transforming-growth-factor-beta and Pleural-Effusion--Malignant

The management of malignant pleural effusions are a common, and sometimes difficult, therapeutic problem. This article reviews the currently available sclerosing agents and discusses novel agents that show promise for the future. Talc, tetracycline and bleomycin are currently the most widely used agents but none is an ideal sclerosant and certain adverse effects are of concern. Novel agents, particularly transforming growth factor (TGF)-beta(2) and metalloproteinase inhibitors, are being investigated, and the increase in knowledge into the mechanisms of pleural fluid formation and pleurodesis will hopefully lead to the development of targeted therapy in the future.

Topics: Adjuvants, Immunologic; Adrenal Cortex Hormones; Animals; Fibrinolytic Agents; Humans; Matrix Metalloproteinase Inhibitors; Pleural Effusion, Malignant; Pleurodesis; Sclerosing Solutions; Transforming Growth Factor beta; Transforming Growth Factor beta2

Malignant pleural effusion (MPE) is defined as the presence of tumor cells in pleural fluid and it is a fatal complication of advanced lung adenocarcinoma (LAC). To understand the immune response to the tumor in MPE, we compared the concentration of immunomodulatory factors in MPE of LAC and pleural effusion of heart failure (HF) patients by ELISA, and the proliferation and cytotoxic phenotype of T cells stimulated in the presence of LAC and HF pleural fluids by cytometry. Platelet factor 4 (PF4), vascular endothelial growth factor (VEGF), transforming growth factor beta (TGF-β) and P-selectin levels were higher in LAC than in HF pleural fluids. However, plasmatic PF4 and P-selectin levels were similar in LAC and HF. VEGF positively correlated with TGF-β and sPD-L1 in LAC but not in HF pleural fluids. LAC pleural fluids also inhibited T lymphocyte proliferation and cytotoxicity and reduced IL-17 production. PF4 levels inversely correlated with T cell function. The high content of PF4 in MPE was associated with poor prognosis. Our findings suggest that an impaired response of T lymphocytes induced by PF4 provides a significant advantage for tumor progression.

Topics: Adenocarcinoma of Lung; Aged; Aged, 80 and over; Female; Heart Failure; Humans; Lung Neoplasms; Lymphocyte Activation; Male; Middle Aged; Platelet Factor 4; Pleural Effusion, Malignant; T-Lymphocytes; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A

Immune dysfunction often occurs in malignant pleural effusion (MPE). In our previous study, TGF-β derived predominantly from macrophages plays an important role in impairing T cell cytotoxicity in MPE. Therefore, we aimed to investigate whether other immunoregulatory cells and factors mediated TGF-β secretion from macrophages, involved in the immunosuppressive microenvironment of MPE, and to provide clues for potential immune therapy for MPE as well. We found that CCL22 level in MPE was significantly higher than that in non-malignant pleural effusion. The high level of CCL22 was closely associated with poor survival in MPE patients with lung cancer. CCL22 was dominantly produced by tumor-associated macrophages (TAMs) in MPE. Meanwhile, TAM-derived TGF-β mediated CCL22 expression in TAMs via c-Fos. CCL22 promoted the recruitment of regulatory T cells (Tregs) in MPE. Lastly, Treg-secreted high level of IL-8 further induced TGF-β production from TAMs, and promoted the immunosuppressive tumor microenvironment in MPE. Our results indicate that macrophage-derived CCL22 plays an important role in the immunosuppressive tumor microenvironment via IL-8 in MPE.

Topics: Cell Line, Tumor; Chemokine CCL22; Humans; Immune Tolerance; Interleukin-8; Lung Neoplasms; Macrophages; Pleural Effusion, Malignant; T-Lymphocytes, Regulatory; THP-1 Cells; Transforming Growth Factor beta; Tumor Microenvironment

We studied the diagnostic value of cytokines, including vascular endothelial growth factor (VEGF), transforming growth factor-β (TGF-β), and interleukin-8 (IL-8), and the ratio of lactate dehydrogenase (LDH) to adenosine deaminase (ADA) in pleural fluid.. Prospective analysis of 44 inpatients or outpatients with pleural fluid, from December 2016 to March 2017 was conducted.. We enrolled patients with malignant pleural effusion (MPE, N = 15), empyema (N = 11), parapneumonic effusion (PPE, N = 7), chronic renal failure (CRF)/chronic heart failure (CHF) (N = 7), and tuberculous pleural effusion (TBPE, N = 4). The pleural fluid values of IL-8 and VEGF were significantly higher in empyema patients than in CRF/CHF or PPE patients. In all patients, the pleural fluid VEGF and IL-8 values were significantly positively correlated (r = 0.405, p = 0.006; r = 0.474, p = 0.047, respectively). TGF-β was elevated in patients with empyema, PPE, TBPE, and MPE. The pleural LDH-to-ADA ratio in patients with MPE or empyema/PPE was significantly higher than in patients with CRF/CHF or TBPE. LDH and ADA levels correlated significantly only in patients with MPE (r = 0.648, p = 0.009) and empyema/PPE (r = 0.978, p < 0.001).. VEGF and IL-8 production in the pleural cavity appear to accelerate the progression of PPE to empyema, by enhancing vascular permeability associated with inflammation. Sequential sampling would be needed to confirm this. The pleural LDH/ADA ratio may be a useful diagnostic tool for discriminating between various pleural effusion etiologies.

Topics: Adenosine Deaminase; Aged; Aged, 80 and over; Biomarkers; Diagnosis, Differential; Empyema, Pleural; Female; Heart Failure; Humans; Interleukin-8; Kidney Failure, Chronic; L-Lactate Dehydrogenase; Male; Middle Aged; Pleural Effusion; Pleural Effusion, Malignant; Pneumonia; Predictive Value of Tests; Prospective Studies; Transforming Growth Factor beta; Tuberculosis; Vascular Endothelial Growth Factor A

Cancer metastatic spread to serous cavity causes malignant pleural effusions (MPEs), indicating dismal prognosis. Tumor microenvironment can implement suppressive activity on host immune responses. Thus, we investigated the prevalence of Tregs and the relationship between them and TGF-

Topics: Cells, Cultured; CTLA-4 Antigen; Forkhead Transcription Factors; Gene Expression Regulation; Glucocorticoid-Induced TNFR-Related Protein; Humans; Interleukin-10; Interleukin-2 Receptor alpha Subunit; Lung Neoplasms; Neoplasm Metastasis; Neoplasm Staging; Pleural Effusion, Malignant; T-Lymphocytes, Regulatory; Transforming Growth Factor beta; Tumor Microenvironment

Malignant pleural effusion (MPE) is an indication of advanced cancer. Immune dysfunction often occurs in MPE. We aimed to identify the reason for impaired T cell activity in MPE from lung cancer patients and to provide clues toward potential immune therapies for MPE. The surface inhibitory molecules and cytotoxic activity of T cells in MPE and peripheral blood (PB) were analyzed using flow cytometry. Levels of inflammatory cytokines in MPE and PB were tested using ELISA. TGF-β expression in tumor-associated macrophages (TAMs) was also analyzed. The effect of TAMs on T cells was verified in vitro. Lastly, changes in T cells were evaluated following treatment with anti-TGF-β antibody. We found that expression levels of Tim-3, PD-1 and CTLA-4 in T cells from MPE were upregulated compared with those from PB, but levels of IFN-γ and Granzyme B were downregulated (p < 0.05). The amount of TGF-β was significantly higher in MPE than in PB (p < 0.05). TGF-β was mainly produced by TAMs in MPE. When T cells were co-cultured with TAMs, expression levels of Tim-3, PD-1 and CTLA-4 were significantly higher than controls, whereas levels of IFN-γ and Granzyme B were significantly decreased, in a dose-dependent manner (p < 0.05). In vitro treatment with anti-TGF-β antibody restored the impaired T cell cytotoxic activity in MPE. Our results indicate that macrophage-derived TGF-β plays an important role in impaired T cell cytotoxicity. It will therefore be valuable to develop therapeutic strategies against TGF-β pathway for MPE therapy of lung cancer.

Topics: CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Humans; Lung Neoplasms; Macrophages; Pleural Effusion, Malignant; Transforming Growth Factor beta

Accumulating evidence shows that an imbalance in regulatory T cells (Tregs)/T helper IL-17-producing cells (Th17) exists in malignant pleural effusion (MPE). However, the cause of this phenomenon in MPE and the underlying mechanism remain uncertain. The percentages of Tregs and Th17 cells in MPE and parapneumonic effusion (PPE) were determined by flow cytometry. Their specific transcription factors, forkhead box P3 (FoxP3) and retinoic acid-related orphan receptor γt (RORγt); related cytokines, interleukin-6 (IL-6), IL-17, IL-10, and transforming growth factor-β1 (TGF‑β1); and chemokines, C-C motif ligand 17 (CCL17) and CCL20, were analyzed by real-time PCR and ELISA, respectively. Compared to patients with PPE, patients with MPE presented a higher percentage of Tregs but a lower frequency of Th17 cells. Foxp3 mRNA expression level in the cells in the pleural effusion was significantly increased in patients with MPE compared to the levels in patients with PPE (MPE vs. PPE: 3.05±0.62 vs. 0.52±0.11, p=0.0012). It was also noted that high levels of IL-10, TGF-β1 and CCL17 were observed in MPE when compared to PPE (MPE vs. PPE: IL-10, 166.3±39.53 vs. 40.38±10.92 pg/ml, p=0.0307; TGF-β1, 10,720±1,274 vs. 1,747±293.2 pg/ml, p<0.0001; CCL17, 341.1±88.22 vs. 119.2±19.80 pg/ml, p=0.0427). Furthermore, a high ratio of Tregs/Th17 cells in MPE was highly correlated to poor survival. An alteration in CCL17 and CCL20 might contribute to the Treg/Th17 imbalance in MPE, which partially predicts a poor prognosis in patients with lung cancer.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Chemokine CCL17; Chemokine CCL20; Female; Forkhead Transcription Factors; Gene Expression Regulation, Neoplastic; Humans; Interleukin-10; Lung Neoplasms; Male; Middle Aged; Nuclear Receptor Subfamily 1, Group F, Member 3; Pleural Effusion; Pleural Effusion, Malignant; Pneumonia; Prognosis; T-Lymphocytes, Regulatory; Th17 Cells; Transforming Growth Factor beta

Topics: Biomarkers; Cytokines; Diagnosis, Differential; Endostatins; Humans; Peptide Hydrolases; Pleural Effusion; Pleural Effusion, Malignant; Predictive Value of Tests; Prognosis; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A

Both regulatory T cells (Tregs) and T helper IL-17-producing cells (Th17 cells) have been found to be involved in human malignancies, however, the possible implication of Tregs in regulating generation and differentiation of Th17 cells in malignant pleural effusion remains to be elucidated.. The numbers of both CD39(+)Tregs and Th17 cells in malignant pleural effusion and peripheral blood from patients with lung cancer were determined by flow cytometry. The regulation and mechanism of Tregs on generation and differentiation of Th17 cells were explored.. Both CD39(+)Tregs and Th17 cells were increased in malignant pleural effusion when compared with blood, and the numbers of CD39(+)Tregs were correlated negatively with those of Th17 cells. It was also noted that high levels of IL-1β, IL-6, and TGF-β1 could be observed in malignant pleural effusion when compared the corresponding serum, and that pleural CD39(+)Tregs could express latency-associated peptide on their surface. When naïve CD4(+) T cells were cocultured with CD39(+)Tregs, Th17 cell numbers decreased as CD39(+)Treg numbers increased, addition of the anti-latency-associated peptide mAb to the coculture reverted the inhibitory effect exerted by CD39(+)Tregs.. Therefore, the above results indicate that CD39(+)Tregs inhibit generation and differentiation of Th17 cells via a latency-associated peptide-dependent mechanism.

Topics: Adult; Aged; Antigens, CD; Apyrase; Cell Differentiation; Cell Proliferation; Cells, Cultured; Coculture Techniques; Flow Cytometry; Humans; Immunophenotyping; Interleukin-1beta; Interleukin-6; Lung Neoplasms; Middle Aged; Peptides; Pleural Effusion, Malignant; Protein Precursors; Signal Transduction; T-Lymphocytes, Regulatory; Th17 Cells; Transforming Growth Factor beta; Transforming Growth Factor beta1

We have studied CD4(+)CD25(high)FOXP3(+) regulatory T-cells (T(regs)) from 51 patients with non-small-cell lung cancer (NSCLC) and 33 healthy donors. Regulatory T-cells were identified by fluorescence-activated cell sorting by using a panel of antibodies and by reverse transcriptase polymerase chain reaction analysis for FOXP3 expression. Functional studies were done to analyze their inhibitory role. Finally, regulatory T-cells were analyzed in malignant pleura effusion (PE) from patients with NSCLC. Patients with NSCLC have increased numbers of CD4(+)CD25(high) FOXP3(+) T(regs) in their peripheral blood and pleura effusion (PE), which express high levels of CTLA-4, GITR. These cells were anergic toward T-cell receptor stimulation and, when cocultured with activated CD4(+)CD25(-) cells, potently suppressed their proliferation and cytokine secretion. Our data suggest that in NSCLC patients, there is an increase of CD4(+)CD25(high)FOXP3(+) regulatory T-cells in the peripheral blood and tumor microenvironment. These T-cells might prevent effective antitumor immune responses, and the increase in frequency of CD4(+)CD25(high)FOXP3(+) Tregs might play a role in the modulation of the immune response against NSCLC and could be important in the design of immunotherapeutic approaches.

Topics: Adult; Aged; Aged, 80 and over; Antigens, CD; Blood Cell Count; Carcinoma, Non-Small-Cell Lung; CD4-Positive T-Lymphocytes; CTLA-4 Antigen; Female; Forkhead Transcription Factors; Gene Expression; Glucocorticoid-Induced TNFR-Related Protein; Humans; Immune Tolerance; Immunophenotyping; Interferon-gamma; Interleukin-2; Interleukin-2 Receptor alpha Subunit; Leukocytes, Mononuclear; Male; Middle Aged; Pleural Effusion, Malignant; Receptors, Nerve Growth Factor; Receptors, Tumor Necrosis Factor; T-Lymphocytes, Regulatory; Transforming Growth Factor beta; Young Adult

Malignant effusions in serous cavities constitute a unique milieu for direct contact of tumor cells with host lymphoid cells in a fluid phase. The aim of this study was to depict agents responsible for suppression of lymphoid cells with putative anti-tumor potential. Pleural effusions drawn from 44 (18 non-malignant and 26 malignant) patients were tested for selected cytokines--interleukin-10 (IL-10), transforming growth factor beta (TGF-beta1) and soluble Fas ligand (sFasL) and nuclear membrane proteins (NMPs) content by ELISA. TCR-zeta expression of T cells and TUNEL reaction for apoptosis were evaluated by three color flow cytometry. Both cytokine concentrations were found to be significantly elevated in malignant pleural effusions (MPE) as compared to non-malignant ones. It was also true for sFasL content. Moreover, NMPs corresponding to decoy cell fragments, were also heightened in MPE. Concentrations of NMPs correlated with the percent of apoptotic (TUNEL+) T CD3+ lymphocytes and inversely correlated with the percent of T cells. The low expression of TCR-zeta chain on T cells corresponded to high concentration of sFasL in MPE. In conclusion, the above data suggest that out of three suppression agents tested, only sFasL appears to show correlation with the downregulation of T cells in MPE.

Topics: Apoptosis; Cytokines; Enzyme-Linked Immunosorbent Assay; Fas Ligand Protein; Female; Flow Cytometry; Humans; Immunosuppression Therapy; In Situ Nick-End Labeling; Interleukin-10; Male; Membrane Glycoproteins; Pleural Effusion, Malignant; T-Lymphocytes; Transforming Growth Factor beta; Transforming Growth Factor beta1

Recent studies have demonstrated high levels of vascular endothelial growth factor (VEGF) in exudative pleural effusions and a possible etiologic role. The factors regulating VEGF accumulation in the pleural space are unknown. Transforming growth factor (TGF)-beta is a potent stimulator of VEGF expression in vitro. We hypothesized that TGF-beta induces VEGF production in pleural tissues, and, hence, the pleural fluid VEGF levels should correlate with the levels of TGF-beta in pleural fluid of different etiologies.. Seventy pleural fluid samples were analyzed. These included 20 malignant, 13 post-coronary artery bypass grafting (CABG), 8 parapneumonic, 11 miscellaneous exudative, and 18 congestive heart failure (CHF) pleural effusions.. Pleural fluid VEGF levels showed good correlation with those of TGF-beta(1) (r = 0.58; p < 0. 0001), TGF-beta(2) (r = 0.43; p < 0.001), and lactate dehydrogenase (r = 0.65; p < 0.001). The levels of TGF-beta(1) and TGF-beta(2) also were correlated (r = 0.60; p < 0.0001). The median levels of TGF-beta(1) (2,480 pg/mL) and TGF-beta(2) (266 pg/mL) in the CHF group were significantly lower than those in the malignant (TGF-beta(1), 4,902 pg/mL; TGF-beta(2), 428 pg/mL), post-CABG (TGF-beta(1), 5,456 pg/mL; TGF-beta(2), 377 pg/mL), parapneumonic (TGF-beta(1), 5,024 pg/mL; TGF-beta(2), 464 pg/mL), and miscellaneous exudate groups (TGF-beta(1), 7,690 pg/mL; TGF-beta(2), 369 pg/mL). There was no significant difference in TGF-beta(1) and TGF-beta(2) levels among the four exudate groups.. VEGF levels in pleural effusions are significantly correlated with the levels of TGF-beta(1) and beta(2) isoforms. VEGF, TGF-beta(1), and TGF-beta(2) levels were all higher in exudative effusions than in effusions secondary to CHF.

Topics: Endothelial Growth Factors; Humans; L-Lactate Dehydrogenase; Lymphokines; Pleural Effusion; Pleural Effusion, Malignant; Protein Isoforms; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

The level of hyaluronic acid (HA) was determined in the pleural fluid of 99 patients, including 19 with malignant mesothelioma, 27 with lung cancer, 1 with breast cancer, 1 with mediastinal tumor and 51 with non-malignant diseases. With a cut-off level at 100 micrograms/ml, the pleural fluid concentration of HA was high in 36.8% of patients (7 of 19) with malignant mesothelioma and 1.3% of patients (1 of 80) with lung cancer and other malignant and non-malignant diseases. The mean concentration of pleural fluid HA was significantly higher in patients with mesothelioma than in those with lung cancer and other malignant and non-malignant diseases. The pre-test probability of MM was 5.9% in this series. The LRs for > or = 100, 50-99 and < or = 49 micrograms/ml are 28.3, 3.3 and 0.5, respectively; these put the post-test probabilities at 64, 17 and 3%, respectively. Indeed, in cases of uncommon disease such as MM, the post-test probability is low even if the cut-off level of HA is > or = 100 micrograms/ml. The discrimination between malignant mesothelioma and lung cancer needs special attention. In these two diseases, the LRs of MM for pleural fluid CEA > 30, 10-30 and < 10 ng/ml were 0.2, 1.9 and 2.4, respectively. The pre-test probability of MM for HA > or = or 100 micrograms/ml is 64%. Furthermore, because the LR for CEA is < 10 ng/ml, the post-test probability is 81%. When the combination of two markers is considered, the high level of HA and the low level of CEA may be useful for the differential diagnosis of MM from pleuritis carcinomatosa.

Topics: Biomarkers, Tumor; Carcinoembryonic Antigen; Diagnosis, Differential; Heart Failure; Humans; Hyaluronic Acid; Lung Neoplasms; Mesothelioma; Pleural Effusion; Pleural Effusion, Malignant; Prospective Studies; Transforming Growth Factor beta; Tuberculosis, Pleural

The detailed mechanisms underlying the formation of malignant effusions are incompletely defined. In order to determine whether transforming growth factor-beta (TGF-beta) would contribute to the formation of malignant effusions, we investigated the effect of TGF-beta on the morphology, growth, and permeability of human mesothelial cells, which are thought to serve as a permeability barrier in the pleuroperitoneal cavities. Treatment of the mesothelial cells with a TGF-beta dose ranging from 0.1 to 10 ng/ml for 96 hr induced distinct morphologic changes in the cells. Each cell increased in size as did the volume of the intercellular spaces. TGF-beta also significantly inhibited the growth of mesothelial cells at a concentration ranging from 0.1 to 10 ng/ml. This growth inhibition was blocked completely by the addition of anti-TGF-beta antibody. Treatment of the mesothelial cells with 2.0 ng/ml TGF-beta significantly increased the permeability of a mesothelial cell monolayer as assessed by a FITC-albumin permeability assay. In our clinical analysis using 10 effusion samples obtained from patients with various types of carcinoma cells, considerable level of TGF-beta could be detected by ELISA, ranged from 0.90 to 8.75 ng/ml. Our data suggest that TGF-beta plays an important role in the formation of malignant effusions through structural and functional damage to the mesothelial cells. Malignant effusions may accumulate in the pleuroperitoneal cavity as a result of the mesothelial cell damage caused by this cytokine which is released from disseminated cancer cells.

Topics: Adult; Aged; Ascites; Cell Division; Cell Membrane Permeability; Cells, Cultured; Epithelial Cells; Female; Growth Inhibitors; Humans; In Vitro Techniques; Male; Middle Aged; Omentum; Pleural Effusion, Malignant; Transforming Growth Factor beta

We investigated the levels of TGF-beta in malignant pleural effusions (MPE) caused by malignant mesothelioma (MESO) or primary lung cancer. TGF-beta levels in MPE caused by MESO were 283.9 +/- 219.2 pm (mean +/- s.d.) and were three to six times higher than those due to primary lung cancers (P < 0.01 or P < 0.05). We also evaluated TGF-beta 1- and beta 2-like activities in MPE using specific polyclonal antibodies. Although TGF-beta 1-like activity could be detected in all cases, TGF-beta 2-like activities were detected in five of seven in MESO and in a few cases with primary lung cancer. These results demonstrate that the levels of total TGF-beta and TGF-beta 2-like activity may be clinically useful to differentiate MESO from primary lung cancer. Our data also suggest that TGF-beta may help further characterize the clinical features of MESO.

Topics: Carcinoma, Small Cell; Carcinoma, Squamous Cell; Humans; Lung Neoplasms; Mesothelioma; Platelet Factor 4; Pleural Effusion, Malignant; Transforming Growth Factor beta; Tuberculosis, Pleural

The expression of TGF-beta-3 was examined in 64 patients with reactive and malignant effusion by immunocytochemistry.. In about half of the patients with malignant effusions (especially breast cancer, gastric cancer, and carcinomas of unknown primary) TGF-beta positive tumor cells could be detected. We could show here for the first time that reactive mesothelial cells could also express TGF-beta. Lymphatic cells were negative in all cases. TGF-beta-3 bioactivity could also be detected in the effusions studied. In our group of patients with far advanced cancer, the expression of TGF-beta had no clear-cut clinical or prognostic correlate. However, the expression of TGF-beta on tumor cells should be interpreted as a marker of tumor progression, taking into account the fibrogenic, angiogenic and immunosuppressive properties of TGF-beta.. Further research is necessary to answer the question if the 3 isoforms of TGF-beta are coordinately expressed and to elucidate the involvement of this cytokine in tumor progression and metastasis.

Topics: Adult; Aged; Ascitic Fluid; Cell Division; Female; Humans; Male; Middle Aged; Neoplasms; Pleural Effusion, Malignant; Transforming Growth Factor beta; Tumor Cells, Cultured