ABSTRACT SESSION TYPE: ILD PostersPRESENTED ON: Wednesday, October 24, 2012 at 01:30 PM - 02:30 P... more ABSTRACT SESSION TYPE: ILD PostersPRESENTED ON: Wednesday, October 24, 2012 at 01:30 PM - 02:30 PMPURPOSE: Recruitment of macrophages in BAL is associated to the development of inflammatory processes during the course of Interstitial Lung Diseases (ILD). However, the mechanisms responsible for macrophage recruitment are poorly defined. Hyaluronan (HA) is a glycosaminoglycan found at high concentrations in BAL from ILD patients. Hyaluronan can regulate different cell functions by interacting with different receptors, among them, CD44 and RHAAM. In the present study we analyzed whether HA was able to induce the chemotaxis of BAL macrophages from ILD patients and the involvement of CD44 and RHAAM in this response.METHODS: Alveolar macrophages were purified from BAL recovered from ILD patients (n=31) by adherence on plastic tissue culture dishes. Migration was measured using a transwell system, in which macrophages were seeded on the upper compartment chamber, and RPMI (diluent), HA (2 μg/mL) or BAL from ILD patients were added into the lower compartment chamber, either in the presence or absence of hialuronidase (5U/mL). To evaluate the role of CD44 and RHAAM in cell migration, macrophages were preincubated with saturating concentrations of specific blocking monoclonal antibodies, before the onset of the migration assay. The results were expressed as a migration index: migration of macrophages in response to HA or BAL/migration of macrophages toward culture medium.RESULTS: We found that both HA and BAL from ILD patients induced the migration of macrophages: migration index = 2.39 ± 0.12 and 2.64 ± 0.43, respectively (p<0.05). Of note, the presence of hialuronidase abrogated the migration response induced by BAL from ILD patients: % inhibition >95, n=4. Treatment of macrophages with antibodies directed to CD44, markedly decreased the migratory response induced by HA: 1,24 ± 0.08 vs 2.40 ± 0.12 for anti-CD44-treated vs untreated macrophages, respectively, p<0.05). Anti-RHAAM antibodies did not mediate any effect.CONCLUSIONS: BAL from patients with ILD induces the migration of macrophages through a mechanism strongly dependent on the presence of HA.CLINICAL IMPLICATIONS: CD44 could be a novel therapeutic target involved in the recruitment of inflammatory cells in the lung of ILD patientsDISCLOSURE: The following authors have nothing to disclose: Glenda Ernst, Carolina Jancic, Auteri Santiago, Fabían Caro, Fernando Galíndez, Jorge Geffner, Silvia Hajos, Pedro GrynblatNo Product/Research Disclosure InformationMaría Ferrer Hospital, Buenos Aires, Argentina.
ABSTRACT SESSION TYPE: ILD PostersPRESENTED ON: Wednesday, October 24, 2012 at 01:30 PM - 02:30 P... more ABSTRACT SESSION TYPE: ILD PostersPRESENTED ON: Wednesday, October 24, 2012 at 01:30 PM - 02:30 PMPURPOSE: Recruitment of macrophages in BAL is associated to the development of inflammatory processes during the course of Interstitial Lung Diseases (ILD). However, the mechanisms responsible for macrophage recruitment are poorly defined. Hyaluronan (HA) is a glycosaminoglycan found at high concentrations in BAL from ILD patients. Hyaluronan can regulate different cell functions by interacting with different receptors, among them, CD44 and RHAAM. In the present study we analyzed whether HA was able to induce the chemotaxis of BAL macrophages from ILD patients and the involvement of CD44 and RHAAM in this response.METHODS: Alveolar macrophages were purified from BAL recovered from ILD patients (n=31) by adherence on plastic tissue culture dishes. Migration was measured using a transwell system, in which macrophages were seeded on the upper compartment chamber, and RPMI (diluent), HA (2 μg/mL) or BAL from ILD patients were added into the lower compartment chamber, either in the presence or absence of hialuronidase (5U/mL). To evaluate the role of CD44 and RHAAM in cell migration, macrophages were preincubated with saturating concentrations of specific blocking monoclonal antibodies, before the onset of the migration assay. The results were expressed as a migration index: migration of macrophages in response to HA or BAL/migration of macrophages toward culture medium.RESULTS: We found that both HA and BAL from ILD patients induced the migration of macrophages: migration index = 2.39 ± 0.12 and 2.64 ± 0.43, respectively (p<0.05). Of note, the presence of hialuronidase abrogated the migration response induced by BAL from ILD patients: % inhibition >95, n=4. Treatment of macrophages with antibodies directed to CD44, markedly decreased the migratory response induced by HA: 1,24 ± 0.08 vs 2.40 ± 0.12 for anti-CD44-treated vs untreated macrophages, respectively, p<0.05). Anti-RHAAM antibodies did not mediate any effect.CONCLUSIONS: BAL from patients with ILD induces the migration of macrophages through a mechanism strongly dependent on the presence of HA.CLINICAL IMPLICATIONS: CD44 could be a novel therapeutic target involved in the recruitment of inflammatory cells in the lung of ILD patientsDISCLOSURE: The following authors have nothing to disclose: Glenda Ernst, Carolina Jancic, Auteri Santiago, Fabían Caro, Fernando Galíndez, Jorge Geffner, Silvia Hajos, Pedro GrynblatNo Product/Research Disclosure InformationMaría Ferrer Hospital, Buenos Aires, Argentina.
There is a large variability between the different normograms of spirometric values, so that we d... more There is a large variability between the different normograms of spirometric values, so that we designed our normogram for Buenos Aires and Gran Buenos Aires. We performed forced spirometry, under American Thoracic Society standardization, in 237 normal subjects (105 females) between 18 to 86 years old, and 144 to 194 cm. We measured Forced Vital Capacity (FVC), forced Expiratory Volume in one second (FEV1), and forced expiratory flow during the middle half of the Forced vital capacity (FEF25-75), in previously calibrated by the explosive decompressor spirometers. Linear regression using height and age was used for each measured value for each sex. The values obtained were in normal distribution, so that we determined the Low Limit of Normality calculating the 95% confidence interval to one tail, and this should replace the common method of the fixed percent of each value to determine the lower limit of normality for a predicted value.
ABSTRACT SESSION TYPE: ILD PostersPRESENTED ON: Wednesday, October 24, 2012 at 01:30 PM - 02:30 P... more ABSTRACT SESSION TYPE: ILD PostersPRESENTED ON: Wednesday, October 24, 2012 at 01:30 PM - 02:30 PMPURPOSE: Recruitment of macrophages in BAL is associated to the development of inflammatory processes during the course of Interstitial Lung Diseases (ILD). However, the mechanisms responsible for macrophage recruitment are poorly defined. Hyaluronan (HA) is a glycosaminoglycan found at high concentrations in BAL from ILD patients. Hyaluronan can regulate different cell functions by interacting with different receptors, among them, CD44 and RHAAM. In the present study we analyzed whether HA was able to induce the chemotaxis of BAL macrophages from ILD patients and the involvement of CD44 and RHAAM in this response.METHODS: Alveolar macrophages were purified from BAL recovered from ILD patients (n=31) by adherence on plastic tissue culture dishes. Migration was measured using a transwell system, in which macrophages were seeded on the upper compartment chamber, and RPMI (diluent), HA (2 μg/mL) or BAL from ILD patients were added into the lower compartment chamber, either in the presence or absence of hialuronidase (5U/mL). To evaluate the role of CD44 and RHAAM in cell migration, macrophages were preincubated with saturating concentrations of specific blocking monoclonal antibodies, before the onset of the migration assay. The results were expressed as a migration index: migration of macrophages in response to HA or BAL/migration of macrophages toward culture medium.RESULTS: We found that both HA and BAL from ILD patients induced the migration of macrophages: migration index = 2.39 ± 0.12 and 2.64 ± 0.43, respectively (p<0.05). Of note, the presence of hialuronidase abrogated the migration response induced by BAL from ILD patients: % inhibition >95, n=4. Treatment of macrophages with antibodies directed to CD44, markedly decreased the migratory response induced by HA: 1,24 ± 0.08 vs 2.40 ± 0.12 for anti-CD44-treated vs untreated macrophages, respectively, p<0.05). Anti-RHAAM antibodies did not mediate any effect.CONCLUSIONS: BAL from patients with ILD induces the migration of macrophages through a mechanism strongly dependent on the presence of HA.CLINICAL IMPLICATIONS: CD44 could be a novel therapeutic target involved in the recruitment of inflammatory cells in the lung of ILD patientsDISCLOSURE: The following authors have nothing to disclose: Glenda Ernst, Carolina Jancic, Auteri Santiago, Fabían Caro, Fernando Galíndez, Jorge Geffner, Silvia Hajos, Pedro GrynblatNo Product/Research Disclosure InformationMaría Ferrer Hospital, Buenos Aires, Argentina.
ABSTRACT SESSION TYPE: ILD PostersPRESENTED ON: Wednesday, October 24, 2012 at 01:30 PM - 02:30 P... more ABSTRACT SESSION TYPE: ILD PostersPRESENTED ON: Wednesday, October 24, 2012 at 01:30 PM - 02:30 PMPURPOSE: Recruitment of macrophages in BAL is associated to the development of inflammatory processes during the course of Interstitial Lung Diseases (ILD). However, the mechanisms responsible for macrophage recruitment are poorly defined. Hyaluronan (HA) is a glycosaminoglycan found at high concentrations in BAL from ILD patients. Hyaluronan can regulate different cell functions by interacting with different receptors, among them, CD44 and RHAAM. In the present study we analyzed whether HA was able to induce the chemotaxis of BAL macrophages from ILD patients and the involvement of CD44 and RHAAM in this response.METHODS: Alveolar macrophages were purified from BAL recovered from ILD patients (n=31) by adherence on plastic tissue culture dishes. Migration was measured using a transwell system, in which macrophages were seeded on the upper compartment chamber, and RPMI (diluent), HA (2 μg/mL) or BAL from ILD patients were added into the lower compartment chamber, either in the presence or absence of hialuronidase (5U/mL). To evaluate the role of CD44 and RHAAM in cell migration, macrophages were preincubated with saturating concentrations of specific blocking monoclonal antibodies, before the onset of the migration assay. The results were expressed as a migration index: migration of macrophages in response to HA or BAL/migration of macrophages toward culture medium.RESULTS: We found that both HA and BAL from ILD patients induced the migration of macrophages: migration index = 2.39 ± 0.12 and 2.64 ± 0.43, respectively (p<0.05). Of note, the presence of hialuronidase abrogated the migration response induced by BAL from ILD patients: % inhibition >95, n=4. Treatment of macrophages with antibodies directed to CD44, markedly decreased the migratory response induced by HA: 1,24 ± 0.08 vs 2.40 ± 0.12 for anti-CD44-treated vs untreated macrophages, respectively, p<0.05). Anti-RHAAM antibodies did not mediate any effect.CONCLUSIONS: BAL from patients with ILD induces the migration of macrophages through a mechanism strongly dependent on the presence of HA.CLINICAL IMPLICATIONS: CD44 could be a novel therapeutic target involved in the recruitment of inflammatory cells in the lung of ILD patientsDISCLOSURE: The following authors have nothing to disclose: Glenda Ernst, Carolina Jancic, Auteri Santiago, Fabían Caro, Fernando Galíndez, Jorge Geffner, Silvia Hajos, Pedro GrynblatNo Product/Research Disclosure InformationMaría Ferrer Hospital, Buenos Aires, Argentina.
ABSTRACT SESSION TYPE: ILD PostersPRESENTED ON: Wednesday, October 24, 2012 at 01:30 PM - 02:30 P... more ABSTRACT SESSION TYPE: ILD PostersPRESENTED ON: Wednesday, October 24, 2012 at 01:30 PM - 02:30 PMPURPOSE: Recruitment of macrophages in BAL is associated to the development of inflammatory processes during the course of Interstitial Lung Diseases (ILD). However, the mechanisms responsible for macrophage recruitment are poorly defined. Hyaluronan (HA) is a glycosaminoglycan found at high concentrations in BAL from ILD patients. Hyaluronan can regulate different cell functions by interacting with different receptors, among them, CD44 and RHAAM. In the present study we analyzed whether HA was able to induce the chemotaxis of BAL macrophages from ILD patients and the involvement of CD44 and RHAAM in this response.METHODS: Alveolar macrophages were purified from BAL recovered from ILD patients (n=31) by adherence on plastic tissue culture dishes. Migration was measured using a transwell system, in which macrophages were seeded on the upper compartment chamber, and RPMI (diluent), HA (2 μg/mL) or BAL from ILD patients were added into the lower compartment chamber, either in the presence or absence of hialuronidase (5U/mL). To evaluate the role of CD44 and RHAAM in cell migration, macrophages were preincubated with saturating concentrations of specific blocking monoclonal antibodies, before the onset of the migration assay. The results were expressed as a migration index: migration of macrophages in response to HA or BAL/migration of macrophages toward culture medium.RESULTS: We found that both HA and BAL from ILD patients induced the migration of macrophages: migration index = 2.39 ± 0.12 and 2.64 ± 0.43, respectively (p<0.05). Of note, the presence of hialuronidase abrogated the migration response induced by BAL from ILD patients: % inhibition >95, n=4. Treatment of macrophages with antibodies directed to CD44, markedly decreased the migratory response induced by HA: 1,24 ± 0.08 vs 2.40 ± 0.12 for anti-CD44-treated vs untreated macrophages, respectively, p<0.05). Anti-RHAAM antibodies did not mediate any effect.CONCLUSIONS: BAL from patients with ILD induces the migration of macrophages through a mechanism strongly dependent on the presence of HA.CLINICAL IMPLICATIONS: CD44 could be a novel therapeutic target involved in the recruitment of inflammatory cells in the lung of ILD patientsDISCLOSURE: The following authors have nothing to disclose: Glenda Ernst, Carolina Jancic, Auteri Santiago, Fabían Caro, Fernando Galíndez, Jorge Geffner, Silvia Hajos, Pedro GrynblatNo Product/Research Disclosure InformationMaría Ferrer Hospital, Buenos Aires, Argentina.
There is a large variability between the different normograms of spirometric values, so that we d... more There is a large variability between the different normograms of spirometric values, so that we designed our normogram for Buenos Aires and Gran Buenos Aires. We performed forced spirometry, under American Thoracic Society standardization, in 237 normal subjects (105 females) between 18 to 86 years old, and 144 to 194 cm. We measured Forced Vital Capacity (FVC), forced Expiratory Volume in one second (FEV1), and forced expiratory flow during the middle half of the Forced vital capacity (FEF25-75), in previously calibrated by the explosive decompressor spirometers. Linear regression using height and age was used for each measured value for each sex. The values obtained were in normal distribution, so that we determined the Low Limit of Normality calculating the 95% confidence interval to one tail, and this should replace the common method of the fixed percent of each value to determine the lower limit of normality for a predicted value.
ABSTRACT SESSION TYPE: ILD PostersPRESENTED ON: Wednesday, October 24, 2012 at 01:30 PM - 02:30 P... more ABSTRACT SESSION TYPE: ILD PostersPRESENTED ON: Wednesday, October 24, 2012 at 01:30 PM - 02:30 PMPURPOSE: Recruitment of macrophages in BAL is associated to the development of inflammatory processes during the course of Interstitial Lung Diseases (ILD). However, the mechanisms responsible for macrophage recruitment are poorly defined. Hyaluronan (HA) is a glycosaminoglycan found at high concentrations in BAL from ILD patients. Hyaluronan can regulate different cell functions by interacting with different receptors, among them, CD44 and RHAAM. In the present study we analyzed whether HA was able to induce the chemotaxis of BAL macrophages from ILD patients and the involvement of CD44 and RHAAM in this response.METHODS: Alveolar macrophages were purified from BAL recovered from ILD patients (n=31) by adherence on plastic tissue culture dishes. Migration was measured using a transwell system, in which macrophages were seeded on the upper compartment chamber, and RPMI (diluent), HA (2 μg/mL) or BAL from ILD patients were added into the lower compartment chamber, either in the presence or absence of hialuronidase (5U/mL). To evaluate the role of CD44 and RHAAM in cell migration, macrophages were preincubated with saturating concentrations of specific blocking monoclonal antibodies, before the onset of the migration assay. The results were expressed as a migration index: migration of macrophages in response to HA or BAL/migration of macrophages toward culture medium.RESULTS: We found that both HA and BAL from ILD patients induced the migration of macrophages: migration index = 2.39 ± 0.12 and 2.64 ± 0.43, respectively (p<0.05). Of note, the presence of hialuronidase abrogated the migration response induced by BAL from ILD patients: % inhibition >95, n=4. Treatment of macrophages with antibodies directed to CD44, markedly decreased the migratory response induced by HA: 1,24 ± 0.08 vs 2.40 ± 0.12 for anti-CD44-treated vs untreated macrophages, respectively, p<0.05). Anti-RHAAM antibodies did not mediate any effect.CONCLUSIONS: BAL from patients with ILD induces the migration of macrophages through a mechanism strongly dependent on the presence of HA.CLINICAL IMPLICATIONS: CD44 could be a novel therapeutic target involved in the recruitment of inflammatory cells in the lung of ILD patientsDISCLOSURE: The following authors have nothing to disclose: Glenda Ernst, Carolina Jancic, Auteri Santiago, Fabían Caro, Fernando Galíndez, Jorge Geffner, Silvia Hajos, Pedro GrynblatNo Product/Research Disclosure InformationMaría Ferrer Hospital, Buenos Aires, Argentina.
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