RESUMO
BACKGROUND/INTRODUCTION: Sarcoidosis is a multi-systemic disorder of unknown etiology, characterized by the presence of non-caseating granulomas in target organs. In 90% of cases, there is thoracic involvement. Fifty to seventy percent of pulmonary sarcoidosis patients will experience acute, self-limiting disease. For the subgroup of patients who develop persistent disease, no targeted therapy is currently available. AIM: To investigate the potential of the single nucleotide polymorphism (SNP), Toll-like receptor 3 Leu412Phe (TLR3 L412F; rs3775291), as a causative factor in the development of and in disease persistence in pulmonary sarcoidosis. To investigate the functionality of TLR3 L412F in vitro in primary human lung fibroblasts from pulmonary sarcoidosis patients. DESIGN: SNP-genotyping and cellular assays, respectively, were used to investigate the role of TLR3 L412F in the development of persistent pulmonary sarcoidosis. METHODS: Cohorts of Irish sarcoidosis patients (n = 228), healthy Irish controls (n = 263) and a secondary cohort of American sarcoidosis patients (n = 123) were genotyped for TLR3 L412F. Additionally, the effect of TLR3 L412F in primary lung fibroblasts from pulmonary sarcoidosis patients was quantitated following TLR3 activation in the context of cytokine and type I interferon production, TLR3 expression and apoptotic- and fibroproliferative-responses. RESULTS: We report a significant association between TLR3 L412F and persistent clinical disease in two cohorts of Irish and American Caucasians with pulmonary sarcoidosis. Furthermore, activation of TLR3 in primary lung fibroblasts from 412 F-homozygous pulmonary sarcoidosis patients resulted in reduced IFN-ß and TLR3 expression, reduced apoptosis- and dysregulated fibroproliferative-responses compared with TLR3 wild-type patients. DISCUSSION/CONCLUSION: This study identifies defective TLR3 function as a previously unidentified factor in persistent clinical disease in pulmonary sarcoidosis and reveals TLR3 L412F as a candidate biomarker.
Assuntos
Polimorfismo de Nucleotídeo Único , Sarcoidose Pulmonar/genética , Receptor 3 Toll-Like/genética , Adolescente , Adulto , Idoso , Estudos de Casos e Controles , Feminino , Predisposição Genética para Doença , Humanos , Irlanda , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Fenótipo , Adulto JovemRESUMO
BACKGROUND: We aimed to investigate whether fluid therapy with a goal of near-maximal stroke volume (SV) guided by oesophageal Doppler (ED) monitoring result in a better outcome than that with a goal of maintaining bodyweight (BW) and zero fluid balance in patients undergoing colorectal surgery. METHODS: In a double-blinded clinical multicentre trial, 150 patients undergoing elective colorectal surgery were randomized to receive fluid therapy after either the goal of near-maximal SV guided by ED (Doppler, D group) or the goal of zero balance and normal BW (Zero balance, Z group). Stratification for laparoscopic and open surgery was performed. The postoperative fluid therapy was similar in the two groups. The primary endpoint was postoperative complications defined and divided into subgroups by protocol. Analysis was performed by intention-to-treat. The follow-up was 30 days. The trial had 85% power to show a difference between the groups. RESULTS: The number of patients undergoing laparoscopic or open surgery and the patient characteristics were similar between the groups. No significant differences between the groups were found for overall, major, minor, cardiopulmonary, or tissue-healing complications (P-values: 0.79; 0.62; 0.97; 0.48; and 0.48, respectively). One patient died in each group. No significant difference was found for the length of hospital stay [median (range) Z: 5.00 (1-61) vs D: 5.00 (2-41); P=0.206]. CONCLUSIONS: Goal-directed fluid therapy to near-maximal SV guided by ED adds no extra value to the fluid therapy using zero balance and normal BW in patients undergoing elective colorectal surgery.
Assuntos
Hidratação/métodos , Intestino Grosso/cirurgia , Cuidados Intraoperatórios/métodos , Volume Sistólico/fisiologia , Equilíbrio Hidroeletrolítico/fisiologia , Idoso , Idoso de 80 Anos ou mais , Método Duplo-Cego , Ecocardiografia Transesofagiana/métodos , Feminino , Objetivos , Humanos , Masculino , Pessoa de Meia-Idade , Monitorização Intraoperatória/métodos , Complicações Pós-Operatórias , Resultado do Tratamento , Ultrassonografia de Intervenção/métodosRESUMO
Sarcoidosis, a systemic granulomatous disease of unknown etiology, likely results from an environmental insult in a genetically susceptible host. In the US, African Americans are more commonly affected with sarcoidosis and suffer greater morbidity than Caucasians. We searched for sarcoidosis susceptibility loci by conducting a genome-wide, sib pair multipoint linkage analysis in 229 African-American families ascertained through two or more sibs with a history of sarcoidosis. Using the Haseman-Elston regression technique, linkage peaks with P-values less than 0.05 were identified on chromosomes 1p22, 2p25, 5p15-13, 5q11, 5q35, 9q34, 11p15 and 20q13 with the most prominent peak at D5S2500 on chromosome 5q11 (P=0.0005). We found agreement for linkage with the previously reported genome scan of a German population at chromosomes 1p and 9q. Based on the multiple suggestive regions for linkage found in our study population, it is likely that more than one gene influences sarcoidosis susceptibility in African Americans. Fine mapping of the linked regions, particularly on chromosome 5q, should help to refine linkage signals and guide further sarcoidosis candidate gene investigation.
Assuntos
Negro ou Afro-Americano/genética , Cardiomiopatias/genética , Predisposição Genética para Doença , Testes Genéticos , Sarcoidose/genética , Cardiomiopatias/etnologia , Cromossomos Humanos , Ligação Genética , Genoma Humano , Humanos , Sarcoidose/etnologiaRESUMO
Progress in our understanding of the scientific basis of granulomatous inflammation in sarcoidosis provides a framework for enlightened treatment decisions. Current evidence supports the concept that the pathogenesis of sarcoidosis involves a highly polarized T-helper 1 (Th1) immune response to pathogenic tissue antigens. Conventional treatment is focused on attenuating granuloma formation with antimalarial drugs that inhibit antigen presentation or with nonspecific anti-inflammatory agents such as glucocorticosteroids, methotrexate, or azathioprine. Anti-tumour necrosis factor (TNF)-alpha agents such as pentoxifylline, thalidomide, etanercept and remicade, have recently shown some successes in sarcoidosis. Designing future therapies depends on improved knowledge of the critical immunological processes operative in different stages of disease.
Assuntos
Granuloma/etiologia , Sarcoidose/tratamento farmacológico , Etanercepte , Granuloma/imunologia , Humanos , Imunoglobulina G/uso terapêutico , Imunossupressores/uso terapêutico , Interleucinas/imunologia , Pentoxifilina/uso terapêutico , Inibidores de Fosfodiesterase/uso terapêutico , Receptores do Fator de Necrose Tumoral/uso terapêutico , Sarcoidose/imunologia , Talidomida/uso terapêutico , Fator de Necrose Tumoral alfa/antagonistas & inibidoresRESUMO
Despite reports of familial clustering of sarcoidosis, little empirical evidence exists that disease risk in family members of sarcoidosis cases is greater than that in the general population. To address this question, we estimated sarcoidosis familial relative risk using data on disease occurrence in 10,862 first- and 17,047 second-degree relatives of 706 age, sex, race, and geographically matched cases and controls who participated in the multicenter ACCESS (A Case-Control Etiology Study of Sarcoidosis) study from 1996 to 1999. Familial relative risk estimates were calculated using a logistic regression technique that accounted for the dependence between relatives. Sibs had the highest relative risk (odds ratio [OR] = 5.8; 95% confidence interval [CI] = 2.1-15.9), followed by avuncular relationships (OR = 5.7; 95% CI = 1.6-20.7), grandparents (OR = 5.2; 95% CI = 1.5-18.0), and then parents (OR = 3.8; 95% CI = 1.2-11.3). In a multivariate model fit to the parents and sibs data, the familial relative risk adjusted for age, sex, relative class, and shared environment was 4.7 (95% CI = 2.3-9.7). White cases had a markedly higher familial relative risk compared with African-American cases (18.0 versus 2.8; p = 0.098). In summary, a significant elevated risk of sarcoidosis was observed among first- and second-degree relatives of sarcoidosis cases compared with relatives of matched control subjects.
Assuntos
Sarcoidose/epidemiologia , Sarcoidose/genética , Adolescente , Adulto , Distribuição por Idade , Idoso , Idoso de 80 Anos ou mais , Ordem de Nascimento , População Negra/genética , Estudos de Casos e Controles , Criança , Análise por Conglomerados , Feminino , Humanos , Incidência , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Linhagem , Vigilância da População , Modelos de Riscos Proporcionais , Risco , Fatores de Risco , Análise de Sobrevida , Estados Unidos/epidemiologia , População Branca/genéticaRESUMO
Sarcoidosis may be affected by sex, race, and age. A Case Control Etiologic Study of Sarcoidosis (ACCESS) enrolled 736 patients with sarcoidosis within 6 mo of diagnosis from 10 clinical centers in the United States. Using the ACCESS sarcoidosis assessment system, we determined organ involvement for the whole group and for subgroups differentiated by sex, race, and age (less than 40 yr or 40 yr and older). The study population was heterogeneous in terms of race (53% white, 44% black), sex (64% female, 36% male), and age (46% < 40 yr old, 54% > or = 40 yr old). Women were more likely to have eye and neurologic involvement (chi(2) = 4.74, p < 0.05 and chi(2) = 4.60, p < 0.05 respectively), have erythema nodosum (chi(2) = 7.28, p < 0.01), and to be age 40 yr or over (chi(2) = 6.07, p < 0.02) whereas men were more likely to be hypercalcemic (chi(2) = 7.38, p < 0.01). Black subjects were more likely to have skin involvement other than erythema nodosum (chi(2) = 5.47, p < 0.05), and eye (chi(2) = 13.8, p < 0.0001), liver (chi(2) = 23.3, p < 0.0001), bone marrow (chi(2) = 18.8, p < 0.001), and extrathoracic lymph node involvement (chi(2) = 7.21, p < 0.01). We conclude that the initial presentation of sarcoidosis is related to sex, race, and age.
Assuntos
Sarcoidose/epidemiologia , Sarcoidose/patologia , Adulto , Distribuição por Idade , Fatores Etários , Idoso , População Negra , Estudos de Casos e Controles , Dispneia/etiologia , Eritema Nodoso/etiologia , Feminino , Volume Expiratório Forçado , Humanos , Hipercalcemia/etiologia , Modelos Lineares , Masculino , Pessoa de Meia-Idade , Modelos de Riscos Proporcionais , Sarcoidose/classificação , Sarcoidose/complicações , Índice de Gravidade de Doença , Caracteres Sexuais , Distribuição por Sexo , Estados Unidos/epidemiologia , Capacidade Vital , População BrancaRESUMO
Because mouse strains susceptible to bleomycin, such as C57BL/ 6J, tend to produce T helper type 1 (Th1) cytokines in response to immune activation, we hypothesized that the inflammatory response to bleomycin is mediated, in part, by local production of the Th1 cytokine interferon-gamma (IFN-gamma). Consistent with this hypothesis, fibrosis-prone C57BL/6J and A/J mice demonstrated significantly elevated expression of IFN-gamma protein (by enzyme-linked immunosorbent assay) in bronchoalveolar lavage fluid at 24 h, and subsequently increased lung inflammation, weight loss, and mortality 10 d after intratracheal bleomycin administration compared with fibrosis-resistant BALB/c mice or saline control mice. To directly determine a role for IFN-gamma in bleomycin toxicity, we exposed C57BL/6J mice with a homozygous null mutation of the IFN-gamma gene (IFN-gamma[-/-]) and wild-type C57BL/6J mice to intratracheal bleomycin. IFN-gamma(-/-) mice demonstrated significantly lower parenchymal inflammation, weight loss, and mortality 10 d after 5 U/kg intratracheal bleomycin administration compared with control mice. At 3 wk after 1.5 U/kg bleomycin exposure, single lung collagen determined by hydroxyproline assay was significantly lower in IFN-gamma(-/-) mice compared with wild-type C57BL/6J mice. Together, these results suggest that IFN-gamma mediates, in part, bleomycin-induced pulmonary inflammation and fibrosis.
Assuntos
Bleomicina , Interferon gama/deficiência , Pneumonia/patologia , Fibrose Pulmonar/patologia , Animais , Bleomicina/administração & dosagem , Peso Corporal/efeitos dos fármacos , Líquido da Lavagem Broncoalveolar/química , Colágeno/metabolismo , Citocinas/genética , Citocinas/metabolismo , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Instilação de Medicamentos , Interferon gama/análise , Interferon gama/genética , Interleucina-4/análise , Pulmão/metabolismo , Pulmão/patologia , Camundongos , Camundongos Endogâmicos , Camundongos Knockout , Pneumonia/induzido quimicamente , Pneumonia/fisiopatologia , Fibrose Pulmonar/induzido quimicamente , Fibrose Pulmonar/fisiopatologia , RNA Mensageiro/metabolismo , Taxa de Sobrevida , TraqueiaRESUMO
Sarcoidosis, a chronic, multisystem disease, impacts quality of life and may increase depression risk. No previous study has reported the depression prevalence among U.S. sarcoid patients. This cross-sectional study examined sociodemographic and disease morbidity factors associated with depression. Patients diagnosed for > or = 1 yr and treated at one of six centers were eligible (n = 176); 154 completed a questionnaire of demographics, treatment, access to medical care, and a short-form Center for Epidemiologic Studies- Depression Scale (CES-D). The primary outcome variable was a CES-D score of > or = 9, indicating clinical depression. The prevalence of depression was 60%. Gender, income, access to medical care, dyspnea on exertion, and number of systems involved were associated with depression. Female sex, decreased access to medical care, and increased dyspnea predicted depression (odds ratio [OR] = 3.33, 11.64, and 2.78, respectively) after adjusting for race, income, and steroid therapy. Despite tertiary care access, patients reported medical care limitation. Health care providers must be sensitive to multiple barriers faced by chronic sarcoid patients; acknowledging depression risk and improving access to medical care will promote better overall health among sarcoid patients. Future studies of sarcoidosis will need to address depression diagnosis and treatment.
Assuntos
Depressão/epidemiologia , Transtorno Depressivo/epidemiologia , Sarcoidose/epidemiologia , Adulto , Idoso , Comorbidade , Estudos Transversais , Depressão/diagnóstico , Depressão/psicologia , Transtorno Depressivo/diagnóstico , Transtorno Depressivo/psicologia , Feminino , Acessibilidade aos Serviços de Saúde , Humanos , Masculino , Pessoa de Meia-Idade , Inventário de Personalidade , Qualidade de Vida , Fatores de Risco , Sarcoidose/diagnóstico , Sarcoidose/psicologia , Papel do Doente , Estados UnidosRESUMO
Granulomatous inflammation develops under the regulatory influence of cytokines produced by local mononuclear phagocytes, T cells, dendritic cells, fibroblasts, and other local cells. In sarcoidosis, granulomatous inflammation is characterized by dominant expression of T helper 1 (Th1) cytokines such as IFN gamma and interleukin (IL)-2 with low levels of expression of T helper 2 (Th2) cytokines such as IL4 and IL5. Recent studies show that the cytokine IL12, the most important regulator of Th1 immune responses currently known, is upregulated at sites of inflammation in sarcoidosis. In particular, enhanced expression of IL12 is seen in sarcoid lung and lymph node, along with dysregulated production of IL12 by stimulated and unstimulated sarcoid alveolar macrophages. The known dependence of granulomatous inflammation on type 1 cytokines (IFN gamma, IL12) in many experimental models of granulomatous disease makes it likely that these cytokines function in a similar fashion in the initiation and maintenance of granulomatous inflammation in sarcoidosis. Whether these same type 1 cytokines drive granulomatous inflammation in patients with extensive fibrocystic lung disease remains unknown. TGF beta, a known inhibitor of IL12 and IFN gamma production, is produced at higher levels by lung cells from those patients who undergo remission of their disease, suggesting that TGF gamma is important in downregulating granulomatous inflammation in sarcoidosis. These studies offer new insight into the molecular mechanisms of granuloma formation in sarcoidosis and provide a framework for developing new therapeutic strategies for the treatment of this disease.
Assuntos
Citocinas/farmacologia , Granuloma/imunologia , Pneumopatias/imunologia , Macrófagos/fisiologia , Sarcoidose/imunologia , Linfócitos T/fisiologia , Fibrose/imunologia , Fibrose/fisiopatologia , Granuloma/fisiopatologia , Humanos , Inflamação/fisiopatologia , Pneumopatias/fisiopatologia , Modelos Teóricos , Sarcoidose/fisiopatologia , Células Th1/fisiologiaRESUMO
Sarcoidosis is recognized to be a multisystem granulomatous disease characterized by activated, cytokine-producing T cells and macrophages at sites of inflammation. The purpose of this article is to review new evidence concerning the role of T cells in sarcoidosis. Recent work on the molecular structure and repertoire of T cell receptor genes in sarcoidosis provide direct evidence that sarcoidosis is characterized by selective expansions of oligoclonal populations of T cells at sites of inflammation, consistent with local antigen-driven immune responses. In addition, data on cytokine production in sarcoidosis indicate that tissue inflammation is dominated by expression of type 1 (T helper 1) cytokines such as interferon-gamma and interleukin-12 that, in keeping with experimental models of granulomatous diseases, likely orchestrate the granulomatous response. These studies offer new insight into the molecular mechanisms of granuloma formation in sarcoidosis and provide a framework for developing new therapeutic strategies for the treatment of this disease.
Assuntos
Receptores de Antígenos de Linfócitos T/imunologia , Sarcoidose Pulmonar/imunologia , Linfócitos T/imunologia , Citocinas/imunologia , HumanosRESUMO
The pathogenesis of sarcoidosis involves activated, cytokine-producing T-cells and macrophages that regulate granulomatous inflammation. At sites of inflammation, T-cells demonstrate reduced surface density of the CD3 component of the T-cell receptor complex, a hallmark of T-cells activated through the T-cell antigen receptor pathway. The stimulus activating these T-cells is unknown. Conventional antigens selectively stimulate T-cells expressing T-cell receptors with specific variable (V) and hypervariable VDJ or VJ region amino acid sequences while superantigens stimulate larger subsets of T-cells based primarily on their expression of specific V beta genes. Recent studies show that subgroups of patients with sarcoidosis are characterized by biased expression of specific V beta, V alpha, or gamma delta + T-cell receptor genes in T-cell subsets from lung, blood and at sites of Kveim-Siltzbach skin reactions. In addition, investigations on the molecular structure of T-cell receptor genes in sarcoidosis provide direct evidence that biased expression of specific alpha beta + or gamma delta + T-cells at sites of inflammation involves selective expansion of oligoclonal populations of T-cells, consistent with an immune response to a conventional antigen (s). Together, these studies provide direct evidence that sarcoidosis is an antigen-driven disorder at sites of granulomatous inflammation. The identification of key, clonally-expanded T-cell populations in sarcoidosis provides a potential tool for determining the specific antigens involved in the pathogenesis of sarcoidosis.
Assuntos
Genes Codificadores dos Receptores de Linfócitos T , Sarcoidose Pulmonar/genética , Humanos , Pulmão/metabolismo , Receptores de Antígenos de Linfócitos T alfa-beta/metabolismo , Receptores de Antígenos de Linfócitos T gama-delta/metabolismo , Sarcoidose Pulmonar/metabolismoRESUMO
Since sarcoidosis was first recognized as a distinct clinical entity, investigators have speculated that a transmissible agent may cause sarcoidosis. Recent attempts at directly isolating infectious organisms or indirectly detecting microbial DNA or RNA from sarcoid tissue have led to inconclusive results. Studies on the immunopathogenic origins of sarcoidosis have provided evidence of persistent antigenic stimulation at sites of inflammation that are associated with dysregulated cytokine production. To date, however, the challenge of defining the cause of sarcoidosis remains unmet.
Assuntos
Sarcoidose/etiologia , Doenças Autoimunes , Beriliose/complicações , Exposição Ambiental , Humanos , Infecções/complicaçõesRESUMO
The immunomodulatory properties of thalidomide are currently being exploited therapeutically in conditions as diverse as erythema nodosum leprosum, chronic graft-vs-host disease, rheumatoid arthritis, and sarcoidosis. The relevant mechanism of action of thalidomide in these diseases remains unclear. The important role recently ascribed to IL-12, a cytokine critical to the development of cellular immune responses, in the pathogenesis of several of these conditions led us to examine whether thalidomide affects the production of IL-12. Thalidomide potently suppressed the production of IL-12 from human PBMC and primary human monocytes in a concentration-dependent manner. Thalidomide-induced inhibition of IL-12 production was additive to that induced by suboptimal inhibiting doses of dexamethasone, and occurred by a mechanism independent of known endogenous inhibitors of IL-12 production. These results suggest that thalidomide may have therapeutic utility in a wide range of immunologic disorders that are characterized by inappropriate cellular immune responses.
Assuntos
Imunossupressores/farmacologia , Interleucina-12/antagonistas & inibidores , Interleucina-12/biossíntese , Talidomida/farmacologia , Anticorpos Monoclonais/farmacologia , Células Cultivadas , Humanos , Indometacina/farmacologia , Interleucina-10/imunologia , Interleucina-12/genética , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/metabolismo , Ativação Linfocitária/efeitos dos fármacos , Ativação Linfocitária/genética , Monócitos/efeitos dos fármacos , Monócitos/metabolismo , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo , Fator de Crescimento Transformador beta/imunologiaRESUMO
Pharmacological control of interleukin-12 (IL-12) production may be a key therapeutic strategy for modulating immunological diseases dominated by type-1 cytokine responses. In this study, we investigated the effects of pentoxifylline on the production of IL-12 by human blood mononuclear cells and primary human monocytes stimulated with heat-killed Staphylococcus aureus Cowan strain I (SAC) or lipopolysaccharide (LPS). Pentoxifylline potently suppressed production of IL-12 in a concentration-dependent manner. In these same experiments, tumour necrosis factor-alpha (TNF-alpha) production was inhibited and IL-10 and prostaglandin E2 (PGE2) production was enhanced by treatment with pentoxifylline. Suppression of IL-12 production by pentoxifylline was found to be independent of several known endogenous inhibitors of IL-12, such as IL-10, transforming growth factor-beta (TGF-beta), IL-4 and PGE2. RNase protection assays revealed that pentoxifylline inhibited accumulation of both IL-12 p40 and p35 mRNA, suggesting a predominant mRNA locus for pentoxifylline-induced IL-12 inhibition. Low levels of pentoxifylline added to the suppression of IL-12 production by suboptimal inhibiting doses of dexamethasone, suggesting that this drug combination may have therapeutic utility. These results provide a firm rationale for the use of pentoxifylline in clinical trials of immunological disorders characterized by inappropriate type-1 immune responses.
Assuntos
Interleucina-12/biossíntese , Leucócitos Mononucleares/efeitos dos fármacos , Pentoxifilina/farmacologia , Técnicas de Cultura de Células , Dexametasona/farmacologia , Relação Dose-Resposta a Droga , Humanos , Indometacina/farmacologia , Interleucina-10/imunologia , Interleucina-12/genética , Leucócitos Mononucleares/imunologia , Monócitos/efeitos dos fármacos , Monócitos/imunologia , RNA Mensageiro/genética , Fator de Crescimento Transformador beta/imunologiaRESUMO
Sarcoidosis is a multisystem disorder characterized by non-caseating granulomas and the accumulation of CD4+ T cells in involved tissues such as the lung. To evaluate the diversity of the CD4+ T-cell repertoire in this disorder, a detailed clonal analysis was performed in five individuals with active sarcoidosis who demonstrated preferential accumulation of T cells expressing the T-cell receptor variable gene family V beta 8 in either the lung or blood. In three individuals, analysis of unselected samples of nucleotide sequences derived from V beta 8+ lung T cells demonstrated degrees of clonality ranging from 11% to 46%, indicating the expansion of limited numbers of V beta 8+ T-cell clones in the lung. Analysis of the corresponding deduced amino acid sequences demonstrated common VDJ junctional amino acid residues in the dominant V beta 8+ T-cell clones derived from two oligoclonal V beta 8+ lung T-cell populations, consistent with an antigen-specific T-cell response. In contrast, analysis of V beta 8+ CD4+ T cells from the blood of an individual with a marked bias for peripheral blood V beta 8+ T cells demonstrated no evidence of oligoclonality, suggesting that the stimulus for circulating biased V beta-specific T cells in sarcoidosis may derive from a different, perhaps superantigenic, origin. Clinical improvement in the disease either in response to treatment with corticosteroids or as a result of spontaneous resolution was associated with a decrease in the proportion of V beta 8-specific T cells in the biased lung and/or blood T-cell compartments. Together, these observations are consistent with a role for this T-cell subset in the clinical manifestations of active granulomatous disease.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Pulmão/imunologia , Receptores de Antígenos de Linfócitos T alfa-beta/genética , Sarcoidose Pulmonar/imunologia , Adulto , Sequência de Aminoácidos , Sequência de Bases , Líquido da Lavagem Broncoalveolar/imunologia , Feminino , Humanos , Masculino , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Receptores de Antígenos de Linfócitos T alfa-beta/metabolismo , Sarcoidose Pulmonar/genéticaRESUMO
Sarcoidosis is a multisystem granulomatous disease of unknown etiology characterized by the expansion of activated oligoclonal CD4+ T cells and macrophages at sites of disease. To investigate the immunopathogenesis of sarcoidosis, we analyzed patterns of cytokine expression in bronchoalveolar lavage cells and fluid from patients with pulmonary sarcoidosis and idiopathic pulmonary fibrosis and from normal volunteers. We found dominant type 1 cytokine expression, with elevated mRNA and protein levels of IFN-gamma, but not IL-4, in sarcoid lung cells and fluid compared with those in normal samples. To define immunoregulatory mechanisms important to this type 1 response, we analyzed the expression of IL-12 and IL-10 in lung cells and fluid. Using semiquantitative PCR, we found significantly higher mRNA expression of the regulated IL-12 p40 subunit, but not IL-10, in sarcoid compared with normal lung cells. Consistent with these observations, strikingly elevated levels of p40 protein were found in sarcoid compared with normal bronchoalveolar lavage fluid. Unstimulated and Staphylococcus aureus-stimulated sarcoid alveolar macrophages produced greater amounts of IL-12 than normal alveolar macrophages when cultured in vitro. We hypothesize that sarcoidosis is a Th1-mediated disease driven by chronic, dysregulated production of IL-12 at sites of disease.
Assuntos
Citocinas/metabolismo , Interleucina-12/metabolismo , Sarcoidose Pulmonar/imunologia , Células Th1/imunologia , Adulto , Idoso , Líquido da Lavagem Broncoalveolar/química , Feminino , Humanos , Interleucina-10/metabolismo , Pulmão/imunologia , Macrófagos Alveolares/imunologia , Masculino , Pessoa de Meia-Idade , Células Th2/imunologiaRESUMO
Sarcoidosis is a multiorgan granulomatous disorder of unknown etiology characterized by noncaseating granulomas in involved tissues. A positive Kveim-Siltzbach reaction is a granulomatous response to an intradermal injection of a suspension of sarcoid tissue extract in individuals with sarcoidosis. The protracted time course and granulomatous features of this reaction have a striking resemblance to the Mitsuda reaction in tuberculous leprosy, which suggests that the Kveim-Siltzbach reaction is a response to an unknown Ag(s). To evaluate whether this reaction is Ag-driven, an analysis of the TCR V beta repertoire in 15 Kveim-Siltzbach reaction sites was performed using a PCR technique and primers specific for 20 V beta gene families. Results of this analysis demonstrated a pattern of V beta expression dominated by expression of V beta 2, V beta 3, V beta 6, or V beta 8 to levels > 20% of total V beta gene expression in nine of 15 individuals. Analysis of paired biopsy and blood specimens revealed a preferential expression of specific V beta genes, such as V beta 3, V beta 5, and V beta 8, at sites of Kveim-Siltzbach reactions to levels four to seven times that of the corresponding peripheral blood. Sequence analysis demonstrated that preferential expression of specific V beta genes at Kveim-Siltzbach reaction sites is oligoclonal. Furthermore, the dominant V beta 8 sequence present at one of the reaction sites contained a sequence motif in the variable-diversity-joining junctional region previously identified in sarcoid lung and blood T cell populations. These results suggest that the Kveim-Siltzbach reaction is characterized by a limited TCR beta-chain repertoire consistent with an Ag-driven T cell immune response.
Assuntos
Teste de Kveim/métodos , Receptores de Antígenos de Linfócitos T alfa-beta/genética , Sarcoidose/imunologia , Sequência de Aminoácidos , Sequência de Bases , Humanos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Receptores de Antígenos de Linfócitos T alfa-beta/imunologia , Sarcoidose/genética , Linfócitos T/imunologiaRESUMO
Sarcoidosis is a granulomatous disease in which activated T cells, responding to an unidentified stimulus, accumulate at sites of disease such as the lung. To evaluate the hypothesis that active sarcoidosis is characterized by a selective activation and expansion of a limited repertoire of T cell receptor (TCR) specific T cells, we analyzed TCR V beta gene expression in lung and blood T cells of patients with active sarcoidosis and, for comparison, normal individuals using polymerase chain reaction amplification of 20 V beta gene families. Analysis of normal bronchoalveolar lavage T cells revealed TCR V beta distributions similar to that of normal blood, providing evidence for a lack of generalized skewing of the T cell repertoire in the normal, noninfected lung. Compared to normal lung and blood, subgroups of individuals with sarcoidosis demonstrated biased expression of one or more V beta genes in either the lung or blood. Five V beta gene families (V beta 5, V beta 8, V beta 15, V beta 16, and V beta 18) were most frequently utilized in a biased fashion by sarcoid lung or blood T cells. Furthermore, dramatic skewing of the T cell repertoire was apparent when sarcoid lung and blood T cells were expanded by short-term culture with IL-2. Sequence analysis demonstrated a bias in V beta gene expression was usually due to expansion of select V beta-specific clones, some of which contained a similar V(D)J junctional region motif. These observations provide evidence for a selective activation and accumulation of antigen-specific V beta-expressing T cells in sarcoidosis.
Assuntos
Ativação Linfocitária/fisiologia , Receptores de Antígenos de Linfócitos T alfa-beta/genética , Sarcoidose Pulmonar/imunologia , Subpopulações de Linfócitos T/imunologia , Adulto , Sequência de Aminoácidos , Sequência de Bases , Líquido da Lavagem Broncoalveolar/citologia , Células Cultivadas , Feminino , Expressão Gênica , Genes Dominantes , Humanos , Interleucina-2/farmacologia , Pulmão/citologia , Masculino , Dados de Sequência Molecular , Receptores de Antígenos de Linfócitos T alfa-beta/análise , Análise de Sequência de DNA , Subpopulações de Linfócitos T/química , Subpopulações de Linfócitos T/efeitos dos fármacosAssuntos
Variação Antigênica/genética , Regulação da Expressão Gênica/genética , Pneumopatias/genética , Pulmão/imunologia , Receptores de Antígenos de Linfócitos T/genética , Sarcoidose/genética , Variação Antigênica/imunologia , Regulação da Expressão Gênica/imunologia , Humanos , Pneumopatias/imunologia , Receptores de Antígenos de Linfócitos T/imunologia , Sarcoidose/imunologiaRESUMO
Evaluation of the T-cells accumulating at sites of disease in active sarcoidosis suggests the accumulation process is not random, evidenced by a bias in the types of T-cells present. To evaluate the concept that this bias extends to the accumulation of T-cells with the preferential use of specific T-cell antigen receptor (TCR) beta-chain constant region elements, beta-chain mRNA transcripts of lung and blood T-cells of normal subjects and patients with pulmonary sarcoidosis were compared for the relative usage of constant region beta 1 or beta 2 elements. Quantitative evaluation of C beta 1 and C beta 2 mRNA transcripts demonstrated a C beta 1/C beta 2 usage in normal blood of 0.63 +/- 0.02, similar to that of normal lung (0.64 +/- 0.06, p greater than 0.7), and in sarcoid blood (0.59 +/- 0.03, p greater than 0.2). In contrast, the lung T-lymphocytes of patients with sarcoidosis reflected a marked bias in the usage of C beta 1 elements (C beta 1/C beta 2: 0.88 +/- 0.06, p less than 0.001 compared with sarcoid blood and normal blood; p less than 0.02 compared with normal lung). Interestingly, a subgroup of these patients (six of 18) showed a markedly exaggerated skewing in the use of C beta 1 elements (C beta 1/C beta 2 ratio greater than 1, i.e., greater than 3 standard deviations above mean), demonstrating heterogeneity among sarcoid patients with regard to specific C beta 1 usage.(ABSTRACT TRUNCATED AT 250 WORDS)