Journal: PLoS ONE

Article Title: Immunotherapy Reduces Allergen-Mediated CD66b Expression and Myeloperoxidase Levels on Human Neutrophils from Allergic Patients

doi: 10.1371/journal.pone.0094558

Figure Lengend Snippet: ( A ) Neutrophils (2×10 6 ) from allergic patients (n = 10) were left untreated (No adds), treated with LPS (1 µg/ml) or an Ag (G 3 ) to which the patient was sensitized (40 µg/ml) for 24 h. ( B ) and ( C ) Neutrophils from allergic patients (n = 10) were incubated with an Ag (G 3 ) to which the patient was sensitized for the indicated doses (for 24 h) or times (at a dose of 40 µg/ml), and CD66b cell-surface expression was analyzed by flow cytometry. The results shown are the means ± SD.

Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated mouse monoclonal antibody (Ab) against human CD66b, and FITC-conjugated IgG isotype control were from Coulter-Izasa (Barcelona, Spain).

Techniques: Incubation, Expressing, Flow Cytometry

Journal: PLoS ONE

Article Title: Immunotherapy Reduces Allergen-Mediated CD66b Expression and Myeloperoxidase Levels on Human Neutrophils from Allergic Patients

doi: 10.1371/journal.pone.0094558

Figure Lengend Snippet: ( A ) Neutrophils (2×10 6 ) from allergic patients (n = 10) sensitized to G 3 but not to D 1 and W 6 , were incubated with 40 µg/ml protein extracts of G 3 , D 1 , and W 6 for 24 h. ( B ) Neutrophils from healthy subject (n = 10) were incubated with 40 µg/ml of T 9 , and G 3 for 24 h. CD66b cell-surface expression was measured by flow cytometry. The values shown are the means ± SD.

Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated mouse monoclonal antibody (Ab) against human CD66b, and FITC-conjugated IgG isotype control were from Coulter-Izasa (Barcelona, Spain).

Techniques: Incubation, Expressing, Flow Cytometry

Journal: PLoS ONE

Article Title: Immunotherapy Reduces Allergen-Mediated CD66b Expression and Myeloperoxidase Levels on Human Neutrophils from Allergic Patients

doi: 10.1371/journal.pone.0094558

Figure Lengend Snippet: ( A ) Neutrophils (2×10 6 ) from allergic patients sensitized to G 3 (n = 10) were treated, where indicated, to elute the Igs from the cell surface, and then incubated with G 3 (40 µg/ml) or LPS (1 µg/ml) for 24 h, and CD66 cell-surface expression was measured by flow cytometry. Specific IgE and IgG for the above Ags were determined in the elution solution by ELISA. ( B ) Neutrophils from allergic patients (n = 5) were incubated with anti-FcεRI (CRA 1), anti-CD23 (9P.25) or anti-galectin-3 (A3A12) antibodies at 5 µg/ml for 24 h, and CD66b cell-surface expression was measured as in A. The values shown are the means ± SD.

Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated mouse monoclonal antibody (Ab) against human CD66b, and FITC-conjugated IgG isotype control were from Coulter-Izasa (Barcelona, Spain).

Techniques: Incubation, Expressing, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: Immunotherapy Reduces Allergen-Mediated CD66b Expression and Myeloperoxidase Levels on Human Neutrophils from Allergic Patients

doi: 10.1371/journal.pone.0094558

Figure Lengend Snippet: CD66b expression is higher on neutrophils from NLP than those from peripheral blood. ( A ) Neutrophils (2×10 6 cells) from healthy subjects (n = 10), non-IT-treated patients with allergic asthma and rhinitis sensitized to G 3 (n = 10), and IT-treated patients with allergic asthma and rhinitis sensitized to G 3 (n = 15) were left untreated (No adds) or treated with G 3 (40 µg/ml) for 24 h. ( B ) Neutrophils purified from peripheral blood and NLF from non-IT treated patients. The levels of CD66b cell-surface expression were measured by flow cytometry. The values shown are the means ± SD.

Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated mouse monoclonal antibody (Ab) against human CD66b, and FITC-conjugated IgG isotype control were from Coulter-Izasa (Barcelona, Spain).

Techniques: Expressing, Purification, Flow Cytometry

Journal: PLoS ONE

Article Title: Immunotherapy Reduces Allergen-Mediated CD66b Expression and Myeloperoxidase Levels on Human Neutrophils from Allergic Patients

doi: 10.1371/journal.pone.0094558

Figure Lengend Snippet: ( A ) CD66b expression was analyzed on cells from NLF in healthy subjects (n = 10), IT-treated allergic patients (n = 15) and non-IT-treated allergic patients (n = 10). ( B ) MPO activity was analyzed in NLF from healthy subjects (n = 10), IT-treated allergic patients (n = 15) and non-IT-treated allergic patients (n = 10). The values shown are the mean ± SD.

Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated mouse monoclonal antibody (Ab) against human CD66b, and FITC-conjugated IgG isotype control were from Coulter-Izasa (Barcelona, Spain).

Techniques: Expressing, Activity Assay

Journal: PLoS ONE

Article Title: Immunotherapy Reduces Allergen-Mediated CD66b Expression and Myeloperoxidase Levels on Human Neutrophils from Allergic Patients

doi: 10.1371/journal.pone.0094558

Figure Lengend Snippet: ( A ) The percentage of CD66b + MPO + cells was analyzed on cells from NLF in healthy subjects (n = 10), IT-treated allergic patients (n = 15) and non-IT-treated allergic patients (n = 10). The values shown are the mean ± SD. ( B ) Three representative dot-blot of the results shown in A.

Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated mouse monoclonal antibody (Ab) against human CD66b, and FITC-conjugated IgG isotype control were from Coulter-Izasa (Barcelona, Spain).

Techniques: Dot Blot