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Evidence for aberrant regulation of the p21Ras pathway in PBMCs of patients with chronic idiopathic urticaria  Ronit Confino-Cohen, MD, Dorit Aharoni,

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Presentation on theme: "Evidence for aberrant regulation of the p21Ras pathway in PBMCs of patients with chronic idiopathic urticaria  Ronit Confino-Cohen, MD, Dorit Aharoni,"— Presentation transcript:

1 Evidence for aberrant regulation of the p21Ras pathway in PBMCs of patients with chronic idiopathic urticaria  Ronit Confino-Cohen, MD, Dorit Aharoni, PhD, Arnon Goldberg, MD, Irena Gurevitch, MD, Andreas Buchs, MD, Mordechai Weiss, MD, Joshua Weissgarten, MD, Micha J. Rapoport, MD  Journal of Allergy and Clinical Immunology  Volume 109, Issue 2, Pages (February 2002) DOI: /mai Copyright © 2002 Mosby, Inc. Terms and Conditions

2 Fig. 1 Increased p21Ras expression in patients with CIU. PBMCs were lysed, and Western blot analysis was performed with specific antibody against p21Ras. A , Representative Western blot demonstrating p21Ras expression in control subjects and patients with CIU. B , Expression of p21Ras was determined by means of photodensitometry in whole-cell lysate of normal control subjects and patients with CIU. Data are means ± SEM of each group and expressed in arbitrary units. Journal of Allergy and Clinical Immunology  , DOI: ( /mai ) Copyright © 2002 Mosby, Inc. Terms and Conditions

3 Fig. 2 Decreased expression of hSOS1 with normal p120GAP expression in patients with CIU. A , Representative Western blot of hSOS1, p120GAP, and p21Ras expression of 2 different patients with CIU with positive skin reactions and their paired control subjects. Reduction in hSOS1 expression is associated with elevated levels of p21Ras within the same patients. B , Expression of hSOS1 and p120GAP in patients with CIU (filled bars) and control subjects (open bars) was determined by means of photodensitometry. Data are means ± SEM of each group and expressed in arbitrary units. Journal of Allergy and Clinical Immunology  , DOI: ( /mai ) Copyright © 2002 Mosby, Inc. Terms and Conditions

4 Fig. 3 hSOS1 M/C distribution. A , Representative blot demonstrating hSOS1 expression in cytoplasm (Cyt) and membrane fractions (M) of nonstimulated and PHA-stimulated PBMCs of normal control subjects and patients with CIU. B , hSOS1 protein level in membrane and cytoplasmic fractions (means ± SEM of each group) were determined by means of photodensitometry, and the M/C ratio was calculated before (open bars) and after (filled bars) stimulation. Journal of Allergy and Clinical Immunology  , DOI: ( /mai ) Copyright © 2002 Mosby, Inc. Terms and Conditions

5 Fig. 4 p120GAP M/C distribution. A , Representative Western blot demonstrating p120GAP expression in cytoplasm (Cyt) and membrane fractions (M) of nonstimulated and PHA-stimulated PBMCs of normal control subjects and patients with CIU with active disease. B , M/C ratio of p120GAP was determined before (open bars) and after (filled bars) stimulation. Protein levels (means ± SEM of each group) were determined by means of photodensitometry and expressed in arbitrary units. Journal of Allergy and Clinical Immunology  , DOI: ( /mai ) Copyright © 2002 Mosby, Inc. Terms and Conditions

6 Fig. 5 Similar MAP-kinase (ERK1 and ERK2) activity in patients with CIU and control subjects. A , Representative Western blot demonstrating the basal ERK1 and ERK2 activity (phosphorylated form) in nonstimulated PBMCs of normal control subjects (lanes 1 , 2 , and 3 ) and patients with CIU (lanes 4 , 5 , and 6 ). B , Representative Western blot of phosphorylated ERK1 and ERK2 in fractions of cytoplasm (Cyt) and membranes (M) before and after stimulation. C , Levels of phosphorylated ERK1 and ERK2 were determined before (open bars) and after (filled bars) PHA stimulation by means of photodensitometry in whole-cell lysates (cytoplasm plus membrane fractions) of patients with CIU and control subjects and expressed in arbitrary units. Data are means ± SEM of each group. Journal of Allergy and Clinical Immunology  , DOI: ( /mai ) Copyright © 2002 Mosby, Inc. Terms and Conditions

7 Fig. 6 In vitro proliferative response to PHA of PBMCs from patients with CIU and normal control subjects. PBMCs were isolated from patients (filled bars) and control subjects (open bars) , as previously described. Maximal proliferative response to PHA ( μg/mL) was assessed after 3 days by means of tritiated thymidine uptake (20-hour pulse). Values ± SEM are expressed in SI units calculated as the maximal proliferation in counts per minute divided by the background unstimulated value of each patient. Journal of Allergy and Clinical Immunology  , DOI: ( /mai ) Copyright © 2002 Mosby, Inc. Terms and Conditions

8 Fig. 7 Aberrant regulation of the p21Ras pathway in lymphocytes of patients with CIU: possible mechanisms leading to autoimmune disease. Genetically determined aberrant p21Ras signaling interferes with thymic T-cell selection (1) , leading to a release of self-aggressive T cells (2) . Alternatively, exposure to an unknown environmental trigger (3) results in an aberrant signaling in B and T cells (4) and disrupts immune balance. T cell-assisted (5) production of anti-FcϵRI autoantibodies (auto Abs) by B cells (6) and a direct interaction of autoimmune T cells with mast cells (7) lead to autoimmune chronic urticaria. Journal of Allergy and Clinical Immunology  , DOI: ( /mai ) Copyright © 2002 Mosby, Inc. Terms and Conditions

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