Lung inflammation involving activated T-cells is a characteristic feature of asthma, sarcoidosis, hypersensitivity pneumonitis and chronic allograft rejection after lung transplantation. T-cell activation triggers intracellular phosphorylations that lead to nuclear translocation of transcription factors in the NF-kB/rel-homology family. In the nucleus of activated T-cells, global changes in chromatin structure allow access of transcription factors to their cognate binding sites. The IL-2 enhancer contains binding sites for NF-kB, AP-1, Oct-I and the purine-box regulator that binds to the antigen receptor response element/NF-AT target site. Purine-box regulator proteins of 45 kDa and 90 kDa were affinity-purified from the nucleus of activated Jurkat T-cells and partial amino acid sequence data were used to clone NP45 and NF9O cDNAs. NF45 and NF90 interact with the DNA-dependent protein kinase, Ku70 and Ku80. NF90 also interacts with dsRNA-activated protein kinase, PKR, and suppresses translation of specific mRNAs. Interleukin enhancer binding factor 3 (ILF3) is a longer form of NF90 that interacts with Protein Arginine Methyltransferase 1. NF45 and NF90 are autoantigens in murine lupus and human autoimmune diseases. Posttranslational modifications of NF45 and NF90/ILF3 during T-cell activation will be characterized and correlated with IL-2 expression. Triptolide is a diterpenoid triepoxide that inhibits NF-kB and IL-2 transcription and T-cell activation through mechanisms that do not involve calcineurin. Triptolide reacts covalently with several nuclear proteins. Immunosuppressive and antiproliferative mechanisms of triptolide will be investigated by characterizing targets of triptolide and triptolide inhibition of transcription in vitro. The developmental phenotypes and immune system functions of mice with targeted disruptions of NF45 and NF90/ILF3 will be determined. The murine NF45 and NF90/ILF3 genes have been sequenced in entirety, targeting vectors generated, and embryonic stem cells screened for homologous recombination. Appropriate ES clones will be injected into blastocysts at the Stanford Transgenic Core facility. Increased understanding of T-cell activation and its modulation by immunosuppressant drugs will guide future therapies for lung inflammation, autoimmune diseases, cancer and AIDS.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI039624-07
Application #
6510493
Study Section
Lung Biology and Pathology Study Section (LBPA)
Program Officer
Adams, Ken
Project Start
1996-04-01
Project End
2006-03-31
Budget Start
2002-04-01
Budget End
2003-03-31
Support Year
7
Fiscal Year
2002
Total Cost
$274,670
Indirect Cost
Name
Stanford University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
800771545
City
Stanford
State
CA
Country
United States
Zip Code
94305
Wu, Ting-Hsuan; Shi, Lingfang; Adrian, Jessika et al. (2018) NF90/ILF3 is a transcription factor that promotes proliferation over differentiation by hierarchical regulation in K562 erythroleukemia cells. PLoS One 13:e0193126
Kiesler, Patricia; Haynes, Paul A; Shi, Lingfang et al. (2010) NF45 and NF90 regulate HS4-dependent interleukin-13 transcription in T cells. J Biol Chem 285:8256-67
Shi, Lingfang; Godfrey, Wayne R; Lin, Joseph et al. (2007) NF90 regulates inducible IL-2 gene expression in T cells. J Exp Med 204:971-7
Shi, Lingfang; Qiu, Daoming; Zhao, Guohua et al. (2007) Dynamic binding of Ku80, Ku70 and NF90 to the IL-2 promoter in vivo in activated T-cells. Nucleic Acids Res 35:2302-10
Zhao, Guohua; Shi, Lingfang; Qiu, Daoming et al. (2005) NF45/ILF2 tissue expression, promoter analysis, and interleukin-2 transactivating function. Exp Cell Res 305:312-23
Shi, Lingfang; Zhao, Guohua; Qiu, Daoming et al. (2005) NF90 regulates cell cycle exit and terminal myogenic differentiation by direct binding to the 3'-untranslated region of MyoD and p21WAF1/CIP1 mRNAs. J Biol Chem 280:18981-9
Graham, Kareem L; Thibault, Donna L; Steinman, Jonathan B et al. (2005) Granzyme B is dispensable for immunologic tolerance to self in a murine model of systemic lupus erythematosus. Arthritis Rheum 52:1684-93
Vaszar, Laszlo T; Nishimura, Toshihiko; Storey, John D et al. (2004) Longitudinal transcriptional analysis of developing neointimal vascular occlusion and pulmonary hypertension in rats. Physiol Genomics 17:150-6
Nishimura, Toshihiko; Vaszar, Laszlo T; Faul, John L et al. (2003) Simvastatin rescues rats from fatal pulmonary hypertension by inducing apoptosis of neointimal smooth muscle cells. Circulation 108:1640-5
Qiu, Daoming; Kao, Peter N (2003) Immunosuppressive and anti-inflammatory mechanisms of triptolide, the principal active diterpenoid from the Chinese medicinal herb Tripterygium wilfordii Hook. f. Drugs R D 4:1-18

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