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GOALS: Our data indicate that ITK plays an important role in gamma delta T cell function. Our objectives are therefore to understand the role of ITK in regulating the activation and function of this cell type using the mouse as a model, in order to better determine how to manipulate the immune response. <P>
OBJECTIVES: <OL> <LI> Determine the role of ITK in regulating the development of "conventional" peripheral gamma delta T cells.<LI>Determine the role of ITK in regulating the development of CD4+ NKT-like T cells. <LI> Determine the role of ITK in regulating the development of skin and mucosa homing gamma delta T cells.</ol> Within each objective there are points where the findings will be written up for publication. These publications will be considered milestones for this project. <P>
SCOPE OF IMPACT: This research will have national and international impact. The findings from the proposed experiments will have an impact on controlling immune responses of humans and animals worldwide. <P>
STAKEHOLDER ENGAGEMENT: This work is of interest to a variety of stakeholders. The results of the work may point to ways of manipulating the immune system of animals and humans, to obtaining healthier animals. Other areas of our work would be of interest to the food safety groups, as our work could point to ways of avoiding food allergies.
NON-TECHNICAL SUMMARY: T cells play a critical part in regulating the immune response in humans and animals. These cells are activated by interacting with foreign antigens, resulting in their activation and generation of an immune response. The two major types of T cells are the alpha beta T cells as well as gamma delta T cells. While alpha beta T cells are generally found in blood and lymphoid organs, gamma delta T cells are found in skin and mucosal areas. These gamma delta T cells play critical roles in immune response to pathogens. Understanding their development and function will allow us to developing strategies to specifically manipulate their activation and function. Effective targeting will depend on intimate knowledge of the molecular mechanisms of the development of gamma delta T cells, their differentiation, and cytokine production. The tyrosine kinase ITK represents a class of enzymes that are tractable to pharmaceutical intervention, and has been implicated in both the development of gamma delta T cells. The rationale behind this project is that a better understanding of the role that ITK plays in gamma delta T cell development and function may lead to the development of ways to manipulate these cells and thus the immune response. The research proposed in this project is significant because it will provide such knowledge.
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APPROACH: METHODS: 1. Mice lacking ITK, those carrying a kinase mutant that can be inhibited with a small molecule inhibitor, or WT mice will be analyzed for presence, percentage and number of gamma delta T cell subsets. These cells will be identified in spleen, blood, lymph nodes, bone marrow, liver and lung, by flow cytometry using antibodies against the gamma delta T cell receptor, as well as other surface markers that identify subsets, including the expression of specific gamma delta T cell receptor. These experiments will determine if gamma delta T cell development is affected by the absence or inhibition of ITK. Follow up experiments will sort these gamma delta T cell subsets and analyze their cytokine production. <P>
2. Mice lacking ITK, those carrying a kinase mutant that can be inhibited with a small molecule inhibitor, or WT mice will be analyzed for presence, percentage and number of CD4+ or NK1.1+ gamma delta T cell subsets. These cells will be identified in spleen, blood, lymph nodes, bone marrow, liver and lung, by flow cytometry using antibodies against the gamma delta T cell receptor, as well as other surface markers that identify subsets, including the expression of specific gamma delta T cell receptor. These experiments will determine if CD4+ or NK1.1+ gamma delta T cell development is affected by the absence or inhibition of ITK. Follow up experiments will sort these CD4+ or NK1.1+ gamma delta T cell subsets and analyze their cytokine production. <P>
3. Mice lacking ITK, those carrying a kinase mutant that can be inhibited with a small molecule inhibitor, or WT mice will be analyzed for presence, percentage and number of skin and mucosa homing gamma delta T cell subsets. These cells will be identified in skin, gut and reproductive tract, by flow cytometry using antibodies against the gamma delta T cell receptor, as well as other surface markers that identify subsets, including the expression of specific gamma delta T cell receptor. These experiments will determine if specific gamma delta T cell development or homing is affected by the absence or inhibition of ITK. Follow up experiments will sort these gamma delta T cell subsets and analyze their cytokine production.