It has become increasingly clear that the workings of the thymus include greater complexity than first appreciated. Not only is this organ a generator of CD4 and CD8 T cells, but it is also a site for development of highly specialized lineages of natural killer T (NKT) cells, intraepithelial T lymphocytes, regulatory T (Treg) cells, and innate-like CD8 T cells. The detailed molecular mechanisms that control the differentiation of these distinct T cell sublineages from common precursors remain to be determined. Here we focus on the role of TOX (thymocyte selection-associated HMG box protein) in this context, a nuclear DNA-binding protein first identified in our laboratory as a regulator of thymic selection events. We produced TOX-deficient mice, demonstrating that TOX is required for development of the canonical CD4 T cell lineage as well as NKT and Treg cell development. Inhibition of the specific stage of T cell development that is affected in these mutant animals has not been reported previously, allowing novel insights into the process. In addition, we have found that TOX plays roles outside the thymus, including an obligatory role in NK cell development and lymph node organogenesis. We propose here three specific aims to delineate the role and mechanism of action of TOX in development of the immune system. First, we will investigate the relationship between TOX and regulation of CD4 and ThPOK expression in the thymus, the latter a dominant CD4 lineage commitment factor. In addition, we propose a novel approach to identify TOX-dependent genes involved in establishment of the CD4 lineage gene program, and will compare the role of TOX in development of different T cell lineages. Second, we will analyze the role of TOX in development of lymph node organogenesis, focusing on the potential for TOX to regulate lymphoid tissue inducer cell development. We also have replicated the inhibition of NK cell development in the absence of TOX in a culture system. We propose detailed analysis of the role for TOX in this context, including whether TOX might function to regulate another nuclear factor that shares some functional properties with TOX. Third, we propose state-of-the-art approaches to identify direct gene targets and protein binding partners of TOX, in an in vivo context of CD4 T cell development. TOX is highly conserved between rodents and humans, as is its expression pattern. Thus, these studies should provide insight into workings of the human immune system as well. TOX defines a small protein subfamily that includes three other members, one of which has been implicated as a breast cancer susceptibility locus, while TOX itself is highly expressed in some cancer cell lines and tumors. Understanding the mechanism of action of TOX is therefore likely to impact both human health and disease.

Public Health Relevance

Development of T lymphocytes in the thymus is a complex process, but one that is key to creating a protective cellular arm of the immune system. This research program is directed at understanding the function of a small family of nuclear proteins, highly conserved in rodents and humans, in regulating this process. This knowledge will aid in future development of methods to reconstitute the immune system where necessary due to disease, and also may have future impact in treatment for breast cancer, since a gene encoding one member of this protein family has been mapped as a disease susceptibility locus.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI054977-08
Application #
8223181
Study Section
Cellular and Molecular Immunology - B Study Section (CMIB)
Program Officer
Prabhudas, Mercy R
Project Start
2003-04-01
Project End
2015-02-28
Budget Start
2012-03-01
Budget End
2013-02-28
Support Year
8
Fiscal Year
2012
Total Cost
$408,375
Indirect Cost
$160,875
Name
Cedars-Sinai Medical Center
Department
Type
DUNS #
075307785
City
Los Angeles
State
CA
Country
United States
Zip Code
90048
Aliahmad, Parinaz; Seksenyan, Akop; Kaye, Jonathan (2012) The many roles of TOX in the immune system. Curr Opin Immunol 24:173-7
Aliahmad, Parinaz; Kadavallore, Asha; de la Torre, Brian et al. (2011) TOX is required for development of the CD4 T cell lineage gene program. J Immunol 187:5931-40
Aliahmad, Parinaz; de la Torre, Brian; Kaye, Jonathan (2010) Shared dependence on the DNA-binding factor TOX for the development of lymphoid tissue-inducer cell and NK cell lineages. Nat Immunol 11:945-52
Aliahmad, Parinaz; Kaye, Jonathan (2008) Development of all CD4 T lineages requires nuclear factor TOX. J Exp Med 205:245-56
Aliahmad, Parinaz; O'Flaherty, Emmett; Han, Peggy et al. (2004) TOX provides a link between calcineurin activation and CD8 lineage commitment. J Exp Med 199:1089-99