observed when MDM were activated for at least 1 h before infection or when they were treated with cytokines only after infection. IFN-gamma activation had a significant but lesser effect than GM-CSF. Activity against C. krusei in the presence of voriconazole was greatest when MDM were activated with IFN-gamma rather than GM-CSF. Our results suggest that cytokines increase the intracellular anticandidal effect of voriconazole and may be useful as therapeutic adjuvants to voriconazole for treatment of infections caused by fluconazole-resistant C. glabrata and C. krusei.
Assuntos
Antifúngicos/farmacologia , Candida glabrata/efeitos dos fármacos , Candida/efeitos dos fármacos , Macrófagos/imunologia , Macrófagos/microbiologia , Pirimidinas/farmacologia , Triazóis/farmacologia , Candida/crescimento & desenvolvimento , Candida glabrata/crescimento & desenvolvimento , Contagem de Colônia Microbiana , Farmacorresistência Fúngica , Fluconazol/farmacologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Humanos , Interferon gama/farmacologia , Monócitos/microbiologia , Especificidade da Espécie , VoriconazolRESUMO
Although antibiotics are known to affect the intracellular growth of Chlamydia pneumoniae in acute infections, their efficacy in therapy for chronic infections, including atherosclerosis, remains debatable. Human monocyte-derived macrophages (MDM) obtained from monocytes of healthy donors were infected with C. pneumoniae AR-39 and treated with levofloxacin (8 microg/mL) immediately after infection (0 hours) or 24 hours after infection. Levofloxacin treatment at 24 hours, but not at 0 hours, resulted in a significant decrease in the number of C. pneumoniae inclusions within the MDM (p < 0.05). Also decreased were concentrations of proinflammatory cytokines tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta, IL-6, and IL-8 in the extracellular medium (p < 0.01). Viable counts in titrations remained similar to those in untreated controls. In summary, levofloxacin administered to MDM at serum-attainable levels 24 hours after C. pneumoniae infection significantly decreased inclusion counts and proinflammatory cytokine production, but did not eliminate the C. pneumoniae infection.
Assuntos
Antibacterianos/farmacologia , Chlamydophila pneumoniae/efeitos dos fármacos , Citocinas/biossíntese , Levofloxacino , Monócitos/efeitos dos fármacos , Ofloxacino/farmacologia , Linhagem Celular , Hepatócitos/microbiologia , Humanos , Monócitos/imunologia , Monócitos/microbiologiaRESUMO
BACKGROUND: Chlamydia pneumoniae is known to cause acute respiratory infection and more recently it has been studied as a pathogen causing inflammatory changes in chronic diseases such as atherosclerosis. This study addresses the antichlamydial effect of levofloxacin and its role in modulation of a proinflammatory cytokine IL-6 production by uninfected and infected HEp-2 cells. METHODS: HEp-2 cell monolayers were infected with previously prepared and frozen aliquots of C.pneumoniae [1 x 10(3) inclusion-forming units (IFU)/ml] by centrifugation for 30 min and incubation at 37 degrees C for 1 h. Infected monolayers were treated with levofloxacin (3 or 8 microg/ml) immediately after infection (0 h) or 24 h after infection. Monolayers were examined daily for 96 h after infection by counting inclusions with fluorescently labeled antichlamydial monoclonal antibody. Aliquots of disrupted monolayers were titrated to determine the numbers of viable C. pneumoniae IFU/ml. IL-6 concentrations in cell supernatants were determined by ELISA assays. RESULTS: Infected HEp-2 cells produced IL-6. Noninfected HEp-2 cells demonstrated modulation of IL-6 production by levofloxacin. No viable C. Pneumoniae were detected in infected HEp-2 cells when the monolayer was treated with levofloxacin immediately after infection (0 h). In contrast, when cells were treated 24 h after infection, a gradual decline in the number of viable C. pneumoniae occurred; by 96 h into the assay >or=98% of C. pneumoniae were killed. IL-6 concentrations were similar in the supernatants of levofloxacin-treated and nontreated HEp-2 cells. CONCLUSIONS: (1). Levofloxacin is effective in eliminating C. pneumoniae from infected HEp-2 cells; (2). although levofloxacin modulates the production of IL-6 in untreated HEp-2 cells, no evidence for such modulation was observed in HEp-2 cells infected with C. pneumoniae. (3). Presence of viable C. pneumoniae may not be necessary for IL-6 production by infected and treated HEp-2 cells.