RESUMO
BACKGROUND: Noninvasive sputum sampling has enabled the identification of biomarkers in asthmatic patients. Studies of discrete cell populations in sputum can enhance measurements compared with whole sputum in which changes in rare cells and cell-cell interactions can be masked. OBJECTIVE: We sought to enrich for sputum-derived human bronchial epithelial cells (sHBECs) and sputum-derived myeloid type 1 dendritic cells (sDCs) to describe transcriptional coexpression of targets associated with a type 2 immune response. METHODS: A case-control study was conducted with patients with mild asthma (asthmatic cases) and healthy control subjects. Induced sputum was obtained for simultaneous enrichment of sHBECs and sDCs by using flow cytometry. Quantitative PCR was used to measure mRNA for sHBEC thymic stromal lymphopoietin (TSLP), IL33, POSTN, and IL25 and downstream targets in sDCs (OX40 ligand [OX40L], CCL17, PPP1R14A, CD1E, CD1b, CD80, and CD86). RESULTS: Final analyses for the study sample were based on 11 control subjects and 13 asthmatic cases. Expression of TSLP, IL33, and POSTN mRNA was increased in sHBECs in asthmatic cases (P = .001, P = .05, and P = .04, respectively). Expression of sDC OX40L and CCL17 mRNA was increased in asthmatic cases (P = .003 and P = .0001, respectively). sHBEC TSLP mRNA expression was strongly associated with sDC OX40L mRNA expression (R = 0.65, P = .001) and less strongly with sDC CCL17 mRNA expression. sHBEC IL33 mRNA expression was associated with sDC OX40L mRNA expression (R = 0.42, P = .04) but not sDC CCL17 mRNA expression. CONCLUSIONS: Noninvasive sampling and enrichment of select cell populations from sputum can further our understanding of cell-cell interactions in asthmatic patients with the potential to enhance endotyping of asthmatic patients.
Assuntos
Asma/genética , Células Dendríticas/metabolismo , Células Epiteliais/metabolismo , Expressão Gênica , Transdução de Sinais/imunologia , Adulto , Antígenos CD1/genética , Antígenos CD1/imunologia , Asma/imunologia , Asma/patologia , Antígeno B7-1/genética , Antígeno B7-1/imunologia , Antígeno B7-2/genética , Antígeno B7-2/imunologia , Estudos de Casos e Controles , Comunicação Celular , Quimiocina CCL17/genética , Quimiocina CCL17/imunologia , Células Dendríticas/imunologia , Células Dendríticas/patologia , Células Epiteliais/imunologia , Células Epiteliais/patologia , Feminino , Perfilação da Expressão Gênica , Humanos , Interleucina-17/genética , Interleucina-17/imunologia , Interleucina-33/genética , Interleucina-33/imunologia , Peptídeos e Proteínas de Sinalização Intracelular , Masculino , Pessoa de Meia-Idade , Proteínas Musculares , Ligante OX40/genética , Ligante OX40/imunologia , Fosfoproteínas Fosfatases/genética , Fosfoproteínas Fosfatases/imunologia , Transdução de Sinais/genética , Escarro/citologiaRESUMO
If maternal atopy and environmental exposure affect prenatal Th cell development, the maternal and fetal immune systems should display common Th1/Th2 phenotypes. To test this hypothesis, we studied maternal and neonatal blood samples from mothers with total serum IgE <300 IU/mL. Basal levels of IFN-γ, IL-4, and eotaxin in paired maternal and fetal sera were tightly correlated. Polyclonal T cell activation in vitro by Staphylococcal exotoxin B induced co-ordinate IFN-γ production from paired maternal and fetal mononuclear cells, accompanied by co-ordinate increases in activated CD4+CD69+ cells that display the CCR4+Th2 and CXCR3+ Th1 phenotypes. Maternal and fetal CD4+CXCR3+ T cells were subsequently identified as the major producers of IFN-γ. The data established that a transplacental nexus exists during normal pregnancy and that fetal Th cell responses may be biased by the maternal immune system.
Assuntos
Sangue Fetal/citologia , Células Th1/imunologia , Células Th2/imunologia , Linfócitos T CD4-Positivos/química , Linfócitos T CD4-Positivos/imunologia , Quimiocina CCL11 , Feminino , Sangue Fetal/química , Sangue Fetal/imunologia , Humanos , Imunoglobulina E/sangue , Recém-Nascido , Interferon gama/sangue , Interleucina-4/sangue , Ativação Linfocitária , Fenótipo , Gravidez , Receptores CCR3/análise , Receptores CCR4/análise , Células Th1/química , Células Th2/químicaRESUMO
HIV-1 envelope (Env) gp120 is an important target for neutralizing antibody (Ab) responses against the virus; however, developing gp120 vaccines that elicit potent and broad neutralizing Abs has proven to be a formidable challenge. Previously, removal of an N-linked glycan at residue 448 by an N to Q mutation (N448Q) has been found to enhance the in vitro antigenicity of neutralizing epitopes in the V3 loop. In this study the mutated gp120 was first compared with wild type gp120 for immunogenicity in mice using a DNA prime and protein boost immunization regimen. The N448Q mutant did not elicit higher titers of anti-gp120 serum Abs and failed to generate anti-V3 Abs. The sera also had no virus-neutralizing activity, even though the mutant induced higher levels of lymphoproliferation and cytokine production. Subsequently, the N448Q mutant was used to construct an immune complex vaccine with the anti-CD4 binding site monoclonal antibody (mAb) 654. The N448Q/654 complex stimulated comparably high levels of serum Abs to gp120 and V3 as the wild type complex. However, Abs against the C1 and C2 regions in the gp120 core were more elevated. Importantly, the mutant complex also elicited higher titers of neutralizing Abs activity than the wild type counterpart. Similar results were achieved with a complex made with gp120 bearing an N448E mutation, confirming the importance of the N448-linked glycan in modulating gp120 immunogenicity. Neutralizing activity was directed to V3 and other undefined neutralizing epitopes. Improved immunogenicity of the immune complexes correlated with alterations in exposure of V3 and other Ab epitopes and their stability against proteases. These data demonstrate the advantage of combining site-specific N-glycan removal and immune complex formation as a novel vaccine strategy to improve immunogenicity of targeted Ab epitopes on critical regions of HIV-1 gp120.
Assuntos
Vacinas contra a AIDS/imunologia , Complexo Antígeno-Anticorpo/imunologia , Proteína gp120 do Envelope de HIV/imunologia , Infecções por HIV/prevenção & controle , Polissacarídeos/química , Vacinas contra a AIDS/administração & dosagem , Animais , Anticorpos Monoclonais/imunologia , Anticorpos Neutralizantes/sangue , Formação de Anticorpos , Especificidade de Anticorpos , Proliferação de Células , Citocinas/imunologia , Epitopos/imunologia , Feminino , Anticorpos Anti-HIV/sangue , Proteína gp120 do Envelope de HIV/administração & dosagem , Proteína gp120 do Envelope de HIV/química , Infecções por HIV/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Testes de Neutralização , Linfócitos T/imunologiaRESUMO
Ambient particulate matter, including diesel exhaust particles (DEP), promotes the development of allergic disorders. DEP increase oxidative stress and influence human bronchial epithelial cell (HBEC)-dendritic cell interactions via cytokines, including thymic stromal lymphopoietin (TSLP). Upregulation of TSLP results in Th2 responses. Using primary culture HBEC and human myeloid dendritic cell (mDC) cocultures, we show in this study that DEP upregulation of Th2 responses occurred via HBEC-dependent mechanisms that resulted from oxidative stress. Moreover, DEP-treated HBEC and ambient particulate matter-treated HBEC upregulated OX40 ligand (OX40L) and the Notch ligand Jagged-1 mRNA and expression on mDC. Upregulation of OX40L as well as Jagged-1 on mDC required HBEC and did not occur in the presence of N-acetylcysteine. Furthermore, OX40L and Jagged-1 upregulation was inhibited when HBEC expression of TSLP was silenced. Thus, DEP treatment of HBEC targeted two distinct pathways in mDC that were downstream of TSLP expression. Upregulation of OX40L and Jagged-1 by mDC resulted in mDC-driven Th2 responses. These studies expand our understanding of the mechanism by which ambient pollutants alter mucosal immunity and promote disorders such as asthma.
Assuntos
Proteínas de Ligação ao Cálcio/biossíntese , Citocinas/fisiologia , Células Dendríticas/imunologia , Peptídeos e Proteínas de Sinalização Intercelular/biossíntese , Proteínas de Membrana/biossíntese , Ligante OX40/biossíntese , Material Particulado/toxicidade , Mucosa Respiratória/imunologia , Regulação para Cima/imunologia , Emissões de Veículos/toxicidade , Brônquios/citologia , Brônquios/efeitos dos fármacos , Brônquios/imunologia , Polaridade Celular/efeitos dos fármacos , Polaridade Celular/imunologia , Células Cultivadas , Técnicas de Cocultura , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/metabolismo , Humanos , Proteína Jagged-1 , Células Mieloides/efeitos dos fármacos , Células Mieloides/imunologia , Células Mieloides/metabolismo , Espécies Reativas de Oxigênio/toxicidade , Receptores Notch/biossíntese , Mucosa Respiratória/citologia , Mucosa Respiratória/efeitos dos fármacos , Proteínas Serrate-Jagged , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/imunologia , Células Estromais/efeitos dos fármacos , Células Estromais/imunologia , Células Estromais/metabolismo , Células Th2/efeitos dos fármacos , Células Th2/imunologia , Células Th2/metabolismo , Timo/citologia , Timo/efeitos dos fármacos , Timo/imunologia , Regulação para Cima/efeitos dos fármacos , Linfopoietina do Estroma do TimoRESUMO
Variation of surface antigen expression is a mechanism used by microbes to adapt to and persist within their host habitats. Helicobacter pylori, a persistent bacterial colonizer of the human stomach, can alter its surface Lewis (Le) antigen expression. We examined H. pylori colonization in mice to test the hypothesis that host phenotype selects for H. pylori (Le) phenotypes. When wild-type and Le(b)-expressing transgenic FVB/N mice were challenged with H. pylori strain HP1, expressing Le(x) and Le(y), we found that bacterial populations recovered after 8 mo from Le(b)-transgenic, but not wild-type, mice expressed Le(b). Changes in Le phenotype were linked to variation of a putative galactosyltransferase gene (beta-(1,3)galT); mutagenesis and complementation revealed its essential role in type I antigen expression. These studies indicate that H. pylori evolves to resemble the host's gastric Le phenotype, and reveal a bacterial genetic locus that is subject to host-driven selection pressure.
Assuntos
Helicobacter pylori/imunologia , Antígenos do Grupo Sanguíneo de Lewis/análise , Antígenos CD15/análise , Oligossacarídeos/fisiologia , Adesinas Bacterianas/análise , Animais , Anticorpos Antibacterianos/sangue , Citometria de Fluxo , Fucosiltransferases/genética , Galactosiltransferases/genética , Lipopolissacarídeos/toxicidade , Camundongos , Camundongos Transgênicos , FenótipoRESUMO
Tuberculids are chronic nodular skin eruptions believed to be a systemic reaction to Mycobacterium tuberculosis. We report on a 6-year-old boy with tender subcutaneous lesions on his legs. A tuberculin skin test resulted in 2.5 cm of induration and an interferon-gamma releasing assay was also markedly positive. A diagnosis of erythema induratum of Bazin was confirmed on skin biopsy. The patient was successfully treated with multi-drug antituberculosis therapy.
Assuntos
Eritema Endurado/diagnóstico , Antituberculosos/uso terapêutico , Biópsia , Criança , Eritema Endurado/tratamento farmacológico , Eritema Endurado/imunologia , Eritema Endurado/microbiologia , Exantema/microbiologia , Humanos , Interferon gama/sangue , Perna (Membro) , Masculino , Mycobacterium tuberculosis/imunologia , Mycobacterium tuberculosis/isolamento & purificação , Reação em Cadeia da PolimeraseRESUMO
We describe a case in which a human immunodeficiency virus (HIV)-positive child presented in severe metabolic acidosis secondary to his candidal sepsis and T-cell lymphoma, a rare finding in pediatric AIDS. Significantly elevated levels of Interleukin-10 (IL-10) were found in the patient's serum, which may have played a role in acute demise.
Assuntos
Candida albicans/isolamento & purificação , Candidíase/complicações , Fungemia/complicações , Infecções por HIV/complicações , Interleucina-10/metabolismo , Linfoma de Células T/complicações , Regulação para Cima , Infecções Oportunistas Relacionadas com a AIDS/microbiologia , Adolescente , Candidíase/microbiologia , Evolução Fatal , Fungemia/microbiologia , Infecções por HIV/imunologia , Humanos , Linfoma Relacionado a AIDS/complicações , MasculinoRESUMO
Human exposure to air pollutants, including ambient particulate matter, has been proposed as a mechanism for the rise in allergic disorders. Diesel exhaust particles, a major component of ambient particulate matter, induce sensitization to neoallergens, but the mechanisms by which sensitization occur remain unclear. We show that diesel exhaust particles upregulate thymic stromal lymphopoietin in human bronchial epithelial cells in an oxidant-dependent manner. Thymic stromal lymphopoietin induced by diesel exhaust particles was associated with maturation of myeloid dendritic cells, which was blocked by anti-thymic stromal lymphopoietin antibodies or silencing epithelial cell-derived thymic stromal lymphopoietin. Dendritic cells exposed to diesel exhaust particle-treated human bronchial epithelial cells induced Th2 polarization in a thymic stromal lymphopoietin-dependent manner. These findings provide new insight into the mechanisms by which diesel exhaust particles modify human lung mucosal immunity.
Assuntos
Poluentes Atmosféricos/farmacologia , Diferenciação Celular/efeitos dos fármacos , Citocinas/metabolismo , Células Dendríticas/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Emissões de Veículos , Acetilcisteína/farmacologia , Anticorpos/farmacologia , Brônquios/citologia , Brônquios/imunologia , Brônquios/metabolismo , Linfócitos T CD4-Positivos/citologia , Linfócitos T CD4-Positivos/efeitos dos fármacos , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/metabolismo , Diferenciação Celular/imunologia , Linhagem Celular Transformada , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Técnicas de Cocultura , Citocinas/genética , Citocinas/imunologia , Citocinas/farmacologia , Células Dendríticas/citologia , Células Dendríticas/metabolismo , Células Epiteliais/metabolismo , Expressão Gênica/efeitos dos fármacos , Humanos , Imunidade nas Mucosas/efeitos dos fármacos , Imunidade nas Mucosas/imunologia , Ativação Linfocitária/efeitos dos fármacos , Ativação Linfocitária/imunologia , Tamanho da Partícula , RNA Interferente Pequeno/genética , Espécies Reativas de Oxigênio/metabolismo , Acetato de Tetradecanoilforbol/farmacologia , Linfopoietina do Estroma do TimoRESUMO
OBJECTIVES: To test the hypothesis that in comparison with those with shorter risk duration, individuals with longer HIV risk duration would have reduced susceptibility to HIV-1 infection as measured by CCR5 expression, and to evaluate whether variation in CCR5 expression could be explained by known genetic polymorphisms. DESIGN AND METHODS: A cross-sectional study of HIV-1 exposed but uninfected men who have sex with men. The risk duration was estimated from self-reported years since first receptive anal intercourse. CCR5 expression on peripheral blood CD4+ monocytes and T cells was determined by flow cytometry. The CCR5-Delta32 mutation and polymorphisms in the CCR5 promoter and CCR2 as well as the copy number of CCL3L1 were analyzed by polymerase chain reaction. Plasma levels of MIP-1alpha (CCL3), MIP-1beta (CCL4) and RANTES (CCL5) were also measured. As risk duration varied with age, analyses were restricted to 67 individuals aged 30-49 years. RESULTS: Multiple linear regression analyses, adjusted for age and race, showed a significant negative association between HIV risk duration and CCR5 expression on monocytes (P = 0.01), and in a separate model, a similar negative association with CCR5 expression on T cells (P = 0.03). Low CCR5 expression was attributable mainly to CCR5-Delta32 heterozygosity and the CCR5-59029G allele. CONCLUSIONS: We confirmed a role for reduced CCR5 expression in HIV-1 resistance. CCR5-Delta32 heterozygosity and the CCR5-59029G allele were significant predictors of low CCR5 expression. Individuals with high CCR5 expression who resisted infection despite long HIV risk duration form an interesting group within which to search for additional mechanisms of resistance to HIV infection.
Assuntos
Soronegatividade para HIV/fisiologia , Homossexualidade Masculina , Receptores CCR5/análise , Comportamento Sexual , Adulto , Linfócitos T CD4-Positivos/química , Estudos Transversais , Genótipo , HIV-1 , Homossexualidade Masculina/psicologia , Humanos , Masculino , Pessoa de Meia-Idade , Monócitos/química , Receptores CCR5/genética , Assunção de Riscos , Fatores de TempoRESUMO
Increased exposure to air pollutants such as diesel exhaust particles (DEP) has been proposed as one mechanism to explain the rise in allergic disorders. However, the immunologic mechanisms by which DEP enhance allergic sensitization and asthma remain unclear. We hypothesized that DEP act as an adjuvant for immature dendritic cell (DC) maturation via its effect on airway epithelial cell-derived microenvironment for DC. Immature monocyte-derived DC (iMDDC) failed to undergo phenotypic (CD80, CD83, CD86) or functional (T cell activation) maturation in response to exposure to DEP (0.001-100 mug/ml). In contrast, primary cultures of human bronchial epithelial cells (HBEC) treated with DEP induced iMDDC phenotypic maturation (2.6 +/- 0.1-fold increase in CD83 expression, n = 4, p < 0.05) and functional maturation (2.6 +/- 0.2-fold increase in T cell activation, n = 4, p < 0.05). Functional maturation of iMDDC was induced by conditioned medium derived from DEP-treated HBEC, and was inhibited in cultures with DEP-treated HBEC and blocking Abs against GM-CSF, or GM-CSF-targeted small interfering RNA. These data suggest that DEP induce Ag-independent DC maturation via epithelial cell-DC interactions mediated by HBEC-derived GM-CSF. Although additional signals may be required for polarization of DC, these data suggest a novel mechanism by which environmental pollutants alter airway immune responses.
Assuntos
Brônquios/citologia , Brônquios/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Células Dendríticas/citologia , Células Dendríticas/efeitos dos fármacos , Mucosa Respiratória/citologia , Mucosa Respiratória/efeitos dos fármacos , Emissões de Veículos , Brônquios/fisiologia , Comunicação Celular/efeitos dos fármacos , Comunicação Celular/fisiologia , Diferenciação Celular/fisiologia , Células Dendríticas/fisiologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/fisiologia , Humanos , Imunofenotipagem , Teste de Cultura Mista de Linfócitos , Monócitos/citologia , Monócitos/efeitos dos fármacos , Monócitos/fisiologia , Tamanho da Partícula , Mucosa Respiratória/fisiologia , SolubilidadeRESUMO
OBJECTIVE: To address the role that alloreactivity may play and better define the window for histoincompatible stem cell transplantation in utero. SUBJECTS, MATERIAL AND METHODS: We studied 9 fetal blood specimens obtained by cardiocentesis during elective abortions in the second trimester by multicolor flow cytometry and in vitro stimulation. RESULTS: Lymphocytes ranged from adult levels (3/9) to >90% leukocytes. Six specimens had T cells within adult range. T cells in the other specimens were reduced, while B cells were conversely elevated. This variability did not correlate with gestational age, or leukocyte composition. Following 4 h of mitogenesis, fetal CD4+ and CD8+ T cells from 1 of 5 specimens showed a response similar to that of maternal T cells, while the other 4 specimens showed a diminished response (0.3 +/- 0.2-fold). This heterogeneity did not correlate with gestational age, or lymphocyte subset distribution. Following 18 h of in vitro mitogenesis, fetal T cells from 2 specimens showed a response similar to that of maternal T cells (0.8 +/- 0.2-fold). Despite that, one specimen gave a 3-fold greater response in a one-way mixed lymphocyte reaction vs. maternal cells compared to the other specimen. CONCLUSION: We determine that fetal immunocompetence differs greatly during the second trimester and assessment of host vs. donor reactivity prior to in utero transplantation is likely to potentiate more favorable outcomes.
Assuntos
Feto/imunologia , Imunocompetência , Segundo Trimestre da Gravidez , Formação de Anticorpos , Contagem de Células Sanguíneas , Feminino , Sangue Fetal , Terapias Fetais , Citometria de Fluxo , Doenças Genéticas Inatas/embriologia , Doenças Genéticas Inatas/cirurgia , Histocompatibilidade , Humanos , Isoantígenos/imunologia , Contagem de Leucócitos , Subpopulações de Linfócitos/citologia , Mitose , Gravidez , Transplante de Células-Tronco , Linfócitos T/citologia , Linfócitos T/imunologiaRESUMO
IFN-alphabeta functions in the transition from innate to adaptive immunity and may impinge on the interaction of Mycobacterium tuberculosis with its host. Infection by M. tuberculosis causes IFN-alphabeta secretion and down-regulation of IFN-alphabeta signaling in human APC and the human monocytic cell line THP-1, which provides a model for these studies. Neutralization of secreted IFN-alphabeta prevents inhibition of IFN-alpha signaling during infection, but several lines of evidence distinguish inhibition due to infection from a negative feedback response to only IFN-alphabeta. First, greater inhibition of IFN-alpha-stimulated STAT-1 tyrosine phosphorylation occurs 3 days postinfection than 1 or 3 days after IFN-alphabeta pretreatment. Second, LPS also induces IFN-alphabeta secretion and causes IFN-alphabeta-dependent down-regulation of IFN-alpha signaling, yet the inhibition differs from that caused by infection. Third, IFN-alpha signaling is inhibited when cells are grown in conditioned medium collected from infected cells 1 day postinfection, but not if it is collected 3 days postinfection. Because IFN-alphabeta is stable, the results with conditioned medium suggest the involvement of an additional, labile substance during infection. Further characterizing signaling for effects of infection, we found that cell surface IFN-alphabeta receptor is not reduced by infection, but that infection increases association of protein tyrosine phosphatase 1c with the receptor and with tyrosine kinase 2. Concomitantly, IFN-alpha stimulation of tyrosine kinase 2 tyrosine phosphorylation and kinase activity decreases in infected cells. Moreover, infection reduces the abundance of JAK-1 and tyrosine-phosphorylated JAK-1. Thus, the distinctive down-regulation of IFN-alpha signaling by M. tuberculosis occurs together with a previously undescribed combination of inhibitory intracellular events.
Assuntos
Regulação para Baixo/imunologia , Retroalimentação , Interferon-alfa/metabolismo , Interferon beta/metabolismo , Mycobacterium tuberculosis/imunologia , Transdução de Sinais/imunologia , Linhagem Celular Tumoral , Membrana Celular/imunologia , Membrana Celular/metabolismo , Membrana Celular/microbiologia , Meios de Cultivo Condicionados/farmacologia , Citoplasma/imunologia , Citoplasma/metabolismo , Citoplasma/microbiologia , Humanos , Interferon-alfa/antagonistas & inibidores , Interferon-alfa/fisiologia , Interferon beta/fisiologia , Peptídeos e Proteínas de Sinalização Intracelular , Janus Quinase 1 , Lipopolissacarídeos/farmacologia , Fosforilação , Proteína Tirosina Fosfatase não Receptora Tipo 6 , Proteínas Tirosina Fosfatases/metabolismo , Proteínas Tirosina Quinases/antagonistas & inibidores , Proteínas Tirosina Quinases/metabolismo , Receptor de Interferon alfa e beta , Receptores de Interferon/metabolismo , TYK2 Quinase , Tirosina/antagonistas & inibidores , Tirosina/metabolismoRESUMO
Opportunistic infections such as pulmonary tuberculosis (TB) increase local HIV-1 replication and mutation. As AIDS progresses, alteration of the HIV-1 gp120 V3 sequence is associated with a shift in viral coreceptor use from CCR5 (CD195) to CXCR4 (CD184). To better understand the effect of HIV/TB coinfection, we screened transcripts from bronchoalveolar lavage cells with high density cDNA arrays and found that CXCR4 mRNA is increased in patients with TB. Surprisingly, CXCR4 was predominately expressed on alveolar macrophages (AM). Mycobacterium tuberculosis infection of macrophages in vitro increased CXCR4 surface expression, whereas amelioration of disease reduced CXCR4 expression in vivo. Bronchoalveolar lavage fluid from TB patients had elevated levels of CCL4 (macrophage inflammatory protein-1beta), CCL5 (RANTES), and CX3CL1 (fractalkine), but not CXCL12 (stromal-derived factor-1alpha). We found that M. tuberculosis infection of macrophages in vitro increased viral entry and RT of CXCR4-using [corrected] HIV-1, but not of CCR5-using [corrected] HIV-1. Lastly, HIV-1 derived from the lung contains CD14, suggesting that they were produced in AM. Our results demonstrate that TB produces a permissive environment for replication of CXCR4-using virus by increasing CXCR4 expression in AM and for suppression of CCR5-using HIV-1 by increasing CC chemokine expression. These changes explain in part why TB accelerates the course of AIDS. CXCR4 inhibitors are a rational therapeutic approach in HIV/TB coinfection.
Assuntos
Quimiocinas CC/biossíntese , Quimiocinas CX3C/biossíntese , HIV-1/imunologia , Macrófagos Alveolares/imunologia , Macrófagos Alveolares/virologia , Mycobacterium tuberculosis/imunologia , Receptores de Quimiocinas/biossíntese , Replicação Viral/imunologia , Sequência de Aminoácidos , Líquido da Lavagem Broncoalveolar/imunologia , Líquido da Lavagem Broncoalveolar/virologia , Linhagem Celular Tumoral , Células Cultivadas , Quimiocinas CC/genética , Quimiocinas CC/fisiologia , Quimiocinas CX3C/genética , Quimiocinas CX3C/fisiologia , Regulação da Expressão Gênica/imunologia , HIV-1/isolamento & purificação , HIV-1/patogenicidade , Humanos , Macrófagos Alveolares/metabolismo , Dados de Sequência Molecular , Infecções Oportunistas/imunologia , Infecções Oportunistas/metabolismo , Infecções Oportunistas/virologia , RNA Mensageiro/biossíntese , Receptores CCR4 , Receptores de Quimiocinas/fisiologia , Especificidade da Espécie , Tuberculose Pulmonar/imunologia , Tuberculose Pulmonar/microbiologia , Tuberculose Pulmonar/virologia , Regulação para Cima/imunologiaRESUMO
Sepsis induces an early inflammatory cascade initiated by the innate immune response. This often results in the development of multisystem organ failure. We examined the role of CD40, a costimulatory molecule that is integral in adaptive immunity, by using a murine model of polymicrobial sepsis. CD40 knockout (KO) mice had delayed death and improved survival after cecal ligation and puncture (CLP). In addition, they had less remote organ injury as manifested by reduced pulmonary capillary leakage. The improvements in survival and remote organ dysfunction in CD40 KO mice were associated with reduced interleukin-6 (IL-6) and IL-10 levels in serum and bronchoalveolar lavage fluid compared to the levels in wild-type (WT) controls. Furthermore, in contrast to WT mice, CD40 KO mice had no induction of the Th1 cytokines IL-12 and gamma interferon in serum or lungs after CLP. The alterations in cytokine production in CD40 KO mice were associated with similar changes in transcription factor activity. After CLP, CD40 KO mice had attenuated activation of nuclear factor kappaB and signal transducer and activator of transcription 3 in both the lung and the liver. Finally, WT mice had increased expression of CD40 on their alveolar macrophages. These data highlight the importance of CD40 activation in the innate immune response during polymicrobial sepsis and the subsequent development of remote organ dysfunction.
Assuntos
Antígenos CD40/fisiologia , Sepse/imunologia , Doença Aguda , Animais , DNA/metabolismo , Proteínas de Ligação a DNA/metabolismo , Feminino , Imunidade Inata , Interleucina-12/biossíntese , Interleucina-6/biossíntese , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , NF-kappa B/metabolismo , Fator de Transcrição STAT3 , Sepse/mortalidade , Transativadores/metabolismoRESUMO
We report a sentinel case of acute eosinophilic pneumonia in a firefighter exposed to high concentrations of World Trade Center dust during the rescue effort from September 11 to 24. The firefighter presented with a Pa(O2) of 53 mm Hg and responded to oxygen and corticosteroids. Computed tomography scan showed patchy ground glass density, thickened bronchial walls, and bilateral pleural effusions. Bronchoalveolar lavage recovered 70% eosinophils, with only 1% eosinophils in peripheral blood. Eosinophils were not degranulated and increased levels of interleukin-5 were measured in bronchoalveolar lavage and serum. Mineralogic analysis counted 305 commercial asbestos fibers/10(6) macrophages including those with high aspect ratios, and significant quantities of fly ash and degraded fibrous glass. Acute eosinophilic pneumonia is a rare consequence of acute high dust exposure. World Trade Center dust consists of large particle-size silicates, but fly ash and asbestos fibers may be found in bronchoalveolar lavage cells.
Assuntos
Poeira/efeitos adversos , Incêndios , Doenças Profissionais/etiologia , Eosinofilia Pulmonar/etiologia , Trabalho de Resgate , Lesão por Inalação de Fumaça/complicações , Terrorismo , Doença Aguda , Administração Oral , Corticosteroides/administração & dosagem , Corticosteroides/uso terapêutico , Adulto , Anti-Inflamatórios/administração & dosagem , Anti-Inflamatórios/uso terapêutico , Amianto Amosita/análise , Líquido da Lavagem Broncoalveolar/citologia , Eosinófilos/citologia , Seguimentos , Vidro/análise , Humanos , Masculino , Microscopia Eletrônica , Cidade de Nova Iorque , Ocupações , Prednisona/administração & dosagem , Prednisona/uso terapêutico , Eosinofilia Pulmonar/diagnóstico , Eosinofilia Pulmonar/terapia , Radiografia Torácica , Lesão por Inalação de Fumaça/etiologia , Fatores de Tempo , Tomografia Computadorizada por Raios X , Resultado do TratamentoRESUMO
HIV-1 replication is markedly upregulated in alveolar macrophages (AM) during pulmonary tuberculosis (TB). This is associated with loss of an inhibitory CCAAT enhancer binding protein beta (C/EBPbeta) transcription factor and activation of nuclear factor (NF)-kappaB. Since the cellular immune response in pulmonary TB requires lymphocyte--macrophage interaction, a model system was developed in which lymphocytes were added to AM. Contact between lymphocytes and AM reduced inhibitory C/EBPbeta, activated NF-kappaB, and enhanced HIV-1 replication. If contact between lymphocytes and macrophages was prevented, inhibitory C/EBPbeta expression was maintained and the HIV-1 long terminal repeat (LTR) was not maximally stimulated although NF-kappaB was activated. Antibodies that cross-linked macrophage expressed B-7, and vascular cell adhesion molecule and CD40 were used to mimic lymphocyte contact. All three cross-linking antibodies were required to abolish inhibitory C/EBPbeta expression. However, the HIV-1 LTR was not maximally stimulated and NF-kappaB was not activated. Maximal HIV-1--LTR stimulation required both lymphocyte-derived soluble factors, and cross-linking of macrophage expressed costimulatory molecules. High level HIV-1--LTR stimulation was also achieved when IL-1beta, IL-6, and TNF-beta were added to macrophages with cross-linked costimulatory molecules. Contact between activated lymphocytes and macrophages is necessary to down-regulate inhibitory C/EBPbeta, thereby derepressing the HIV-1 LTR. Lymphocyte-derived cytokines activate NF-kappaB, further enhancing the HIV-1 LTR.