RESUMO
Pneumocystis pneumonia (PCP) is a major cause of morbidity and mortality in patients with HIV infection. CD4(+) T lymphocytes are critical for host defense against this infection, but in the absence of CD4(+) T lymphocytes, CD8(+) T lymphocytes may provide limited host defense. The cytokine interleukin-7 (IL-7) functions to enhance lymphocyte proliferation, survival, and recruitment of immune cells to sites of infection. However, there is little known about the role of IL-7 in PCP or its potential use as an immunotherapeutic agent. We hypothesized that treatment with recombinant human IL-7 (rhIL-7) would augment host defense against Pneumocystis and accelerate pathogen clearance in CD4-depleted mice. Control and CD4-depleted mice were infected with Pneumocystis, and rhIL-7 was administered via intraperitoneal injection. Our studies indicate that endogenous murine IL-7 is part of the normal host response to Pneumocystis murina and that administration of rhIL-7 markedly enhanced clearance of Pneumocystis in CD4-depleted mice. Additionally, we observed increased recruitment of CD8(+) T lymphocytes to the lungs and decreased apoptosis of pulmonary CD8(+) T lymphocytes in rhIL-7-treated animals compared to those in untreated mice. The antiapoptotic effect of rhIL-7 was associated with increased levels of Bcl-2 protein in T lymphocytes. rhIL-7 immunotherapy in CD4-depleted mice also increased the number of gamma interferon (IFN-γ)-positive CD8(+) central memory T lymphocytes in the lungs. We conclude that rhIL-7 has a potent therapeutic effect in the treatment of murine Pneumocystis pneumonia in CD4-depleted mice. This therapeutic effect is mediated through enhanced recruitment of CD8(+) T cells and decreased apoptosis of lung T lymphocytes, with a preferential action on central memory CD8(+) T lymphocytes.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Interleucina-7/uso terapêutico , Depleção Linfocítica , Pneumonia por Pneumocystis/imunologia , Animais , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Feminino , Interferon gama/imunologia , Pulmão/imunologia , Ativação Linfocitária/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Pneumocystis/imunologia , Pneumocystis/patogenicidade , Pneumonia por Pneumocystis/tratamento farmacológico , Pneumonia por Pneumocystis/microbiologia , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Proteínas Recombinantes/uso terapêuticoRESUMO
Bacterial pneumonia is an increasing complication of HIV infection and inversely correlates with the CD4(+) lymphocyte count. Interleukin (IL)-17 is a cytokine produced principally by CD4(+) T cells, which induces granulopoiesis via granulocyte colony-stimulating factor (G-CSF) production and induces CXC chemokines. We hypothesized that IL-17 receptor (IL-17R) signaling is critical for G-CSF and CXC chemokine production and lung host defenses. To test this, we used a model of Klebsiella pneumoniae lung infection in mice genetically deficient in IL-17R or in mice overexpressing a soluble IL-17R. IL-17R-deficient mice were exquisitely sensitive to intranasal K. pneumoniae with 100% mortality after 48 h compared with only 40% mortality in controls. IL-17R knockout (KO) mice displayed a significant delay in neutrophil recruitment into the alveolar space, and had greater dissemination of K. pneumoniae compared with control mice. This defect was associated with a significant reduction in steady-state levels of G-CSF and macrophage inflammatory protein (MIP)-2 mRNA and protein in the lung in response to the K. pneumoniae challenge in IL-17R KO mice. Thus, IL-17R signaling is critical for optimal production of G-CSF and MIP-2 and local control of pulmonary K. pneumoniae infection. These data support impaired IL-17R signaling as a potential mechanism by which deficiency of CD4 lymphocytes predisposes to bacterial pneumonia.
Assuntos
Quimiocinas CXC/metabolismo , Fator Estimulador de Colônias de Granulócitos/metabolismo , Pulmão/metabolismo , Neutrófilos/citologia , Receptores de Interleucina/metabolismo , Proteínas Recombinantes/metabolismo , Transdução de Sinais , Animais , Líquido da Lavagem Broncoalveolar , Infecções por Klebsiella/imunologia , Klebsiella pneumoniae/isolamento & purificação , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Receptores de Interleucina/genética , Receptores de Interleucina-17 , Proteínas Recombinantes/genéticaRESUMO
Host defenses are profoundly compromised in HIV-infected hosts due to progressive depletion of CD4+ T lymphocytes. Moreover, deficient CD4+ T lymphocytes impair vaccination approaches to prevent opportunistic infection. Therefore, we investigated a CD4+ T cell-independent vaccine approach to a prototypic AIDS-defining infection, Pneumocystis carinii (PC) pneumonia. Here, we demonstrate that bone marrow-derived dendritic cells (DCs) expressing the murine CD40 ligand, when pulsed ex vivo by PC antigen, elicited significant titers of anti-PC IgG in CD4-deficient mice. Vaccinated animals demonstrated significant protection from PC infection, and this protection was the result of an effective humoral response, since adoptive transfer of CD4-depleted splenocytes or serum conferred this protection to CD4-deficient mice. Western blot analysis of PC antigen revealed that DC-vaccinated, CD4-deficient mice predominantly reacted to a 55-kDa PC antigen. These studies show promise for advances in CD4-independent vaccination against HIV-related pathogens.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Vacinas Fúngicas/administração & dosagem , Pneumocystis/imunologia , Pneumonia por Pneumocystis/prevenção & controle , Transferência Adotiva , Animais , Sequência de Bases , Primers do DNA , Ativação Linfocitária , Masculino , Camundongos , Camundongos Endogâmicos BALB CRESUMO
Tumor necrosis factor alpha has potent immunomodulatory and antitumor activity, but its therapeutic applications may be limited by its significant host toxicity. We showed that liposome-encapsulated recombinant human tumor necrosis factor alpha (rHuTNF-alpha) retained full anticellular activity in vitro. We then assessed the immunomodulatory and toxic effects of two different doses of i.v. free or liposome-encapsulated rHuTNF-alpha in normal rats. Both free and liposome-encapsulated rHuTNF-alpha significantly enhanced alveolar macrophage- and blood monocyte-mediated interleukin 1 release and tumor cell lysis, as well as natural killer cell cytotoxicity, when compared to buffer-treated controls. However, administration of rHuTNF-alpha in liposomes substantially reduced tumor necrosis factor alpha-mediated toxicity. Animals receiving liposome-encapsulated rHuTNF-alpha showed significantly less tissue injury, gastric retention, and circulating leukocyte shifts than animals receiving free rHuTNF-alpha. In addition, liposome-based delivery significantly increased lung and liver uptake of rHuTNF-alpha. Therefore, liposome-encapsulated rHuTNF-alpha retains immunomodulatory activity, significantly reduces toxic inflammatory effects, and may allow targeting of tumor necrosis factor alpha to selected organs after i.v. administration.
Assuntos
Adjuvantes Imunológicos/farmacologia , Fator de Necrose Tumoral alfa/farmacologia , Animais , Portadores de Fármacos , Células Matadoras Naturais/efeitos dos fármacos , Células Matadoras Naturais/imunologia , Lipossomos , Masculino , Ratos , Ratos Endogâmicos F344 , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/farmacologia , Proteínas Recombinantes/toxicidade , Distribuição Tecidual , Fator de Necrose Tumoral alfa/administração & dosagem , Fator de Necrose Tumoral alfa/toxicidadeRESUMO
SETTING: The ability of chemokines such as macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, and regulated-upon-activation, normal T cell expressed and secreted (RANTES), to attract and activate T cells and monocytes, the building blocks of the granuloma, suggests that these chemokines may have a role in modulating immune responses to Mycobacterium tuberculosis infection. OBJECTIVE: We hypothesized that the chemokine receptor 5 (CCR5) ligands, MIP-1alpha, MIP-1beta and RANTES, are virulence correlates in M. tuberculosis infection and are indispensable to granuloma formation. DESIGN: The ability of virulent (H37Rv) and avirulent (H37Ra) strains of M. tuberculosis to induce chemokine production in vivo and in vitro was determined at protein and mRNA levels. We also compared bacterial burden, and granuloma numbers and size in H37Rv-infected CCR5-/- or wild-type C57BL/6 mice. RESULTS: In vivo, lung mRNA and protein measurements of MIP-1alpha, MIP-1beta and RANTES indicate significantly higher (p<0.05) values (days 14-28) in the H37Rv-infected than the H37Ra-infected mice. This is consistent with a higher infection burden of the virulent strain. However, in vitro alveolar macrophage stimulation by H37Rv or H37Ra yielded no significant differences in production of the three chemokines at all time points. Histological analysis of granulomas did not show any significant differences in granuloma numbers, size and M. tuberculosis growth in CCR5-/- compared to wild-type mice. CONCLUSIONS: The production of the CCR5 ligands, MIP-1alpha, MIP-1beta, and RANTES, does not clearly correlate with virulence of M. tuberculosis. These ligands and their receptors may not be indispensable to the development of granulomas in murine tuberculosis.
Assuntos
Mycobacterium tuberculosis/patogenicidade , Receptores CCR5/imunologia , Tuberculose/imunologia , Animais , Células Cultivadas , Quimiocina CCL3 , Quimiocina CCL4 , Quimiocina CCL5/biossíntese , Quimiocina CCL5/genética , Regulação da Expressão Gênica , Granuloma/imunologia , Granuloma/microbiologia , Granuloma/patologia , Ligantes , Pulmão/imunologia , Proteínas Inflamatórias de Macrófagos/biossíntese , Proteínas Inflamatórias de Macrófagos/genética , Macrófagos Alveolares/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , RNA Mensageiro/genética , Tuberculose/microbiologia , VirulênciaRESUMO
Alcohol (EtOH) is a well-documented immunosuppressant. Acute EtOH-induced immunosuppression is partially due to suppression of tumor necrosis factor alpha (TNF-alpha) secretion. We investigated the mechanism of acute EtOH-induced TNF-alpha suppression in two monocytic cell lines, Mono Mac 6 and DRM. EtOH inhibited TNF-alpha secretion in a dose-dependent manner. However, TNF-alpha transcription was not affected by EtOH. Enzyme-linked immunosorbent assay and confocal microscopy showed that EtOH treatment increased cell-associated TNF-alpha. Ectodomain shedding of TNF-alpha from the cell surface is mediated by TNF-alpha converting enzyme (TACE). In contrast with TNF-alpha, EtOH did not inhibit interleukin-8 (IL-8) secretion, which does not require shedding. Furthermore, TNF p75 receptor shedding, a biomarker for TACE activity, was inhibited by EtOH in both cell lines. EtOH also inhibited TNF p75 receptor shedding in TACE-reconstituted fibroblasts, suggesting that EtOH inhibits the shedding process. These data show that acute EtOH exposure can posttranscriptionally suppress TNF-alpha production, resulting in specific defects in immune defense.
Assuntos
Etanol/metabolismo , Imunossupressores/metabolismo , Metaloendopeptidases/metabolismo , Processamento de Proteína Pós-Traducional/efeitos dos fármacos , Fator de Necrose Tumoral alfa/metabolismo , Proteínas ADAM , Proteína ADAM17 , Antígenos CD/metabolismo , Linhagem Celular , Etanol/farmacologia , Fibroblastos/citologia , Citometria de Fluxo/métodos , Expressão Gênica/efeitos dos fármacos , Imunossupressores/farmacologia , Interleucina-8/metabolismo , Líquido Intracelular/metabolismo , Metaloendopeptidases/genética , Microscopia Confocal/métodos , Monócitos/efeitos dos fármacos , Monócitos/metabolismo , RNA Mensageiro , Receptores do Fator de Necrose Tumoral/metabolismo , Receptores Tipo II do Fator de Necrose Tumoral , Transfecção , Fator de Necrose Tumoral alfa/genéticaRESUMO
E1-deleted adenoviral vectors are increasingly being utilized for in vivo gene transfer. The potential use of these vectors is limited by transient expression of the transgene and a markedly reduced rate of transduction following readministration, presumably due to a host immune response to the vector. We hypothesized that CD4+ lymphocytes are necessary to generate an immune response to these vectors and that administration of a depleting anti-CD4 antibody (GK1.5) might prolong transgene expression in vivo. We found that pretreatment of mice with a single injection (transient depletion) or weekly injections of GK1.5 (persistent depletion), markedly prolonged expression of an adenovirus-encoded tumor necrosis factor (TNF) inhibitor or luciferase gene compared to controls. Moreover, mice treated with GK1.5 showed no antiadenoviral antibody response to repeat administration of the vector and a second adenoviral transgene could be expressed in these animals. However, control mice developed a significant neutralizing antibody response that prevented transgene expression with administration of a second adenovirus. These findings demonstrate that manipulation of the host immune response may expand potential applications of gene transfer utilizing adenoviral vectors.
Assuntos
Adenovírus Humanos/genética , Linfócitos T CD4-Positivos/imunologia , Vetores Genéticos/genética , Transgenes , Proteínas E1 de Adenovirus/genética , Adenovírus Humanos/imunologia , Animais , Anticorpos Monoclonais/administração & dosagem , Anticorpos Monoclonais/imunologia , Formação de Anticorpos , Sequência de Bases , Primers do DNA , Deleção de Genes , Expressão Gênica , Vetores Genéticos/imunologia , Humanos , Fígado/imunologia , Depleção Linfocítica , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Recombinação Genética , Fatores de TempoRESUMO
Previous data from our laboratory and others have demonstrated a critical role for the CD4+ T lymphocyte in in vivo immune responses to recombinant adenoviral vectors. In rodent models, this subset of T cells is required for T cell proliferation, subsequent cytotoxic T cell generation, and production of anti-adenoviral antibodies by B cells. Both depleting and nondepleting anti-CD4 antibodies can attenuate these immune responses to recombinant adenovirus. On the basis of these data, we hypothesized that a nondepleting CDR-engrafted anti-human CD4 antibody (OKT4A) with cross-reactivity to rhesus macaques would attenuate both T and B cell responses to intrapulmonary administration of recombinant adenovirus and permit prolonged reporter gene expression and permit secondary gene transfer. Juvenile rhesus macaques were treated with PBS or OKT4A antibody (10 mg/kg) daily beginning 1 day prior to and up to 11 days after gene transfer. OKT4A resulted in significant attenuation of lymphocyte recruitment into the lung, lymphocyte-proliferative responses to both adenovirus capsid proteins and transgene protein, and adenovirus-induced interferon-gamma elaboration in whole blood and hilar lymph nodes. However, OKT4A was ineffective in attenuating adenovirus-induced IL-4 production in whole blood or hilar lymph nodes, generating neutralizing anti-adenoviral antibodies, or permitting secondary gene transfer. As all the monkeys in this protocol had baseline-detectable anti-adenoviral antibodies by ELISA that were nonneutralizing, analogous to most patients with cystic fibrosis, we postulate that anti-CD4 did not block the proliferation of memory B cells. Moreover, these data suggest that for transient immunomodulation to be successful, strategies need to focus specifically on B cell activation independent of CD4+ T cell help.
Assuntos
Adenoviridae/genética , Adenoviridae/imunologia , Antígenos CD4/imunologia , Técnicas de Transferência de Genes , Pulmão/imunologia , Macaca mulatta , Animais , Linfócitos B/imunologia , Antígenos CD4/genética , Antígenos CD4/metabolismo , Regulação da Expressão Gênica , Vetores Genéticos/farmacocinética , Humanos , Interferon gama/metabolismo , Interleucina-4/metabolismo , Pulmão/patologia , Pulmão/virologia , Linfonodos/imunologia , Linfonodos/metabolismo , Pneumonia/genética , Pneumonia/patologia , Linfócitos T/imunologiaRESUMO
E1-deleted adenoviral vectors are efficient vectors for somatic cell gene therapy, but transgene expression is limited in part by a cytotoxic T cell response directed against virally transduced cells. Moreover, the development of a neutralizing antibody response limits secondary gene transfer with these vectors. Therapy with a depleting anti-CD4 antibody permits prolonged transgene expression in the lung and liver of mice. Furthermore, transient depletion of CD4+ lymphocytes blocks neutralizing antibody production and therefore allows repeat administration and expression of E1-deleted recombinant adenovirus. In this study, we investigated the efficacy of a novel nondepleting anti-CD4 antibody (RIB 5/2) in a model of lung-directed gene therapy in outbred rats. Treatment with RIB 5/2 permitted prolonged reporter gene expression and reduced adenovirus-induced peribronchial and alveolar inflammation in the lung. Moreover administration of RIB 5/2 blocked the development of an anti-adenoviral neutralizing antibody response in the lung and permitted secondary administration and expression of a recombinant adenovirus. These data support the role of immunomodulation in prolonging in vivo transgene expression by recombinant adenovirus.
Assuntos
Adenovírus Humanos/genética , Anticorpos Monoclonais , Antígenos CD4/imunologia , Regulação da Expressão Gênica/imunologia , Vetores Genéticos/genética , Pulmão/imunologia , Proteínas E1 de Adenovirus , Adenovírus Humanos/imunologia , Animais , Anticorpos Antivirais/análise , Anticorpos Antivirais/sangue , Líquido da Lavagem Broncoalveolar/imunologia , Genes Reporter/genética , Vetores Genéticos/administração & dosagem , Vetores Genéticos/imunologia , Humanos , Luciferases/genética , Pulmão/metabolismo , Subpopulações de Linfócitos , Masculino , Testes de Neutralização , Ratos , Ratos Sprague-Dawley , Linfócitos T Citotóxicos/imunologia , beta-Galactosidase/genéticaRESUMO
The mechanism of amiodarone-induced pulmonary toxicity is unknown. Two cases of amiodarone pulmonary toxicity are presented in which abnormal inclusion bodies containing whorls of membrane were seen on electron microscopy of extrapulmonary tissues. These cytoplasmic lysosomal inclusion bodies were observed in lymphocytes, plasma cells, granulocytes, tissue macrophages, and hepatocytes. These widespread histopathologic changes in extrapulmonary tissues and in a variety of cell types are similar to more extensively investigated findings in animal models that are thought to represent a drug-induced lysosomal storage disease, phospholipidosis.
Assuntos
Amiodarona/efeitos adversos , Benzofuranos/efeitos adversos , Leucócitos/ultraestrutura , Fígado/ultraestrutura , Pneumopatias/induzido quimicamente , Pulmão/ultraestrutura , Linfonodos/ultraestrutura , Biópsia por Agulha , Seguimentos , Humanos , Corpos de Inclusão/ultraestrutura , Leucócitos/efeitos dos fármacos , Fígado/efeitos dos fármacos , Pulmão/efeitos dos fármacos , Pneumopatias/diagnóstico por imagem , Pneumopatias/patologia , Linfonodos/efeitos dos fármacos , Lisossomos/ultraestrutura , Masculino , Microscopia Eletrônica , Pessoa de Meia-Idade , Radiografia , Taquicardia/tratamento farmacológicoAssuntos
Linfócitos T CD4-Positivos/imunologia , Hospedeiro Imunocomprometido/imunologia , Pneumonia por Pneumocystis/imunologia , Aerossóis , Animais , Modelos Animais de Doenças , Interferon gama/administração & dosagem , Pulmão/imunologia , Pulmão/patologia , Ativação de Macrófagos/efeitos dos fármacos , Ativação de Macrófagos/imunologia , Macrófagos Alveolares/efeitos dos fármacos , Macrófagos Alveolares/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Pneumonia por Pneumocystis/patologia , Pneumonia por Pneumocystis/terapia , Fatores de TempoRESUMO
In July 1979, a 72-year-old white woman presented to the Bernalillo County Medical Center Emergency Department with complaints of shortness of breath and wheezing. She had been asthmatic since childhood. Current management included bronchodilator therapy and continuous low flow oxygen. An apparently curative left radical nephrectomy had been performed in 1978 for renal cell carcinoma. Her evaluation in the emergency room included multiple attempts at right radial artery puncture for blood gases. The arterial blood gases were obtained only after repeated efforts by several individuals. The patient was discharged from the emergency room after receiving subcutaneous terbutaline and intravenous aminophylline. Approximately one week later, she noticed a swelling on her right wrist at the site of the punctures. Over the ensuing three weeks, the lesion doubled in size and became painful. In chest clinic, one month after emergency room visit, we found a 1 cm by 1 cm raised erythematous tender nodule on the right wrist overlying the radial artery pulse. We did not hear a bruit, the lesion did not feel fluctuant, and attempts to aspirate fluid were unsuccessful. Because we thought the lesion represented local infection, we began oral antibiotic therapy. The lesion was unchanged after 1 1/2 weeks of therapy and was excised. Pathologic examination showed a well-defined cutaneous nodule with histology diagnostic of a keratocanthoma. The patient developed a recurrence of the tumor at the excision site a few weeks later and required a wide excision, also under local anesthesia. She has since remained clinically free of recurrent tumor.
Assuntos
Coleta de Amostras Sanguíneas , Ceratoacantoma/etiologia , Punções/efeitos adversos , Dermatopatias/etiologia , Idoso , Gasometria , Feminino , HumanosRESUMO
Previous work from our laboratory has shown that chronic alcohol consumption in mice creates immunosuppression sufficient to permit infection with the opportunistic pathogen Pneumocystis carinii. Host defense against P. carinii is critically dependent upon host T lymphocytes. In these experiments, we address the effect of chronic alcohol consumption on recruitment of T lymphocytes into infected lung tissue and on lymphocytes in host lymphoid tissue. We find that mice administered alcohol in drinking water and then inoculated with P. carinii show significantly decreased recruitment of CD4+ and CD8+ T lymphocytes into lung tissue in comparison with control mice. Additional experiments show significant depletion of CD4+ lymphocytes in spleens from alcohol mice and decreased numbers of activated T lymphocytes. Analysis of surface expression of the adhesion molecules LFA-1, VLA-4, and ICAM-1 show no significant differences in lymphocytes from alcohol-consuming mice, and lymphocyte chemotaxis in vitro is also unaltered. We conclude that chronic consumption of alcohol impairs lung recruitment of lymphocytes in response to an infectious challenge. This impaired lymphocyte recruitment may be a consequence of depletion of T lymphocytes in host lymphoid tissue. Impaired recruitment of lymphocytes may explain the increased morbidity and mortality of pulmonary infections in alcoholic subjects.
Assuntos
Alcoolismo/imunologia , Inibição de Migração Celular , Etanol/farmacologia , Pulmão/imunologia , Pneumonia por Pneumocystis/imunologia , Baço/imunologia , Linfócitos T/efeitos dos fármacos , Animais , Linfócitos T CD4-Positivos/efeitos dos fármacos , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/efeitos dos fármacos , Linfócitos T CD8-Positivos/imunologia , Quimiotaxia de Leucócito/efeitos dos fármacos , Quimiotaxia de Leucócito/imunologia , Feminino , Tolerância Imunológica/efeitos dos fármacos , Tolerância Imunológica/imunologia , Contagem de Linfócitos , Camundongos , Camundongos Endogâmicos BALB C , Linfócitos T/imunologiaRESUMO
Autofluorescence has limited quantitative description by flow cytometry of certain alveolar macrophage surface antigens. Autofluorescence is most significant when macrophages are excited by blue light and emission is monitored in the green-yellow light range. Because of recent advances in flow cytometry and in the development of fluorescent labeling compounds, such as allophycocyanin, autofluorescence can be minimized by the use of red excitation and emission wavelengths. We have developed a three-step labeling technique that discretely identifies the minor subpopulation of normal rat alveolar macrophages that express Class II major histocompatibility (Ia) antigens. Using this technique, we observed that alveolar macrophage Ia antigen expression increases at least fourfold after intravenous administration of bacillus Calmette-Guerin to previously primed rats. Additionally, we observed that in vitro treatment of alveolar macrophages by exposure to gamma interferon results in an increase in Ia antigen expression. This new method is significant because flow cytometry is a powerful tool with which to analyze quickly, accurately, and reproducibly alveolar macrophage surface antigens.
Assuntos
Antígenos de Superfície/análise , Citometria de Fluxo , Macrófagos/imunologia , Ficocianina , Pigmentos Biológicos , Alvéolos Pulmonares/citologia , Animais , Vacina BCG/farmacologia , Contagem de Células , Formaldeído/farmacologia , Interferon gama/farmacologia , Polímeros/farmacologia , Valores de Referência , Coloração e RotulagemRESUMO
This study was undertaken to investigate the effects of pulmonary inflammation induced by bacillus Calmette-Guérin (BCG) on the density distribution of lavaged alveolar macrophages. We sought to determine macrophage cytotoxicity and interleukin-1 elaboration in density-defined subpopulations of macrophages during tissue inflammation. At all time points after intravenously administered BCG, lavaged alveolar macrophages contained increased percentages of higher density cells. Alveolar macrophage cytotoxicity against the rat sarcoma cell line XC increased maximally 2 to 6 days after intravenous administration of BCG before declining on Day 13. Macrophage interleukin-1 elaboration increased maximally 14 days after administration of BCG before declining on Day 23. Additionally, macrophage cytotoxicity and interleukin-1 elaboration were increased above normal in cells from each of five density fractions. We conclude that a subpopulation of higher density macrophages, probably recently derived from blood monocytes, accumulates in inflammatory sites. Cellular activation increases the cytotoxicity and interleukin-1 elaboration by macrophages in all density-defined subpopulations and obscures the relationship between cellular density and function that is present in normal animals.
Assuntos
Citotoxicidade Imunológica , Interleucina-1/imunologia , Ativação de Macrófagos , Macrófagos/imunologia , Pneumonia/imunologia , Animais , Líquido da Lavagem Broncoalveolar/citologia , Líquido da Lavagem Broncoalveolar/imunologia , Testes Imunológicos de Citotoxicidade , Interleucina-1/análise , Interleucina-2/análise , Masculino , Mycobacterium bovis/patogenicidade , Pneumonia/etiologia , Alvéolos Pulmonares/imunologia , Ratos , Ratos Endogâmicos F344 , Organismos Livres de Patógenos Específicos , Fatores de TempoRESUMO
Respiratory infections remain a significant public health problem and are presently the 6th leading cause of death in the United States. Antibiotic-resistant organisms are encountered increasingly in both community-acquired and nosocomial infections. Despite progress in antibiotic development, biological-response modifiers may have increasing application to augment pulmonary host defenses against either drug-resistant infections or in high-risk hosts. Toward this end, gene therapy proposes to deliver biologicals as nucleic acids rather than protein. Gene therapy has the potential advantage of targeting the biological to specific cells or tissue compartments, and a more favorable pharmacokinetic profile. Data on gene delivery and efficacy in preclinical models of pulmonary infection are presented and discussed.
Assuntos
Terapia Genética , Pulmão/imunologia , Pneumonia/terapia , Infecções Respiratórias/terapia , Animais , Técnicas de Transferência de Genes , Humanos , Interferon gama/genética , Interleucina-12/genética , Interleucina-17/genética , Pneumonia/imunologia , Infecções Respiratórias/imunologia , Fator de Necrose Tumoral alfa/genéticaRESUMO
The recruitment of specific subsets of CD4(+) T lymphocytes to the lungs in response to Pneumocystis carinii was investigated. For mice inoculated with P. carinii, an ELISPOT assay was used to calculate the numbers of lymph node and lung tissue CD4(+) cells that secreted interferon (IFN)-gamma (Th1 cytokine) and interleukin (IL)-4 (Th2 cytokine) after concanavalin A stimulation. An ELISA was used to assay culture supernatants for cytokine concentrations. Precursor frequency of both IFN-gamma- and IL-4-secreting cells was increased in lymph nodes at 1 week, whereas increases in Th1 and Th2 cells in lung tissue were delayed 3 weeks before declining. The frequency of IL-4-secreting cells always was greater than the frequency of IFN-gamma secreting cells. These results demonstrate an early T lymphocyte response in draining lymph nodes, followed by later recruitment of Th1 and Th2 lymphocytes into lung tissue. The overall CD4(+) T cell response to P. carinii involves both Th1 and Th2 subsets, but the response is Th2 dominant in both lymph node and lung tissue.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Pneumonia por Pneumocystis/imunologia , Animais , Citocinas/metabolismo , Feminino , Pulmão/imunologia , Linfonodos/imunologia , Ativação Linfocitária , Camundongos , Camundongos Endogâmicos BALB CRESUMO
We investigated the effect of intratracheal (i.t.) lipopolysaccharide (LPS) on alveolar macrophage release of nitric oxide. Mice received i.t. LPS at doses ranging from 1 to 100 micrograms/100 g body weight and were killed at serial intervals for bronchoalveolar lavage. Control mice received i.t. phosphate-buffered saline. We found that after i.t. LPS, there was an early (1 to 3 days) influx of neutrophils followed by a later (5 to 7 days) influx of macrophages into the lungs. Alveolar macrophages lavaged from mice given i.t. LPS did not spontaneously release nitric oxide (measured as nitrite), but the capacity of these cells to release nitric oxide in vitro in response to interferon-gamma (IFN-gamma) or LPS was markedly upregulated. Alveolar macrophages lavaged from mice given i.t. LPS but not i.t. phosphate-buffered saline also expressed mRNA for inducible nitric oxide synthase as measured by semiquantitative reverse-transcription polymerase chain reaction. To investigate possible mechanisms for cellular priming for increased nitric oxide release after i.t. LPS, mice were depleted of CD4+ lymphocytes with an anti-CD4 antibody. Alveolar macrophages from CD4-depleted mice given i.t. LPS released significantly less nitric oxide in vitro in comparison to macrophages from nondepleted mice. Additional mice were treated with neutralizing doses of anti-tumor necrosis factor or anti-IFN-gamma antibody before i.t. LPS. Pretreatment with each cytokine antibody decreased but did not eliminate macrophage priming for nitric oxide release after i.t. LPS. We conclude that intratracheal LPS induces mRNA for nitric oxide synthase in alveolar macrophages, priming the cells for increased release of nitric oxide in vitro.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Lipopolissacarídeos/farmacologia , Macrófagos Alveolares/metabolismo , Óxido Nítrico/metabolismo , Aminoácido Oxirredutases/análise , Aminoácido Oxirredutases/genética , Animais , Sequência de Bases , Líquido da Lavagem Broncoalveolar/citologia , Linfócitos T CD4-Positivos , Escherichia coli/química , Radicais Livres , Interferon gama/imunologia , Interferon gama/metabolismo , Intubação Intratraqueal , Depleção Linfocítica , Macrófagos Alveolares/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Óxido Nítrico Sintase , Reação em Cadeia da Polimerase , Organismos Livres de Patógenos Específicos , Fator de Necrose Tumoral alfa/imunologia , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Cytomegalovirus (CMV) infection in patients with the acquired immunodeficiency syndrome (AIDS) can present as either disseminated disease, pneumonitis, retinitis, gastroenteritis, neuropathy, or a subclinical infection. We report a patient whose initial manifestation of CMV infection was severe central airways obstruction due to necrotizing tracheitis. At bronchoscopy, the lesion appeared deeply ulcerated, distinctly different from previously described airway lesions in patients with AIDS. Mucosal biopsies showed characteristic intranuclear and intracytoplasmic inclusions and cultures yielded only CMV. The patient responded partially to ganciclovir, steroids, and antibiotics against suspected anaerobic superinfection but died as a result of central nervous system disease believed due to toxoplasmosis or lymphoma. CMV infection of the upper airway should be considered in the patient with AIDS presenting with atypical cough or stridor and ulcerated endobronchial lesions.