Natural Killer T (NKT) cells have been implicated in the regulation of immune responses associated with a broad range of diseases, including autoimmunity, allergy, infectious disease and cancer. The study of the role of NKT cells in disease relies upon two similar but not equivalent mouse models, both of which have NKT cell deficiencies. In one model, the CD1d-encoding genes have been deleted, thereby preventing the development of any CD1d-reactive T cells, including, but not limited to, NKT cells. Another model directly targeted the TRAJ18 gene segment, which in combination with TRAV11 gene is absolutely required to form the NKT T cell receptor (TCR) with the appropriate antigenic specificity. Highlighting the importance of such mouse model, TRAJ18-deificient mice (J?18-/-) have been cited in over 700 publications since their production in 1997. Using high-throughput sequencing, we recently made the observation that the J?18-/- mouse has a significantly reduced conventional T cell repertoire in addition to a deficiency in NKT cells. As such, the NKT cell field urgently requires a new mouse model to permit the uninterrupted study of NKT cell biology. We propose to generate a new mouse strain in which the J?18 gene segment will be specifically and uniquely modified to disrupt the generation of NKT cells without significantly altering the conventional T cell compartment using transcription activator-like effector nucleases (TALENs).

Public Health Relevance

Natural Killer T (NKT) cells are a type of immune cell that play an important role in our health. Numerous studies in humans and mice have suggested that if you do not have a normal population of NKT cells, you have an increased susceptibility to autoimmune diseases and cancers. In addition, NKT cells are known to play important roles during infection with bacterial, viral, protozoan, and fungal pathogens. NKT cells are very similar between mice and humans, and mouse models of NKT cell deficiencies are useful tools to study their role in disease. One of the primary mouse models to study NKT cells has been used heavily to test the role of NKT cells in a variety of diseases. As a result, the original paper describing the creation of these mice has been cited over 700 times. Recently, we discovered that these mice, in addition to lacking NKT cells, have a significant problem in another population of immune cells. This problem was not easy to find until scientists had the ability to perform DNA sequencing on an extensive scale. As such, any study using this mouse model to evaluate the biological role of NKT cells will need to be re-evaluated. Since this mouse model is in such high demand and disease studies coming from the use of these mice have tremendous potential, we propose to make a new mouse strain in which the NKT cells will be specifically and uniquely removed without affecting the immune cells, which are compromised in the original mouse model. To do this, we will use a cutting edge technology, permitting us to rapidly produce a replacement mouse model for NKT cell research.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Small Research Grants (R03)
Project #
5R03AI105468-02
Application #
8601043
Study Section
Cellular and Molecular Immunology - B Study Section (CMIB)
Program Officer
Miller, Lara R
Project Start
2012-12-26
Project End
2014-11-30
Budget Start
2013-12-01
Budget End
2014-11-30
Support Year
2
Fiscal Year
2014
Total Cost
$39,625
Indirect Cost
$14,625
Name
National Jewish Health
Department
Type
DUNS #
076443019
City
Denver
State
CO
Country
United States
Zip Code
80206