RESUMEN

Information related to the clinical characteristics and isolated microbes associated with lung abscesses comparing immunocompromised (IC) to non-immunocompromised (non-IC) patients is limited. A retrospective review for 1984-1996 identified 34 consecutive adult cases of lung abscess (representing 0.2% of all cases of pneumonia), including 10 non-IC and 24 IC patients. Comparison of age, gender, tobacco use, pre-existing pulmonary disease or recognized aspiration risk factors were not significantly different between the two groups. Upper lobe involvement accounted for the majority of cases, although multi-lobe involvement was limited to IC patients. There were no differences in the need for surgical intervention, and mortality was very low for both groups. Anaerobes were the most frequent isolates for non-IC patients (30%), whereas aerobes were the most frequent isolate for IC patients (63%). Importantly, certain organisms were exclusively isolated in the IC group and multiple isolates were obtained only from the IC patients.Thus, comparing non-IC to IC patients, clinical characteristics may be similar whereas important differences may exist in the microbiology associated with lung abscess. These findings have important implications for the clinical management of these patient groups, and support a strategy to aggressively identify microbial agents in abscess material.

Asunto(s)

Huésped Inmunocomprometido , Absceso Pulmonar/microbiología , Adulto , Anciano , Bacterias Aerobias/aislamiento & purificación , Bacterias Anaerobias/aislamiento & purificación , Distribución de Chi-Cuadrado , Femenino , Humanos , Pulmón/diagnóstico por imagen , Absceso Pulmonar/diagnóstico por imagen , Absceso Pulmonar/inmunología , Masculino , Persona de Mediana Edad , Radiografía , Estudios Retrospectivos , Factores de Riesgo , Fumar

RESUMEN

One hallmark of innate immunity apparently conserved from primitive life forms through to humans is the ability of the host to recognize pathogen-associated molecular patterns (PAMPs). Since macrophage pattern recognition receptors are not well defined in Drosophila, we set out to identify such receptors. Our findings reveal that Drosophila macrophages express multiple pattern recognition receptors and that the Drosophila scavenger receptor, dSR-CI, is one such receptor capable of recognizing both gram-negative and gram-positive bacteria, but not yeast. Our data indicate that scavenger receptor bacterial recognition is conserved from insects to humans and may represent one of the most primitive forms of microbial recognition.

Asunto(s)

Proteínas de Drosophila/fisiología , Drosophila melanogaster/fisiología , Bacterias Gramnegativas/metabolismo , Bacterias Grampositivas/metabolismo , Macrófagos/fisiología , Receptores Inmunológicos/fisiología , Secuencia de Aminoácidos , Animales , Células CHO , Células COS , Candida/metabolismo , Células Cultivadas/microbiología , Células Cultivadas/fisiología , Chlorocebus aethiops , Cricetinae , Cricetulus , Drosophila melanogaster/embriología , Drosophila melanogaster/microbiología , Escherichia coli/metabolismo , Evolución Molecular , Proteínas de Insectos/biosíntesis , Proteínas de Insectos/genética , Macrófagos/efectos de los fármacos , Macrófagos/microbiología , Datos de Secuencia Molecular , Fagocitosis/efectos de los fármacos , Polimorfismo Genético , Receptores Depuradores , Proteínas Recombinantes de Fusión/biosíntesis , Receptores Depuradores de Clase C , Especificidad de la Especie , Staphylococcus aureus/metabolismo

RESUMEN

Phagocytosis of extracellular organisms in the alveolar spaces of the lungs represents the first-line of host defense against pulmonary pathogens. Disruption of this process is likely to interfere with the generation of appropriate specific immune responses, and lead to a delayed or inefficient clearance of the pathogen. Pneumocystis carinii, an opportunistic pathogen in immunodeficient individuals, is cleared from the lung by alveolar macrophages. In the absence of specific anti-Pneumocystis antibodies, phagocytosis is dependent on the non-opsonic macrophage mannose receptor (MR). Recent studies have demonstrated that alveolar macrophage MR activity is downregulated in individuals infected with HIV, and that functional MR is shed from the macrophage cell surface. Here we report that P. carinii enhances the formation of soluble MR by macrophages in vitro. Soluble MR was detected in cell-free alveolar fluid from humans infected with HIV and/or P. carinii, but not in alveolar fluid from healthy controls. Soluble MR was found in association with extracellular clumps of P. carinii in the lungs of mice with P. carinii pneumonia, and was associated with P. carinii organisms purified from these mice. When purified P. carinii organisms were incubated with soluble MR-containing supernatants, they were phagocytosed less readily by alveolar macrophages than were control organisms. Our results suggest that P. carinii organisms enhance the shedding of MR from the surface of alveolar macrophages, and that the resultant soluble MR binds to intra-alveolar organisms, thereby interfering with their non-opsonic uptake via the macrophage cell surface MR.

Asunto(s)

Lectinas Tipo C , Macrófagos Alveolares/metabolismo , Lectinas de Unión a Manosa , Pneumocystis/patogenicidad , Neumonía por Pneumocystis/microbiología , Receptores de Superficie Celular/metabolismo , Infecciones Oportunistas Relacionadas con el SIDA/inmunología , Infecciones Oportunistas Relacionadas con el SIDA/metabolismo , Infecciones Oportunistas Relacionadas con el SIDA/microbiología , Adulto , Animales , Líquido del Lavado Bronquioalveolar/química , Línea Celular , Infecciones por VIH/metabolismo , VIH-1 , Humanos , Pulmón/microbiología , Macrófagos Alveolares/inmunología , Macrófagos Alveolares/microbiología , Receptor de Manosa , Ratones , Ratones SCID , Fagocitosis , Pneumocystis/inmunología , Pneumocystis/metabolismo , Neumonía por Pneumocystis/inmunología , Neumonía por Pneumocystis/metabolismo , Solubilidad

RESUMEN

The alveolar macrophage (AM) oxidative burst response is an important component of microbicidal effector cell function against a variety of potential pathogens in the lungs, although the role against Pneumocystis carinii has not been fully investigated. The goals of this study were to characterize the P. carinii-mediated oxidative burst of AMs from healthy individuals, and to examine the oxidative burst of AMs from human immunodeficiency virus (HIV)-infected persons. For healthy individuals, the AM oxidative burst (measured as hydrogen peroxide [H(2)O(2)] production) increased in a time- and concentration-dependent manner in response to P. carinii or to the major surface glycoprotein of P. carinii, gp-A (0.01 to 10 microg/ml), required physical contact of P. carinii with AMs, and was not dependent on organism viability. Enzymatic removal of the surface-associated molecules of P. carinii reduced the oxidative burst to 43% of control (P = 0.01). Blocking the AM mannose receptor reduced the P. carinii-mediated oxidative burst response to 37% of control (P = 0.01). Compared with AMs from healthy individuals, P. carinii-mediated H(2)O(2) production was significantly reduced in AMs from asymptomatic HIV-positive (HIV+) persons with CD4+ counts < 200 cells/mm(3) (249+/-43 relative fluorescence units [RFU] versus 130+/-44 RFU; mean +/- standard error of the mean, P = 0.038) and HIV+ persons with active P. carinii pneumonia (78+/-40 RFU; P = 0.014), but preserved for HIV+ persons with CD4+ counts > 200 cells/mm(3). Importantly, H2O2 production in response to phorbol myristate acetate or serum-opsonized zymosan particles was preserved in all groups studied. Thus, AM oxidative burst, mediated in part via P. carinii gp-A and AM mannose receptor may represent an important host response to P. carinii. A specific impairment of P. carinii-mediated AM oxidative burst in persons with advanced HIV infection may contribute to the pathogenesis of P. carinii pneumonia.

Asunto(s)

Infecciones por VIH/inmunología , Macrófagos Alveolares/inmunología , Pneumocystis/inmunología , Estallido Respiratorio , Animales , Líquido del Lavado Bronquioalveolar , Estudios de Casos y Controles , Infecciones por VIH/patología , Seronegatividad para VIH/inmunología , Humanos , Masculino , Ratas , Ratas Sprague-Dawley , Especies Reactivas de Oxígeno

RESUMEN

STUDY OBJECTIVES: Encompassing periods preceding and following major advances in the diagnosis and management of HIV-related Pneumocystis carinii pneumonia (PCP), the purpose of this study was to determine whether management and outcome patterns of non-HIV PCP parallel the management and outcomes of AIDS-related PCP. DESIGN: Retrospective review of medical records. SETTING: A 375-bed tertiary-care urban teaching hospital and referral center. PATIENTS: All adult patients with morphologically confirmed PCP from 1985 to 1995. MEASUREMENTS AND RESULTS: From 1985 to 1995, 638 confirmed cases of PCP were identified, including 605 cases in 442 HIV-positive persons (HIV + PCP), and 33 cases in 33 non-HIV patients (non-HIV PCP). For HIV + PCP cases, a peak of 104 cases occurred in 1987, with a gradual decline to 23 in 1995. The proportion of cases requiring hospitalization declined from a peak of 91.6% in 1987 to a low of 51.6% in 1992. ICU admission was required for 6.3 to 8.2%, and mechanical ventilation for 4.7 to 5.7%. Overall mortality improved from 11.7 to 6.6%, although mortality for intubated patients remained at 50 to 60%. For the non-HIV PCP cases, 97% occurred from 1989 to 1995 with similar annual frequency, 97% required hospitalization, 69% required ICU admission, and 66% required intubation. Overall mortality was 39%, and mortality for intubated patients was 59%. CONCLUSIONS: Despite major advances in diagnosis and management, PCP remains a significant problem in non-HIV-infected patients, and respiratory failure remains associated with a high mortality rate for patients with both HIV + PCP and non-HIV PCP.

Asunto(s)

Infecciones Oportunistas Relacionadas con el SIDA/terapia , Huésped Inmunocomprometido , Inmunosupresores/uso terapéutico , Neumonía por Pneumocystis/terapia , Respiración Artificial , Infecciones Oportunistas Relacionadas con el SIDA/diagnóstico , Infecciones Oportunistas Relacionadas con el SIDA/mortalidad , Adulto , ADN Viral/análisis , Femenino , VIH/genética , Hospitalización/estadística & datos numéricos , Humanos , Incidencia , Masculino , Persona de Mediana Edad , Neumonía por Pneumocystis/diagnóstico , Neumonía por Pneumocystis/mortalidad , Reacción en Cadena de la Polimerasa , Recurrencia , Estudios Retrospectivos , Tasa de Supervivencia , Resultado del Tratamiento , Población Urbana

RESUMEN

STUDY OBJECTIVES: To assess the potential use of peripheral blood CD4 + T-lymphocyte counts (CD4 + counts) as a clinically useful biological marker to identify specific immunocompromised patients (without HIV infection) at high risk for Pneumocystis carinii pneumonia (PCP). DESIGN: Prospective observational study. SETTING: Three hundred seventy-five-bed tertiary-care urban referral teaching hospital, and 250-bed community-based referral hospital. PATIENTS: One hundred seventy-one consecutive confirmed HIV-seronegative hospitalized and ambulatory adults, including 22 patients with active PCP, 8 patients with bacterial pneumonia, 24 persons in two groups considered at high clinical risk, 38 persons in two groups considered at low or undefined risk, and 79 persons in four groups considered not at risk for PCP (including healthy individuals). MEASUREMENTS AND RESULTS: Compared to counts in healthy individuals, median CD4 + counts were significantly decreased in patients with active PCP (61 cells/microL vs 832 cells/microL; p = 0.001) where 91% of patients had a CD4 + count < 300 cells/microL at the time of PCP diagnosis. Median CD4 + counts were also reduced in the high clinical risk groups of recent organ transplant recipients (117 cells/microL; p = 0.007), 64% with < 300 cells/microL, and patients receiving chemotherapy (221 cells/microL; p<0.01), 80% with < 300 cells/microL. For the low or undefined clinical risk groups, the median CD4 + counts were not significantly reduced, although 39 to 46% of individuals receiving long-term corticosteroid therapy (alone or in combination with other agents) had CD4 + counts < 300 cells/microL. Median CD4 + counts in individuals considered not at risk for PCP were similar to those in healthy subjects. Compared to counts in patients with active PCP, median CD4 + counts were significantly higher in bacterial pneumonia patients (486 cells/microL; p<0.05), but similar to those in healthy subjects. CONCLUSIONS: These data suggest that for immunosuppressed persons without HIV infection (especially in low or undefined PCP risk groups), CD4 + counts may be a useful clinical marker to identify specific individuals at particularly high clinical risk for PCP and may help to guide chemoprophylaxis.

Asunto(s)

Linfocitos T CD4-Positivos/inmunología , Huésped Inmunocomprometido/inmunología , Neumonía por Pneumocystis/inmunología , Adulto , Anciano , Anciano de 80 o más Años , Biomarcadores , Recuento de Linfocito CD4 , Femenino , VIH/inmunología , Anticuerpos Anti-VIH/análisis , Infecciones por VIH/inmunología , Humanos , Masculino , Persona de Mediana Edad , Neumonía por Pneumocystis/sangre , Pronóstico , Estudios Prospectivos , Sensibilidad y Especificidad

RESUMEN

STUDY OBJECTIVE: To examine the safety of bedside percutaneous dilatational tracheostomy in obese patients. DESIGN: Case series of consecutive obese patients (body mass index > or = 27 kg/m(2)) with acute respiratory failure in a medical, cardiac, or surgical ICU unit who required tracheostomy for failure to wean and continued mechanical ventilatory support. RESULTS: Thirteen obese patients were identified and consented to the procedure. Bedside percutaneous dilatational tracheostomy was successfully performed in the ICU for all 13 patients. Procedural complications were limited to paratracheal tracheostomy tube placement in one patient, with immediate identification and appropriate correction. Postprocedural complications were limited to a cuff leak in one patient. CONCLUSION: Bedside percutaneous tracheostomy can be safely performed in obese patients.

Asunto(s)

Cuidados Críticos , Obesidad/fisiopatología , Insuficiencia Respiratoria/terapia , Traqueostomía , Enfermedad Aguda , Adulto , Anciano , Anciano de 80 o más Años , Índice de Masa Corporal , Dilatación , Femenino , Humanos , Masculino , Persona de Mediana Edad , Insuficiencia Respiratoria/etiología , Insuficiencia Respiratoria/fisiopatología , Desconexión del Ventilador

RESUMEN

Local TNF-alpha production in different organs may affect HIV replication and pathogenesis. Alveolar macrophages (AMs) obtained by bronchoalveolar lavage from asymptomatic HIV-seropositive and HIV-seronegative individuals did not spontaneously release TNF-alpha, but LPS stimulation of these cells significantly increased TNF-alpha production. We tested whether NF-kappa B affects TNF-alpha production by AMs using N-tosyl-l -phenylalanine chloromethylketone (TPCK) or N-benzoyl-l -tyrosine ethyl ester (BTEE), which inhibit the degradation of I kappa B, or tricyclodecan-9-yl-xanthogenate-potassium (D609), which inhibits phospholipase C. Alveolar macrophages were exposed to LPS alone and with the chemical protease inhibitors TPCK, BTEE, and D609. NF-kappa B DNA binding induced by LPS treatment of AMs was inhibited by TPCK, BTEE, and D609. These agents also inhibited TNF-alpha mRNA and TNF-alpha protein production. After 24 h, the levels of TNF-alpha mRNA reached equilibrium, as assessed by RT-PCR. The levels of NF-kappa B mRNA remained constant under all conditions. The levels of I kappa B-alpha mRNA were similar after 30, 60, and 180 min, but the I kappa B-beta mRNA concentration was initially low and increased over time under all conditions. I kappa B-alpha and I kappa B-beta protein production was not affected by the chemical protease inhibitors. Our data show that TNF-alpha production by LPS-stimulated AMs from asymptomatic HIV-seropositive and -seronegative individuals is regulated via the phospholipase C pathway and by NF-kappa B DNA binding activity without obvious changes in I kappa B-alpha or I kappa B-beta protein concentrations.

Asunto(s)

Seropositividad para VIH/inmunología , Proteínas I-kappa B , Macrófagos Alveolares/inmunología , Macrófagos Alveolares/metabolismo , FN-kappa B/fisiología , Factor de Necrosis Tumoral alfa/biosíntesis , Adulto , Adhesión Celular/genética , Adhesión Celular/inmunología , Proteínas de Unión al ADN/metabolismo , Femenino , Humanos , Masculino , Persona de Mediana Edad , Inhibidor NF-kappaB alfa , FN-kappa B/genética , FN-kappa B/metabolismo , Reacción en Cadena de la Polimerasa , Etiquetado in Situ Primed , Estudios Prospectivos , ARN Mensajero/antagonistas & inhibidores , ARN Mensajero/biosíntesis , Inhibidores de Serina Proteinasa/farmacología , Transducción de Señal/efectos de los fármacos , Transducción de Señal/inmunología , Clorometilcetona de Tosilfenilalanila/farmacología , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Factor de Necrosis Tumoral alfa/genética , Fosfolipasas de Tipo C/antagonistas & inhibidores , Tirosina/análogos & derivados , Tirosina/farmacología

RESUMEN

The relationship of serum human immunodeficiency virus-1 (HIV-1) RNA levels to HIV-1 RNA levels in other compartments, such as the lungs, is not well characterized. The purpose of this study was to determine the viral burden of HIV-1 in the lungs by comparing HIV-1 RNA in cell-free bronchoalveolar lavage fluid (BALF) with that in serum. Specimens were examined from 77 HIV-seropositive adults (CD4(+) cell counts: 0 to 700 cells/mm(3); 48% receiving prescribed antiretroviral agents), comprising 43 asymptomatic individuals who were compared with 34 persons with active lung disease caused by Pneumocystis carinii (n = 26), bacteria (n = 3), Mycobacterium avium complex (n = 2), Nocardia sp. (n = 1), Aspergillus sp. (n = 1), or pulmonary Kaposi's sarcoma (n = 1). For serum HIV-1 RNA, the proportion of subjects with detectable levels and the mean values were similar for asymptomatic individuals and persons with active lung disease (85% versus 86%, respectively) (6.64 x 10(4) versus 1. 81 x 10(5) HIV-1 RNA copies/ml; p = 0.13). In contrast, HIV-1 RNA in BALF was more often detected (16% versus 62%; p = 0.001), and mean values were higher (1.04 x 10(5) versus 3.31 x 10(6) HIV-1 RNA copies/ml; p = 0.032), in subjects with active lung disease than in asymptomatic subjects, independent of early or advanced clinical stages of HIV-related disease. For both study groups, HIV-1 RNA levels in BALF exceeded those in serum in 56% of cases by up to 66-fold, and did not correlate with local levels of tumor necrosis factor-alpha, granulocyte-macrophage colony-stimulating factor, or interleukin-16. HIV-1 proviral DNA in cells from BALF was detected in up to 86% of subjects, more frequently in persons with advanced HIV disease (p = 0.0496), and often involved > 10% of BALF cells, but did not correlate with HIV-1 RNA detected in BALF. These data provide evidence for active HIV-1 replication in the lungs. HIV-1 replication is compartmentalized relative to serum, may be restricted, is independent of HIV-1 proviral DNA and clinical stage of HIV, and may be influenced by pulmonary disease such as P. carinii pneumonia or by other local or lung-specific factors. The lungs represent a large reservoir for HIV-1, and may present a source of persistent HIV-1 replication even during periods of apparent clinical latency of HIV-1 infection.

Asunto(s)

Infecciones Oportunistas Relacionadas con el SIDA/virología , Infecciones por VIH/virología , VIH-1/fisiología , Pulmón/virología , Neumonía por Pneumocystis/virología , Replicación Viral , Adulto , Anciano , Líquido del Lavado Bronquioalveolar/virología , Recuento de Linfocito CD4 , Citocinas/análisis , ADN Viral/análisis , Femenino , Infecciones por VIH/complicaciones , Infecciones por VIH/inmunología , VIH-1/aislamiento & purificación , Humanos , Neoplasias Pulmonares/complicaciones , Neoplasias Pulmonares/virología , Masculino , Persona de Mediana Edad , Infección por Mycobacterium avium-intracellulare/virología , Neumonía Bacteriana/virología , ARN Viral/análisis , ARN Viral/sangre , Sarcoma de Kaposi/complicaciones , Sarcoma de Kaposi/virología , Tuberculosis Pulmonar/virología

RESUMEN

Elevated concentrations of ambient air particles can result in increased mortality and morbidity, especially in people with preexisting pulmonary disease. We postulate that in the inflammatory milieu of diseased lungs, alveolar macrophages (AMs) may be primed for enhanced responses to stimuli such as inhaled air particles. To test this hypothesis in vitro, we first cultured normal AMs with or without lipopolysaccharide (LPS). We then incubated the cells with particle suspensions (urban air particles (UAP, Washington, D.C.), residual oil fly ash (ROFA), concentrated respirable-size (PM2.5) air particles (CAPs), and inert TiO2) and compared rat and human AM production of the critical proinflammatory mediator, tumor necrosis factor (TNF). LPS priming amplified TNF production by both rat and human AMs in response to UAP and CAPs but not inert TiO2. There were also differences observed between rat and human AM responses to particle suspensions. Striking changes seen only in rat were cytotoxic effects of ROFA and diminished particle uptake in response to LPS priming. The potency of CAPs samples (which are collected on separate days) varied when comparing one day's sample with another. When centrifuged, the majority of bioactivity seen in particle suspensions (TNF release) remained within the pelleted fraction while the supernatant showed minimal bioactivity. The data suggest that AMs activated by extant pulmonary inflammation may promote further inflammation by an enhanced cytokine response to inhaled ambient air particles.

Asunto(s)

Contaminantes Atmosféricos/toxicidad , Lipopolisacáridos/farmacología , Pulmón/metabolismo , Macrófagos Alveolares/fisiología , Factor de Necrosis Tumoral alfa/metabolismo , Animales , Femenino , Citometría de Flujo , Humanos , Técnicas In Vitro , Pulmón/efectos de los fármacos , Macrófagos Alveolares/efectos de los fármacos , Tamaño de la Partícula , Fagocitosis/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Especificidad de la Especie

RESUMEN

Alveolar macrophages (AMs) avidly bind and ingest unopsonized environmental particles and bacteria through scavenger-type receptors (SRs). AMs from mice with a genetic deletion of the major macrophage SR (types AI and AII; SR-/-) showed no decrease in particle binding compared with SR+/+ mice, suggesting that other SRs are involved. To identify these receptors, we generated a monoclonal antibody (mAb), PAL-1, that inhibits hamster AM binding of unopsonized particles (TiO2, Fe2O3, and latex beads; 66 +/- 5, 77 +/- 2, and 85 +/- 2% inhibition, respectively, measured by flow cytometry). This antibody identifies a protein of approximately 70 kD on the AM surface (immunoprecipitation) that is expressed by AMs and other macrophages in situ. A cDNA clone encoding the mAb PAL-1-reactive protein isolated by means of COS cell expression was found to be 84 and 77% homologous to mouse and human scavenger receptor MARCO mRNA, respectively. Transfection of COS cells with MARCO cDNA conferred mAb-inhibitable TiO2 binding. Hamster MARCO also mediates AM binding of unopsonized bacteria (67 +/- 5 and 47 +/- 4% inhibition of Escherichia coli and Staphylococcus aureus binding by mAb PAL-1). A polyclonal antibody to human MARCO identified the expected approximately 70-kD band on Western blots of lysates of normal bronchoalveolar lavage (BAL) cells (>90% AMs) and showed strong immunolabeling of human AMs in BAL cytocentrifuge preparations and within lung tissue specimens. In normal mouse AMs, the anti-MARCO mAb ED31 also showed immunoreactivity and inhibited binding of unopsonized particles (e.g., TiO2 approximately 40%) and bacteria. The novel function of binding unopsonized environmental dusts and pathogens suggests an important role for MARCO in the lungs' response to inhaled particles.

Asunto(s)

Macrófagos Alveolares/metabolismo , Proteínas de la Membrana , Receptores Inmunológicos/fisiología , Receptores de Lipoproteína , Secuencia de Aminoácidos , Animales , Anticuerpos Monoclonales/inmunología , Secuencia de Bases , Células COS , Clonación Molecular , Cricetinae , ADN Complementario , Escherichia coli/metabolismo , Humanos , Ratones , Datos de Secuencia Molecular , Pruebas de Precipitina , Cuarzo/metabolismo , Receptores Inmunológicos/genética , Receptores Inmunológicos/metabolismo , Receptores Depuradores , Receptores Depuradores de Clase B , Staphylococcus aureus/metabolismo , Titanio/metabolismo

RESUMEN

Alveolar macrophages (AM) are important host-defense cells and targets of human immunodeficiency virus type 1 (HIV-1) infection. However, the receptors mediating HIV-1 entry into AM are not completely characterized. We observed that, in addition to CD4 receptors, AM from healthy adults expressed low levels of CCR5, CCR3, and CXCR4 chemokine receptors by flow cytometry, and specific messenger RNA was detected for all three receptors by reverse transcriptase/polymerase chain reaction. The macrophage monocytotropic (M-tropic; YU2) and dual-tropic (89.6) HIV-1 env-pseudotypes entered AM efficiently, as expected given CCR3 and CCR5 expression. However, the T-lymphocytotropic (T-tropic; HXB2) pseudotype did not enter AM despite expression of the appropriate chemokine coreceptor CXCR4. Incubation of AM with regulated on activation, normal T cells expressed and secreted (RANTES) significantly impaired entry of the M-tropic (YU2) HIV-1 pseudotype, whereas SDF-1beta or eotaxin did not impair entry. The entry of simian immunodeficiency virus (SIV) pbj1.9 env-pseudotype into AM was not blocked by RANTES, SDF-1beta, or eotaxin. The competence of these chemokine receptors for virus entry was confirmed in Cf2Th canine thymocytes cotransfected with the human CD4 and chemokine receptors. Entry of the M-tropic (YU2) HIV-1 pseudotype was shown to be mediated by either CCR3 or CCR5, the T-tropic (HXB2) HIV-1 pseudotype by CXCR4, and the dual-tropic (89.6) HIV-1 or the SIVpbj1. 9 pseudotype by CCR5, CCR3, or CXCR4. Our data indicate that the mechanisms for HIV-1 entry are both receptor-specific and cell type-specific, and that chemokine receptor expression on AM does not fully explain cell susceptibility to different virus isolates.

Asunto(s)

Antígenos CD4/fisiología , VIH-1/fisiología , Macrófagos Alveolares/virología , Receptores de Quimiocina/metabolismo , Adulto , Animales , Secuencia de Bases , Línea Celular , Cloranfenicol O-Acetiltransferasa/genética , Cartilla de ADN , Perros , Femenino , Citometría de Flujo , Humanos , Macrófagos Alveolares/metabolismo , Masculino , Virus de la Inmunodeficiencia de los Simios/fisiología , Especificidad de la Especie

RESUMEN

The innate immune system evolved to protect the host in the early phases of an infectious challenge. The soluble mannose binding protein, and the cell surface mannose receptor are two key pattern recognition molecules of innate immunity. The ligand binding specificity of these molecules enables them to differentiate 'self' from 'non-self'. These pattern recognition capabilities are coupled to effector functions, which enable them to interact with other molecules of the immune system. In this way, these pattern recognition molecules are able to serve as a link between the innate and adaptive immune systems.

Asunto(s)

Proteínas Portadoras/fisiología , Inmunidad , Infecciones/inmunología , Lectinas Tipo C , Receptores de Superficie Celular/fisiología , Animales , Activación de Complemento , Humanos , Receptor de Manosa , Lectinas de Unión a Manosa

RESUMEN

The macrophage mannose receptor, a pattern recognition molecule and component of innate immunity, mediates binding and phagocytosis of Pneumocystis carinii and likely represents an important clearance mechanism in the lungs of immunocompetent hosts. The purpose of this study was to examine the ability of alveolar macrophages from HIV-infected individuals to bind and phagocytose P. carinii, and to investigate the role of the macrophage mannose receptor in mediating this interaction. Compared with healthy individuals, alveolar macrophage phagocytosis of P. carinii from HIV+ persons was reduced up to 74% (P = 0.02), primarily reflecting a reduction in the number of organisms associated with each macrophage (P = 0.019). Furthermore, macrophages from HIV+ individuals demonstrated up to an 80% (P < 0.05) reduction in mannose receptor surface expression and endocytosis. Mannose receptor affinity was unaltered, and mRNA levels were modestly reduced (P < 0.05). Cells from HIV+ individuals with CD4(+) counts < 200 cells/mm3 (representing individuals at high clinical risk for P. carinii pneumonia) demonstrated the lowest levels of P. carinii phagocytosis and mannose receptor endocytosis. In vitro HIV infection of alveolar macrophages from healthy individuals reduced mannose receptor endocytosis to 53.2% (P < 0.05) and P. carinii binding and phagocytosis to 67.4% (P < 0.05) of control. Our studies suggest that HIV infection may alter innate immunity in the lungs, and that impaired alveolar macrophage mannose receptor-mediated binding and phagocytosis of P. carinii may contribute to the susceptibility of HIV-infected individuals to this opportunistic pulmonary pathogen.

Asunto(s)

Seropositividad para VIH/fisiopatología , VIH-1 , Lectinas Tipo C , Macrófagos Alveolares/microbiología , Macrófagos Alveolares/fisiología , Lectinas de Unión a Manosa , Fagocitosis , Pneumocystis/fisiología , Receptores de Superficie Celular/biosíntesis , Líquido del Lavado Bronquioalveolar/citología , Broncoscopía , Recuento de Linfocito CD4 , Adhesión Celular , Regulación hacia Abajo , Seronegatividad para VIH , Seropositividad para VIH/inmunología , Seropositividad para VIH/microbiología , Humanos , Receptor de Manosa , ARN Mensajero/biosíntesis , Receptores de Superficie Celular/genética , Valores de Referencia , Transcripción Genética

Asunto(s)

Bacteriemia/microbiología , Choque Séptico/microbiología , Infecciones Estafilocócicas , Staphylococcus/aislamiento & purificación , Adulto , Bacteriemia/diagnóstico , Femenino , Humanos , Choque Séptico/diagnóstico , Infecciones Estafilocócicas/diagnóstico

RESUMEN

Surfactant protein-A (SP-A) levels are increased in Pneumocystis carinii pneumonia, but the role of SP-A in the pathogenesis of P. carinii pneumonia is not completely understood. This study investigated the effect of SP-A on the in vitro binding and phagocytosis of P. carinii by normal human alveolar macrophages (AM). Determination of binding and phagocytosis was done with a fluorescence-based assay, utilizing fluorescein isothiocyanate (FITC)-labeled P. carinii. Binding and phagocytosis of P. carinii to AM correlated inversely with the levels of SP-A present on the surface of the organisms (r = -0.6323, P = 0.0086; and r = -0.9827, P < 0.0001, respectively). The addition of exogenous SP-A to organisms with low surface-associated SP-A reduced P. carinii binding by 30% (P < 0.05) and reduced phagocytosis by 20% (P < 0.05), whereas this effect was reversed with ethylenediamine tetraacetic acid (EDTA) or anti-SP-A antibody. Furthermore, binding and phagocytosis were enhanced after enzymatic removal of P. carinii surface-associated SP-A, and this effect was reversed with the addition of exogenous SP-A. The observed inhibitory effect of SP-A on P. carinii binding and phagocytosis reflected binding of SP-A to the organisms rather than a direct effect of SP-A on the macrophages. These data suggest that increased levels of SP-A may contribute to the pathogenesis of P. carinii pneumonia through binding to the surface of the organism and interfering with AM recognition of this opportunistic pulmonary pathogen.

Asunto(s)

Macrófagos Alveolares/fisiología , Pneumocystis/metabolismo , Proteolípidos/metabolismo , Surfactantes Pulmonares/metabolismo , Candida albicans/metabolismo , Células Cultivadas , Ácido Edético/farmacología , Glicoproteínas/inmunología , Glicoproteínas/metabolismo , Glicoproteínas/farmacología , Humanos , Hidrolasas/farmacología , Inmunoglobulina G/farmacología , Macrófagos Alveolares/efectos de los fármacos , Fagocitosis/efectos de los fármacos , Proteolípidos/inmunología , Proteolípidos/farmacología , Proteína A Asociada a Surfactante Pulmonar , Proteínas Asociadas a Surfactante Pulmonar , Surfactantes Pulmonares/inmunología , Surfactantes Pulmonares/farmacología , Factores de Tiempo

RESUMEN

STUDY OBJECTIVE: To examine the use of adjunctive corticosteroids in cases of severe Pneumocystis carinii pneumonia (PCP) in non-HIV-infected adult patients. DESIGN: Retrospective review of medical records. SETTING: Tertiary care urban teaching hospital. PATIENTS: Review identified 31 consecutive histologically confirmed primary cases of adult non-HIV-related PCP. Complete records were available for 30 patients, including 20 male and 10 female patients with a mean age of 58.3+/-15 years (+/-SD). Underlying conditions included organ transplantation (n=13), long-term immunosuppressive therapy (n=9), and chemotherapy for malignancy (n=8). All patients had documented PO2 <65 mm Hg or arterial oxygen saturation <90% on room air. INTERVENTIONS: Following the identification of P carinii, in addition to trimethoprim-sulfamethoxazole or pentamidine therapy, 16 patients received increased steroids (> or =60 mg prednisone daily equivalent; increased high-dose steroid group), whereas 14 patients were maintained on a regimen of low doses (< or =30 mg prednisone equivalent daily) or had steroid therapy tapered (low-dose steroid group). RESULTS: The increased high-dose steroid group demonstrated a shorter required duration for mechanical ventilation (6.3+/-6 days vs 18.0+/-21 days; p=0.047), a shorter duration of ICU admission (8.5+/-7 days vs 15.8+/-8 days; p=0.025), and a shorter duration of supplemental oxygen use (10.0+/-4 vs 32.2+/-33; p=0.05). The hospital duration to discharge for the nine survivors in each group favored the use of corticosteroids (15.4+/-5 days vs 36.3+/-33 days; p=0.077). Similar rates were observed for intubation (75% vs 57%; p=0.442) and in-hospital mortality (44% vs 36%; p=0.722). CONCLUSIONS: These preliminary data suggest that high-dose adjunctive corticosteroids may accelerate recovery in cases of severe adult non-HIV PCP.

Asunto(s)

Glucocorticoides/uso terapéutico , Infecciones Oportunistas/tratamiento farmacológico , Neumonía por Pneumocystis/tratamiento farmacológico , Prednisona/uso terapéutico , Antiinfecciosos/uso terapéutico , Antifúngicos/uso terapéutico , Quimioterapia Combinada , Femenino , Glucocorticoides/administración & dosificación , Infecciones por VIH , Humanos , Intubación Intratraqueal , Masculino , Persona de Mediana Edad , Infecciones Oportunistas/complicaciones , Infecciones Oportunistas/epidemiología , Infecciones Oportunistas/microbiología , Pentamidina/uso terapéutico , Neumonía por Pneumocystis/complicaciones , Neumonía por Pneumocystis/epidemiología , Prednisona/administración & dosificación , Respiración Artificial , Insuficiencia Respiratoria/epidemiología , Insuficiencia Respiratoria/terapia , Estudios Retrospectivos , Resultado del Tratamiento , Combinación Trimetoprim y Sulfametoxazol/uso terapéutico

RESUMEN

Legionella micdadei (Pittsburgh pneumonia agent) is the second most common cause of Legionella pneumonia, and occurs predominantly in immunocompromised hosts. L micdadei is the cause of nosocomial pneumonia in renal transplant recipients, but has not been described in other adult solid organ transplant recipients. This report describes the first case of L micdadei pneumonia in an adult liver transplant recipient on immunosuppressive therapy. Importantly, this case highlights the difficulties in establishing the diagnosis, as the Legionella urinary antigen is negative, and special culture conditions are required. Furthermore, this case illustrates several atypical clinical features of L micdadei pneumonia in a transplant recipient, including a community acquired mode of transmission, occurrence several years after organ transplantation, and lung abcess formation. The patient was successfully treated with limited surgical resection and quinolone antimicrobial monotherapy.

Asunto(s)

Legionelosis/complicaciones , Trasplante de Hígado , Absceso Pulmonar/complicaciones , Neumonía Bacteriana/complicaciones , Complicaciones Posoperatorias , Adulto , Antiinfecciosos/uso terapéutico , Líquido del Lavado Bronquioalveolar/microbiología , Ciprofloxacina/uso terapéutico , Diabetes Mellitus Tipo 2/complicaciones , Hepatitis B/complicaciones , Humanos , Inmunosupresores/uso terapéutico , Masculino , Tacrolimus/uso terapéutico , Tomografía Computarizada por Rayos X

RESUMEN

Pneumocystis carinii lipids are similar to host lipids, but it is not known if some of these lipids are acquired from host cells. The ability of P. carinii to incorporate a fluorescent fatty acid analogue (Bodipy-C12) was analyzed, the metabolism of the incorporated lipid by P. carinii was characterized, and lipid transfer from human alveolar epithelial cells (A549) to P. carinii was investigated. Both P. carinii and A549 cells incorporated exogenous Bodipy-C12 in a concentration-dependent manner. Biochemical analysis of labeled P. carinii revealed incorporation of Bodipy-C12 into complex lipid classes. Incubation of unlabeled P. carinii with Bodipy-C12-labeled A549 cells demonstrated lipid transfer to P. carinii, a process facilitated by attachment. These data suggest that P. carinii can incorporate and modify an exogenous fluorescent lipid. The observed transfer of lipid from A549 cells to P. carinii provides important insight into the interaction of this organism with alveolar epithelial cells.

Asunto(s)

Lípidos de la Membrana/metabolismo , Pneumocystis/metabolismo , Animales , Compuestos de Boro , Células Cultivadas , Epitelio/parasitología , Ácidos Grasos/metabolismo , Humanos , Alveolos Pulmonares/parasitología , Ratas

RESUMEN

OBJECTIVE: The purpose of this study was to investigate the in vitro processing of Cryptococcus neoformans by human alveolar macrophages from HIV-seropositive individuals compared with HIV-seronegative individuals. DESIGN AND METHODS: Bronchalveolar lavage (BAL) was performed on smoking and nonsmoking HIV-seropositive and seronegative volunteers. Lavage cells from the four groups were challenged in vitro with C. neoformans and assessed for fungal binding, phagocytosis, and growth inhibition. RESULTS: The results indicated that BAL cells from the smoking HIV-infected group had increased fungistatic activity compared with HIV-seronegative smokers (mean percentage growth inhibition +/- SD, 77.5 +/- 14.2 versus 59.1 +/- 16.6%; P = 0.015). However, late-staged HIV-infected patients (Centers for Disease Control and Prevention class C3) were found to have decreased fungistasis compared with early stage A patients (63.8 +/- 11.1 versus 83.0 +/- 2.2%; P < 0.05). BAL cells recovered from HIV- seronegative smoking volunteers demonstrated reduced fungistatic activity when compared with BAL cells from HIV- seronegative nonsmokers (56.8 +/- 8.8 versus 83.0 +/- 2.2%; P < 0.001). Smoking also induced a decrease in internalization of C. neoformans by alveolar macrophages as assessed by confocal laser microscopy in both HIV-seronegative and HIV-infected groups. CONCLUSION: We conclude that BAL cells from early stage HIV-1-infected individuals do not have an intrinsic defect in fungistasis of C. neoformans. In fact, it appears that BAL cells from HIV-seropositive people are activated for fungistasis in early HIV infection, although fungistatic activity declines as the disease progresses. Incidentally noted was the finding that smoking decreases the internalization of C. neoformans in both HIV-infected and HIV-seronegative individuals, suggesting the possibility that smoking might enhance the susceptibility to cryptococcosis.

Asunto(s)

Cryptococcus neoformans , Infecciones por VIH/inmunología , Macrófagos Alveolares/inmunología , Fagocitosis , Células Cultivadas , Cryptococcus neoformans/crecimiento & desarrollo , Cryptococcus neoformans/inmunología , Infecciones por VIH/patología , Humanos