Regulation of immune responsiveness in vivo by disrupting an early T-cell signaling event using a cell-permeable peptide

• Published in: PloS one 2013-05, Vol.8 (5), p.e63645
• Main Authors: Guimond, David M, Cam, Nicholas R, Hirve, Nupura, Duan, Wei, Lambris, John D, Croft, Michael, Tsoukas, Constantine D
• Format: Article
• Language: English
• Subjects: Activation; Allergens; Allergies; Amino Acid Sequence; Analysis; Animal models; Animals; Antigens; Asthma; Bacteria; Bacterial diseases; Bacterial infections; Biology; Bronchial Hyperreactivity - complications; Bronchial Hyperreactivity - immunology; Bronchial Hyperreactivity - pathology; Bronchioles - drug effects; Bronchioles - metabolism; Bronchioles - pathology; Cell-Penetrating Peptides - chemistry; Cell-Penetrating Peptides - pharmacology; Cellular signal transduction; Cytokines; Cytokines - biosynthesis; Eosinophilia - complications; Eosinophilia - immunology; Eosinophilia - pathology; Experiments; Female; Goblet Cells - drug effects; Goblet Cells - metabolism; Goblet Cells - pathology; Health aspects; Hypersensitivity; Immune response; Immune system; Immunity - drug effects; Immunology; Infiltration; Inflammation; Inflammatory response; Inhibition; Intracellular; Itk protein; Kinases; Laboratory animals; Leukocytes; Lungs; Lymph nodes; Lymph Nodes - pathology; Lymphocytes; Lymphocytes - drug effects; Lymphocytes - metabolism; Lymphocytes T; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Molecular Sequence Data; Mucus; Mucus - metabolism; Ovalbumin; Ovalbumin - immunology; Parasitic diseases; Pathogenesis; Peptides; Permeability; Pneumonia - complications; Pneumonia - immunology; Pneumonia - pathology; Proline; Protein-Tyrosine Kinases; Respiratory tract; Signal transduction; Signal Transduction - drug effects; Signal Transduction - immunology; Signaling; SLP-76 protein; T cell receptors; T cells; Th2 Cells - drug effects; Th2 Cells - immunology
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0063645

The inducible T cell kinase (ITK) regulates type 2 (Th2) cytokines that provide defense against certain parasitic and bacterial infections and are involved in the pathogenesis of lung inflammation such as allergic asthma. Activation of ITK requires the interaction of its SH3 domain with the poly-proline region of its signaling partner, the SH2 domain containing leukocyte phosphoprotein of 76 kilodaltons (SLP-76). The specific disruption of the ITK-SH3/SLP-76 poly-proline interaction in vitro by a cell-permeable competitive inhibitor peptide (R9-QQP) interferes with the activation of ITK and the transduction of its cellular functions in T lymphocytes. In the present investigation, we assessed the effects of R9-QQP treatment on the induction of an in vivo immune response as represented by lung inflammation in a murine model of allergic asthma. We found that mice treated with R9-QQP and sensitized and challenged with the surrogate allergen ovalbumin (OVA) display significant inhibition of lung inflammation in a peptide-specific manner. Thus, parameters of the allergic response, such as airway hyper-responsiveness, suppression of inflammatory cell infiltration, reduction of bronchial mucus accumulation, and production of relevant cytokines from draining lymph nodes were significantly suppressed. These findings represent the first demonstration of the biological significance of the interaction between ITK and SLP-76 in the induction of an immune response in a whole animal model and specifically underscore the significance of the ITK-SH3 domain interaction with the poly-proline region of SLP-76 in the development of an inflammatory response. Furthermore, the experimental approach of intracellular peptide-mediated inhibition might be applicable to the study of other important intracellular interactions thus providing a paradigm for dissecting signal transduction pathways.