Characterization of Monocytes from Peripheral Blood and Pleural Effusion of patients with Active Pulmonary Tuberculosis

BALBOA L, ROMERO M, YOKOBORI N, SCHIERLOH P, MUSELLA R, ABBATE E, S DE LA BARRERA, SASIAIN M, ALEMAN M.

Keystone, Colorado

Congreso; Tuberculosis: Biology, Pathology and Therapy. Keystone Symposia; 2009

Global Health Series

Monocytes/macrophages (Mo/Mö) are the main target for Mycobacterium tuberculosis (Mtb) and their innate capacity to deal with Mtb defines the early progression of the infection. In such chronic disease, circulating Mo arriving to the site of infection are the best candidates for the renewal of Mö and dendritic cells in order to establish granuloma formation and to sustain T-lymphocyte activation in the lymph nodes. The aim of this work was to perform a phenotypic and functional characterization of peripheral blood (PB) Mo as well as Mo present in pleural effusions (PE) from patients with tuberculosis (Mo-TB). As control, circulating Mo from healthy individuals (Mo-N), and Mo present in PB and PE of pulmonary cancer as inflammatory but non-infectious disease were evaluated. order to establish granuloma formation and to sustain T-lymphocyte activation in the lymph nodes. The aim of this work was to perform a phenotypic and functional characterization of peripheral blood (PB) Mo as well as Mo present in pleural effusions (PE) from patients with tuberculosis (Mo-TB). As control, circulating Mo from healthy individuals (Mo-N), and Mo present in PB and PE of pulmonary cancer as inflammatory but non-infectious disease were evaluated. progression of the infection. In such chronic disease, circulating Mo arriving to the site of infection are the best candidates for the renewal of Mö and dendritic cells in order to establish granuloma formation and to sustain T-lymphocyte activation in the lymph nodes. The aim of this work was to perform a phenotypic and functional characterization of peripheral blood (PB) Mo as well as Mo present in pleural effusions (PE) from patients with tuberculosis (Mo-TB). As control, circulating Mo from healthy individuals (Mo-N), and Mo present in PB and PE of pulmonary cancer as inflammatory but non-infectious disease were evaluated. order to establish granuloma formation and to sustain T-lymphocyte activation in the lymph nodes. The aim of this work was to perform a phenotypic and functional characterization of peripheral blood (PB) Mo as well as Mo present in pleural effusions (PE) from patients with tuberculosis (Mo-TB). As control, circulating Mo from healthy individuals (Mo-N), and Mo present in PB and PE of pulmonary cancer as inflammatory but non-infectious disease were evaluated. tuberculosis (Mtb) and their innate capacity to deal with Mtb defines the early progression of the infection. In such chronic disease, circulating Mo arriving to the site of infection are the best candidates for the renewal of Mö and dendritic cells in order to establish granuloma formation and to sustain T-lymphocyte activation in the lymph nodes. The aim of this work was to perform a phenotypic and functional characterization of peripheral blood (PB) Mo as well as Mo present in pleural effusions (PE) from patients with tuberculosis (Mo-TB). As control, circulating Mo from healthy individuals (Mo-N), and Mo present in PB and PE of pulmonary cancer as inflammatory but non-infectious disease were evaluated. order to establish granuloma formation and to sustain T-lymphocyte activation in the lymph nodes. The aim of this work was to perform a phenotypic and functional characterization of peripheral blood (PB) Mo as well as Mo present in pleural effusions (PE) from patients with tuberculosis (Mo-TB). As control, circulating Mo from healthy individuals (Mo-N), and Mo present in PB and PE of pulmonary cancer as inflammatory but non-infectious disease were evaluated. progression of the infection. In such chronic disease, circulating Mo arriving to the site of infection are the best candidates for the renewal of Mö and dendritic cells in order to establish granuloma formation and to sustain T-lymphocyte activation in the lymph nodes. The aim of this work was to perform a phenotypic and functional characterization of peripheral blood (PB) Mo as well as Mo present in pleural effusions (PE) from patients with tuberculosis (Mo-TB). As control, circulating Mo from healthy individuals (Mo-N), and Mo present in PB and PE of pulmonary cancer as inflammatory but non-infectious disease were evaluated. order to establish granuloma formation and to sustain T-lymphocyte activation in the lymph nodes. The aim of this work was to perform a phenotypic and functional characterization of peripheral blood (PB) Mo as well as Mo present in pleural effusions (PE) from patients with tuberculosis (Mo-TB). As control, circulating Mo from healthy individuals (Mo-N), and Mo present in PB and PE of pulmonary cancer as inflammatory but non-infectious disease were evaluated. ö) are the main target for Mycobacterium tuberculosis (Mtb) and their innate capacity to deal with Mtb defines the early progression of the infection. In such chronic disease, circulating Mo arriving to the site of infection are the best candidates for the renewal of Mö and dendritic cells in order to establish granuloma formation and to sustain T-lymphocyte activation in the lymph nodes. The aim of this work was to perform a phenotypic and functional characterization of peripheral blood (PB) Mo as well as Mo present in pleural effusions (PE) from patients with tuberculosis (Mo-TB). As control, circulating Mo from healthy individuals (Mo-N), and Mo present in PB and PE of pulmonary cancer as inflammatory but non-infectious disease were evaluated. order to establish granuloma formation and to sustain T-lymphocyte activation in the lymph nodes. The aim of this work was to perform a phenotypic and functional characterization of peripheral blood (PB) Mo as well as Mo present in pleural effusions (PE) from patients with tuberculosis (Mo-TB). As control, circulating Mo from healthy individuals (Mo-N), and Mo present in PB and PE of pulmonary cancer as inflammatory but non-infectious disease were evaluated. progression of the infection. In such chronic disease, circulating Mo arriving to the site of infection are the best candidates for the renewal of Mö and dendritic cells in order to establish granuloma formation and to sustain T-lymphocyte activation in the lymph nodes. The aim of this work was to perform a phenotypic and functional characterization of peripheral blood (PB) Mo as well as Mo present in pleural effusions (PE) from patients with tuberculosis (Mo-TB). As control, circulating Mo from healthy individuals (Mo-N), and Mo present in PB and PE of pulmonary cancer as inflammatory but non-infectious disease were evaluated. order to establish granuloma formation and to sustain T-lymphocyte activation in the lymph nodes. The aim of this work was to perform a phenotypic and functional characterization of peripheral blood (PB) Mo as well as Mo present in pleural effusions (PE) from patients with tuberculosis (Mo-TB). As control, circulating Mo from healthy individuals (Mo-N), and Mo present in PB and PE of pulmonary cancer as inflammatory but non-infectious disease were evaluated. (Mtb) and their innate capacity to deal with Mtb defines the early progression of the infection. In such chronic disease, circulating Mo arriving to the site of infection are the best candidates for the renewal of Mö and dendritic cells in order to establish granuloma formation and to sustain T-lymphocyte activation in the lymph nodes. The aim of this work was to perform a phenotypic and functional characterization of peripheral blood (PB) Mo as well as Mo present in pleural effusions (PE) from patients with tuberculosis (Mo-TB). As control, circulating Mo from healthy individuals (Mo-N), and Mo present in PB and PE of pulmonary cancer as inflammatory but non-infectious disease were evaluated. order to establish granuloma formation and to sustain T-lymphocyte activation in the lymph nodes. The aim of this work was to perform a phenotypic and functional characterization of peripheral blood (PB) Mo as well as Mo present in pleural effusions (PE) from patients with tuberculosis (Mo-TB). As control, circulating Mo from healthy individuals (Mo-N), and Mo present in PB and PE of pulmonary cancer as inflammatory but non-infectious disease were evaluated. ö and dendritic cells in order to establish granuloma formation and to sustain T-lymphocyte activation in the lymph nodes. The aim of this work was to perform a phenotypic and functional characterization of peripheral blood (PB) Mo as well as Mo present in pleural effusions (PE) from patients with tuberculosis (Mo-TB). As control, circulating Mo from healthy individuals (Mo-N), and Mo present in PB and PE of pulmonary cancer as inflammatory but non-infectious disease were evaluated. Methods: Mononuclear cells were isolated from PB and PE by Ficoll-Hypaque. For phenotypic analysis a three-colour flow cytometric analysis was performed in order to evaluate marker expression in the basis of CD14+/CD16- and CD14+/CD16+ profile. Therefore, FITC, PE or PE-Cy5 conjugated monoclonal antibodies against CD14, CD16, CD11b, CD54, CCR5, TLR2, and TLR4 molecules were used. Besides, phagocytosis assays using Mtb-FITC was carried out. phenotypic analysis a three-colour flow cytometric analysis was performed in order to evaluate marker expression in the basis of CD14+/CD16- and CD14+/CD16+ profile. Therefore, FITC, PE or PE-Cy5 conjugated monoclonal antibodies against CD14, CD16, CD11b, CD54, CCR5, TLR2, and TLR4 molecules were used. Besides, phagocytosis assays using Mtb-FITC was carried out. : Mononuclear cells were isolated from PB and PE by Ficoll-Hypaque. For phenotypic analysis a three-colour flow cytometric analysis was performed in order to evaluate marker expression in the basis of CD14+/CD16- and CD14+/CD16+ profile. Therefore, FITC, PE or PE-Cy5 conjugated monoclonal antibodies against CD14, CD16, CD11b, CD54, CCR5, TLR2, and TLR4 molecules were used. Besides, phagocytosis assays using Mtb-FITC was carried out. Results: Patients with TB have a higher number of CD14+/CD16+ Mo population compared with N-Mo (p<0.0001, n=30) with a higher expression of the Mtb receptors: CD14 (p<0.006) and CD11b (p<0.001), and a higher expression of a chemokine receptor involved in homing to inflamed tissue: CCR5 (p<0.003) as well as CD54 (p<0.05). In addition this Mo subset expressed higher levels of TLR2 and TLR4 (p<0.05), suggesting that these pre-activated state of Mo could be reflected in a strongly response to Mtb and migration to the site of infection. Accordingly, CD14+/CD16+ Mo showed a higher ability to bind and phagocytose Mtb-FITC than CD14+/CD16- Mo, and moreover, it was the predominantly subset in PE from TB and not in PE from pulmonary cancer. compared with N-Mo (p<0.0001, n=30) with a higher expression of the Mtb receptors: CD14 (p<0.006) and CD11b (p<0.001), and a higher expression of a chemokine receptor involved in homing to inflamed tissue: CCR5 (p<0.003) as well as CD54 (p<0.05). In addition this Mo subset expressed higher levels of TLR2 and TLR4 (p<0.05), suggesting that these pre-activated state of Mo could be reflected in a strongly response to Mtb and migration to the site of infection. Accordingly, CD14+/CD16+ Mo showed a higher ability to bind and phagocytose Mtb-FITC than CD14+/CD16- Mo, and moreover, it was the predominantly subset in PE from TB and not in PE from pulmonary cancer. : Patients with TB have a higher number of CD14+/CD16+ Mo population compared with N-Mo (p<0.0001, n=30) with a higher expression of the Mtb receptors: CD14 (p<0.006) and CD11b (p<0.001), and a higher expression of a chemokine receptor involved in homing to inflamed tissue: CCR5 (p<0.003) as well as CD54 (p<0.05). In addition this Mo subset expressed higher levels of TLR2 and TLR4 (p<0.05), suggesting that these pre-activated state of Mo could be reflected in a strongly response to Mtb and migration to the site of infection. Accordingly, CD14+/CD16+ Mo showed a higher ability to bind and phagocytose Mtb-FITC than CD14+/CD16- Mo, and moreover, it was the predominantly subset in PE from TB and not in PE from pulmonary cancer. Conclusion: In patients with TB, circulating Mo have a state of activation that leds to a high capacity to bind Mtb and to migrate into the site of infection. The trafficking and the fate of those subpopulations once in the lungs remain to be established. leds to a high capacity to bind Mtb and to migrate into the site of infection. The trafficking and the fate of those subpopulations once in the lungs remain to be established. : In patients with TB, circulating Mo have a state of activation that leds to a high capacity to bind Mtb and to migrate into the site of infection. The trafficking and the fate of those subpopulations once in the lungs remain to be established.