transforming-growth-factor-beta and Tuberculosis--Pulmonary

Topics: Antigens, Bacterial; CD4-Positive T-Lymphocytes; Cytokines; Interferon-gamma; Interleukin-2; Lymphocyte Activation; Monocytes; Th1 Cells; Th2 Cells; Transforming Growth Factor beta; Tuberculosis, Pulmonary

Topics: Animals; CD8-Positive T-Lymphocytes; DNA, Viral; HIV; HIV Core Protein p24; HIV Infections; Humans; Macrophages, Alveolar; Mice; Monocytes; Mycobacterium tuberculosis; Rabbits; Smoking; Transforming Growth Factor beta; Tuberculosis, Pulmonary; Tumor Necrosis Factor-alpha

Tuberculosis (TB) disease is usually marked by inflammation which is closely linked to haemostasis both in health and disease. Close monitoring of haemostatic response to inflammatory changes during treatment is important to improve TB management. Here we studied associations between haemostatic markers and inflammatory cytokines in 60 TB-infected individuals, aged 18-65 years who received anti-TB therapy. They were recruited before commencement of therapy and followed up till completion of therapy after 6-months. The TNF-α, IL-6, IL-2 (pro-inflammatory cytokines) and P-selectin, GP IIb/IIIa, thrombopoietin (haemostatic variables) were significantly increased at 2 month into therapy compared to pre-treatment values and decreased at 6 month into therapy. Also at 6 month into therapy in comparison to 2-month into therapy, there were significant increase in IL-10 and TGF-β (anti-inflammatory cytokines) as well as a significant decline in PF-4. There were significant positive correlations between GP IIb/IIIa and TNF-α, IL-6 and PSEL, IL-6 and TPO, PF4 and TGF-β. Conclusively, the changes in the TNF-α, IL-6, IL-2 aligned with changes in the levels of P-selectin, GP IIb/IIIa, and TPO in the course of TB therapy. This may suggest that the levels of inflammatory cytokines are linked to the levels of these haemostatic variables in TB individuals.

Topics: Cytokines; Hemostasis; Hemostatics; Humans; Inflammation; Interleukin-2; Interleukin-6; Mycobacterium tuberculosis; P-Selectin; Platelet Membrane Glycoprotein IIb; Transforming Growth Factor beta; Tuberculosis; Tuberculosis, Pulmonary; Tumor Necrosis Factor-alpha

Long non-coding RNAs (lncRNAs) play crucial roles in the progression and pathogenesis of cancer. Right now, less is known about the association between the expression of lncRNAs and the pathogenesis of pulmonary tuberculosis (PTB).. In present study, the expression profiles of lncRNAs were investigated by transcriptome sequencing from PTB patients vs. healthy individuals.. A total of 449 differentially expressed (DE) (fold change ≥2, false discovery rate ≤ 0.05) lncRNAs were screened out from the PTB patients. Lnc-HNRNPU-1:7 and lnc-FAM76B-4:1 was found the most upregulated lncRNAs and downregulated lncRNAs in PTB patients, respectively. GO annotation and KEGG analysis were used to explore the potential roles of these DE lncRNAs. The JAK/STAT and TGF-β signaling pathways related to PTB pathogenesis were enriched in PTB patients. The co-expressed of a few lncRNAs and mRNAs on chromosome were shown by cis-regulatory gene analysis. Trans analysis indicated that STAT1, STAT2 and TAF7 transcription factors regulated the expression of lncRNA and mRNA. The constructed lncRNA ceRNA network suggested that lncRNAs regulating mRNAs expression may mediate by sponged miRNAs.. We comprehensively analyzed the expression profiles of lncRNAs in PTB patients, thus providing new clues for exploring the regulatory mechanisms of dysregulated lncRNAs in the pathogenesis of PTB.

Topics: Gene Regulatory Networks; Humans; Janus Kinases; MicroRNAs; RNA, Long Noncoding; RNA, Messenger; STAT Transcription Factors; TATA-Binding Protein Associated Factors; Transcription Factor TFIID; Transcriptome; Transforming Growth Factor beta; Tuberculosis, Pulmonary

Little is known about how tuberculosis (TB) impairs dendritic cell (DC) function and anti-TB immune responses. We previously showed that the B and T lymphocyte attenuator (BTLA), an immune inhibitory receptor, is involved in TB pathogenesis. Here, we examined whether BTLA expression in TB affects phenotypic and functional aspects of DCs. Active TB patients exhibited higher expression of BTLA in myeloid dendritic cells (mDCs) and plasmacytoid DCs (pDCs) subsets compared with healthy controls (HCs). BTLA expression was similarly high in untreated TB, TB relapse, and sputum-bacillus positive TB, but anti-TB therapy reduced TB-driven increases in frequencies of BTLA

Topics: Adolescent; Adult; Aged; Cell Differentiation; Dendritic Cells; Female; Humans; Interferon-alpha; Interleukin-12; Interleukin-4; Lymphocyte Activation; Male; Middle Aged; Receptors, Immunologic; T-Lymphocytes, Regulatory; Th2 Cells; Transforming Growth Factor beta; Tuberculosis, Pulmonary; Young Adult

Aging influences the susceptibility and prognosis to various infectious diseases including tuberculosis (TB). Despite the impairment of T-cell function and immunity in older individuals, the mechanism for the higher incidence of TB in the elderly remains largely unknown. Here, we evaluated the age-associated immune alterations, particularly in effector and Treg responses in pulmonary TB patients. We also evaluated the impact of redox status and its modulation with N-acetyl-cysteine (NAC) in elderly TB. Higher frequency of Treg cells and reduced IFN-γ positive T cells were observed among older TB patients. The elevated number of Treg cells correlated tightly with bacillary load (i.e. disease severity); which declined significantly in response to successful anti-tubercular treatment. We could rescue Myobacterium tuberculosis-specific effector T cell (Th1) responses through various in vitro approaches, for example, Treg cell depletion and co-culture experiments, blocking experiments using antibodies against IL-10, TGF-β, and programmed death-1 (PD-1) as well as NAC supplementation. We report old age-associated enrichment of Treg cells and suppression of M. tuberculosis-specific effector T (Th1) cell immune responses. Monitoring these immune imbalances in older patients may assist in immune potentiation through selectively targeting Treg cells and/or optimizing redox status by NAC supplementation.

Topics: Acetylcysteine; Adolescent; Adult; Age Factors; Aged; Cytokines; Female; Humans; Male; Middle Aged; Oxidation-Reduction; Oxidative Stress; Programmed Cell Death 1 Receptor; T-Lymphocytes, Regulatory; Th1 Cells; Transforming Growth Factor beta; Tuberculosis, Pulmonary; Young Adult

Tuberculosis (TB) caused by Mycobacterium tuberculosis is a health problem worldwide. Patients with pulmonary TB show a neuro-immune-endocrine imbalance characterized by an impaired cellular immunity together with increased plasma levels of cortisol, pro- and anti-inflammatory cytokines and markedly decreased dehydroepiandrosterone (DHEA) levels. Extending these findings, we now investigated the immune-endocrine profile of TB patients undergoing specific treatment. Patients (n = 24) were bled at diagnosis (T0), 2, 4, 6 months after treatment initiation and 3 months following its completion. At T0, TB patients showed increased plasma levels of interleukin-6 (IL-6), C reactive protein, interferon-gamma (IFN-γ) and transforming growth factor beta (TGF-β). These mediators decreased during treatment, reaching levels similar to those from healthy controls (n = 26). Specific treatment led to an increased lymphoproliferative response along with clinical improvement. Newly diagnosed patients had low levels of DHEA, with increased cortisol amounts and cortisol/DHEA ratio, which normalized upon specific treatment. As regards glucocorticoid receptors (GR), TB patients at diagnosis presented a reduced mRNA GRα/GRβ ratio in their peripheral blood mononuclear cells. Furthermore, multivariate analysis showed that cortisol/DHEA ratio was positively associated with inflammatory mediators for which this ratio may constitute a disease biomarker. Anti-mycobacterial treatment results in a better immune-endocrine scenario for the control of physiopathological processes accompanying disease development and hence implied in clinical recovery.

Topics: Adult; Antitubercular Agents; C-Reactive Protein; Case-Control Studies; Dehydroepiandrosterone; Ethambutol; Female; Gene Expression Regulation; Humans; Hydrocortisone; Interferon-gamma; Interleukin-6; Isoniazid; Leukocytes, Mononuclear; Male; Middle Aged; Mycobacterium tuberculosis; Pyrazinamide; Receptors, Glucocorticoid; Rifampin; Transforming Growth Factor beta; Treatment Outcome; Tuberculosis, Pulmonary

We have reported previously that T cells from patients with multi-drug-resistant tuberculosis (MDR-TB) express high levels of interleukin (IL)-17 in response to the MDR strain M (Haarlem family) of Mycobacterium tuberculosis (M. tuberculosis). Herein, we explore the pathways involved in the induction of Th17 cells in MDR-TB patients and healthy tuberculin reactors [purified protein derivative healthy donors (PPD

Topics: Adult; Cells, Cultured; Drug Resistance, Multiple, Bacterial; Female; Humans; Immunologic Memory; Interferon-gamma; Interleukin-17; Interleukin-23; Male; Middle Aged; Mycobacterium tuberculosis; Signal Transduction; Species Specificity; Th17 Cells; Toll-Like Receptor 2; Transforming Growth Factor beta; Tuberculosis, Multidrug-Resistant; Tuberculosis, Pulmonary; Young Adult

Role of lncRNAs in human adaptive immune response to TB infection is largely unexplored. To address this issue, here we characterized lncRNA expression profile in primary human B cell response to TB infection using microarray assay. Several lncRNAs and mRNAs were chosen for RT-qPCR validation. Bioinformatics prediction was applied to delineate function of the deregulated mRNAs. We found that 844 lncRNAs and 597 mRNAs were differentially expressed between B cell samples from individuals with or without TB. KEGG pathway analysis for the deregulated mRNAs indicated a number of pathways, such as TB, TLR signaling pathway and antigen processing and presentation. Moreover, corresponding to the dysregulation of many lncRNAs, we also found that their adjacent protein-coding genes were also deregulated. Functional annotation for the corresponding mRNAs showed that these lncRNAs were mainly associated with TLR signaling, TGF-β signaling. Interestingly, SOCS3, which is a critical negative regulator of cytokine response to TB infection and its nearby lncRNA XLOC_012582, were highly expressed in active TB B cells. Subsequent RT-qPCR results confirmed the changes. Whether upregulated XLOC_012582 causes SOCS3 overexpression and is eventually involved in the context of exacerbations of active TB represents an interesting issue that deserves to be further explored. Taken together, for the first time, we identified a set of deregulated lncRNAs in active TB B cells and their functions were predicted. Such findings provided novel insight into the pathogenesis of TB and further studies should focus on the function and pathogenic mechanisms of the lncRNAs involved in active TB.

Topics: Adaptive Immunity; Adult; Antigen Presentation; B-Lymphocytes; Case-Control Studies; Computational Biology; Female; Gene Expression Profiling; Gene Expression Regulation; Gene Ontology; Humans; Male; Microarray Analysis; Middle Aged; Molecular Sequence Annotation; Mycobacterium tuberculosis; Primary Cell Culture; RNA, Long Noncoding; RNA, Messenger; Signal Transduction; Suppressor of Cytokine Signaling 3 Protein; Toll-Like Receptors; Transforming Growth Factor beta; Tuberculosis, Pulmonary

Mycobacterium tuberculosis (M. tb) is the etiological agent of pulmonary tuberculosis (TB); this disease remains a worldwide health problem. Yin-Yang-1 (YY1) plays a major role in the maintenance and progression of some pulmonary diseases, including pulmonary fibrosis. However, the role of YY1 in TB remains unknown. The aim of this study was to elucidate the role of YY1 in the regulation of CCL4 and its implication in TB. We determined whether YY1 regulates CCL4 using reporter plasmids, ChIP and siRNA assays. Immunohistochemistry and digital pathology were used to measure the expression of YY1 and CCL4 in a mouse model of TB. A retrospective comparison of patients with TB and control subjects was used to measure the expression of YY1 and CCL4 using tissue microarrays. Our results showed that YY1 regulates the transcription of CCL4; moreover, YY1, CCL4 and TGF-β were overexpressed in the lung tissues of mice with TB during the late stages of the disease and the tissues of TB patients. The expression of CCL4 and TGF-β correlated with YY1 expression. In conclusion, YY1 regulates CCL4 transcription; moreover, YY1 is overexpressed in experimental and human TB and is positively correlated with CCL4 and TGF-β expression. Therefore, treatments that decrease YY1 expression may be a new therapeutic strategy against TB.

Topics: Animals; Cell Line; Chemokine CCL4; Chromatin Immunoprecipitation; Disease Models, Animal; Disease Progression; Gene Expression Regulation; Host-Pathogen Interactions; Humans; Immunohistochemistry; Lung; Male; Mice, Inbred BALB C; Mycobacterium tuberculosis; Retrospective Studies; RNA Interference; Signal Transduction; Time Factors; Tissue Array Analysis; Transcription, Genetic; Transfection; Transforming Growth Factor beta; Tuberculosis, Pulmonary; YY1 Transcription Factor

Tuberculosis (TB) is a major health problem requiring an appropriate cell immune response (IR) to be controlled. Since regulatory T cells (Tregs) are relevant in IR regulation, we analyzed Tregs variations throughout the course of TB treatment and its relationship with changes in immune-endocrine mediators dealing with disease immunopathology. The cohort was composed of 41 adult patients, 20 of them completing treatment and follow-up. Patients were bled at diagnosis (T0) and at 2 (T2), 4 (T4), 6 (T6), and 9 months following treatment initiation. Twenty-four age- and sex-matched healthy controls (HCo) were also included. Tregs (flow cytometry) from TB patients were increased at T0 (versus HCo P < 0.05), showing even higher values at T2 (versus T0 P < 0.01) and T4 (versus T0 P < 0.001). While IL-6, IFN-γ, TGF-β (ELISA), and Cortisol (electrochemiluminescence, EQ) were augmented, DHEA-S (EQ) levels were diminished at T0 with respect to HCo, with cytokines and Cortisol returning to normal values at T9. Tregs correlated positively with IFN-γ (R = 0.868, P < 0.05) at T2 and negatively at T4 (R = -0.795, P < 0.05). Lowered levels of proinflammatory cytokines together with an increased frequency of Tregs of patients undergoing specific treatment might reflect a downmodulatory effect of these cells on the accompanying inflammation.

Topics: Adult; Antitubercular Agents; CD4 Antigens; Dehydroepiandrosterone Sulfate; Female; Flow Cytometry; Forkhead Transcription Factors; Humans; Hydrocortisone; Interferon-gamma; Interleukin-2 Receptor alpha Subunit; Interleukin-6; Lung; Male; Middle Aged; T-Lymphocytes, Regulatory; Transforming Growth Factor beta; Tuberculosis, Pulmonary; Young Adult

Interleukin-37 (IL-37), a member of the IL-1 family, primarily functions as an anti-inflammatory cytokine, reducing inflammation and suppressing the immune response. However, the expression and role of IL-37 in tuberculosis (TB) remains unknown. We aimed to measure serum levels of IL-37 and several important cytokines in 25 patients with active TB and to analyse their association with disease activity. We found that IL-37 levels decreased in patients with TB and recovered after treatment. IL-37 levels negatively correlated with the serum concentration of IFN-γ and IL-12 but positively correlated with IL-10 and TGF-β levels. After IL-37, secretion was blocked in peripheral blood mononuclear cells from active patients with TB, IFN-γ and IL-10 production was significantly upregulated; this was not observed in healthy donors or patients after treatment. IL-37 knockdown significantly enhanced the phagocytic activity of THP1-derived macrophages towards Mycobacterium tuberculosis (M. tb). M1/M2 polarization-associated markers were detected simultaneously, and IL-37 induced a phenotypic shift in THP1-derived macrophages towards a high CD206(+) and low CD86(+) macrophage subtype. Furthermore, this phenotypic shift was accompanied by upregulated mRNA levels of arginase 1, TGF-β and IL-10, which are characteristic hallmarks of M2 macrophages. In conclusion, our results suggest that increased levels of IL-37 in patients with TB are associated with IFN-γ, IL-12, IL-10 and TGF-β levels and that IL-37 plays a pathological role in TB infection by inhibiting the production of pro-inflammatory cytokines and inducing macrophages towards an M2-like phenotype. Thus, IL-37 may be a novel research target to understand the pathogenesis of TB infection.

Topics: Antitubercular Agents; Arginase; B7-2 Antigen; Cell Differentiation; Cells, Cultured; Ethambutol; Female; Humans; Interferon-gamma; Interleukin-1; Interleukin-10; Interleukin-12 Subunit p35; Isoniazid; Lectins, C-Type; Macrophages; Male; Mannose Receptor; Mannose-Binding Lectins; Middle Aged; Mycobacterium tuberculosis; Nitric Oxide; Phagocytosis; Phenotype; Pyrazinamide; Receptors, Cell Surface; Rifampin; RNA Interference; RNA, Messenger; RNA, Small Interfering; Transcriptional Activation; Transforming Growth Factor beta; Tuberculosis, Pulmonary; Up-Regulation

In patients with infiltrative pulmonary tuberculosis, the increase in IL-6 secretion, the decrease in TGFβ production (in case of drug resistance of the causative agent), and unchanged level of IL-1β secretion by mononuclear leukocytes in vitro were associated with increased number of CD4(+)CD161(+)IL-17A(+) Th17 lymphocytes in the peripheral blood. In patients with disseminated drug-resistant pulmonary tuberculosis, TGFβ hyperproduction promoted differentiation of CD4(+)CD25(+)FoxP3(+) Treg lymphocytes with immunosuppressive activity.

Topics: Adult; CD4 Antigens; Cell Differentiation; Female; Forkhead Transcription Factors; Humans; Interleukin-2 Receptor alpha Subunit; Male; Middle Aged; NK Cell Lectin-Like Receptor Subfamily B; Statistics, Nonparametric; T-Lymphocytes, Regulatory; Th17 Cells; Transforming Growth Factor beta; Tuberculosis, Pulmonary

We studied possible mechanisms of immunosuppression mediated by regulatory T cells that promotes suppression of antigen-specific immune response to Mycobacterium tuberculosis in patients with pulmonary tuberculosis and eosinophilia. It was shown that the number of CD4(+)CD25(+)Foxp3(+) regulatory T cells with immunosuppressive activity (Treg) increased in the peripheral blood of patients with disseminated destructive forms of pulmonary tuberculosis with multiple resistance of the causative agent to antituberculosis substances and eosinophilia. These changes were accompanied by imbalance in secretion of Treg-associated cytokines (in vitro) manifested in hyperproduction of TGFβ and IL-10 and decreased production of IL-2.

Topics: Adolescent; Adult; CD4-Positive T-Lymphocytes; Eosinophilia; Female; Forkhead Transcription Factors; Humans; Interleukin-10; Interleukin-2; Interleukin-2 Receptor alpha Subunit; Male; Middle Aged; Mycobacterium tuberculosis; T-Lymphocytes, Regulatory; Transforming Growth Factor beta; Tuberculosis, Pulmonary; Young Adult

During intracellular residence in macrophages, mycobacterial lipids, namely phosphatidylinositol mannosides (PIM) and lipoarabinomannans, are expelled in the lung milieu to interact with host cells. PIM include a group of essential lipid components of Mycobacterium tuberculosis (M. tb) cell wall. Given that PIM function as mycobacterial adhesins for binding to host cells, the present study explored the consequences of interaction of M. tb PIM with alveolar epithelial cells (AEC). A 24-h PIM exposure at a concentration of 10 μg/mL to AEC conferred cytolysis to AEC via induction of apoptosis, suggesting their potential to alter alveolar epithelium integrity. The results also reflected that type I like AEC are more sensitive to cytolysis than type II AEC. PIM-treated AEC exhibited significant production of reactive oxygen species (ROS) and an immunosuppressive cytokine transforming growth factor-β (TGF-β) in the culture supernatants. Although AEC displayed constitutive mRNA transcripts for toll-like receptors (TLR2 and 4) as well as chemokines (IL-8 and MCP-1), no significant change in their expression was observed upon PIM treatment. Collectively, these results offer insights into PIM potential as M. tb virulence factor that might promote mycobacterial dissemination by causing cytolysis of AEC via increased production of ROS and TGF-β.

Topics: Adhesins, Bacterial; Antigens, Bacterial; Apoptosis; Cell Line; Chemokines; Cytokines; Epithelial Cells; Host-Pathogen Interactions; Humans; Immunomodulation; Mycobacterium tuberculosis; Phosphatidylinositols; Pulmonary Alveoli; Reactive Oxygen Species; RNA, Messenger; Toll-Like Receptors; Transforming Growth Factor beta; Tuberculosis, Pulmonary; Virulence

Circulating T follicular helper (Tfh) cells represent a distinct subset of CD4+ T cells and are important in immunity to infections. Although they have been shown to play a role in experimental models of tuberculosis infection, their role in human tuberculosis remains unexplored.. To determine the distribution of circulating Tfh cells in human TB, we measured the frequencies of Tfh cells ex vivo and following TB - antigen or polyclonal stimulation in pulmonary TB (PTB; n = 30) and latent TB (LTB; n = 20) individuals, using the markers CXCR5, PD-1 and ICOS.. We found that both ex vivo and TB - antigen induced frequencies of Tfh cell subsets was significantly lower in PTB compared to LTB individuals. Similarly, antigen induced frequencies of Tfh cells expressing IL-21 was also significantly lower in PTB individuals and this was reflected in diminished circulating levels of IL-21 and IFNγ. This was not accompanied by diminished frequencies of activated or memory B cell subsets. Finally, the diminution in frequency of Tfh cells in PTB individuals was dependent on IL-10, CTLA-4 and PD-L1 in vitro.. Thus, PTB is characterized by adiminution in the frequency of Tfh cell subsets.

Topics: Antigens, Bacterial; CD3 Complex; CTLA-4 Antigen; Humans; Immunologic Memory; Inducible T-Cell Co-Stimulator Protein; Interleukin-10; Interleukins; Latent Tuberculosis; Lymphocyte Count; Plasma Cells; Programmed Cell Death 1 Receptor; T-Lymphocytes, Helper-Inducer; Transforming Growth Factor beta; Tuberculosis, Pulmonary

The nervous, endocrine and immune systems play a crucial role in maintaining homeostasis and interact with each other for a successful defensive strategy against injurious agents. However, the situation is different in long-term diseases with marked inflammation, in which defensive mechanisms become altered. In the case of tuberculosis (TB), this is highlighted by several facts: an imbalance of plasma immune and endocrine mediators, that results in an adverse environment for mounting an adequate response against mycobacteria and controlling inflammation; the demonstration that dehidroepiandrosterone (DHEA) secretion by a human adrenal cell line can be inhibited by culture supernatants from Mycobacterium tuberculosis-stimulated peripheral blood mononuclear cells - PBMC - of TB patients, with this effect being partly reverted when neutralizing transforming growth factor-β in such supernantants; the in vitro effects of adrenal steroids on the specific immune response of PBMC from TB patients, that is a cortisol inhibition of mycobacterial antigen-driven lymphoproliferation and interferon-γ production as well as a suppression of TGF-β production in DHEA-treated PBMC; and lastly the demonstration that immune and endocrine compounds participating in the regulation of energy sources and immune activity correlated with the consumption state of TB patients. Collectively, immune-endocrine disturbances of TB patients are involved in critical components of disease pathology with implications in the impaired clinical status and unfavorable disease outcome. This article is part of a Special Issue entitled 'Neuroinflammation in neurodegeneration and neurodysfunction'.

Topics: Cytokines; Dehydroepiandrosterone; Humans; Inflammation; Neuroimmunomodulation; Stress, Psychological; Transforming Growth Factor beta; Tuberculosis, Pulmonary

The article deals with the characteristics of sup-population composition of T-regulator: cells with suppressor activity and production of immunoregulatory cytokines suppressing immune response (IL-10, TGF-beta) in patients with infiltrative tuberculosis of lungs. It is proved that the leading role in formation of immunodepression and tuberculin anergy under infiltrative tuberculosis of lungs is played by T-regulatory, cells with phenotype CD4+CD25+Foxp3+. It is demonstrated that the immunodepression mediated by cytokine production is connected with increasing of both basal and BCG-induced secretion of IL-10 on the background of decrease of level of production of TGF-beta by mononuclear leukocytes in vitro.

Topics: Adolescent; Adult; Clonal Anergy; Female; Humans; Interleukin-10; Male; Middle Aged; T-Lymphocytes, Regulatory; Transforming Growth Factor beta; Tuberculin; Tuberculosis, Pulmonary

Supernatants (SN) from cultures of peripheral blood mononuclear cells (PBMC) of tuberculosis (TB) patients inhibit dehydroepiandrosterone (DHEA) secretion by the adrenal cell line NCI-H295R. To analyze whether TGF-β is involved in this effect, SN of PBMC from healthy controls or patients with severe TB infections, stimulated or not with Mycobacterium tuberculosis (Mtb SN), were added to adrenal cells under basal conditions or following stimulation with forskolin. Cortisol and DHEA concentrations were evaluated in supernatants of the adrenal cells cultured with or without the addition of anti-TGF-β. Treatment with Mtb SN from TB inhibited DHEA production, and this effect was reversed when SN were treated with anti-TGF-β. The increase in cortisol production induced by SN from TB patients was not affected by TGF-β neutralization. Mediators released during the anti-TB immune response differentially modulate steroid production by adrenal cells, and TGF-β is a cytokine implicated in the inhibition of DHEA production observed in TB.

Topics: Adrenal Glands; Adult; Case-Control Studies; Cell Line; Colforsin; Culture Media, Conditioned; Dehydroepiandrosterone; Female; Host-Pathogen Interactions; Humans; Hydrocortisone; In Vitro Techniques; Leukocytes, Mononuclear; Male; Middle Aged; Mycobacterium tuberculosis; Neutralization Tests; Transforming Growth Factor beta; Tuberculosis, Pulmonary

Different cytokines have been suggested to be involved in the pathogenesis of pulmonary tuberculosis (TB). The frequencies of Mycobacterium tuberculosis (MTB) specific CD4(+) and CD8(+) T cells, CD4(+)CD25(+) Forkhead Box Protein (FoxP)3(+) T cells, interleukin (IL)-6, interferon (IFN)-γ, Tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-β and IL-10 were assessed in HIV-negative, pulmonary tuberculosis (TB) patients (n=30) and in healthy controls (n=23) in Gabon. Peripheral blood mononuclear cells (PBMC) were stimulated with purified protein derivative (PPD) and early secretory antigenic target-6 (ESAT-6). In patients, a pronounced pro-inflammatory cytokine response with highly significant increased levels of IL-6 and TNF-α accompanied by increased TGF-β was detectable. Differences in IFN-γ responses between patients and healthy individuals were less pronounced than expected. FoxP3 expression did not differ between groups. A distinct cytokine pattern is associated with active pulmonary TB in patients from Central Africa.

Topics: Adolescent; Adult; Aged; Antigens, Bacterial; Bacterial Proteins; Body Temperature; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cell Count; Cytokines; Female; Forkhead Transcription Factors; Gabon; Gene Expression; Hemoglobins; Humans; Interferon-gamma; Interleukin-10; Interleukin-2; Interleukin-6; Leukocytes, Mononuclear; Male; Middle Aged; T-Lymphocyte Subsets; Transforming Growth Factor beta; Tuberculin; Tuberculosis, Pulmonary; Tumor Necrosis Factor-alpha; Young Adult

Although it has been recognized that ectopic follicle-like B cell aggregate formation is common in the lungs of patients with tuberculosis, the role of infiltrated B cells in human tuberculosis remains to be elucidated. In the present study, we showed that ectopic B cell aggregate formation was associated with containment of Mycobacterium tuberculosis. The area ratio of ectopic B cell aggregates was correlated with localized IL-17 mRNA expression and peripheral TGF-β and IL-6 mRNA expression. Depletion of B cells from pleural fluid mononuclear cells resulted in significantly diminished M. tuberculosis antigen-specific IL-17 and IL-22 production, but not in IFN-γ secretion. Therefore, ectopic lung B cell formation is important for containment of M. tuberculosis, and up-regulation of IL-17 and IL-22 responses may be an important mechanism underlying the protective role B cells in human tuberculosis.

Topics: Adolescent; Adult; Antigens, Bacterial; B-Lymphocytes; Cell Aggregation; Cytokines; Female; Humans; Interleukin-17; Interleukin-22; Interleukin-6; Interleukins; Male; Middle Aged; Mycobacterium tuberculosis; RNA, Messenger; Transforming Growth Factor beta; Tuberculosis, Pulmonary; Up-Regulation

By applying the current immunological and molecular genetic studies, the authors revealed a genetically determined imbalance in the pro- duction of immunoregulatory cytokines by monoluclear leukocytes in pulmonary tuberculosis. T allele and TT genotype of IFNG gene poly- morphism +874A/T were found to be an immunogenetic factor that had a protective effect against susceptibility to pulmonary tuberculosis. Susceptibility to tuberculosis infection is associated with A allele and with AA and AT genotypes of IFNG gene polymorphism +874A/T and the CTpolymorphic variant G509Tof the TGFB gene. The maximum risk for pulmonary tuberculosis is associated with a combination of the AA genotypes of the polymorphic site +874A/Tof the IFNG gene and TT polymorphism G509T of the TGFB gene (AA/TT).

Topics: Adult; Cytokines; Female; Genetic Predisposition to Disease; Haplotypes; Humans; Interferon-gamma; Male; Middle Aged; Polymorphism, Single Nucleotide; Transforming Growth Factor beta; Tuberculosis, Pulmonary

Human tuberculosis (TB) principally involves the lungs, where local immunity impacts on the load of Mycobacterium tuberculosis (M.tb). Because concomitants of local Th1 immunity are still under-explored in humans, we characterized immune responses in bronchoalveolar cells (BACs) and systemically in peripheral blood mononuclear cells (PBMCs) in persons with active pulmonary TB and in healthy community controls. PPD- and live M.tb-induced IFN-gamma-production were observed in CD4(+), CD8(+), gammadeltaTCR(+), and CD56(+) alveolar T cell subpopulations and NK cells (CD3(-)CD56(+)). IFN-gamma-producing CD4(+) T cells (mostly CD45RO(+)) were more abundant (p<0.05). M.tb-induced IL-12p70, but interestingly also IL-4, was increased (p<0.05) in BACs from TB patients. Constitutive expression of IL-12Rbeta1 and IL-12Rbeta2 mRNA in BACs and PBMCs and IFN-gammaR1 in BACs was similar in both study groups. Data were normalized to account for differences in proportions of alveolar T cells and macrophages in the study groups. IFN-gamma-production and its induction by IL-12R engagement occur virtually unimpaired in the bronchoalveolar spaces of patients with pulmonary TB. The reasons for the apparent failure to control M. tuberculosis growth during active pulmonary TB disease is unknown but could be the expression of locally acting immunosuppressive mechanisms that subvert the antimycobacterial effects of IFN-gamma.

Topics: Adult; Bacteriological Techniques; Bronchi; Bronchoalveolar Lavage Fluid; Case-Control Studies; CD4-Positive T-Lymphocytes; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Gene Expression; Humans; Immunologic Memory; Interferon-gamma; Interleukin-10; Interleukin-12; Interleukin-4; Interleukin-9; Lung; Lymphocyte Activation; Male; Middle Aged; Mycobacterium tuberculosis; Pulmonary Alveoli; Reverse Transcriptase Polymerase Chain Reaction; Transforming Growth Factor beta; Tuberculosis, Pulmonary

Cytokine messenger RNA (mRNA) expression was investigated in the spleen and lung digest cells of bacillus Calmette-Guérin (BCG)-vaccinated and non-vaccinated guinea pigs following low-dose, pulmonary exposure to virulent Mycobacterium tuberculosis. After purified protein derivative (PPD) stimulation, the levels of lung cell interferon-gamma (IFN-gamma), tumour necrosis factor-alpha (TNF-alpha) and spleen cell interleukin-12 (IL-12) p40 mRNAs were significantly increased in the non-vaccinated M. tuberculosis-infected guinea pigs compared to the BCG-vaccinated guinea pigs. In contrast, the expression of anti-inflammatory transforming growth factor-beta and IL-10 mRNAs was significantly enhanced in the spleens of BCG-vaccinated animals. Despite the presence of protective cytokine mRNA expression, the non-vaccinated guinea pigs had significantly higher lung and spleen bacterial burdens. In contrast, BCG-vaccinated guinea pigs controlled the bacterial multiplication in their lungs and spleens, indicating that both protective as well as anti-inflammatory cytokine responses are associated with a reduction in bacteria. In addition, lung digest cells from non-vaccinated guinea pigs contained a significantly higher percentage of neutrophils, CD3(+) and CD8(+) T cells, while the percentage of macrophages was increased in the BCG-vaccinated animals. Total and purified lung digest T cells co-cultured with lung macrophages (LMøs) proliferated poorly after PPD stimulation in both non-vaccinated and BCG-vaccinated animals while robust proliferation to PPD was observed when T cells were co-cultured with peritoneal macrophages (PMøs). Macrophages within the lung compartment appear to regulate the response of T cells irrespective of the vaccination status in guinea pigs. Taken together, our results suggest that type I cytokine mRNA expression is not associated with vaccine-induced protection in the low-dose guinea pig model of tuberculosis.

Topics: Animals; BCG Vaccine; Cell Proliferation; Concanavalin A; Disease Models, Animal; Guinea Pigs; Interferon-gamma; Interleukin-10; Interleukin-12 Subunit p40; Lung; Macrophages, Alveolar; Macrophages, Peritoneal; Mitogens; Mycobacterium tuberculosis; RNA, Messenger; Spleen; Stem Cells; T-Lymphocytes; Transcription, Genetic; Transforming Growth Factor beta; Tuberculin; Tuberculosis, Pulmonary; Tumor Necrosis Factor-alpha; Vaccination

Failure of mice to produce IL-10 has no effect on the bacterial burden of Mycobacterium tuberculosis infection in the lungs over the first 4-5 months of the disease. We show here that after 185 days of the infection, IL-10 gene disrupted (IL-10 KO) mice showed evidence of bacterial regrowth, began to show signs of wasting, and were moribund. We assessed the status of the acquired immune response and compared the lung lymphocyte cell populations, as well as the expression of Th1 (IL-12 and IFNgamma) and immunosuppressive (TGFbeta) cytokines, between IL-10 KO and wild type mice. The results demonstrated that at 60 days of the infection in the absence of IL-10 there was an increased expression of Th1 type immunity and an overall lack of control of the inflammatory responses. After 185 days of the infection, in the absence of IL-10 there was excessive pulmonary inflammation and increased expression of inflammatory cytokine TNFalpha. These results imply therefore that IL-10 plays a central role in dampening of the Th1 response and protection against chronic lung inflammation in the M. tuberculosis infected lung, and the complete removal of this regulatory component eventually leads to disease progression.

Topics: Animals; Cytokines; Disease Progression; Female; Flow Cytometry; Immune Tolerance; Interleukin-10; Lung; Lymphocyte Activation; Lymphocyte Subsets; Mice; Mice, Knockout; Mycobacterium tuberculosis; Th1 Cells; Transforming Growth Factor beta; Tuberculosis, Pulmonary

Studies of patients with active tuberculosis (TB) and infected healthy individuals have shown that interferon (IFN)-gamma is present in sites of Mycobacterium tuberculosis infection in comparable levels. This suggests that there is a deficiency in the macrophage response to IFN-gamma in TB patients. We used recombinant human IFN-gamma to stimulate adherent monocyte-derived macrophages from three groups of people: patients with active tuberculosis (TBP), their healthy household contacts (HHC) and healthy uninfected controls from the community (CC). We then evaluated the ability of the macrophages to inhibit the growth of M. tuberculosis H37Rv as well as their cytokine profile at early in infection (48 h). After IFN-gamma treatment, macrophages of healthy individuals (HHC and CC) controlled M. tuberculosis growth and produced mainly nitric oxide (NO) and interleukin (IL)-12p70, whereas TBP macrophages did not kill M. tuberculosis. Additionally, TBP macrophages produced low levels of NO and IL-12p70 and high levels of tumour necrosis factor (TNF)-alpha and IL-10. Transforming growth factor (TGF)-beta levels were similar among all three groups. M. tuberculosis infection had little effect on the cytokine response after IFN-gamma stimulus, but infection alone induced more IL-10 and TGF-beta in TBP macrophages. There were no differences in Stat1 nuclear translocation and DNA binding between the groups. However, the phosphorylated Stat1 and c-Jun (AP-1) in nuclear protein extracts was diminished in TBP macrophages compared to macrophages of healthy individuals. These results indicate an impairment of Stat1-dependent and Stat1-independent IFN-gamma signalling in macrophages of people with active tuberculosis, suggesting a different molecular regulation that could impact macrophage functionality and disease outcome.

Topics: Adult; Blotting, Western; Case-Control Studies; Cell Nucleus; Colony-Forming Units Assay; Electrophoretic Mobility Shift Assay; Humans; Interferon-gamma; Interleukin-10; JNK Mitogen-Activated Protein Kinases; Macrophages; Middle Aged; Mycobacterium tuberculosis; Nitric Oxide; Recombinant Proteins; STAT1 Transcription Factor; STAT5 Transcription Factor; Statistics, Nonparametric; Transforming Growth Factor beta; Tuberculosis, Pulmonary; Tumor Suppressor Proteins; Young Adult

This study investigated interleukin-4 (IL-4), IL-4 delta 2, transforming growth factor beta (TGF-beta), TGF-beta RII, Foxp3, GATA-3, T-bet, and gamma interferon (IFN-gamma) transcription in peripheral blood samples of adult pulmonary tuberculosis patients prior to and after 1 week of therapy. Twenty patients with positive results for sputum culture for Mycobacterium tuberculosis were enrolled and treated with directly observed short-course antituberculosis chemotherapy. Early treatment response was assessed. At the end of the intensive phase of treatment (month 2), 12 patients remained sputum culture positive (slow responders) and 8 converted to a negative culture (fast responders). Only the expression levels of IL-4 (4-fold decrease) and IL-4 delta 2 (32-fold increase) changed significantly during the first week of therapy in the 20 patients. No baseline differences were present between the responder groups, but fast responders had significantly higher IL-4 transcripts than slow responders at week 1. Fast responders showed a 19-fold upregulation and slow responders a 47-fold upregulation of IL-4 delta 2 at week 1. Only slow responders also showed a significant decrease in IL-4 expression at week 1. There were no significant differences in expression of TGF-beta, TGF-beta RII, Foxp3, IFN-gamma, and GATA-3 between the groups. These data show that differential IL-4-related gene expression in the early stage of antituberculosis treatment accompanies differential treatment responses and may hold promise as a marker for treatment effect.

Topics: Adolescent; Adult; Antitubercular Agents; Female; Forkhead Transcription Factors; GATA3 Transcription Factor; Gene Expression Regulation; Humans; Interferon-gamma; Interleukin-4; Male; Middle Aged; Mycobacterium tuberculosis; Receptors, Transforming Growth Factor beta; RNA, Messenger; Sputum; T-Box Domain Proteins; Time Factors; Transforming Growth Factor beta; Treatment Outcome; Tuberculosis, Pulmonary

To evaluate the pattern of pro-inflammatory cytokines, anti-inflammatory cytokines and the acute phase response (APR) as markers of the response to treatment of pulmonary tuberculosis.. Twenty-eight patients with pulmonary tuberculosis were evaluated at three time points: pretreatment (T0), treatment month 3 (T3) and treatment month 6 (T6). Levels of interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), interleukine-10 (IL-10) and transforming growth factor-beta (TGF-beta) were determined using ELISA in the supernatant of peripheral blood mononuclear cell and monocyte culture. Levels of total protein, albumin, globulins, C-reactive protein (CRP), alpha-1-acid glycoprotein (AAG) and erythrocyte sedimentation rate (ESR) were also determined. All of these parameters were also evaluated, only once, in a group of healthy controls.. In relation to controls, patients presented cytokine levels and APR that were higher at T0, lower at T3 and either lower (TNF-alpha, IL-10, TGF-beta, AAG and ESR) or normal (IFN-gamma and CRP) at T6.. For individuals with negative smear sputum microscopy, CRP, AAG and ESR are potential markers of pulmonary tuberculosis and of the need for treatment; CRP (T0 > T3 > T6 = reference) can also be a marker of treatment response. In the patients, the Th0 profile (IFN-gamma, IL-10, TNF-alpha and TGF-beta), inducer of and protector against inflammation, predominated at T0, whereas the Th2 profile (IL-10, TNF-alpha and TGF-beta), protecting against the harmful pro-inflammatory effect of the remaining TNF-alpha, predominated at T6. The behavior of IFN-gamma (T0 > T3 > T6 = controls) suggests its use as a marker of treatment response.

Topics: Adult; Aged; Biomarkers; Blood Sedimentation; C-Reactive Protein; Cell Culture Techniques; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interferon-gamma; Interleukin-10; Male; Middle Aged; Time Factors; Transforming Growth Factor beta; Tuberculosis, Pulmonary; Tumor Necrosis Factor-alpha; Young Adult

Levels of IL-12p40, TNFalpha, TGFbeta, IFNgamma and IL-10 mRNA were assessed by laser capture microdissection followed by quantitative real-time PCR in the pulmonary granulomas of unimmunized and BCG-vaccinated guinea pigs infected by aerosol with virulent Mycobacterium tuberculosis. Lesions microdissected from unimmunized guinea pigs were overwhelmed by the pro-inflammatory TNFalpha mRNA at both 3 and 6 weeks post infection, indicating the struggle to control the mounting infection. The cytokine profile of granulomas from vaccinated guinea pigs shifted from type 1 cytokine mRNA (IFNgamma and IL-12p40) at 3 weeks to a predominantly anti-inflammatory environment (TGFbeta mRNA) at 6 weeks. The relative proportions of cytokine mRNA transcripts in the periphery of the granuloma were different from the centre, reflecting differences in cell composition and architecture. Moreover, analysis of the individual lung lobes at 6 weeks post infection suggests that heterogeneity exists in the cytokine profile between the lobes of the lung.

Topics: Animals; BCG Vaccine; Cytokines; Gene Expression Profiling; Granuloma; Guinea Pigs; Interferon-gamma; Interleukin-10; Interleukin-12; Interleukin-12 Subunit p40; Lung; Microdissection; Mycobacterium tuberculosis; RNA, Messenger; Time Factors; Transforming Growth Factor alpha; Transforming Growth Factor beta; Tuberculosis, Pulmonary; Virulence

CD4(+) CD25(+) regulatory T cells produce the anti-inflammatory cytokines transforming growth factor (TGF)-beta or interleukin (IL)-10. Regulatory T cells have been recognized to suppress autoimmunity and promote self-tolerance. These cells may also facilitate pathogen persistence by down-regulating the host defence response during infection with Mycobacterium tuberculosis. We evaluated TGF-beta(+) and IL-10(+) lung CD4(+) CD25(+) T cells in a murine model of M. tuberculosis. BALB/c mice were infected with approximately 50 colony-forming units of M. tuberculosis H37Rv intratracheally. At serial times post-infection, lung cells were analysed for surface marker expression (CD3, CD4, CD25) and intracellular IL-10, TGF-beta, and interferon (IFN)-gamma production (following stimulation in vitro with anti-CD3 and anti-CD28 antibodies). CD4(+) lung lymphocytes were also selected positively after lung digestion, and stimulated in vitro for 48 h with anti-CD3 and anti-CD28 antibodies in the absence and presence of anti-TGF-beta antibody, anti-IL-10 antibody or rmTGF-beta soluble receptor II/human Fc chimera (TGFbetasrII). Supernatants were assayed for elicited IFN-gamma and IL-2. Fluorescence activated cell sorter analyses showed that TGF-beta- and IL-10-producing CD4(+) CD25(+) T cells are present in the lungs of infected mice. Neutralization of TGF-beta and IL-10 each resulted in increases in elicited IFN-gamma, with the greatest effect seen when TGFbetasrII was used. Elicited IL-2 was not affected significantly by TGF-beta neutralization. These results confirm the presence of CD4(+) CD25(+) TGF-beta(+) T cells in murine pulmonary tuberculosis, and support the possibility that TGF-beta may contribute to down-regulation of the host response.

Topics: Animals; CD4-Positive T-Lymphocytes; Cells, Cultured; Cytokines; Immune Tolerance; Interleukin-10; Interleukin-2 Receptor alpha Subunit; Lung; Mice; Mice, Inbred BALB C; T-Lymphocyte Subsets; Transforming Growth Factor beta; Tuberculosis, Pulmonary

Tuberculosis (TB) has different clinical presentations. Pulmonary TB affects only the lungs and exhibits variable anti-mycobacterial immune responses. Pleural TB is a localized disease with a strong immune response. Miliary TB is a disseminated form with poor immune response. Cytokines play a pivotal role in anti-mycobacterial response and may determine the type of TB. Thus, gene polymorphisms associated with cytokine production may be associated with clinical presentations of TB. In this study, 54 tuberculin-negative healthy controls, 81 tuberculin-positive healthy controls, 140 patients with pulmonary TB, 30 with pleural TB and 20 with miliary TB were studied. Single nucleotide polymorphisms were typed for tumour necrosis factor-alpha, interferon-gamma (IFN-gamma), transforming growth factor-beta1, interleukin-10 (IL-10) and interleukin-6 by sequence-specific primer polymerase chain reaction (SSP-PCR). Allelic, genotypic and haplotypic associations with clinical forms of TB were evaluated. IL-10 -1082 A/A genotype and IFNgamma+874 T allele were associated with pleural TB. Seventy-five extended genotypes were found; two differed between patients and controls, and two between groups of patients. Results suggest that IL-10 low-producer polymorphism and IFN-gamma high-producer polymorphism are associated with pleural TB.

Topics: Adolescent; Adult; Aged; Case-Control Studies; Colombia; Cytokines; Female; Humans; Interferon-gamma; Interleukin-10; Interleukin-6; Male; Middle Aged; Polymorphism, Genetic; Transforming Growth Factor beta; Tuberculosis, Miliary; Tuberculosis, Pleural; Tuberculosis, Pulmonary; Tumor Necrosis Factor-alpha

Active tuberculosis (TB) is associated with prolonged suppression of Mycobacterium tuberculosis (MTB)-specific immune responses, but mechanisms involved are understood incompletely. We investigated a potential role for CD4+CD25+ regulatory T cells in depressed anti-MTB immunity by evaluating serially CD4 cell phenotype and interferon (IFN)-gamma production by mononuclear cells from patients with TB. At diagnosis, frequencies of CD4+CD25+ T cells were increased in blood from TB patients compared to healthy purified protein derivative (PPD)-positive controls (with a history of prior TB exposure), and remained elevated at completion of therapy (6 months). By contrast, expression of another activation marker, CD69, by CD4 T cells was increased at diagnosis, but declined rapidly to control levels with treatment. Among CD4+CD25+ T cells from TB patients at diagnosis those expressing high levels of CD25, probably representing regulatory T cells, were increased 2.9-fold when compared to control subjects, while MTB-stimulated IFN-gamma levels in whole blood supernatants were depressed. A role for CD4+CD25+ T cells in depressed IFN-gamma production during TB was substantiated in depletion experiments, where CD25+-depleted CD4 T cells produced increased amounts of IFN-gamma upon MTB stimulation compared to unseparated T cells. At follow-up, IFN-gamma production improved most significantly in blood from TB patients with high baseline frequencies of CD4+CD25+ T cells (more than threefold higher than controls for both total and CD25hi+ CD4 T cells), who also had a significant drop in frequencies of both total and 'regulatory' CD4+CD25+ T cells in response to treatment. Expansion of CD4+CD25+ regulatory T cells during active TB may play a role in depressed T cell IFN-gamma production.

Topics: Adult; Antigens, Bacterial; Antigens, CD; Antigens, Differentiation, T-Lymphocyte; CD8-Positive T-Lymphocytes; Female; Humans; Interferon-gamma; Interleukin-10; Lectins, C-Type; Lung; Male; Monocytes; Mycobacterium tuberculosis; Phenotype; Receptors, Interleukin-2; T-Lymphocytes, Regulatory; Transforming Growth Factor beta; Tuberculin; Tuberculosis, Pulmonary; Tumor Necrosis Factor-alpha

Transforming growth factor-beta (TGF-beta) and prostaglandins (PG) regulate the cell-mediated immune response, so it has been proposed that they affect the progression of pulmonary tuberculosis. Here we report that the administration of soluble betaglycan, a potent TGF-beta antagonist, and niflumic acid, a PG synthesis inhibitor, during the chronic phase of experimental murine tuberculosis enhanced Th1 and decreased Th2 cytokines, increased the expression of iNOS and reduced pulmonary inflammation, fibrosis and bacillary load. This immunotherapeutic approach resulted in significant control of the disease comparable to that achieved by anti-microbial treatment alone. Importantly, the combination of immunotherapy and anti-microbials resulted in an accelerated clearance of bacilli from the lung. These results confirm that TGF-beta and PG have a central pathophysiological role in the progression of pulmonary tuberculosis in the mouse and suggest that the addition of immunotherapy to conventional anti-microbial drugs might result in improved treatment of the disease.

Topics: Animals; Antitubercular Agents; Colony Count, Microbial; Cyclooxygenase Inhibitors; Cytokines; Disease Models, Animal; Hypersensitivity, Delayed; Immunotherapy; Lung; Male; Mice; Mice, Inbred BALB C; Niflumic Acid; Nitric Oxide Synthase Type II; Prostaglandin Antagonists; Proteoglycans; Receptors, Transforming Growth Factor beta; Th1 Cells; Th2 Cells; Transforming Growth Factor beta; Tuberculosis, Pulmonary

Tuberculosis (TB) is the main cause of death by infection diseases worldwide. Considering that NO, TNF-alpha and TGF-beta participate a great deal in TB immunopathogenesis, we wished to analyse whether these mediators showed some relationship with the degree of pulmonary affectation. The sample comprised 29 TB (HIV-), inpatients with mild-moderate (n = 10) or advanced (n = 19) newly-diagnosed disease, together with 12 healthy controls HCo. Serum nitrite was assessed by reducing nitrate to nitrite, and further measured by the Griess reaction. Levels of TNF-alpha and TGF-beta were determined by ELISA (R&D Systems). Serum levels of TNF-alpha were significantly higher in the advanced TB cases if compared with HCo, (p < 0.05 ) and from values of Mild-Moderate TB patients (p < 0.05). Serum levels of TGF-beta from advanced TB patients have increased values if compared with Hco (p < 0.005) and Mild-Moderate patients (p < 0.05). These values were also significantly different from Mild-Moderate cases + HCo (p = 0.01) Advanced TB patients had significantly reduced nitrite levels compared with those of Mild-Moderate patients and HCo (p < 0.002). Taken as a whole NO-derived metabolites in TB patients (M-M and Advanced cases) remained lower than values in HCo (p = 0.005) A negative correlation was found when comparing the two cytokines with nitrites(r = -0.44 ).TGF-beta and TNF-alpha were positively correlated (r = 0.44, p < 0.01), 0.44, p < 0.01. In synthesis, the inverse correlation found between both cytokines concentrations and NO levels in TB patients may be viewed as a consequence of a more predominant TGF-beta effect.

Topics: Adult; Humans; Mycobacterium tuberculosis; Nitric Oxide; Severity of Illness Index; Transforming Growth Factor beta; Tuberculosis, Pulmonary; Tumor Necrosis Factor-alpha

The role of TGF-beta TNF-alpha FasL and Bcl-2 in apoptosis of CD4 T-cells during active TB was studied. Coculture of PBMC from TB patients with neutralizing antibodies to TGF-beta or TNF-alpha decreased spontaneous (P < or = 0.05) and MTB-induced (P < or = 0.02) T-cell apoptosis by 50-90%, but effects were not additive. Interestingly, only levels of TGF-beta in supernatants correlated with rates of spontaneous and MTB-induced apoptosis. FasL surface and mRNA expression were higher in unstimulated and MTB-stimulated PBMC from patients than controls, and neutralization of FasL abrogated apoptosis of T-cells from patients only. Intracellular Bcl-2 protein was lower among unstimulated CD4 T-cells from patients than those from controls (P < or = 0.02), and MTB stimulation reduced intracellular Bcl-2 content in CD4 T-cells from patients only (P < or = 0.001). These findings may indicate that, during TB, predisposition of CD4 T-cells to apoptosis may involve both low expression of Bcl-2, and excessive expression of TGF-beta TNF-alpha and FasL.

Topics: Adolescent; Adult; Apoptosis; CD4-Positive T-Lymphocytes; Down-Regulation; Fas Ligand Protein; fas Receptor; Female; Humans; Male; Middle Aged; Mycobacterium tuberculosis; Proto-Oncogene Proteins c-bcl-2; Receptors, Tumor Necrosis Factor, Type II; Transforming Growth Factor beta; Tuberculosis, Pulmonary; Tumor Necrosis Factor-alpha; Up-Regulation

The aim of the present study was to determine the immune response profile that differentiates patients with newly diagnosed (non-treated) pulmonary tuberculosis from multidrug-resistant (MDR) ones, as well as from healthy, tuberculin positive individuals.. Lymphocytes proliferative response to non-specific mitogen (PHA) and PPD were evaluated by 3H thymidine incorporation and cytokines were quantified using an ELISA assay.. Patients with active disease showed a diminished proliferative response to PHA and PPD, while multidrug-resistant patients showed a diminished proliferative response to PHA, but a normal response to PPD. The cytokine production of newly diagnosed patients was characterized by a diminished production of IFNgamma and normal production of transforming growth factor (TGFbeta), while MDR patients revealed a normal production of IFNgamma accompanied by an increase in TGFbeta.. The production of significant amounts of TGFbeta in MDR patients leads to a poor immune response and may contribute to the resistance of tuberculosis patients to drugs.

Topics: Adult; Brazil; Case-Control Studies; Drug Resistance, Multiple; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunity, Cellular; Interferon-gamma; Leukocytes, Mononuclear; Lymphocyte Activation; Lymphocytes; Male; Middle Aged; Mitogens; Mycobacterium tuberculosis; Transforming Growth Factor beta; Tuberculin; Tuberculosis, Pulmonary

Immune factors influencing progression to active tuberculosis (TB) remain poorly defined. In this study, we investigated the expression of immunoregulatory cytokines and receptors by using lung bronchoalveolar lavage cells obtained from patients with pulmonary TB, patients with other lung diseases (OLD patients), and healthy volunteers (VOL) by using reverse transcriptase PCR, a transforming growth factor beta (TGF-beta) bioactivity assay, and an enzyme immunoassay. TB patients were significantly more likely than OLD patients to coexpress TGF-beta receptor I (RI) and RII mRNA, as well as interleukin-10 (IL-10) mRNA (thereby indicating the state of active gene transcription in the alveolar cells at harvest). In contrast, gamma interferon (IFN-gamma) and IL-2 mRNA was seen in both TB and OLD patients. Likewise, significantly elevated pulmonary steady-state protein levels of IL-10, IFN-gamma, and bioactive TGF-beta were found in TB patients versus those in OLD patients and VOL. These data suggest that the combined production of the immunosuppressants IL-10 and TGF-beta, as well as coexpression of TGF-beta RI and RII (required for cellular response to TGF-beta), may act to down-modulate host anti-Mycobacterium tuberculosis immunity and thereby allow uncontrolled bacterial replication and overt disease. Delineating the underlying mechanisms of M. tuberculosis-triggered expression of these immune elements may provide a molecular-level understanding of TB immunopathogenesis.

Topics: Activin Receptors, Type I; Adult; Base Sequence; Bronchoalveolar Lavage Fluid; Case-Control Studies; DNA, Complementary; Female; Gene Expression; Humans; Immune Tolerance; Interleukin-10; Lung Diseases; Male; Middle Aged; Protein Serine-Threonine Kinases; Receptor, Transforming Growth Factor-beta Type I; Receptor, Transforming Growth Factor-beta Type II; Receptors, Transforming Growth Factor beta; RNA, Messenger; Transforming Growth Factor beta; Tuberculosis, Pulmonary

Following infection with Mycobacterium tuberculosis, host cytokine responses influence disease manifestation. Differences in cytokine expression likely determine whether tuberculosis (TB) progresses, resolves, or becomes latent. In particular, the balance between Th(1) and Th(2) cytokine responses influences the expression of disease in individuals with pulmonary TB.. Since the cytokine microenvironment in pulmonary TB remains suboptimally defined, we utilized quantitative immunohistochemistry to compare the expression of Th(1) cytokines [interferon-gamma (IFNgamma) and interleukin-12 (IL-12)] and Th(2) cytokines [IL-4, IL-10, transforming growth factor-beta (TGFbeta)] in surgically resected lungs of seven TB patients and four control subjects. We also quantified IFNgamma-inducible protein 10 (IP-10) expression, a CXC chemokine for macrophages and T cells.. Morphometric analyses revealed increased IFNgamma, IL-12, IP-10, and TGFbeta in granulomas and in pneumonitis areas of TB lungs. In contrast, IL-10 and IL-4 expressions were globally reduced in TB lung tissues compared to controls.. Th(1) cytokines and TGFbeta are increased while Th(2) cytokines are decreased in well-formed pulmonary granulomas of TB patients compared to controls.

Topics: B-Lymphocytes; Chemokine CXCL10; Chemokines, CXC; Cytokines; Humans; Immunohistochemistry; Interferon-gamma; Interleukin-10; Interleukin-12; Interleukin-4; Lung; Macrophages; Th1 Cells; Th2 Cells; Transforming Growth Factor beta; Tuberculosis, Pulmonary

The present study was designed to evaluate the distribution of epithelioid cells, myofibroblasts, and TGF-beta1 in the formation of granuloma caused by Mycobacterium avium intracellulare complex (MAC) lung infection. A retrospective study was performed for 9 cases of positive MAC culture in which lung resections were performed between January 1989 and August 1999. Resected lung specimens were evaluated histologically and immunohistochemically for CD68 (stain for monocytes and macrophages, and epithelioid cells) and alpha-smooth muscle actin as well as vimentin (stain for myofibroblasts), and TGF-beta1 was performed. When granuloma was initially formed, no myofibroblasts were found, but as caseous necrosis appeared, the thin epithelioid cell layer was detected and the outer myofibroblast layer gradually became thick. In the cavitary wall, the layer of epithelioid cells and multinucleated giant cells surrounded necrosis, and was associated with the outer layer of myofibroblasts. In addition, the anti-TGF-beta1 antibody stained the cytoplasm of epithelioid cells and multinucleated giant cells, preceding the advent of myofibroblasts. In summary, our present study evaluated distributions of epithelioid cells, myofibroblasts, and TGF-beta along with the morphogenesis of granuloma, and clearly demonstrated the immunohistochemical difference between granuloma with caseous necrosis and granulomas without caseous necrosis.

Topics: Actins; Adult; Aged; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Epithelioid Cells; Female; Fibroblasts; Granuloma; Humans; Immunohistochemistry; Lung; Male; Middle Aged; Mycobacterium avium-intracellulare Infection; Necrosis; Occupational Diseases; Peptide Fragments; Pneumonectomy; Retrospective Studies; Transforming Growth Factor beta; Transforming Growth Factor beta1; Tuberculosis, Pulmonary; Vimentin

Pulmonary tuberculosis is an infectious disease caused by Mycobacterium tuberculosis (MTB). This microorganism is capable of inducing a delayed hypersensitivity in the lung, with subsequent expression of the disease. This reaction depends on the presence of different cytokines that exert specific functions. To study the variability of different cytokine responses after MTB antigenic challenge, we used antigens derived from MTB to stimulate the monocytes from both normal healthy contact and the patients with active tuberculosis. We found in the resting state monocytes from healthy contact secreted higher amounts of IL-12 than those from patients. After stimulation with MTB antigen, the secretion of IL-12 did not increase in normal healthy contact, but in patients with tuberculosis the secretion increased. After MTB antigen stimulation, monocytes from patients with active tuberculosis secreted a higher amount of TNF-alpha. In summary, the patterns of monocyte-derived cytokine secretion upon mycobacterial antigen challenge were different in these two groups.

Topics: Adult; Antigens, Bacterial; Cells, Cultured; Female; Humans; Interleukin-12; Male; Middle Aged; Monocytes; Mycobacterium tuberculosis; Transforming Growth Factor beta; Transforming Growth Factor beta1; Tuberculosis, Pulmonary; Tumor Necrosis Factor-alpha

Transforming growth factor-beta (TGF-beta) plays an important role in many diseases, influencing as it does such processes as immune responses, fibrosing processes, and angiogenesis. Recently, polymorphisms have been described for TGF-beta that are associated with the risk of several diseases. In this study, we investigated whether TGF-beta 1 polymorphism has an effect on sarcoidosis and tuberculosis.. TGF-beta 1 Codon 10 T869C polymorphism was investigated in 110 healthy control subjects, 104 sarcoidosis patients, and 101 tuberculosis patients.. The TGF-beta genotype was determined using polymerase chain reaction restriction fragment length polymorphism.. We found no significant differences in TGF-beta genotypes between sarcoidosis patients and healthy controls or tuberculosis patients and controls. The long axis of the tuberculin skin test was larger in the CC type compared with the CT type. However, there was no association between the TGF-beta genotype and the roentgenographic stage, the disappearance of shadows, or organ involvement in sarcoidosis, nor any association between genotype, the extent or type of roentgenographic shadow, or detected volume of tubercle bacilli in tuberculosis.. From the results, we believe that TGF-beta polymorphisms on the whole do not have a strong influence on disease onset or clinical progression in sarcoidosis and tuberculosis, although this polymorphism might have an effect on the immune response in a tuberculosis host.

Topics: Adult; Aged; Codon; Disease Progression; Female; Genotype; Humans; Male; Middle Aged; Polymorphism, Genetic; Sarcoidosis; Severity of Illness Index; Skin Tests; Transforming Growth Factor beta; Tuberculosis, Pulmonary

Levels of tumour necrosis factor (TNF)-alpha, transforming growth factor (TGF)-beta, and interleukin (IL)-10 in plasma of pulmonary tuberculosis (TB) patients and healthy contacts and plasma and pleural fluid of patients with tuberculous pleuritis were examined by enzyme immunoassay. Plasma TNF-alpha and IL-10 were elevated to significant levels in healthy contacts. High levels of TGF-beta and IL-10 were also detected in plasma from TB patients and healthy contacts. Pleural fluid contained all three cytokines with the level of IL-10 being highest followed by TGF-beta and TNF-alpha. Plasma of tuberculous pleuritis patients also had detectable levels of the three cytokines. Increased levels of TNF-alpha in plasma of contacts and to some extent pleural fluid of pleuritis patients, is perhaps to limit the infection, while elevated IL-10 in plasma of TB patients and contacts and pleural fluid would perhaps modulate excess proinflammation. Elevated TGF-beta in TB patients suggests its role in the immunopathogenesis.

Topics: Adolescent; Adult; Female; HIV Seronegativity; Humans; Interleukin-10; Male; Middle Aged; Organ Specificity; Pleural Effusion; Transforming Growth Factor beta; Tuberculosis, Pleural; Tuberculosis, Pulmonary; Tumor Necrosis Factor-alpha

The expression of transforming growth factor (TGF-beta 1), tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) were assessed in lung tissues from patients with tuberculosis. Vimentin, a constitutively expressed cellular protein, was present in 12 of 19 tissue sections indicating adequate preservation of tissue proteins in these cases. Immunohistochemical studies for cytokines were done in the vimentin positive sections only. TGF-beta 1 was localized to mononuclear phagocytes of tuberculous lung lesions in 4 of 12 tuberculosis patients. TNF-alpha, IFN-gamma, and IL-4 were absent in sections from all tuberculosis patients. The failure to detect the latter cytokines may indicate that these molecules may not be expressed at the site of disease, or are not a feature of the late stages of tuberculous granulomas. TGF beta-1, although not universally expressed, may be involved in the development and/or consequences of tuberculous granuloma formation. These data substantiate further the role of TGF-beta 1 in the immunopathology of tuberculosis.

Topics: Antibodies, Monoclonal; Case-Control Studies; Humans; Interferon-gamma; Interleukin-4; Phagocytes; Transforming Growth Factor beta; Tuberculosis, Pulmonary; Tumor Necrosis Factor-alpha; Vimentin

Immune parameters were compared in four groups of Ugandan subjects: HIV-and HIV+ adult patients with active pulmonary TB (HIV- PTB n = 38; HIV+ PTB n = 28), patients with HIV infection only (n = 26) and PPD+ healthy controls (n = 25). Compared with healthy controls, CD4 and CD8 T cells from patients with HIV and/or PTB expressed more activation markers (HLA-DR, CD38); their CD8 T cells expressed more CD95 (pre-apoptosis) and less CD28 (co-stimulatory receptor). Peripheral blood mononuclear cells (PBMC) of patients with either HIV or PTB were impaired in interferon-gamma (IFN-gamma) production upon antigenic stimulation. PTB (with or without HIV) was characterized by monocytosis, granulocytosis, increased transforming growth factor-beta 1 production and PPD-induced apoptosis. In vivo CD4 T cell depletion, in vitro increased spontaneous CD4 T cell apoptosis and defects in IFN-gamma responses upon mitogenic stimulation were restricted to HIV+ subjects (with or without PTB). Overlapping and distinctive immune alterations, associated with PTB and HIV, might explain mutual unfavourable influences of both diseases.

Topics: Adolescent; Adult; AIDS-Related Opportunistic Infections; Apoptosis; CD4 Lymphocyte Count; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cell Division; Cells, Cultured; Cytokines; Female; Humans; Interferon-gamma; Leukocytes, Mononuclear; Lymphocyte Activation; Male; Middle Aged; Mitogens; Pokeweed Mitogens; Sputum; Transforming Growth Factor beta; Transforming Growth Factor beta1; Tuberculin; Tuberculosis, Pulmonary; Tumor Necrosis Factor-alpha; Uganda

Earlier studies in patients with pulmonary TB have revealed a higher production of Th1 cell type cytokines in moderate TB, with predominant Th2-like responses in advanced disease. Given the influence of IL-12 in T cell differentiation, as well as the roles of transforming growth factor-beta (TGF-beta), nitric oxide and tumour necrosis factor-alpha (TNF-alpha) in the immune response against intracellular pathogens, we decided to analyse the interferon-gamma (IFN-gamma), IL-4, IL-12, TGF-beta, TNF-alpha and nitrite concentrations in culture supernatants of PBMC from TB patients showing different degrees of lung involvement. The sample population comprised 18 untreated TB patients with either moderate (n = 9) or advanced (n = 9) disease and 12 age- and sex-matched healthy controls (total population (patients and controls) 12 women, 18 men, aged 37 +/- 13 years (mean +/- s.d.)). PBMC were stimulated with whole sonicate from Mycobacterium tuberculosis and the supernatants were collected on day 4 for measurement of cytokine and nitrite levels. Antigen-stimulated IFN-gamma, TGF-beta and TNF-alpha production was found to be significantly increased in TB patients, both moderate and advanced, compared with the controls. Levels of IFN-gamma were significantly higher in moderate disease than advanced cases, whereas advanced cases showed significantly higher IL-12, TGF-beta and TNF-alpha concentrations when compared with cases of moderate TB. Nitrite levels were also increased in TB patients and the increase was statistically significant when advanced cases were compared with controls. These findings may contribute to a clearer picture of the net effect of cytokine interactions in TB, essential for a better understanding of the immunopathological mechanisms underlying the distinct clinical forms of the disease.

Topics: Adolescent; Adult; Aged; Cells, Cultured; Cytokines; Female; Humans; Interferon-gamma; Interleukin-12; Interleukin-4; Leukocytes, Mononuclear; Male; Middle Aged; Mycobacterium tuberculosis; Nitrites; Transforming Growth Factor beta; Tuberculosis, Pulmonary; Tumor Necrosis Factor-alpha

Pediatric tuberculosis (TB) differs from adult TB in many features. To date, cytokine expression has not been studied in children with TB. The relative amounts of the various cytokines released at the site of infection may be important determinants of TB disease development and pathology. We determined cytokine transcripts in bronchoalveolar cells (BACs) recovered from 9 children presenting with TB and from 9 children with pulmonary diseases other than TB. An RT-PCR-based method was developed to quantify the mRNAs encoding six cytokines (IFN- gamma, IL-12, TNF- alpha, IL-10, IL-4, TGF- beta 1) known to play key roles in mycobacterial infections. Expression of mRNA encoding TGF- beta, TNF- alpha and IFN- gamma was statistically significantly higher in BACs from children with TB than in BACs from children with other pulmonary diseases; whereas the levels of mRNA transcription for TGF- beta is high, the levels of mRNA transcription for IFN- gamma and TNF- alpha remain low. All children had low levels of mRNA for IL-12(p40). IL-4 was barely detectable in all cases. Children with miliary TB had high levels of IL-10 transcripts and low levels of mRNA encoding TGF- beta. The immunosuppressive cytokines TGF- beta and IL-10, are overproduced in children with non-miliary TB and miliary TB respectively and are probably involved in the progression of the disease. These data suggest that Th1 responses are reduced in children with TB.

Topics: Adolescent; Bronchoalveolar Lavage Fluid; Case-Control Studies; Child; Child, Preschool; Cytokines; Female; Humans; Infant; Interferon-gamma; Interleukin-10; Interleukin-12; Interleukin-4; Male; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Statistics, Nonparametric; Transforming Growth Factor beta; Tuberculosis, Pulmonary; Tumor Necrosis Factor-alpha

Immunological and clinical profiles were evaluated in 2 groups: human immunodeficiency virus (HIV)-uninfected and HIV-infected patients, with newly diagnosed pulmonary tuberculosis (TB), and tuberculin-skin-test-reactive healthy control subjects. HIV-uninfected patients with TB were also followed up longitudinally during and after chemotherapy. At the time of diagnosis, purified protein derivative (PPD)-stimulated production of interferon (IFN)-gamma by peripheral blood mononuclear cells from TB patients was depressed, compared with that of healthy control subjects, whereas levels of transforming growth factor (TGF)-beta and interleukin (IL)-10 were increased. In longitudinal studies, PPD stimulated production of IL-10 and TGF-beta returned to baseline by 3 months, whereas IFN-gamma production remained depressed for at least 12 months. These data indicate that the immunosuppression of TB is not only immediate and apparently dependent (at least in part) on immunosuppressive cytokines early during the course of Mycobacterium TB infection but is also long lasting, presumably relating to a primary abnormality in T-cell function.

Topics: Adolescent; Adult; Antibodies, Bacterial; Antitubercular Agents; Coculture Techniques; Cytokines; HIV Infections; Humans; Interferon-gamma; Interleukin-10; Longitudinal Studies; Middle Aged; Mycobacterium tuberculosis; T-Lymphocytes; Transforming Growth Factor beta; Tuberculin; Tuberculin Test; Tuberculosis, Pulmonary

Besides the classical major histocompatibility complex (MHC) class I and MHC class II molecules, human CD1 molecules have been shown to present mycobacterial antigens in vitro. In this study, in vivo treatment of mice with anti-CD1 monoclonal antibodies resulted in exacerbated tuberculosis at very early time points. In CD1-modulated mice, Mycobacterium tuberculosis-specific production of the type 1 cytokines, IL-12, TNF, and IFN-gamma as well as of TGF beta was reduced. These findings suggest an antigen-presenting role of CD1 molecules in tuberculosis.

Topics: Animals; Antibodies, Monoclonal; Antigens, CD1; Colony Count, Microbial; Cytokines; Histocompatibility Antigens Class I; Histocompatibility Antigens Class II; Interferon-gamma; Interleukin-12; Lung; Mice; Mice, Inbred C57BL; Mice, Knockout; Time Factors; Transforming Growth Factor beta; Tuberculosis, Pulmonary; Tumor Necrosis Factor-alpha

TGF-beta 1 has been implicated as an important mediator of immuno-suppression in clinical tuberculosis.. The objective was to determine the role of TGF-beta 1 in experimental pulmonary tuberculosis in the guinea pig.. Groups of guinea pigs, maintained on either a low protein (LP) diet or an isocaloric high protein (HP) diet, were challenged via the respiratory route with virulent Mycobacterium tuberculosis H37Rv. Ten days post-infection, guinea pigs were given daily intraperitoneal injections of recombinant human TGF-beta 1 (rhTGF-beta 1 tau for 10 consecutive days). Following the treatment, guinea pigs were euthanized, and PPD-induced proliferation of peripheral blood mononuclear cells (PBMCs) was assessed and disease resistance measured by recovery of mycobacteria from the lungs and spleens. In a second set of experiments, groups of HP and LP guinea pigs were vaccinated with attenuated M. tuberculosis H37Ra. Six weeks later, the effects of rhTGF-beta 1 on lymphoproliferation and cytokine production were determined.. Protein deficiency significantly impaired host anti-tuberculosis resistance, as expected. Treatment with rhTGF-beta 1 significantly increased mycobacterial loads in the tissues of guinea pigs and decreased the PPD-induced proliferation of PBMCs from both LP and HP guinea pigs. PPD-driven lymphoproliferation, TNF-alpha and IFN production were significantly suppressed in vaccinated, protein-deficient guinea pigs, and rhTGF-beta 1 further inhibited lymphoproliferation and cytokine production.. Both in vivo and in vitro results indicate that TGF-beta 1 exerts immunosuppressive activity and exacerbates the progression of experimental pulmonary tuberculosis in both normally nourished and protein-deficient guinea pigs.

Topics: Animals; Cytokines; Dietary Proteins; Guinea Pigs; Humans; Immunosuppression Therapy; Protein Deficiency; Recombinant Proteins; T-Lymphocytes; Transforming Growth Factor beta; Tuberculosis, Pulmonary

Topics: Animals; BCG Vaccine; Disease Models, Animal; Drug Resistance, Microbial; Guinea Pigs; Mycobacterium tuberculosis; Protein Deficiency; Recombinant Proteins; Transforming Growth Factor beta; Tuberculosis, Pulmonary

Local cellular immune responses may affect presentation and outcome in tuberculosis (TB). To investigate this hypothesis, we performed bronchoalveolar lavage (BAL) on 30 patients with untreated pulmonary tuberculosis and assessed the type of cellular inflammatory response and cytokine production. We then correlated BAL findings and cytokine production with clinical findings. We also performed BAL on a subset of patients to examine changes in cytokine production by BAL cells over time. We found that at presentation patients with less clinically and radiographically advanced TB (smear-negative, noncavitary disease) had a local immune response characterized by a predominance of lymphocytes. Furthermore, BAL cells from these patients secreted interferon (IFNgamma), and not Interleukin-4, suggesting a Th 1-type lymphocytic response. In patients with smear-positive and/or cavitary disease, macrophages or polymorphonuclear leukocytes were the predominant BAL cell type, but with treatment and clinical improvement these patients went on to recruit IFNgamma producing cells to the lung. We conclude that the type of cellular immune response that occurs locally in the lung may affect presentation and outcome in pulmonary TB, and an understanding of the development of this response may lead to insights into pathogenesis and novel therapies for TB.

Topics: Adult; Aged; AIDS-Related Opportunistic Infections; Antitubercular Agents; Bronchoalveolar Lavage Fluid; Cell Movement; Cytokines; Female; Follow-Up Studies; Humans; Immunity, Cellular; Interferon-gamma; Interleukin-1; Interleukin-4; Lung; Lymphocytes; Macrophages; Male; Middle Aged; Neutrophils; Radiography; Th1 Cells; Transforming Growth Factor beta; Treatment Outcome; Tuberculosis, Pulmonary; Tumor Necrosis Factor-alpha

The effect of exogenous transforming growth factor beta (TGF-beta) on Mycobacterium bovis BCG-induced tumor necrosis factor alpha (TNF-alpha) production by human mononuclear cells was studied. It was found that TNF-alpha production by human cells stimulated with BCG was significantly inhibited by TGF-beta. The specificity of the observed inhibition was demonstrated, since the addition of an anti-TGF-beta neutralizing monoclonal antibody completely reversed the inhibitory effect. Furthermore, the suppressive effect of TGF-beta on TNF-alpha secretion in this system was not due to a direct cytotoxic effect, since cell viability was comparable in the presence or absence of TGF-beta. Interestingly, our results demonstrated comparative suppressive effects of TGF-beta and interleukin-10 on BCG-induced TNF-alpha secretion. Together, the data demonstrate, for the first time, that TGF-beta inhibits BCG-induced TNF-alpha secretion by human cells.

Topics: Antibodies, Monoclonal; BCG Vaccine; Cytotoxicity, Immunologic; Humans; In Vitro Techniques; Interleukin-10; Leukocytes, Mononuclear; Mycobacterium bovis; Neutralization Tests; Transforming Growth Factor beta; Tuberculosis, Pulmonary; Tumor Necrosis Factor-alpha

A mouse model of pulmonary tuberculosis induced by the intratracheal instillation of live and virulent mycobacteria strain H37-Rv was used to examine the relationship of the histopathological findings with the local kinetics production and cellular distribution of tumour necrosis factor-alpha (TNF-alpha), interleukin-1 alpha (IL-1 alpha) and transforming growth factor-beta (TGF-beta). The histopathological and immunological studies showed two phases of the disease: acute or early and chronic or advanced. The acute phase was characterized by inflammatory infiltrate in the alveolar-capillary interstitium, blood vessels and bronchial wall with formation of granulomas. During this acute phase, which lasted from 1 to 28 days, high percentages of TNF-alpha and IL-1 alpha immunostained activated macrophages were observed principally in the interstium-intralveolar inflammatory infiltrate and in granulomas. Electron microscopy studies of these cells, showed extensive rough endoplasmic reticulum, numerous lysosomes and occasional mycobacteria. Double labelling with colloid gold showed that TNF-alpha and IL-1 alpha were present in the same cells, but were confined to separate vacuoles near the Golgi area, and mixed in larger vacuoles near to cell membrane. The concentration of TNF-alpha and IL-1 alpha as well as their respective mRNAs were elevated in the early phase, particularly at day 3 when the bacillary count decreased. A second peak was seen at days 14 and 21-28 when granulomas appeared and evolved to full maturation. In contrast, TGF-beta production and numbers of immunoreactive cells were low in comparison with the advanced phase of the disease. The chronic phase was characterized by histopathological changes indicative of more severity (i.e. pneumonia, focal necrosis and extensive interstitial fibrosis) with a decrease in the TNF-alpha and IL-1 alpha production that coincided with the highest level of TGF-beta. The bacillary counts were highest as the macrophages became large, vacuolated foamy cells, and containing numerous bacilli with immunoreactivity to mycobacterial lipids and lipoarabinomannan (LAM). These macrophages displayed poor and scarce TNF-alpha and IL-1 alpha immunostaining but still strong immunoreactivity to TGF-beta. These cytokine production kinetics and the spatial relationship between immunostained cells and lung lesions corroborate the important role of TNF-alpha and IL-1 alpha in the constitution of granulomas and immune protection dur

Topics: Animals; Immunoenzyme Techniques; Interleukin-1; Lung; Macrophages; Male; Mice; Mice, Inbred BALB C; Mycobacterium tuberculosis; Polymerase Chain Reaction; Transcription, Genetic; Transforming Growth Factor beta; Tuberculosis, Pulmonary; Tumor Necrosis Factor-alpha

In humans, tuberculosis is associated with suppression of T-cell responses to antigens of Mycobacterium tuberculosis. Recently, the macrophage product, transforming growth factor-beta (TGF-beta) has been implicated in suppression of T-cell proliferation and cytokine production during tuberculosis. We studied the effect of TGF-beta on production of IL-12, and on the augmentation of M. tuberculosis-induced IFN gamma production by IL-12, in patients with pulmonary tuberculosis and by M. tuberculosis. Induction of IL-12 p35, but not IL-12 p40, by M. tuberculosis in monocytes was dependent on prior priming of the cells with IFN gamma. Expression of both IL-12 p40 and p35, however, was suppressed by TGF-beta. Further, TGF-beta interfered with the bioactivity of IL-12 in the enhancement of M. tuberculosis-induced IFN gamma mRNA expression and cytokine production. However, in mononuclear cells from patients with tuberculosis the main effect of TGF-beta on IL-12 appeared to be counter action to IL-12 induced IFN gamma production in response to M. tuberculosis.

Topics: Gene Expression Regulation; Interferon-gamma; Interleukin-12; Monocytes; Mycobacterium tuberculosis; Recombinant Fusion Proteins; RNA, Messenger; T-Lymphocytes; Transforming Growth Factor beta; Tuberculosis, Pulmonary

In tuberculosis, Mycobacterium tuberculosis (MTB)-stimulated T-cell responses are depressed transiently, whereas antibody levels are increased. Lymphoproliferative responses of peripheral blood mononuclear cells (PBMCs) from Pakistani tuberculosis (TB) patients to both mycobacterial and candidal antigens were suppressed by approximately 50% when compared to healthy purified protein derivative (PPD)-positive household contacts. Production of interferon gamma (IFN-gamma) in response to PPD also was depressed by 78%. Stimulation with PPD and the 30-kDa alpha antigen of MTB (30-kDa antigen) induced greater secretion of transforming growth factor beta (TGF-beta), but not interleukin 10 (IL-10) or tumor necrosis factor alpha (TNF-alpha), by PBMCs from TB patients compared to healthy contacts. The degree of suppression correlated with the duration of treatment; patients treated for <1 month had significantly lower T-cell blastogenesis and IFN-gamma production and higher levels of TGF-beta than did patients treated for >1 month. Neutralizing antibody to TGF-beta normalized lymphocyte proliferation in response to PPD, partially restored blastogenesis to candidal antigen, and significantly increased PPD-stimulated production of IFN-gamma in TB patients but not in contacts. Neutralizing antibody to IL-10 augmented, but did not normalize, T-cell responses to both PPD and candida in TB patients and candidal antigen in contacts. TGF-beta, produced in response to MTB antigens, therefore plays a prominent role in down-regulating potentially protective host effector mechanisms and looms as an important mediator of immunosuppression in TB.

Topics: Antigens, Bacterial; Antigens, Fungal; Base Sequence; Candida; Case-Control Studies; DNA Primers; Humans; Immune Tolerance; In Vitro Techniques; Interferon-gamma; Leukocytes, Mononuclear; Lymphocyte Activation; Molecular Sequence Data; Neutralization Tests; RNA, Messenger; Transforming Growth Factor beta; Tuberculin; Tuberculin Test; Tuberculosis, Pulmonary

The expression of TGF-beta, a molecule that affects both immune responsiveness and wound healing, was examined in blood monocytes and granulomatous lesions from patients with active pulmonary tuberculosis. The spontaneous release of TGF-beta was higher in culture supernatants of monocytes from patients as compared with those of healthy subjects by an ELISA (p < 0.0005). TGF-beta activity was also confirmed in a bioassay in supernatants from patients. Next, freshly isolated monocytes from patients with tuberculosis and matched subjects were examined for TGF-beta activity. Cytosmears of monocytes were stained with an Ab against TGF-beta 1 (anti-LC) or isotype-specific Ab by using an alkaline-phosphatase anti-alkaline phosphatase method. In contrast to monocytes from healthy individuals, 60 to 70% of monocytes from patients demonstrated cytoplasmic staining for TGF-beta (n = 3). Upon hypotonic lysis, monocytes from patients with tuberculosis contained immunoreactive TGF-beta (n = 3). By Northern blot analysis, monocytes from three of seven patients with tuberculosis had increased expression of TGF-beta mRNA as compared with concurrently examined monocytes from healthy subjects. Within the granulomas of lung sections from two patients with untreated tuberculosis, TGF-beta immunoreactivity was identified in the Langhan's giant cells mainly and to a lesser extent the epithelioid cells using anti-LC Ab and the peroxidase-anti-peroxidase technique. Thus, both blood monocytes and lung granuloma macrophages from patients with active tuberculosis express TGF-beta. Excess activity of this cytokine in blood monocytes may underlie the depressed T cell responses of patients with tuberculosis. Moreover, within the infected tissues excess TGF-beta activity may interfere with anti-mycobacterial mechanisms and effective granuloma formation.

Topics: Gene Expression; Humans; Lung; Macrophages, Alveolar; Monocytes; RNA, Messenger; Transforming Growth Factor beta; Tuberculoma; Tuberculosis, Pulmonary