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TL1A/DR3 axis involvement in the inflammatory cytokine network during pulmonary sarcoidosis.

Facco M, Cabrelle A, Calabrese F, Teramo A, Cinetto F, Carraro S, Martini V, Calzetti F, Tamassia N, Cassatella MA, Semenzato G, Agostini C - Clin Mol Allergy (2015)

Bottom Line: TNF-like ligand 1A (TL1A), a recently recognized member of the TNF superfamily, and its death domain receptor 3 (DR3), firstly identified for their relevant role in T lymphocyte homeostasis, are now well-known mediators of several immune-inflammatory diseases, ranging from rheumatoid arthritis to inflammatory bowel diseases to psoriasis, whereas no data are available on their involvement in sarcoidosis, a multisystemic granulomatous disease where a deregulated T helper (Th)1/Th17 response takes place.Our results demonstrated a significant higher expression, both at protein and mRNA levels, of TL1A and DR3 in pulmonary T cells and alveolar macrophages of patients with active sarcoidosis as compared to patients with the inactive form of the disease and to controls.Additionally, zymography assays showed that TL1A is able to increase the production of matrix metalloproteinase 9 by sarcoid alveolar macrophages characterized, in patients with the active form of the disease, by reduced mRNA levels of the tissue inhibitor of metalloproteinase (TIMP)-1.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Hematology and Clinical Immunology Branch, Padua University School of Medicine, Padua, Italy ; Venetian Institute of Molecular Medicine, Padua, Italy.

ABSTRACT

Background: TNF-like ligand 1A (TL1A), a recently recognized member of the TNF superfamily, and its death domain receptor 3 (DR3), firstly identified for their relevant role in T lymphocyte homeostasis, are now well-known mediators of several immune-inflammatory diseases, ranging from rheumatoid arthritis to inflammatory bowel diseases to psoriasis, whereas no data are available on their involvement in sarcoidosis, a multisystemic granulomatous disease where a deregulated T helper (Th)1/Th17 response takes place.

Methods: In this study, by flow cytometry, real-time PCR, confocal microscopy and immunohistochemistry analyses, TL1A and DR3 were investigated in the pulmonary cells and the peripheral blood of 43 patients affected by sarcoidosis in different phases of the disease (29 patients with active sarcoidosis, 14 with the inactive form) and in 8 control subjects.

Results: Our results demonstrated a significant higher expression, both at protein and mRNA levels, of TL1A and DR3 in pulmonary T cells and alveolar macrophages of patients with active sarcoidosis as compared to patients with the inactive form of the disease and to controls. In patients with sarcoidosis TL1A was strongly more expressed in the lung than the blood, i.e., at the site of the involved organ. Additionally, zymography assays showed that TL1A is able to increase the production of matrix metalloproteinase 9 by sarcoid alveolar macrophages characterized, in patients with the active form of the disease, by reduced mRNA levels of the tissue inhibitor of metalloproteinase (TIMP)-1.

Conclusions: These data suggest that TL1A/DR3 interactions are part of the extended and complex immune-inflammatory network that characterizes sarcoidosis during its active phase and may contribute to the pathogenesis and to the progression of the disease.

No MeSH data available.


Related in: MedlinePlus

A. Gelatin zymograms of MMP-9 activity in the BAL fluid (a), and in freshly isolated BAL cellular components (alveolar macrophages and T cells) (b), of two representative cases of patients with active and inactive sarcoidosis. c, d) MMP-9 activity of alveolar macrophages (c) and lung T cells (d) obtained from the BAL of two representative cases of patients with active and inactive sarcoidosis, cultured in medium alone and with the cytokine TL1A. B) MMP-9 activity of alveolar macrophages obtained from the lung of patients affected by active sarcoidosis (black columns) and inactive sarcoidosis (grey columns), freshly isolated, cultured in medium alone and with TL1A. Zymographic band densities from all samples were quantified by densitometry. Data are expressed as mean ± SD. D) MMP-9 and TIMP-1 mRNA expression in alveolar macrophages obtained from the lung of patients affected by active sarcoidosis (black columns) and inactive sarcoidosis (grey columns), freshly isolated, cultured in medium alone and with TL1A. Expression of MMP-9 and TIMP-1 mRNAs was normalized on GAPDH mRNA. Data are expressed as mean ± SD
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Fig4: A. Gelatin zymograms of MMP-9 activity in the BAL fluid (a), and in freshly isolated BAL cellular components (alveolar macrophages and T cells) (b), of two representative cases of patients with active and inactive sarcoidosis. c, d) MMP-9 activity of alveolar macrophages (c) and lung T cells (d) obtained from the BAL of two representative cases of patients with active and inactive sarcoidosis, cultured in medium alone and with the cytokine TL1A. B) MMP-9 activity of alveolar macrophages obtained from the lung of patients affected by active sarcoidosis (black columns) and inactive sarcoidosis (grey columns), freshly isolated, cultured in medium alone and with TL1A. Zymographic band densities from all samples were quantified by densitometry. Data are expressed as mean ± SD. D) MMP-9 and TIMP-1 mRNA expression in alveolar macrophages obtained from the lung of patients affected by active sarcoidosis (black columns) and inactive sarcoidosis (grey columns), freshly isolated, cultured in medium alone and with TL1A. Expression of MMP-9 and TIMP-1 mRNAs was normalized on GAPDH mRNA. Data are expressed as mean ± SD

Mentions: As shown in Fig. 4a, panel a, BAL fluids from patients with active sarcoidosis were characterized by a stronger MMP-9 activity (12.5 ± 5.8) as compared to those obtained from inactive sarcoidosis (3.6 % ± 1.3; p < 0.05). AM and T cell subset isolation (Fig. 4a, panel b) demonstrated that the source responsible of MMP-9 activity was represented by AMs (2.8 ± 0.35 and 22.5 ± 3.8 in AMs from inactive and active sarcoidosis, respectively; p < 0.01 vs active disease). Successively, purified AMs and T lymphocytes were cultured in presence (and absence) of TL1A to investigate its possible effects on MMP-9 production and activity. Figure 4a (panel c and d), and 4B showed that MMP-9 activity referable to lung T cells was not up-regulated by TL1A/DR3 interactions, whereas AMs obtained from patients with inactive sarcoidosis presented an appreciable increase of the metalloproteinase activity (3.56 ± 0.27 and 6.40 ± 1.75, in absence and presence of TL1A, respectively; p < 0.05). We did not detect any relevant change in MMP-9 activity of AMs derived from patients with active sarcoidosis in presence of TL1A (22.10 ± 4.03 and 22.73 ± 4.46, in absence and presence of TL1A, respectively; p: not significant), probably due to an already achieved peak of MMP-9 production and release by these strongly activated macrophages.Fig. 4


TL1A/DR3 axis involvement in the inflammatory cytokine network during pulmonary sarcoidosis.

Facco M, Cabrelle A, Calabrese F, Teramo A, Cinetto F, Carraro S, Martini V, Calzetti F, Tamassia N, Cassatella MA, Semenzato G, Agostini C - Clin Mol Allergy (2015)

A. Gelatin zymograms of MMP-9 activity in the BAL fluid (a), and in freshly isolated BAL cellular components (alveolar macrophages and T cells) (b), of two representative cases of patients with active and inactive sarcoidosis. c, d) MMP-9 activity of alveolar macrophages (c) and lung T cells (d) obtained from the BAL of two representative cases of patients with active and inactive sarcoidosis, cultured in medium alone and with the cytokine TL1A. B) MMP-9 activity of alveolar macrophages obtained from the lung of patients affected by active sarcoidosis (black columns) and inactive sarcoidosis (grey columns), freshly isolated, cultured in medium alone and with TL1A. Zymographic band densities from all samples were quantified by densitometry. Data are expressed as mean ± SD. D) MMP-9 and TIMP-1 mRNA expression in alveolar macrophages obtained from the lung of patients affected by active sarcoidosis (black columns) and inactive sarcoidosis (grey columns), freshly isolated, cultured in medium alone and with TL1A. Expression of MMP-9 and TIMP-1 mRNAs was normalized on GAPDH mRNA. Data are expressed as mean ± SD
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Fig4: A. Gelatin zymograms of MMP-9 activity in the BAL fluid (a), and in freshly isolated BAL cellular components (alveolar macrophages and T cells) (b), of two representative cases of patients with active and inactive sarcoidosis. c, d) MMP-9 activity of alveolar macrophages (c) and lung T cells (d) obtained from the BAL of two representative cases of patients with active and inactive sarcoidosis, cultured in medium alone and with the cytokine TL1A. B) MMP-9 activity of alveolar macrophages obtained from the lung of patients affected by active sarcoidosis (black columns) and inactive sarcoidosis (grey columns), freshly isolated, cultured in medium alone and with TL1A. Zymographic band densities from all samples were quantified by densitometry. Data are expressed as mean ± SD. D) MMP-9 and TIMP-1 mRNA expression in alveolar macrophages obtained from the lung of patients affected by active sarcoidosis (black columns) and inactive sarcoidosis (grey columns), freshly isolated, cultured in medium alone and with TL1A. Expression of MMP-9 and TIMP-1 mRNAs was normalized on GAPDH mRNA. Data are expressed as mean ± SD
Mentions: As shown in Fig. 4a, panel a, BAL fluids from patients with active sarcoidosis were characterized by a stronger MMP-9 activity (12.5 ± 5.8) as compared to those obtained from inactive sarcoidosis (3.6 % ± 1.3; p < 0.05). AM and T cell subset isolation (Fig. 4a, panel b) demonstrated that the source responsible of MMP-9 activity was represented by AMs (2.8 ± 0.35 and 22.5 ± 3.8 in AMs from inactive and active sarcoidosis, respectively; p < 0.01 vs active disease). Successively, purified AMs and T lymphocytes were cultured in presence (and absence) of TL1A to investigate its possible effects on MMP-9 production and activity. Figure 4a (panel c and d), and 4B showed that MMP-9 activity referable to lung T cells was not up-regulated by TL1A/DR3 interactions, whereas AMs obtained from patients with inactive sarcoidosis presented an appreciable increase of the metalloproteinase activity (3.56 ± 0.27 and 6.40 ± 1.75, in absence and presence of TL1A, respectively; p < 0.05). We did not detect any relevant change in MMP-9 activity of AMs derived from patients with active sarcoidosis in presence of TL1A (22.10 ± 4.03 and 22.73 ± 4.46, in absence and presence of TL1A, respectively; p: not significant), probably due to an already achieved peak of MMP-9 production and release by these strongly activated macrophages.Fig. 4

Bottom Line: TNF-like ligand 1A (TL1A), a recently recognized member of the TNF superfamily, and its death domain receptor 3 (DR3), firstly identified for their relevant role in T lymphocyte homeostasis, are now well-known mediators of several immune-inflammatory diseases, ranging from rheumatoid arthritis to inflammatory bowel diseases to psoriasis, whereas no data are available on their involvement in sarcoidosis, a multisystemic granulomatous disease where a deregulated T helper (Th)1/Th17 response takes place.Our results demonstrated a significant higher expression, both at protein and mRNA levels, of TL1A and DR3 in pulmonary T cells and alveolar macrophages of patients with active sarcoidosis as compared to patients with the inactive form of the disease and to controls.Additionally, zymography assays showed that TL1A is able to increase the production of matrix metalloproteinase 9 by sarcoid alveolar macrophages characterized, in patients with the active form of the disease, by reduced mRNA levels of the tissue inhibitor of metalloproteinase (TIMP)-1.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Hematology and Clinical Immunology Branch, Padua University School of Medicine, Padua, Italy ; Venetian Institute of Molecular Medicine, Padua, Italy.

ABSTRACT

Background: TNF-like ligand 1A (TL1A), a recently recognized member of the TNF superfamily, and its death domain receptor 3 (DR3), firstly identified for their relevant role in T lymphocyte homeostasis, are now well-known mediators of several immune-inflammatory diseases, ranging from rheumatoid arthritis to inflammatory bowel diseases to psoriasis, whereas no data are available on their involvement in sarcoidosis, a multisystemic granulomatous disease where a deregulated T helper (Th)1/Th17 response takes place.

Methods: In this study, by flow cytometry, real-time PCR, confocal microscopy and immunohistochemistry analyses, TL1A and DR3 were investigated in the pulmonary cells and the peripheral blood of 43 patients affected by sarcoidosis in different phases of the disease (29 patients with active sarcoidosis, 14 with the inactive form) and in 8 control subjects.

Results: Our results demonstrated a significant higher expression, both at protein and mRNA levels, of TL1A and DR3 in pulmonary T cells and alveolar macrophages of patients with active sarcoidosis as compared to patients with the inactive form of the disease and to controls. In patients with sarcoidosis TL1A was strongly more expressed in the lung than the blood, i.e., at the site of the involved organ. Additionally, zymography assays showed that TL1A is able to increase the production of matrix metalloproteinase 9 by sarcoid alveolar macrophages characterized, in patients with the active form of the disease, by reduced mRNA levels of the tissue inhibitor of metalloproteinase (TIMP)-1.

Conclusions: These data suggest that TL1A/DR3 interactions are part of the extended and complex immune-inflammatory network that characterizes sarcoidosis during its active phase and may contribute to the pathogenesis and to the progression of the disease.

No MeSH data available.


Related in: MedlinePlus