A role for mitogen kinase kinase 3 in pulmonary inflammation validated from a proteomic approach
| Autor: | Claire A. Sand, Yanira Riffo-Vasquez, Tatjana Holand, Domenico Spina, K.B. Bacon, C. Rohlff, Francis Woisin, Clive P. Page, Brian J. O'Connor, Michael S. Marber |
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| Rok vydání: | 2013 |
| Předmět: |
Pulmonary and Respiratory Medicine
Adult Male Proteomics Lipopolysaccharide Neutrophils MAP Kinase Kinase 3 Blotting Western Inflammation CD8-Positive T-Lymphocytes Peripheral blood mononuclear cell chemistry.chemical_compound Mice Young Adult Western blot medicine Eosinophilia Animals Humans Pharmacology (medical) Mice Knockout medicine.diagnostic_test business.industry Kinase Biochemistry (medical) Pneumonia Neutrophilia Asthma Mice Inbred C57BL Disease Models Animal chemistry Gene Expression Regulation Case-Control Studies Immunology Leukocytes Mononuclear medicine.symptom business CD8 |
| Zdroj: | Pulmonary pharmacologytherapeutics. 27(2) |
| ISSN: | 1522-9629 |
| Popis: | Proteomics is a powerful tool to ascertain which proteins are differentially expressed in the context of disease. We have used this approach on inflammatory cells obtained from patients with asthma to ascertain whether novel drugs targets could be illuminated and to investigate the role of any such target in a range of in vitro and in vivo models of inflammation. A proteomic study was undertaken using peripheral blood mononuclear cells from mild asthmatic subjects compared with healthy subjects. The analysis revealed an increased expression of the intra- cellular kinase, mitogen activated protein kinase (MKK3), and the function of this protein was investi- gated further in preclinical models of inflammation using MKK3 knockout mice. We describe a 3.65 fold increase in the expression of MKK3 in CD8 þ T lymphocytes obtained from subjects with asthma compared with healthy subjects using a proteomic approach which we have confirmed in CD8 þ , but not in CD4 þ T lymphocytes or human bronchial epithelial cells from asthmatic patients using a Western blot technique. In wild type mice, bacterial lipopolysaccharide (LPS) caused a significant increase in MKK3 expression and significantly reduced airway neutrophilia in MKK3 � /� mice (median, 25, 75% percentile; wild/LPS; 5.3 (0.7e9.9) � 10 5 cells/mL vs MKK3 � /� /LPS; 0 (0e1.9) � 10 5 cells/mL, P < 0.05). In contrast, eosinophilia in sensitized wild type mice challenged with allergen (0.5 (0.16e0.65) � 10 5 cells/mL) was significantly increased in MKK3 � /� mice (2.2 (0.9e3.5) � 10 5 cells/mL, P < 0.05). Our results suggest that asthma is associated with MKK3 over-expression in CD8 þ cells. We have also demonstrated that MKK3 may be critical for airway neutrophilia, but not eosinophilia, suggesting that this may be a target worthy of further consideration in the context of diseases associated with neutrophil activation such as severe asthma and COPD. |
| Databáze: | OpenAIRE |
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