60 results on '"Gordon S. Duncan"'
Search Results
2. Cholinergic control of Th17 cell pathogenicity in experimental autoimmune encephalomyelitis
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Robert Nechanitzky, Duygu Nechanitzky, Parameswaran Ramachandran, Gordon S. Duncan, Chunxing Zheng, Christoph Göbl, Kyle T. Gill, Jillian Haight, Andrew C. Wakeham, Bryan E. Snow, Vivian Bradaschia-Correa, Milan Ganguly, Zhibin Lu, Mary E. Saunders, Richard A. Flavell, and Tak W. Mak
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Cell Biology ,Molecular Biology - Published
- 2022
3. IDH2 and TET2 mutations synergize to modulate T Follicular Helper cell functional interaction with the AITL microenvironment
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Julie Leca, Franҫois Lemonnier, Cem Meydan, Jonathan Foox, Samah El Ghamrasni, Diana-Laure Mboumba, Gordon S. Duncan, Jerome Fortin, Takashi Sakamoto, Chantal Tobin, Kelsey Hodgson, Jillian Haight, Logan K. Smith, Andrew J. Elia, Daniel Butler, Thorsten Berger, Laurence de Leval, Christopher E. Mason, Ari Melnick, Philippe Gaulard, and Tak W. Mak
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Cancer Research ,Oncology ,Animals ,Humans ,Mice ,Dioxygenases/genetics ,DNA-Binding Proteins/genetics ,Immunoblastic Lymphadenopathy/genetics ,Isocitrate Dehydrogenase/genetics ,Lymphoma, T-Cell/genetics ,Mutation ,T Follicular Helper Cells/pathology ,T-Lymphocytes, Helper-Inducer ,Tumor Microenvironment/genetics ,Angioimmunoblastic T cell lymphoma ,Idh2 ,T follicular helper cells ,Tet2 ,cytokines ,epigenetics ,germinal center B cells ,preclinical mouse model ,therapeutic agents ,tumor microenvironment - Abstract
Angioimmunoblastic T cell lymphoma (AITL) is a peripheral T cell lymphoma that originates from T follicular helper (Tfh) cells and exhibits a prominent tumor microenvironment (TME). IDH2 and TET2 mutations co-occur frequently in AITL, but their contribution to tumorigenesis is poorly understood. We developed an AITL mouse model that is driven by Idh2 and Tet2 mutations. Malignant Tfh cells display aberrant transcriptomic and epigenetic programs that impair TCR signaling. Neoplastic Tfh cells bearing combined Idh2 and Tet2 mutations show altered cross-talk with germinal center B cells that promotes B cell clonal expansion while decreasing Fas-FasL interaction and reducing B cell apoptosis. The plasma cell count and angiogenesis are also increased in the Idh2-mutated tumors, implying a major relationship between Idh2 mutation and the characteristic AITL TME. Our mouse model recapitulates several features of human IDH2-mutated AITL and provides a rationale for exploring therapeutic targeting of Tfh-TME cross-talk for AITL patients.
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- 2023
4. Glutathione Restricts Serine Metabolism to Preserve Regulatory T Cell Function
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Melanie Grusdat, Ying Chen, Dennis Das Gupta, Tak W. Mak, Rashi Halder, Sophie Farinelle, Christian Jäger, Johannes Meiser, Luana Guerra, Lynn Bonetti, Gordon S. Duncan, Anouk Ewen, Lisa Schlicker, Paul Wilmes, Karsten Hiller, Jillian Haight, Myriam P. Merz, Yannic Nonnenmacher, Michael Lohoff, Rabia Taskesen, Vasilis Vasiliou, Markus Ollert, Davide G. Franchina, Carole Binsfeld, Isaac S. Harris, Christiane B. Knobbe-Thomsen, Oliver Hunewald, Henry Kurniawan, Dirk Brenner, Jonathan D. Turner, Leticia Soriano Baguet, Catherine Dostert, and HZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany.
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0301 basic medicine ,one carbon metabolism ,Physiology ,Regulatory T cell ,chemical and pharmacologic phenomena ,T-Lymphocytes, Regulatory ,Article ,Serine ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,FoxP3 ,medicine ,cancer ,Animals ,Serine import ,glutathione ,Molecular Biology ,PI3K/AKT/mTOR pathway ,glutamate cysteine ligase ,serine metabolism ,Effector ,autoimmunity ,FOXP3 ,ROS ,hemic and immune systems ,Cell Biology ,Glutathione ,Cell biology ,Treg ,030104 developmental biology ,medicine.anatomical_structure ,GCLC ,chemistry ,diet ,030217 neurology & neurosurgery - Abstract
Regulatory T cells (Tregs) maintain immune homeostasis and prevent autoimmunity. Serine stimulates glutathione (GSH) synthesis and feeds into the one-carbon metabolic network (1CMet) essential for effector T cell (Teff) responses. However, serine's functions, linkage to GSH, and role in stress responses in Tregs are unknown. Here, we show, using mice with Treg-specific ablation of the catalytic subunit of glutamate cysteine ligase (Gclc), that GSH loss in Tregs alters serine import and synthesis and that the integrity of this feedback loop is critical for Treg suppressive capacity. Although Gclc ablation does not impair Treg differentiation, mutant mice exhibit severe autoimmunity and enhanced anti-tumor responses. Gclc-deficient Tregs show increased serine metabolism, mTOR activation, and proliferation but downregulated FoxP3. Limitation of cellular serine in vitro and in vivo restores FoxP3 expression and suppressive capacity of Gclc-deficient Tregs. Our work reveals an unexpected role for GSH in restricting serine availability to preserve Treg functionality. Regulatory T cells (Tregs) rely on oxidative metabolism, which triggers the generation of reactive oxygen species (ROS). Accumulating ROS are controlled by the antioxidant glutathione (GSH). Kurniawan et al. reveal an unexpected subset-specific role of GSH in serine metabolism and Treg function.
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- 2020
5. Ibrutinib Therapy Downregulates Toso, the Fcr for IgM, Expression in CLL Patients
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Mariela Sivina, William G. Wierda, Andreas Rosenwald, Michael J. Keating, Ekaterina Kim, Elena Hartmann, Alessandra Ferrajoli, Alicia Vaca, Jan A. Burger, Gordon S. Duncan, Takashi Sakamoto, and Tak W. Mak
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Immunoglobulin gene ,biology ,business.industry ,Immunology ,B-cell receptor ,breakpoint cluster region ,Cell Biology ,Hematology ,Biochemistry ,chemistry.chemical_compound ,medicine.anatomical_structure ,chemistry ,Immunoglobulin M ,Ibrutinib ,biology.protein ,Cancer research ,Medicine ,Rituximab ,business ,IGHV@ ,B cell ,medicine.drug - Abstract
Background: Toso, the FcR receptor (FcmR) for IgM, is a type I transmembrane protein belonging to the immunoglobulin gene superfamily. Toso is expressed in lymphocytes, and plays an important role in B cell development and survival. In normal B cells, Toso expression increases after B cell receptor (BCR) stimulation, and Toso enhances BCR signaling induced cell survival, NK-kB pathway activation, and BCL-xL expression. CLL B cells express high levels of Toso, especially in IgHV unmutated patients. Upon binding of IgM to Toso on CLL cells, the Toso-IgM complex is internalized and undergoes lysosomal degradation. Given the importance of BCR signaling in CLL pathogenesis and treatment, we performed a series of preclinical and correlative studies to examine the cross talk between Toso and BCR signaling, effects of Toso on CLL cell survival and how Toso is affected by BCR signaling inhibition with ibrutinib. Methods and Results: Toso gene expression (FAIM3) was analyzed in serial samples from eight CLL patients treated with ibrutinib plus rituximab using Affymetrics HG U133 plus 2.0 oligonucleotide arrays at baseline, and after 1 and 4 weeks of continuous ibrutinib-based therapy. We noted that the relative mean Toso gene expression declined during ibrutinib therapy (-0.26 ± 0.12 after 2 weeks and -0.3 ± 0.18 after 4 weeks). Accordingly, Toso surface expression on CLL cells, assessed by flow cytometry, also significantly decreased after 1 and 4 weeks of ibrutinib therapy (Figure 1). We next measured plasma levels of soluble Toso (sToso), which is encoded by an alternative spliced Toso transcript, in serial samples from 35 CLL ibrutinib treated patients using an enzyme-linked immunosorbent assay (ELISA). We noted that baseline sToso levels in CLL patients were significantly higher than in 6 normal controls, i.e. 70 ± 12.1 arbitrary units (AU, n=35) versus 18 ± 8 AU (n=6, p Conclusions: These studies demonstrate that ibrutinib therapy downregulates Toso (FAIM3) gene expression, as well as cell surface and soluble Toso expression. IgHV unmutated patients have higher levels of soluble Toso and the effect on Toso activation on viability and CCL3 and CCL4 secretion is more pronounced in this group of patients. These findings corroborate the close relation between BCR signaling and Toso in CLL. Disclosures Wierda: Cyclcel: Research Funding; Miragen: Research Funding; Oncternal Therapeutics Inc.: Research Funding; Loxo Oncology Inc.: Research Funding; Janssen: Research Funding; Xencor: Research Funding; Acerta Pharma Inc: Research Funding; Pharmacyclics LLC: Research Funding; Genentech: Research Funding; AbbVie: Research Funding; GSK/Novartis: Research Funding; Gilead Sciences: Research Funding; Juno Therapeutics: Research Funding; KITE pharma: Research Funding; Sunesis: Research Funding. Burger:Janssen Pharmaceuticals: Consultancy, Honoraria; Aptose Biosciences, Inc: Research Funding; Gilead Sciences: Research Funding; Pharmacyclics, an AbbVie company: Research Funding; AstraZeneca: Honoraria; BeiGene: Research Funding.
- Published
- 2019
6. Idh1 protects murine hepatocytes from endotoxin-induced oxidative stress by regulating the intracellular NADP+/NADPH ratio
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Dave Cescon, Dirk Brenner, Evan F. Lind, Annick You-Ten, Andrew J. Elia, Momoe Itsumi, Philipp A. Lang, Masato Sasaki, Andrew Wakeham, Isaac S. Harris, Alisha R. Elford, Gordon S. Duncan, Pamela S. Ohashi, Tak W. Mak, Rob A. Cairns, Wanda Y. Li, Satoshi Inoue, Iok In Christine Chio, Jillian Haight, Kiichi Murakami, J Ye, Samia Afzal, Kazuo Yamamoto, and Bryan E. Snow
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IDH1 ,Lipopolysaccharide ,Biology ,medicine.disease_cause ,Proinflammatory cytokine ,Mice ,chemistry.chemical_compound ,medicine ,Animals ,Molecular Biology ,Cells, Cultured ,Mice, Knockout ,chemistry.chemical_classification ,Original Paper ,Reactive oxygen species ,Reverse Transcriptase Polymerase Chain Reaction ,Cell Biology ,Flow Cytometry ,Molecular biology ,Isocitrate Dehydrogenase ,Endotoxins ,Oxidative Stress ,Isocitrate dehydrogenase ,chemistry ,Biochemistry ,Hepatocytes ,Tumor necrosis factor alpha ,NADP ,Oxidative stress ,Intracellular - Abstract
Isocitrate dehydrogenase-1 (Idh1) is an important metabolic enzyme that produces NADPH by converting isocitrate to α-ketoglutarate. Idh1 is known to reduce reactive oxygen species (ROS) induced in cells by treatment with lipopolysaccharide (LPS) in vitro. Here, we used Idh1-deficient knockout (Idh1 KO) mice to investigate the role of Idh1 in antioxidant defense in vivo. Idh1 KO mice showed heightened susceptibility to death induced by LPS and exhibited increased serum levels of inflammatory cytokines such as tumor necrosis factor-α and interleukin-6. The serum of LPS-injected Idh1 KO mice also contained elevated levels of AST, a marker of inflammatory liver damage. Furthermore, after LPS injection, livers of Idh1 KO mice showed histological evidence of elevated oxidative DNA damage compared with livers of wild-type (WT) mice. Idh1 KO livers showed a faster and more pronounced oxidative stress than WT livers. In line with that, Idh1 KO hepatocytes showed higher ROS levels and an increase in the NADP(+)/NADPH ratio when compared with hepatocytes isolated from WT mice. These results suggest that Idh1 has a physiological function in protecting cells from oxidative stress by regulating the intracellular NADP(+)/NADPH ratio. Our findings suggest that stimulation of Idh1 activity may be an effective therapeutic strategy for reducing oxidative stress during inflammatory responses, including the early stages of septic shock.
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- 2015
7. K48-linked KLF4 ubiquitination by E3 ligase Mule controls T-cell proliferation and cell cycle progression
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Tak W. Mak, Zhenyue Hao, Sara R. Hamilton, Juan Carlos Zúñiga-Pflücker, Jillian Haight, Satoshi Inoue, Wanda Y. Li, Gloria H. Y. Lin, Gordon S. Duncan, Yi Sheng, Dirk Brenner, Alisha R. Elford, Yi Liang, Yu-Wen Su, Jennifer Sylvester, Annick You-Ten, Pamela S. Ohashi, Housheng Hansen He, Andrew Wakeham, Bryan E. Snow, and Carmen Dominguez
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Male ,0301 basic medicine ,Science ,T-Lymphocytes ,Ubiquitin-Protein Ligases ,T cell ,Amino Acid Motifs ,Kruppel-Like Transcription Factors ,General Physics and Astronomy ,Lymphocytic Choriomeningitis ,Article ,General Biochemistry, Genetics and Molecular Biology ,Kruppel-Like Factor 4 ,Mice ,03 medical and health sciences ,Downregulation and upregulation ,medicine ,Animals ,Humans ,Lymphocytic choriomeningitis virus ,B cell ,Cell Proliferation ,Mice, Knockout ,Multidisciplinary ,biology ,Cell growth ,Tumor Suppressor Proteins ,T-cell receptor ,Ubiquitination ,General Chemistry ,3. Good health ,Cell biology ,Ubiquitin ligase ,Mice, Inbred C57BL ,030104 developmental biology ,medicine.anatomical_structure ,KLF4 ,biology.protein ,Female ,CD8 - Abstract
T-cell proliferation is regulated by ubiquitination but the underlying molecular mechanism remains obscure. Here we report that Lys-48-linked ubiquitination of the transcription factor KLF4 mediated by the E3 ligase Mule promotes T-cell entry into S phase. Mule is elevated in T cells upon TCR engagement, and Mule deficiency in T cells blocks proliferation because KLF4 accumulates and drives upregulation of its transcriptional targets E2F2 and the cyclin-dependent kinase inhibitors p21 and p27. T-cell-specific Mule knockout (TMKO) mice develop exacerbated experimental autoimmune encephalomyelitis (EAE), show impaired generation of antigen-specific CD8+ T cells with reduced cytokine production, and fail to clear LCMV infections. Thus, Mule-mediated ubiquitination of the novel substrate KLF4 regulates T-cell proliferation, autoimmunity and antiviral immune responses in vivo., The E3 ligase Mule has been previously reported to be essential for B cell development and function by modulating p53 ubiquitination and degradation. Here Hao et al. identify KLF4 as a novel ubiquitination target of Mule and show it controls T cell proliferation and autoimmunity.
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- 2017
8. 2-Methoxyestradiol inhibits experimental autoimmune encephalomyelitis through suppression of immune cell activation
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Anne Brüstle, Thomas Mock, Michael W. Tusche, Andrew J. Elia, Dirk Brenner, Gordon S. Duncan, Christiane B. Knobbe, Peter H. Krammer, Mark R. Bray, and Tak W. Mak
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CD4-Positive T-Lymphocytes ,Encephalomyelitis, Autoimmune, Experimental ,Multiple Sclerosis ,Encephalomyelitis ,medicine.medical_treatment ,Autoimmunity ,Biology ,Lymphocyte Activation ,Mice ,NFAT Pathway ,medicine ,Animals ,Humans ,2-Methoxyestradiol ,Lymphocytes ,Multidisciplinary ,Estradiol ,NFATC Transcription Factors ,Experimental autoimmune encephalomyelitis ,NF-kappa B ,NFAT ,Biological Sciences ,medicine.disease ,NFKB1 ,Tubulin Modulators ,Mice, Inbred C57BL ,Transcription Factor AP-1 ,Cytokine ,Immunology ,Cancer research ,Cytokines ,Interleukin 17 ,Signal Transduction ,medicine.drug - Abstract
The endogenous metabolite of estradiol, 2-Methoxyestradiol (2ME2), is an antimitotic and antiangiogenic cancer drug candidate that also exhibits disease-modifying activity in animal models of rheumatoid arthritis (RA). We found that 2ME2 dramatically suppresses development of mouse experimental autoimmune encephalomyelitis (EAE), a rodent model of multiple sclerosis (MS). 2ME2 inhibits in vitro lymphocyte activation, cytokine production, and proliferation in a dose-dependent fashion. 2ME2 treatment of lymphocytes specifically reduced the nuclear translocation and transcriptional activity of nuclear factor of activated T-cells (NFAT) c1, whereas NF-κB and activator protein 1 (AP-1) activation were not adversely affected. We therefore propose that 2ME2 attenuates EAE through disruption of the NFAT pathway and subsequent lymphocyte activation. By extension, our findings provide a molecular rationale for the use of 2ME2 as a tolerable oral immunomodulatory agent for the treatment of autoimmune disorders such as MS in humans.
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- 2012
9. ARIH2 is essential for embryogenesis, and its hematopoietic deficiency causes lethal activation of the immune system
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Simon Preston, Yongkai Ow, Jesse G. Toe, Amy E. Lin, Hamish W Scott, Andrew Wakeham, Tak W. Mak, James P Cooney, Gregor Ebert, Jennifer Silvester, Raymond H. Kim, Samuel D. Saibil, Pamela S. Ohashi, Annick You-Ten, Arda Shahinian, Marc Pellegrini, Masato Sasaki, Dilan Dissanayake, and Gordon S. Duncan
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Transcriptional Activation ,Multiple Organ Failure ,Ubiquitin-Protein Ligases ,Immunology ,Embryonic Development ,Mice ,Immune system ,Ubiquitin ,Transcription (biology) ,Animals ,Humans ,Immunology and Allergy ,Molecular Targeted Therapy ,Transcription factor ,Cells, Cultured ,Mice, Knockout ,biology ,NF-kappa B ,Ubiquitination ,Dendritic Cells ,NFKB1 ,Hematopoiesis ,Cell biology ,Ubiquitin ligase ,Mice, Inbred C57BL ,Disease Models, Animal ,Haematopoiesis ,Immune System ,biology.protein ,Stem cell - Abstract
The E3 ligase ARIH2 has an unusual structure and mechanism of elongating ubiquitin chains. To understand its physiological role, we generated gene-targeted mice deficient in ARIH2. ARIH2 deficiency resulted in the embryonic death of C57BL/6 mice. On a mixed genetic background, the lethality was attenuated, with some mice surviving beyond weaning and then succumbing to an aggressive multiorgan inflammatory response. We found that in dendritic cells (DCs), ARIH2 caused degradation of the inhibitor IκBβ in the nucleus, which abrogated its ability to sequester, protect and transcriptionally coactivate the transcription factor subunit p65 in the nucleus. Loss of ARIH2 caused dysregulated activation of the transcription factor NF-κB in DCs, which led to lethal activation of the immune system in ARIH2-sufficent mice reconstituted with ARIH2-deficient hematopoietic stem cells. Our data have therapeutic implications for targeting ARIH2 function.
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- 2012
10. Glutathione Primes T Cell Metabolism for Inflammation
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Karsten Hiller, Anne Brüstle, Momoe Itsumi, Vasilis Vasiliou, Isaac S. Harris, Dirk Brenner, Tak W. Mak, Christian Jäger, Carsten Bindslev-Jensen, Maureen A. Cox, Catherine Dostert, Carole Binsfeld, Ying Chen, Philipp A. Lang, Michael Lohoff, Bärbel Camara, Yannic Nonnenmacher, Olaf Pinkenburg, Chiara Gorrini, Zhenyue Hao, Melanie Grusdat, Markus Ollert, and Gordon S. Duncan
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0301 basic medicine ,Glutamine ,T-Lymphocytes ,Myc ,chemistry.chemical_compound ,0302 clinical medicine ,Immunology and Allergy ,metabolic reprogramming ,glutathione ,reactive oxygen species ,Mice, Knockout ,0303 health sciences ,TOR Serine-Threonine Kinases ,NFAT ,ROS ,glycolysis ,Glutathione ,Cell biology ,GCLC ,Infectious Diseases ,Biochemistry ,030220 oncology & carcinogenesis ,mTOR ,Signal transduction ,Glycolysis ,Signal Transduction ,Encephalomyelitis, Autoimmune, Experimental ,Glutamate-Cysteine Ligase ,Immunology ,Immunoblotting ,T cells ,Biology ,Gclc ,Proto-Oncogene Proteins c-myc ,03 medical and health sciences ,GSH ,Animals ,Transcription factor ,PI3K/AKT/mTOR pathway ,030304 developmental biology ,Inflammation ,Glutaminolysis ,NFATC Transcription Factors ,Mice, Inbred C57BL ,030104 developmental biology ,chemistry ,Energy Metabolism ,Reactive Oxygen Species ,metabolism - Abstract
Activated T cells produce reactive oxygen species (ROS), which trigger the antioxidative glutathione (GSH) response necessary to buffer rising ROS and prevent cellular damage. We report that GSH is essential for T cell effector functions through its regulation of metabolic activity. Conditional gene targeting of the catalytic subunit of glutamate cysteine ligase (Gclc) blocked GSH production specifically in murine T cells. Gclc-deficient T cells initially underwent normal activation but could not meet their increased energy and biosynthetic requirements. GSH deficiency compromised the activation of mammalian target of rapamycin-1 (mTOR) and expression of NFAT and Myc transcription factors, abrogating the energy utilization and Myc-dependent metabolic reprogramming that allows activated T cells to switch to glycolysis and glutaminolysis. In vivo, T-cell-specific ablation of murine Gclc prevented autoimmune disease but blocked antiviral defense. The antioxidative GSH pathway thus plays an unexpected role in metabolic integration and reprogramming during inflammatory T cell responses.
- Published
- 2016
11. Characterization of dsRNA-induced pancreatitis model reveals the regulatory role of IFN regulatory factor 2 ( Irf2 ) in trypsinogen5 gene transcription
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Koon Jiew Chua, Toshifumi Matsuyama, Ichiro Katayama, Tomoki Nakashima, Hideki Hayashi, Tohru Kimura, Yoshinao Kubo, Shizuo Akira, Tomoko Kohno, Kiyoshi Yasui, Tak W. Mak, Yuhua Ma, Hiroyuki Murota, Gordon S. Duncan, Takashi Suematsu, Kazuo Yamamoto, and Isao Shimokawa
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Transcription, Genetic ,Mice, Transgenic ,Acinar Cells ,Biology ,Cathepsin B ,Ca2+-binding proteins ,Gene product ,Mice ,Animals ,Humans ,TRIF ,Gene ,Glycoproteins ,RNA, Double-Stranded ,IPS-1 ,Messenger RNA ,Multidisciplinary ,Prostatic Secretory Proteins ,MDA5 ,Biological Sciences ,Molecular biology ,HEK293 Cells ,Poly I-C ,Pancreatitis ,Trypsin Inhibitor, Kazal Pancreatic ,TLR3 ,Trypsinogen ,Trypsin Inhibitors ,IRF2 ,Interferon Regulatory Factor-2 ,HeLa Cells ,Interferon regulatory factors - Abstract
Mice deficient for interferon regulatory factor (Irf)2 (Irf2(-/-) mice) exhibit immunological abnormalities and cannot survive lymphocytic choriomeningitis virus infection. The pancreas of these animals is highly inflamed, a phenotype replicated by treatment with poly(I:C), a synthetic double-stranded RNA. Trypsinogen5 mRNA was constitutively up-regulated about 1,000-fold in Irf2(-/-) mice compared with controls as assessed by quantitative RT-PCR. Further knockout of IFNα/β receptor 1(Ifnar1) abolished poly(I:C)-induced pancreatitis but had no effect on the constitutive up-regulation of trypsinogen5 gene, indicating crucial type I IFN signaling to elicit the inflammation. Analysis of Ifnar1(-/-) mice confirmed type I IFN-dependent transcriptional activation of dsRNA-sensing pattern recognition receptor genes MDA5, RIG-I, and TLR3, which induced poly(I:C)-dependent cell death in acinar cells in the absence of IRF2. We speculate that Trypsin5, the trypsinogen5 gene product, leaking from dead acinar cells triggers a chain reaction leading to lethal pancreatitis in Irf2(-/-) mice because it is resistant to a major endogenous trypsin inhibitor, Spink3., This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1116273108/-/DCSupplemental., Proceedings of the National Academy of Sciences of the United States of America, 108(46), pp.18766-18771; 2011
- Published
- 2011
12. Nfil3/E4bp4 is required for the development and maturation of NK cells in vivo
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Koichi Hamada, Shintaro Kamizono, Pamela S. Ohashi, Koichi Akashi, Gerard Grosveld, Gordon S. Duncan, Tak W. Mak, Akira Morimoto, Evan F. Lind, Jillian Haight, A. Thomas Look, and Markus G. Seidel
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Immunology ,Mice ,03 medical and health sciences ,Interleukin 21 ,0302 clinical medicine ,MHC class I ,Animals ,Immunology and Allergy ,Cell Lineage ,030304 developmental biology ,Mice, Knockout ,0303 health sciences ,Lymphokine-activated killer cell ,biology ,Cell growth ,Janus kinase 3 ,NFIL3 ,Brief Definitive Report ,Natural killer T cell ,Cell biology ,Killer Cells, Natural ,Mice, Inbred C57BL ,Basic-Leucine Zipper Transcription Factors ,biology.protein ,Interleukin 12 ,030215 immunology - Abstract
Nuclear factor interleukin-3 (Nfil3; also known as E4-binding protein 4) is a basic region leucine zipper transcription factor that has antiapoptotic activity in vitro under conditions of growth factor withdrawal. To study the role of Nfil3 in vivo, we generated gene-targeted Nfil3-deficient (Nfil3−/−) mice. Nfil3−/− mice were born at normal Mendelian frequency and were grossly normal and fertile. Although numbers of T cells, B cells, and natural killer (NK) T cells were normal in Nfil3−/− mice, a specific disruption in NK cell development resulted in severely reduced numbers of mature NK cells in the periphery. This defect was NK cell intrinsic in nature, leading to a failure to reject MHC class I–deficient cells in vivo and reductions in both interferon γ production and cytolytic activity in vitro. Our results confirm the specific and essential requirement of Nfil3 for the development of cells of the NK lineage.
- Published
- 2009
13. Fas Receptor Expression in Germinal-Center B Cells Is Essential for T and B Lymphocyte Homeostasis
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Claire Hong, Jane Seagal, Stefano Casola, Klaus Rajewsky, Zhenyue Hao, Geoffrey A. Wood, Yu-Wen Su, Jennifer Liepa, Jillian Haight, Tak W. Mak, Gordon S. Duncan, Andrew E. H. Elia, Nien Jung Chen, Andrew Wakeham, and Wanda Y. Li
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T-Lymphocytes ,Immunology ,Naive B cell ,B-cell receptor ,Somatic hypermutation ,Cell Communication ,Biology ,Lymphocyte Activation ,Article ,Mice ,03 medical and health sciences ,0302 clinical medicine ,CD28 Antigens ,Antigens, CD ,medicine ,Animals ,Homeostasis ,Immunology and Allergy ,CTLA-4 Antigen ,fas Receptor ,CD40 Antigens ,Memory B cell ,B cell ,Cell Proliferation ,030304 developmental biology ,Mice, Knockout ,B-Lymphocytes ,0303 health sciences ,Germinal center ,Cell Differentiation ,Germinal Center ,Molecular biology ,Mice, Mutant Strains ,Mice, Inbred C57BL ,B-1 cell ,Infectious Diseases ,medicine.anatomical_structure ,CELLIMMUNO ,B7-1 Antigen ,Cytokines ,CD80 ,030215 immunology - Abstract
SummaryFas is highly expressed in activated and germinal center (GC) B cells but can potentially be inactivated by misguided somatic hypermutation. We employed conditional Fas-deficient mice to investigate the physiological functions of Fas in various B cell subsets. B cell-specific Fas-deficient mice developed fatal lymphoproliferation due to activation of B cells and T cells. Ablation of Fas specifically in GC B cells reproduced the phenotype, indicating that the lymphoproliferation initiates in the GC environment. B cell-specific Fas-deficient mice also showed an accumulation of IgG1+ memory B cells expressing high amounts of CD80 and the expansion of CD28-expressing CD4+ Th cells. Blocking T cell-B cell interaction and GC formation completely prevented the fatal lymphoproliferation. Thus, Fas-mediated selection of GC B cells and the resulting memory B cell compartment is essential for maintaining the homeostasis of both T and B lymphocytes.
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- 2008
14. Beyond tumor necrosis factor receptor: TRADD signaling in toll-like receptors
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Juan Carlos Zúñiga-Pflücker, Huey-Lan Huang, Gordon S. Duncan, Tak W. Mak, Yu-Wen Su, Zhenyue Hao, Hien Chau, Kelly A. Pike, Kazuo Yamamoto, Renée F. de Pooter, Wen-Jye Lin, Iok In Christine Chio, Wen Chen Yeh, Nien Jung Chen, David F. Katz, and Andrew Wakeham
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T-Lymphocytes ,Apoptosis ,Mice ,Animals ,Death domain ,Inflammation ,Mice, Knockout ,Toll-like receptor ,Multidisciplinary ,Tumor Necrosis Factor-alpha ,Chemistry ,NF-kappa B ,Biological Sciences ,Germinal Center ,TRADD ,TNF Receptor-Associated Death Domain Protein ,Toll-Like Receptor 3 ,Cell biology ,Toll-Like Receptor 4 ,TLR3 ,TLR4 ,Cancer research ,Tumor necrosis factor alpha ,Tumor necrosis factor receptor 1 ,Signal transduction ,Signal Transduction - Abstract
Tumor necrosis factor receptor 1-associated death domain protein (TRADD) is the core adaptor recruited to TNF receptor 1 (TNFR1) upon TNFα stimulation. In cells from TRADD-deficient mice, TNFα-mediated apoptosis and TNFα-stimulated NF-κB, JNK, and ERK activation are defective. TRADD is also important for germinal center formation, DR3-mediated costimulation of T cells, and TNFα-mediated inflammatory responses in vivo . TRADD deficiency does not enhance IFNγ-induced signaling. Importantly, TRADD has a novel role in TLR3 and TLR4 signaling. TRADD participates in the TLR4 complex formed upon LPS stimulation, and TRADD-deficient macrophages show impaired cytokine production in response to TLR ligands in vitro . Thus, TRADD is a multifunctional protein crucial both for TNFR1 signaling and other signaling pathways relevant to immune responses.
- Published
- 2008
15. Generation and Characterization of B7-H4/B7S1/B7x-Deficient Mice
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Tina D. Walker, Manel Jordana, Elissa K. Deenick, Andrew Wakeham, Gordon S. Duncan, Yoshiyuki Miyazaki, Seng Wang, Beata U. Gajewska, Pamela S. Ohashi, Annick Itie, Hitoshi Okada, Woong-Kyung Suh, Elizabeth C. Cates, Tak W. Mak, Hiroki Yoshida, Tania H. Watts, and Wojciech Dawicki
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medicine.medical_treatment ,Inflammation ,Biology ,Autoimmune Diseases ,Mice ,Immune system ,In vivo ,medicine ,Animals ,Lymphocytic choriomeningitis virus ,Cytotoxic T cell ,IL-2 receptor ,Receptor ,Molecular Biology ,Cell Proliferation ,Leishmania major ,Mice, Knockout ,Articles ,Cell Biology ,Th1 Cells ,V-Set Domain-Containing T-Cell Activation Inhibitor 1 ,In vitro ,Cell biology ,Mice, Inbred C57BL ,Cytokine ,Influenza A virus ,Gene Targeting ,Immunology ,B7-1 Antigen ,medicine.symptom ,T-Lymphocytes, Cytotoxic - Abstract
Members of the B7 family of cosignaling molecules regulate T-cell proliferation and effector functions by engaging cognate receptors on T cells. In vitro and in vivo blockade experiments indicated that B7-H4 (also known as B7S1 or B7x) inhibits proliferation, cytokine production, and cytotoxicity of T cells. B7-H4 binds to an unknown receptor(s) that is expressed on activated T cells. However, whether B7-H4 plays nonredundant immune regulatory roles in vivo has not been tested. We generated B7-H4-deficient mice to investigate the roles of B7-H4 during various immune reactions. Consistent with its inhibitory function in vitro, B7-H4-deficient mice mounted mildly augmented T-helper 1 (Th1) responses and displayed slightly lowered parasite burdens upon Leishmania major infection compared to the wild-type mice. However, the lack of B7-H4 did not affect hypersensitive inflammatory responses in the airway or skin that are induced by either Th1 or Th2 cells. Likewise, B7-H4-deficient mice developed normal cytotoxic T-lymphocyte reactions against viral infection. Thus, B7-H4 plays a negative regulatory role in vivo but the impact of B7-H4 deficiency is minimal. These results suggest that B7-H4 is one of multiple negative cosignaling molecules that collectively provide a fine-tuning mechanism for T-cell-mediated immune responses.
- Published
- 2006
16. Lipocalin 2-deficient mice exhibit increased sensitivity toEscherichia coliinfection but not to ischemia-reperfusion injury
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Atsushi Togawa, Gordon S. Duncan, Andrew Wakeham, Andrew J. Elia, Tak W. Mak, Annick You-Ten, Hannah E. H. Fong, Carol C. Cheung, and Thorsten Berger
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medicine.medical_specialty ,Apoptosis ,Siderocalin ,Lipocalin ,Biology ,Kidney ,Mice ,Lipocalin-2 ,In vivo ,Internal medicine ,Escherichia coli ,medicine ,Animals ,Cells, Cultured ,Escherichia coli Infections ,Mice, Knockout ,Oncogene Proteins ,Multidisciplinary ,integumentary system ,Acute-phase protein ,Kidney metabolism ,Biological Sciences ,medicine.disease ,Immunity, Innate ,Lipocalins ,Endocrinology ,medicine.anatomical_structure ,Reperfusion Injury ,Immunology ,Reperfusion injury ,Acute-Phase Proteins - Abstract
Diverse functions have been reported for lipocalin 2. To investigate these functionsin vivo, we generated gene-targeted lipocalin 2-deficient mice (Lcn2−/−mice).In vitrostudies have suggested that lipocalin 2 is important for cellular apoptosis induced by IL-3 withdrawal, and for the induction of kidney differentiation during embryogenesis. Analysis ofLcn2−/−mice showed normal cell death upon IL-3 withdrawal and normal kidney development. However, we found thatLcn2−/−mice exhibited an increased susceptibility to bacterial infections, in keeping with the proposed function of lipocalin 2 in iron sequestration. Neutrophils isolated fromLcn2−/−mice showed significantly less bacteriostatic activity compared with WT controls. The bacteriostatic property of the WT neutrophils was abolished by the addition of exogenous iron, indicating that the main function of lipocalin 2 in the antibacterial innate immune response is to limit this essential element. Another important function ascribed to lipocalin 2 has been its protective role against kidney ischemia-reperfusion injury. We analyzedLcn2−/−mice using a mouse model for severe renal failure and could not detect any significant differences compared with their WT littermates.
- Published
- 2006
17. Cutting Edge: Tissue Inhibitor of Metalloproteinase 3 Regulates TNF-Dependent Systemic Inflammation
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Tak W. Mak, Rama Khokha, Fazilat F. Mohammed, Gordon S. Duncan, David Smookler, and Zamaneh Kassiri
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chemistry.chemical_classification ,Innate immune system ,Immunology ,Biology ,Tissue inhibitor of metalloproteinase ,Pharmacology ,Systemic inflammation ,Molecular biology ,Enzyme ,chemistry ,Immunity ,medicine ,Immunology and Allergy ,Tumor necrosis factor alpha ,medicine.symptom ,Receptor ,Gene - Abstract
Host response to infectious agents must be rapid and powerful. One mechanism is the release of presynthesized membrane-bound TNF. TNF shedding is mediated by TNF-α converting enzyme, which is selectively inhibited by the tissue inhibitor of metalloproteinase 3 (TIMP3). We show that loss of TIMP3 impacts innate immunity by dysregulating cleavage of TNF and its receptors. Cultured timp3−/− macrophages release more TNF in response to LPS than wild-type macrophages. In timp3−/− mice, LPS causes serum levels of TNF and its receptors to rise more rapidly and remain higher compared with wild-type mice. The altered kinetics of ligand and receptor shedding enhances TNF signaling in timp3−/− mice, indicated by elevated serum IL-6. Physiologically, timp3−/− mice are more susceptible to LPS-induced mortality. Ablation of the TNF receptor gene p55 (Tnfrsf1a) or treatment with a synthetic metalloproteinase inhibitor rescues timp3−/− mice. Thus, TIMP3 is essential for normal innate immune function.
- Published
- 2006
18. Integral role of IRF-5 in the gene induction programme activated by Toll-like receptors
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Yusuke Ohba, Hideyuki Yanai, Kenya Honda, Shin Ichi Kano, Hideo Negishi, Seiji Kondo, Tak W. Mak, Akinori Takaoka, Tadatsugu Taniguchi, Gordon S. Duncan, and Tatsuaki Mizutani
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Lipopolysaccharides ,medicine.medical_treatment ,Receptors, Cell Surface ,Biology ,Proinflammatory cytokine ,Mice ,medicine ,Animals ,RNA, Messenger ,Receptors, Immunologic ,Promoter Regions, Genetic ,Transcription factor ,Adaptor Proteins, Signal Transducing ,Inflammation ,TNF Receptor-Associated Factor 6 ,Toll-like receptor ,Membrane Glycoproteins ,Multidisciplinary ,Innate immune system ,Toll-Like Receptors ,Acquired immune system ,Antigens, Differentiation ,Shock, Septic ,Up-Regulation ,Cell biology ,Cytokine ,Interferon Regulatory Factors ,Myeloid Differentiation Factor 88 ,Immunology ,Cytokines ,Signal transduction ,Gene Deletion ,Signal Transduction ,Transcription Factors - Abstract
The activation of Toll-like receptors (TLRs) is central to innate and adaptive immunity. All TLRs use the adaptor MyD88 for signalling, but the mechanisms underlying the MyD88-mediated gene induction programme are as yet not fully understood. Here, we demonstrate that the transcription factor IRF-5 is generally involved downstream of the TLR-MyD88 signalling pathway for gene induction of proinflammatory cytokines, such as interleukin-6 (IL-6), IL-12 and tumour-necrosis factor-alpha. In haematopoietic cells from mice deficient in the Irf5 gene (Irf5-/- mice), the induction of these cytokines by various TLR ligands is severely impaired, whereas interferon-alpha induction is normal. We also provide evidence that IRF-5 interacts with and is activated by MyD88 and TRAF6, and that TLR activation results in the nuclear translocation of IRF-5 to activate cytokine gene transcription. Consistently, Irf5-/- mice show resistance to lethal shock induced by either unmethylated DNA or lipopolysaccharide, which correlates with a marked decrease in the serum levels of proinflammatory cytokines. Thus, our study identifies IRF-5 as a new, principal downstream regulator of the TLR-MyD88 signalling pathway and a potential target of therapeutic intervention to control harmful immune responses.
- Published
- 2005
19. Enhanced TCR-induced Apoptosis in Interferon Regulatory Factor 4–deficient CD4+ Th Cells
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Gordon S. Duncan, Hans-Willi Mittrücker, Tak W. Mak, Frank Sommer, Magda Huber, David A. Ferrick, Anne Brüstle, Michael Lohoff, and Bärbel Casper
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CD4-Positive T-Lymphocytes ,Th cell ,Time Factors ,medicine.medical_treatment ,Immunology ,Receptors, Antigen, T-Cell ,Apoptosis ,Mice, Transgenic ,Biology ,Mice ,IRF4 ,In Situ Nick-End Labeling ,medicine ,Animals ,Immunology and Allergy ,T helper cell ,fas Receptor ,Annexin A5 ,Coloring Agents ,Interleukin 4 ,Leishmania major ,Mice, Inbred BALB C ,Cell growth ,Brief Definitive Report ,IL-4 ,Interleukin ,T-Lymphocytes, Helper-Inducer ,Flow Cytometry ,Fas receptor ,Molecular biology ,DNA-Binding Proteins ,Mice, Inbred C57BL ,Cytokine ,medicine.anatomical_structure ,CD4 Antigens ,Interferon Regulatory Factors ,CD95 ,Cytokines ,Cell Division ,Transcription Factors ,Interferon regulatory factors - Abstract
Transcription factors of the interferon regulatory factor (IRF) family contribute to the regulation of cell proliferation and apoptosis. Here, we show that CD4(+) T helper (Th) cells lacking IRF4 (IRF4(-/-)) are highly sensitive to apoptosis. After infection of IRF4(-/-) mice with the protozoan parasite Leishmania major, the lesion-draining lymph nodes developed the prototypic lymphadenopathy of wild-type mice after 4 wk, but demonstrated almost total loss of cellularity and enhanced apoptosis after 7 wk. In vitro, activation of IRF4(-/-) CD4(+) Th cells led to greatly increased apoptosis compared with wild-type cells. Coculture of IRF4(-/-) and IRF4(+/+) CD4(+) cells did not increase survival of IRF4(-/-) CD4(+) cells, indicating that the enhanced rate of IRF4(-/-) Th cell apoptosis was neither transferable nor due to lack of a cytokine. Enhanced CD4(+) cell apoptosis was also observed after anti-CD95 mAb treatment, despite normal CD95 expression. Removal of endogenous cytokines, notably interleukin (IL)-4, led to increased and equally high levels of IRF4(-/-) and IRF4(+/+) cell apoptosis, whereas the protective activity of exogenous IL-4 was reduced in IRF4(-/-) CD4(+) cells despite normal expression of the IL-4 receptor. Therefore, IRF4 is central in protecting CD4(+) cells against proapoptotic stimuli.
- Published
- 2004
20. The Inducible Costimulator Plays the Major Costimulatory Role in Humoral Immune Responses in the Absence of CD28
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Hitoshi Okada, Bernhard Odermatt, Andrew Wakeham, Woong-Kyung Suh, Tak W. Mak, Suzanne Plyte, Arda Shahinian, Pamela S. Ohashi, Nancy N. Berg-Brown, Anna Tafuri, and Gordon S. Duncan
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Antigens, Differentiation, T-Lymphocyte ,CD4-Positive T-Lymphocytes ,medicine.medical_treatment ,T cell ,Immunology ,chemical and pharmacologic phenomena ,Biology ,Antibodies, Viral ,Vesicular stomatitis Indiana virus ,Inducible T-Cell Co-Stimulator Protein ,Mice ,Th2 Cells ,Immune system ,CD28 Antigens ,Antigen ,Agammaglobulinemia ,Rhabdoviridae Infections ,medicine ,Animals ,Immunology and Allergy ,Crosses, Genetic ,B cell ,Mice, Knockout ,Germinal center ,CD28 ,hemic and immune systems ,Germinal Center ,Immunoglobulin Class Switching ,Clone Cells ,Immunoglobulin A ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Cytokine ,Immunoglobulin M ,Immunoglobulin G ,Cytokines ,Cell Division - Abstract
CD28 plays crucial costimulatory roles in T cell proliferation, cytokine production, and germinal center response. Mice that are deficient in the inducible costimulator (ICOS) also have defects in cytokine production and germinal center response. Because the full induction of ICOS in activated T cells depends on CD28 signal, the T cell costimulatory capacity of ICOS in the absence of CD28 has remained unclear. We have clarified this issue by comparing humoral immune responses in wild-type, CD28 knockout (CD28 KO), and CD28-ICOS double-knockout (DKO) mice. DKO mice had profound defects in Ab responses against environmental Ags, T-dependent protein Ags, and vesicular stomatitis virus that extended far beyond those observed in CD28 KO mice. However, DKO mice mounted normal Ab responses against a T-independent Ag, indicating that B cell function itself was normal. Restimulated CD4+ DKO T cells that had been primed in vivo showed decreased proliferation and reduced IL-4 and IL-10 production compared with restimulated CD4+ T cells from CD28 KO mice. Thus, in the absence of CD28, ICOS assumes the major T cell costimulatory role for humoral immune responses. Importantly, CD28-mediated ICOS up-regulation is not essential for ICOS function in vivo.
- Published
- 2004
21. Survivin Loss in Thymocytes Triggers p53-mediated Growth Arrest and p53-independent Cell Death
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Hitoshi Okada, Maria Ciofani, Woong-Kyung Suh, Gordon S. Duncan, Andrew Wakeham, Chris Bakal, Robert Rottapel, Juan Carlos Zúñiga-Pflücker, Andrew E. H. Elia, Arda Shahinian, and Tak W. Mak
- Subjects
Cell cycle checkpoint ,Survivin ,T-Lymphocytes ,Immunology ,Thymus Gland ,Biology ,Article ,Inhibitor of Apoptosis Proteins ,03 medical and health sciences ,Mice ,0302 clinical medicine ,thymus ,Immunology and Allergy ,Animals ,neoplasms ,development ,030304 developmental biology ,mitosis ,Mice, Knockout ,0303 health sciences ,Early embryonic stage ,Cell Death ,Cell growth ,Reverse Transcriptase Polymerase Chain Reaction ,Cell Cycle ,Cell cycle ,Flow Cytometry ,Cell biology ,Neoplasm Proteins ,Thymocyte ,Kinetics ,pre–T cell ,030220 oncology & carcinogenesis ,Cancer research ,Signal transduction ,Tumor Suppressor Protein p53 ,Microtubule-Associated Proteins ,CD8 ,Cell Division - Abstract
Because survivin-null embryos die at an early embryonic stage, the role of survivin in thymocyte development is unknown. We have investigated the role by deleting the survivin gene only in the T lineage and show here that loss of survivin blocks the transition from CD4− CD8− double negative (DN) thymocytes to CD4+ CD8+ double positive cells. Although the pre–T cell receptor signaling pathway is intact in survivin-deficient thymocytes, the cells cannot respond to its signals. In response to proliferative stimuli, cycling survivin-deficient DN cells exhibit cell cycle arrest, a spindle formation defect, and increased cell death. Strikingly, loss of survivin activates the tumor suppressor p53. However, the developmental defects caused by survivin deficiency cannot be rescued by p53 inactivation or introduction of Bcl-2. These lines of evidence indicate that developing thymocytes depend on the cytoprotective function of survivin and that this function is tightly coupled to cell proliferation but independent of p53 and Bcl-2. Thus, survivin plays a critical role in early thymocyte development.
- Published
- 2004
22. Differential Requirement for Malt1 in T and B Cell Antigen Receptor Signaling
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Jürgen Ruland, Tak W. Mak, Andrew Wakeham, and Gordon S. Duncan
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T cell ,Immunology ,Regulator ,Receptors, Antigen, T-Cell ,Receptors, Antigen, B-Cell ,Biology ,Mice ,medicine ,Animals ,Immunology and Allergy ,Receptor ,B cell ,T-cell receptor ,NF-kappa B ,Lymphoma, B-Cell, Marginal Zone ,Paracaspase ,BCL10 ,Cell biology ,Neoplasm Proteins ,MALT1 ,medicine.anatomical_structure ,Infectious Diseases ,Mucosa-Associated Lymphoid Tissue Lymphoma Translocation 1 Protein ,Caspases ,Cancer research ,Signal Transduction - Abstract
The translocation t(11;18)(q21;q21) involving MALT1 is the most common chromosomal abnormality in lymphomas of mucosa-associated lymphoid tissue. Although the paracaspase MALT1 can bind to BCL10, the physiological function of MALT1 is unknown. Using mouse models, we show that Malt1 is essential for T cell activation, proliferation, and IL-2 production in response to TCR ligation and strictly required for signal-specific NF-kappaB activation induced by the TCR but not TNF-alpha or IL-1 signaling. Malt1 operates downstream of Bcl10, controls the catalytic activity of the canonical IKK complex, and regulates the signaling of Jnk and p38 MAP kinases. In contrast to Bcl10 disruption, however, inactivation of Malt1 has only mild effects on B cell activation and does not cause defects during neurodevelopment. Thus, Malt1 is an essential regulator of Bcl10 signaling that is differentially required depending on cellular context.
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- 2003
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23. Caspase-3 regulates cell cycle in B cells: a consequence of substrate specificity
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Tak W. Mak, Liwei Lu, Caren Furlonger, Denis Bouchard, Razqallah Hakem, Takehiko Sasaki, Christopher J. Paige, Minna Woo, Anne Hakem, Gillian E. Wu, and Gordon S. Duncan
- Subjects
Cyclin-Dependent Kinase Inhibitor p21 ,Cell division ,Immunology ,Apoptosis ,Lymphocyte Activation ,Substrate Specificity ,Mice ,B cell homeostasis ,Cyclin-dependent kinase ,Cyclins ,Proliferating Cell Nuclear Antigen ,medicine ,Animals ,Immunology and Allergy ,Antigen-presenting cell ,Caspase ,B cell ,Mice, Knockout ,B-Lymphocytes ,biology ,Caspase 3 ,Cell Cycle ,Cell cycle ,Flow Cytometry ,Immunohistochemistry ,Molecular biology ,Cyclin-Dependent Kinases ,Cell biology ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Bromodeoxyuridine ,Caspases ,biology.protein ,Spleen - Abstract
Caspases are important for apoptosis but are also involved in mammalian cell survival and cell division. Here we report that caspase-3 is a negative regulator of B cell cycling. Mice deficient in caspase-3 (Casp3-/- mice) have increased numbers of splenic B cells that show normal apoptosis but enhanced proliferation in vivo and hyperproliferation after mitogenic stimulation in vitro. Cdkn1a encodes p21 (also called Waf1 or Cip1), a cyclin-dependent kinase (CDK) inhibitor. Although expression of p21 was increased, CDK activities and proliferating cell nuclear antigen (PCNA) were increased in Casp3-/- B cells. Using Casp3-/-Cdkn1a-/- mice, we show that the hyperproliferation of Casp3-/- B cells is abolished when Cdkn1a is also deleted. Our genetic and biochemical data demonstrate that caspase-3 is essential in the regulation of B cell homeostasis.
- Published
- 2003
24. Severe impairment of interleukin-1 and Toll-like receptor signalling in mice lacking IRAK-4
- Author
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Tak W. Mak, Teiji Wada, Josef M. Penninger, Wen Chen Yeh, Douglas G. Millar, Gordon S. Duncan, Andrew Wakeham, Shyun Li, Annick Itie, Nobutaka Suzuki, Christine Mirtsos, Holger Wesche, Shinobu Suzuki, Hidetoshi Takada, and Pamela S. Ohashi
- Subjects
Lipopolysaccharides ,Staphylococcus aureus ,Ratón ,medicine.medical_treatment ,Receptors, Cell Surface ,macromolecular substances ,Biology ,Ligands ,Nitric Oxide ,p38 Mitogen-Activated Protein Kinases ,Interferon-gamma ,Mice ,Interleukin-1 Receptor-Associated Kinases ,medicine ,Animals ,Arenaviridae Infections ,Drosophila Proteins ,Lymphocytic choriomeningitis virus ,Cells, Cultured ,B-Lymphocytes ,Toll-like receptor ,Membrane Glycoproteins ,Multidisciplinary ,Innate immune system ,Interleukin-6 ,Tumor Necrosis Factor-alpha ,Macrophages ,musculoskeletal, neural, and ocular physiology ,Toll-Like Receptors ,JNK Mitogen-Activated Protein Kinases ,NF-kappa B ,Receptors, Interleukin-1 ,Interleukin ,Staphylococcal Infections ,IRAK4 ,Immunity, Innate ,Killer Cells, Natural ,Cytokine ,Signalling ,nervous system ,Immunology ,Mitogen-Activated Protein Kinases ,Protein Kinases ,Gene Deletion ,Interleukin-1 ,Signal Transduction - Abstract
Toll-like receptors (TLRs), which recognize pathogen-associated molecular patterns, and members of the pro-inflammatory interleukin-1 receptor (IL-1R) family, share homologies in their cytoplasmic domains called Toll/IL-1R/plant R gene homology (TIR) domains. Intracellular signalling mechanisms mediated by TIRs are similar, with MyD88 (refs 5-8) and TRAF6 (refs 9, 10) having critical roles. Signal transduction between MyD88 and TRAF6 is known to involve the serine-threonine kinase IL-1 receptor-associated kinase 1 (IRAK-1) and two homologous proteins, IRAK-2 (ref. 12) and IRAK-M. However, the physiological functions of the IRAK molecules remain unclear, and gene-targeting studies have shown that IRAK-1 is only partially required for IL-1R and TLR signalling. Here we show by gene-targeting that IRAK-4, an IRAK molecule closely related to the Drosophila Pelle protein, is indispensable for the responses of animals and cultured cells to IL-1 and ligands that stimulate various TLRs. IRAK-4-deficient animals are completely resistant to a lethal dose of lipopolysaccharide (LPS). In addition, animals lacking IRAK-4 are severely impaired in their responses to viral and bacterial challenges. Our results indicate that IRAK-4 has an essential role in innate immunity.
- Published
- 2002
25. Toso controls encephalitogenic immune responses by dendritic cells and regulatory T cells
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Sara R. Hamilton, Michael St. Paul, Bryan E. Snow, Colin Reardon, Anne Brüstle, Aline Pfefferle, Gloria H. Y. Lin, Tak W. Mak, Michael W. Tusche, Christiane B. Knobbe-Thomsen, Gordon S. Duncan, Karl S. Lang, Pamela S. Ohashi, Philipp A. Lang, Dirk Brenner, and Syed O. Gilani
- Subjects
Encephalomyelitis, Autoimmune, Experimental ,Time Factors ,Medizin ,Inflammation ,Biology ,medicine.disease_cause ,T-Lymphocytes, Regulatory ,Autoimmunity ,Mice ,Immune system ,medicine ,Animals ,Humans ,Receptor ,Cell Proliferation ,Mice, Knockout ,Multidisciplinary ,Innate immune system ,Multiple sclerosis ,Experimental autoimmune encephalomyelitis ,Membrane Proteins ,Cell Differentiation ,Dendritic Cells ,Th1 Cells ,Biological Sciences ,medicine.disease ,Immunohistochemistry ,Mice, Inbred C57BL ,HEK293 Cells ,Gene Expression Regulation ,Apoptosis ,Immunology ,Cytokines ,Th17 Cells ,medicine.symptom ,Carrier Proteins - Abstract
The ability to mount a strong immune response against pathogens is crucial for mammalian survival. However, excessive and uncontrolled immune reactions can lead to autoimmunity. Unraveling how the reactive versus tolerogenic state is controlled might point toward novel therapeutic strategies to treat autoimmune diseases. The surface receptor Toso/Faim3 has been linked to apoptosis, IgM binding, and innate immune responses. In this study, we used Toso-deficient mice to investigate the importance of Toso in tolerance and autoimmunity. We found that Toso(-/-) mice do not develop severe experimental autoimmune encephalomyelitis (EAE), a mouse model for the human disease multiple sclerosis. Toso(-/-) dendritic cells were less sensitive to Toll-like receptor stimulation and induced significantly lower levels of disease-associated inflammatory T-cell responses. Consistent with this observation, the transfer of Toso(-/-) dendritic cells did not induce autoimmune diabetes, indicating their tolerogenic potential. In Toso(-/-) mice subjected to EAE induction, we found increased numbers of regulatory T cells and decreased encephalitogenic cellular infiltrates in the brain. Finally, inhibition of Toso activity in vivo at either an early or late stage of EAE induction prevented further disease progression. Taken together, our data identify Toso as a unique regulator of inflammatory autoimmune responses and an attractive target for therapeutic intervention.
- Published
- 2014
26. Platelet-endothelial cell adhesion molecule-1 (PECAM-1)–deficient mice demonstrate a transient and cytokine-specific role for PECAM-1 in leukocyte migration through the perivascular basement membrane
- Author
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Richard Thompson, Sussan Nourshargh, Karen E. Noble, Tak W. Mak, Gordon S. Duncan, Karen Y. Larbi, and Ann Dewar
- Subjects
Leukocyte migration ,Immunology ,Leukocyte Rolling ,Granulocyte ,Biology ,Biochemistry ,Basement Membrane ,Neutrophil Activation ,Mice ,Venules ,Leukocyte Trafficking ,Cell Adhesion ,Leukocytes ,medicine ,Animals ,Cell adhesion ,Mice, Knockout ,Basement membrane ,Venule ,Tumor Necrosis Factor-alpha ,Cell adhesion molecule ,Cell Biology ,Hematology ,Cell biology ,Mice, Inbred C57BL ,Platelet Endothelial Cell Adhesion Molecule-1 ,Chemotaxis, Leukocyte ,Microscopy, Electron ,medicine.anatomical_structure ,cardiovascular system ,Interleukin-1 - Abstract
Studies with neutralizing antibodies have indicated roles for platelet-endothelial cell adhesion molecule-1 (PECAM-1) in leukocyte migration through the endothelium and the perivascular basement membrane. Because some of these findings have been contentious, this study aimed to explore the role of PECAM-1 in leukocyte migration by analyzing leukocyte responses in interleukin 1beta (IL-1beta)- and tumor necrosis factor-alpha (TNFalpha)-activated cremasteric venules of PECAM-1-deficient mice using intravital and electron microscopy. Although no differences in levels of leukocyte rolling flux or firm adhesion were observed, a delay in leukocyte transmigration in response to IL-1beta, but not TNFalpha, was detected in PECAM-1-deficient mice. Electron microscopy indicated that this delay occurred at the level of perivascular basement membrane. To address the cytokine specificity of PECAM-1 dependence, in vitro experiments demonstrated that TNFalpha, but not IL-1beta, could induce rapid adhesion of murine neutrophils to protein-coated surfaces, suggesting that TNFalpha elicited leukocyte transmigration in wild-type mice via direct stimulation of leukocytes. In summary, the results suggest a regulatory role for PECAM-1 in leukocyte migration through the perivascular basement membrane, a role that appears to be cytokine-specific and associated with the ability of the cytokine to stimulate rapid neutrophil adhesion.
- Published
- 2001
27. Bcl10 Is a Positive Regulator of Antigen Receptor–Induced Activation of NF-κ B and Neural Tube Closure
- Author
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Tak W. Mak, Linh T. Nguyen, Pamela S. Ohashi, Gordon S. Duncan, Andrew E. H. Elia, Sue Plyte, Andrew Wakeham, Denis Bouchard, Douglas G. Millar, Ivan del Barco Barrantes, and Jürgen Ruland
- Subjects
MAPK/ERK pathway ,Central Nervous System ,T-Lymphocytes ,CARD11 ,Antineoplastic Agents ,Apoptosis ,Lymphocyte proliferation ,Biology ,Lymphocyte Activation ,General Biochemistry, Genetics and Molecular Biology ,chemistry.chemical_compound ,Mice ,Antigen ,Animals ,Neural Tube Defects ,Enzyme Inhibitors ,Adaptor Proteins, Signal Transducing ,Etoposide ,Nucleic Acid Synthesis Inhibitors ,Mice, Knockout ,Protein Synthesis Inhibitors ,B-Lymphocytes ,Immunity, Cellular ,Biochemistry, Genetics and Molecular Biology(all) ,NF-kappa B ,Tyrosine phosphorylation ,B-Cell CLL-Lymphoma 10 Protein ,Staurosporine ,Molecular biology ,In vitro ,Neoplasm Proteins ,MALT1 ,Receptors, Antigen ,chemistry ,Antibody Formation ,Cancer research ,Genes, Lethal ,Cisplatin ,Anisomycin ,Cell Division ,Signal Transduction - Abstract
Bcl10, a CARD-containing protein identified from the t(1;14)(p22;q32) breakpoint in MALT lymphomas, has been shown to induce apoptosis and activate NF- κ B in vitro. We show that one-third of bcl10 −/− embryos developed exencephaly, leading to embryonic lethality. Surprisingly, bcl10 −/− cells retained susceptibility to various apoptotic stimuli in vivo and in vitro. However, surviving bcl10 −/− mice were severely immunodeficient and bcl10 −/− lymphocytes are defective in antigen receptor or PMA/Ionomycin-induced activation. Early tyrosine phosphorylation, MAPK and AP-1 activation, and Ca 2+ signaling were normal in mutant lymphocytes, but antigen receptor–induced NF- κ B activation was absent. Thus, Bcl10 functions as a positive regulator of lymphocyte proliferation that specifically connects antigen receptor signaling in B and T cells to NF- κ B activation.
- Published
- 2001
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28. ICOS is essential for effective T-helper-cell responses
- Author
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Gordon S. Duncan, Louis-Martin Boucher, Friedhelm Bladt, Tom Horan, Manel Jordana, Arda Shahinian, Vera S. F. Chan, Tak W. Mak, Josef M. Penninger, Bernhard Odermatt, John Whoriskey, Andrew Wakeham, Pamela S. Ohashi, Alexandra Ho, Tony Pawson, Steve K. Yoshinaga, Annick Itie, Denis Bouchard, and Anna Tafuri
- Subjects
ICOS LIGAND ,Multidisciplinary ,medicine.anatomical_structure ,Antigen ,Immunoglobulin class switching ,Immunology ,medicine ,CD28 ,T lymphocyte ,T helper cell ,Inducible T-Cell Co-Stimulator Protein ,Biology ,Inducible T-Cell Co-Stimulator Ligand - Abstract
The outcome of T-cell responses after T-cell encounter with specific antigens is modulated by co-stimulatory signals, which are required for both lymphocyte activation and development of adaptive immunity1,2,3. ICOS4,5, an inducible co-stimulator with homology to CD28, is expressed on activated, but not resting T cells, and shows T-cell co-stimulatory function in vitro. ICOS binds specifically to its counter-receptor B7RP-1 (refs 5,6,7), but not to B7-1 or B7-2. Here we provide in vivo genetic evidence that ICOS delivers a co-stimulatory signal that is essential both for efficient interaction between T and B cells and for normal antibody responses to T-cell-dependent antigens. To determine the physiological function of ICOS, we generated and characterized gene-targeted ICOS-deficient mice. In vivo, a lack of ICOS results in severely deficient T-cell-dependent B-cell responses. Germinal centre formation is impaired and immunoglobulin class switching, including production of allergy-mediating IgE, is defective. ICOS-deficient T cells primed in in vivo and restimulated in vitro with specific antigen produce only low levels of interleukin-4, but remain fully competent to produce interferon-γ.
- Published
- 2001
29. Deficiency in the Transcription Factor Interferon Regulatory Factor (Irf)-2 Leads to Severely Compromised Development of Natural Killer and T Helper Type 1 Cells
- Author
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Andreas Pahl, Martin Röllinghoff, Hans-Willi Mittrücker, Michael Lohoff, David A. Ferrick, Tak W. Mak, Stefan Prechtl, Edgar Schmitt, Susi Bischof, and Gordon S. Duncan
- Subjects
CD4-Positive T-Lymphocytes ,Male ,Interferon Regulatory Factor 2 ,Cellular differentiation ,Immunology ,Leishmaniasis, Cutaneous ,Biology ,Nitric Oxide ,Th1 ,Mice ,Interleukin 21 ,Immune system ,Bone Marrow ,Interferon ,medicine ,Animals ,Immunology and Allergy ,Lymphocyte Count ,Leishmania major ,Interleukin-15 ,Mice, Knockout ,Leishmania ,Mice, Inbred BALB C ,natural killer cells ,Cell Differentiation ,Th1 Cells ,Interleukin-12 ,Cell biology ,DNA-Binding Proteins ,Killer Cells, Natural ,Mice, Inbred C57BL ,Repressor Proteins ,Disease Models, Animal ,Interleukin 15 ,interferon regulatory factor ,Interleukin 12 ,Female ,Original Article ,Disease Susceptibility ,Interferon Regulatory Factor-2 ,interleukin 15 ,Transcription Factors ,Interferon regulatory factors ,medicine.drug - Abstract
Interferon (IFN) regulatory factor (IRF)-2 was originally described as an antagonist of IRF-1–mediated transcriptional regulation of IFN-inducible genes. IRF-1−/− mice exhibit defective T helper type 1 (Th1) cell differentiation. We have used experimental leishmaniasis to show that, like IRF-1−/− mice, IRF-2−/− mice are susceptible to Leishmania major infection due to a defect in Th1 differentiation. Natural killer (NK) cell development is compromised in both IRF-1−/− and IRF-2−/− mice, but the underlying mechanism differs. NK (but not NK+ T) cell numbers are decreased in IRF-2−/− mice, and the NK cells that are present are immature in phenotype. Therefore, like IRF-1, IRF-2 is required for normal generation of Th1 responses and for NK cell development in vivo. In this particular circumstance the absence of IRF-2 cannot be compensated for by the presence of IRF-1 alone. Mechanistically, IRF-2 may act as a functional agonist rather than antagonist of IRF-1 for some, but not all, IFN-stimulated regulatory element (ISRE)-responsive genes.
- Published
- 2000
30. PECAM-1 (CD31) Expression Modulates Bleeding Time in Vivo
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Sonali Patil, Tak W. Mak, Joseph A. Madri, Donnasue Graesser, Gordon S. Duncan, Peter J. Newman, and Sepi Mahooti
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Male ,Heterozygote ,Bleeding Time ,Time Factors ,Angiogenesis ,Integrin ,Short Communications ,Biology ,Pathology and Forensic Medicine ,Mice ,In vivo ,Animals ,Platelet ,Mice, Knockout ,Platelet Count ,Cell adhesion molecule ,Homozygote ,Hematopoietic Stem Cell Transplantation ,Hematopoietic Stem Cells ,In vitro ,Cell biology ,Platelet Endothelial Cell Adhesion Molecule-1 ,Endothelial stem cell ,Haematopoiesis ,embryonic structures ,Immunology ,cardiovascular system ,biology.protein ,Female ,tissues - Abstract
PECAM-1 is a 130-kd member of the Ig superfamily present on endothelial cells, platelets, polymorphonuclear leukocytes, monocytes, and lymphocytes. Its expression begins early in development and persists through adulthood. PECAM-1 functions as an adhesion and signaling molecule between adjacent endothelial cells and between endothelial cells and circulating blood elements. Antibodies directed against PECAM-1 have been shown to affect angiogenesis, endothelial cell migration, and polymorphonuclear leukocyte transmigration. Furthermore, its dimerization is associated with the modulation of integrin affinity. Antibody inhibition studies suggest that PECAM-1 plays a role in modulating thrombosis; however, recent in vitro aggregation studies performed on platelets harvested from PECAM-1-deficient mice revealed no abnormalities. In this report we demonstrate prolonged in vivo bleeding times in PECAM-1-deficient mice. This abnormality was not corrected when wild-type hematopoietic precursors were engrafted into marrow-ablated PECAM-1-deficient mice. Furthermore, normal bleeding times were observed when marrow-ablated wild-type mice were engrafted with hematopoietic precursors harvested from PECAM-1-deficient mice. These studies are consistent with a role for PECAM-1 in modulating thrombosis in the vasculature, which is potentially mediated by endothelial cell PECAM-1 expression.
- Published
- 2000
31. T-cell co-stimulation through B7RP-1 and ICOS
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Gary Elliott, Grace Shih, Ming Zhang, Gwyneth Van, Pauline Campbell, Christine L. Shaklee, Tak W. Mak, John Whoriskey, Ulla Sarmiento, Jane Guo, Tadahiko Kohno, David Chang, Ariela Hui, Marco A. Coccia, Tom Horan, Tianang Dai, Anna Tafuri-Bladt, Laura Chiu, Giorgio Senaldi, Sheila Scully, David W. Brankow, Susan McCabe, Gordon S. Duncan, Steven Kiyoshi Yoshinaga, Sanjay D. Khare, and Arda Shahinian
- Subjects
Antigens, Differentiation, T-Lymphocyte ,DNA, Complementary ,Recombinant Fusion Proteins ,T-Lymphocytes ,T cell ,Molecular Sequence Data ,Gene Expression ,Mice, Transgenic ,CHO Cells ,Biology ,Dermatitis, Contact ,Ligands ,Lymphocyte Activation ,Inducible T-Cell Co-Stimulator Protein ,Inducible T-Cell Co-Stimulator Ligand ,Mice ,Immune system ,Co-stimulation ,Cricetinae ,medicine ,Animals ,Humans ,Amino Acid Sequence ,Cells, Cultured ,Mice, Inbred BALB C ,Multidisciplinary ,COS cells ,Sequence Homology, Amino Acid ,T-cell receptor ,CD28 ,Acquired immune system ,Cell biology ,Mice, Inbred C57BL ,ICOS LIGAND ,medicine.anatomical_structure ,COS Cells ,Immunology ,B7-1 Antigen ,Female ,Lymph Nodes ,Protein Binding - Abstract
T-cell activation requires co-stimulation through receptors such as CD28 and antigen-specific signalling through the T-cell antigen receptor. Here we describe a new murine costimulatory receptor-ligand pair. The receptor, which is related to CD28 and is the homologue of the human protein ICOS, is expressed on activated T cells and resting memory T cells. The ligand, which has homology to B7 molecules and is called B7-related protein-1 (B7RP-1), is expressed on B cells and macrophages. ICOS and B7RP-I do not interact with proteins in the CD28-B7 pathway, and B7RP-1 co-stimulates T cells in vitro independently of CD28. Transgenic mice expressing a B7RP-1-Fc fusion protein show lymphoid hyperplasia in the spleen, lymph nodes and Peyer's patches. Presensitized mice treated with B7RP-1-Fc during antigen challenge show enhanced hypersensitivity. Therefore, B7RP-1 exhibits co-stimulatory activities in vitro and in vivo. ICOS and B7RP-1 define a new and distinct receptor-ligand pair that is structurally related to CD28-B7 and is involved in the adaptive immune response.
- Published
- 1999
32. In Vivo Evidence That Caspase-3 Is Required for Fas-Mediated Apoptosis of Hepatocytes
- Author
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Minna Woo, Anne Hakem, Andrew J. Elia, Razqallah Hakem, Gordon S. Duncan, Bruce J. Patterson, and Tak W. Mak
- Subjects
Immunology ,Immunology and Allergy - Abstract
Caspase-3 is essential for Fas-mediated apoptosis in vitro. We investigated the role of caspase-3 in Fas-mediated cell death in vivo by injecting caspase-3-deficient mice with agonistic anti-Fas Ab. Wild-type controls died rapidly of fulminant hepatitis, whereas the survival of caspase-3−/− mice was increased due to a delay in hepatocyte cell death. Bcl-2 expression in the liver was dramatically decreased in wild-type mice following anti-Fas injection, but was unchanged in caspase-3−/− mice. Hepatocytes from anti-Fas-injected wild-type, but not caspase-3−/−, mice released cytochrome c into the cytoplasm. Western blotting confirmed the lack of caspase-3-mediated cleavage of Bcl-2. Presumably the presence of intact Bcl-2 in caspase-3−/− hepatocytes prevents the release of cytochrome c from the mitochondria, a required step for the mitochondrial death pathway. We also show by Western blot that Bcl-xL, caspase-9, caspase-8, and Bid are processed by caspase-3 in injected wild-type mice but that this processing does not occur in caspase-3−/− mice. This study thus provides novel in vivo evidence that caspase-3, conventionally known for its downstream effector function in apoptosis, also modifies Bcl-2 and other upstream proteins involved in the regulation of Fas-mediated apoptosis.
- Published
- 1999
33. TRAF2 Deficiency Results in Hyperactivity of Certain TNFR1 Signals and Impairment of CD40-Mediated Responses
- Author
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Gordon S. Duncan, Linh T. Nguyen, Arda Shahinian, Michael W. Marino, Michelle Ng, Pamela S. Ohashi, Tak W. Mak, Christine Mirtsos, Wen-Chen Yeh, and Daniel E. Speiser
- Subjects
Male ,medicine.medical_specialty ,TRAF2 ,Immunology ,Nitric Oxide Synthase Type II ,Stimulation ,Nitric Oxide ,Receptors, Tumor Necrosis Factor ,Vesicular stomatitis Indiana virus ,Negative regulator ,Nitric oxide ,chemistry.chemical_compound ,Mice ,Mediator ,Antigens, CD ,Internal medicine ,medicine ,Animals ,Immunology and Allergy ,CD40 Antigens ,Cells, Cultured ,Mice, Knockout ,Mice, Inbred BALB C ,CD40 ,biology ,Tumor Necrosis Factor-alpha ,NF-kappa B ,Proteins ,TNF Receptor-Associated Factor 2 ,Immunoglobulin Class Switching ,Interleukin-12 ,Immunoglobulin Isotypes ,Mice, Inbred C57BL ,Endocrinology ,Phenotype ,Infectious Diseases ,chemistry ,Receptors, Tumor Necrosis Factor, Type I ,biology.protein ,Macrophages, Peritoneal ,Tumor necrosis factor alpha ,Female ,Nitric Oxide Synthase ,Tumor necrosis factor receptor ,Cell Division ,Spleen ,Signal Transduction - Abstract
Tumor necrosis factor (TNF) receptor–associated factor 2 (TRAF2) can interact with various members of the TNF receptor family. Previously, we reported that TRAF2-deficient mice die prematurely and have elevated serum TNF levels. In this study, we demonstrate that TRAF2-deficient macrophages produce increased amounts of nitric oxide (NO) and TNF in response to TNF stimulation. Furthermore, we could enhance the survival of TRAF2-deficient mice by eliminating either TNF or TNFR1. Using these double-knockout mice, we show that in the absence of TRAF2, the T helper–dependent antibody response, CD40-mediated proliferation, and NF-κB activation are defective. These data demonstrate two important roles of TRAF2, one as a negative regulator of certain TNFR1 signals and the other as a positive mediator of CD40 signaling.
- Published
- 1999
- Full Text
- View/download PDF
34. Genetic Evidence for Functional Redundancy of Platelet/Endothelial Cell Adhesion Molecule-1 (PECAM-1): CD31-Deficient Mice Reveal PECAM-1-Dependent and PECAM-1-Independent Functions
- Author
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Gordon S. Duncan, David P. Andrew, Hiroaki Takimoto, Stephen A. Kaufman, Hiroki Yoshida, Jason Spellberg, José Luis de la Pompa, Andrew Elia, Andrew Wakeham, Barbara Karan-Tamir, William A. Muller, Giorgio Senaldi, Mark M. Zukowski, and Tak W. Mak
- Subjects
Immunology ,Immunology and Allergy - Abstract
Platelet/endothelial cell adhesion molecule-1 (PECAM-1; CD31), a member of the Ig superfamily, is expressed strongly at endothelial cell-cell junctions, on platelets, and on most leukocytes. CD31 has been postulated to play a role in vasculogenesis and angiogenesis, and has been implicated as a key mediator of the transendothelial migration of leukocytes. To further define the physiologic role of CD31, we used targeted gene disruption of the CD31 gene in embryonic stem cells to generate CD31-deficient mice. CD31-deficient mice (CD31KO) are viable and born at the expected Mendelian frequency, remain healthy, and exhibit no obvious vascular developmental defects. In response to inflammatory challenge, polymorphonuclear leukocytes of CD31KO mice are arrested between the vascular endothelium and the basement membrane of inflammatory site mesenteric microvessels, confirming a role for CD31 in the migration of neutrophils through the subendothelial extracellular matrix. Normal numbers of leukocytes are recovered from inflammatory sites in CD31KO mice, however, suggesting that the defect in leukocyte migration across basal lamina observed in the absence of CD31 may be compensated for by the use of other adhesion molecules, or possibly an increased rate of migration. Homing of T lymphocytes in vivo is normal, and CD31KO mice are able to mount a cutaneous hypersensitivity response normally. In addition, CD31-mediated homophilic adhesion does not appear to play a role in platelet aggregation in vitro. This study provides genetic evidence that CD31 is involved in transbasement membrane migration, but does not play an obligatory role in either vascular development or leukocyte migration.
- Published
- 1999
35. Differential Requirement for Caspase 9 in Apoptotic Pathways In Vivo
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José Luis de la Pompa, Ritsuko Yoshida, David Kägi, Andrew E. H. Elia, Razqallah Hakem, Stephen Kaufman, Minna Woo, Julia Potter, Wilson Khoo, Josef M. Penninger, Anne Hakem, Tak W. Mak, Maria S. Soengas, Gordon S. Duncan, Jeffrey T. Henderson, and Scott W. Lowe
- Subjects
Apoptosis ,Cytochrome c Group ,Mice, Inbred Strains ,Thymus Gland ,Lymphocyte Activation ,Dexamethasone ,General Biochemistry, Genetics and Molecular Biology ,Cell Line ,Membrane Potentials ,Mice ,03 medical and health sciences ,Prosencephalon ,0302 clinical medicine ,Animals ,APAF1 ,Caspase ,030304 developmental biology ,Cerebral Cortex ,Mice, Knockout ,Caspase-9 ,0303 health sciences ,biology ,Biochemistry, Genetics and Molecular Biology(all) ,Stem Cells ,Cytochrome c ,Gene Expression Regulation, Developmental ,Fibroblasts ,Embryo, Mammalian ,Embryonic stem cell ,Molecular biology ,Caspase 9 ,Mitochondria ,3. Good health ,Cell biology ,Enzyme Activation ,Mice, Inbred C57BL ,Cysteine Endopeptidases ,Gamma Rays ,Organ Specificity ,Cell culture ,Caspases ,030220 oncology & carcinogenesis ,biology.protein ,Stem cell ,Spleen ,Signal Transduction - Abstract
Mutation of Caspase 9 (Casp9) results in embryonic lethality and defective brain development associated with decreased apoptosis. Casp9 −/− embryonic stem cells and embryonic fibroblasts are resistant to several apoptotic stimuli, including UV and γ irradiation. Casp9−/− thymocytes are also resistant to dexamethasone- and γ irradiation–induced apoptosis, but are surprisingly sensitive to apoptosis induced by UV irradiation or anti-CD95. Resistance to apoptosis is accompanied by retention of the mitochondrial membrane potential in mutant cells. In addition, cytochrome c is translocated to the cytosol of Casp9−/− ES cells upon UV stimulation, suggesting that Casp9 acts downstream of cytochrome c. Caspase processing is inhibited in Casp9−/− ES cells but not in thymocytes or splenocytes. Comparison of the requirement for Casp9 and Casp3 in different apoptotic settings indicates the existence of at least four different apoptotic pathways in mammalian cells.
- Published
- 1998
- Full Text
- View/download PDF
36. The Transcription Factor Interferon Regulatory Factor 1 (IRF-1) Is Important during the Maturation of Natural Killer 1.1+ T Cell Receptor–α/β+ (NK1+ T) Cells, Natural Killer Cells, and Intestinal Intraepithelial T Cells
- Author
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Tak W. Mak, Toshifumi Matsuyama, Pamela S. Ohashi, Toshiaki Ohteki, Gordon S. Duncan, and Hiroki Yoshida
- Subjects
Receptors, Antigen, T-Cell, alpha-beta ,Immunology ,Biology ,Mice ,Interleukin 21 ,T-Lymphocyte Subsets ,Animals ,Antigens, Ly ,Immunology and Allergy ,Cytotoxic T cell ,Lectins, C-Type ,RNA, Messenger ,IL-2 receptor ,Antigens ,Intestinal Mucosa ,Interleukin 3 ,Interleukin-15 ,ZAP70 ,Brief Definitive Report ,Proteins ,Receptors, Antigen, T-Cell, gamma-delta ,Phosphoproteins ,Natural killer T cell ,Molecular biology ,DNA-Binding Proteins ,Killer Cells, Natural ,Mice, Inbred C57BL ,Gene Expression Regulation ,Interleukin 15 ,Antigens, Surface ,Interleukin 12 ,Brief Definitive Reports ,Interferon Regulatory Factor-1 ,NK Cell Lectin-Like Receptor Subfamily B ,Transcription Factors - Abstract
In contrast to conventional T cells, natural killer (NK) 1.1+ T cell receptor (TCR)-alpha/beta+ (NK1+T) cells, NK cells, and intestinal intraepithelial lymphocytes (IELs) bearing CD8-alpha/alpha chains constitutively express the interleukin (IL)-2 receptor (R)beta/15Rbeta chain. Recent studies have indicated that IL-2Rbeta/15Rbeta chain is required for the development of these lymphocyte subsets, outlining the importance of IL-15. In this study, we investigated the development of these lymphocyte subsets in interferon regulatory factor 1-deficient (IRF-1-/-) mice. Surprisingly, all of these lymphocyte subsets were severely reduced in IRF-1-/- mice. Within CD8-alpha/alpha+ intestinal IEL subset, TCR-gamma/delta+ cells and TCR-alpha/beta+ cells were equally affected by IRF gene disruption. In contrast to intestinal TCR-gamma/delta+ cells, thymic TCR-gamma/delta+ cells developed normally in IRF-1-/- mice. Northern blot analysis further revealed that the induction of IL-15 messenger RNA was impaired in IRF-1-/- bone marrow cells, and the recovery of these lymphocyte subsets was observed when IRF-1-/- cells were cultured with IL-15 in vitro. These data indicate that IRF-1 regulates IL-15 gene expression, which may control the development of NK1+T cells, NK cells, and CD8-alpha/alpha+ IELs.
- Published
- 1998
37. Essential contribution of caspase 3/CPP32 to apoptosis and its associated nuclear changes
- Author
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Razqallah Hakem, Tak W. Mak, Arda Shahinian, Giorgio Senaldi, Stephen Kaufman, Tamara Howard, Mila E. McCurrach, Minna Woo, Anne Hakem, Maria S. Soengas, Wilson Khoo, Scott W. Lowe, David Kägi, and Gordon S. Duncan
- Subjects
Cytotoxicity, Immunologic ,Male ,Programmed cell death ,CD3 Complex ,Neutrophils ,Ultraviolet Rays ,T-Lymphocytes ,Longevity ,Gene Expression ,Apoptosis ,Bone Marrow Cells ,Mice, Inbred Strains ,Caspase 3 ,Embryonic and Fetal Development ,Mice ,Osmotic Pressure ,Tumor Cells, Cultured ,Genetics ,Animals ,fas Receptor ,Caspase ,Cell Nucleus ,B-Lymphocytes ,Cell Death ,biology ,Stem Cells ,Intrinsic apoptosis ,Embryo, Mammalian ,Embryonic stem cell ,Mice, Mutant Strains ,Cell biology ,Mice, Inbred C57BL ,Cysteine Endopeptidases ,UVB-induced apoptosis ,Caspases ,Mutation ,biology.protein ,Female ,Stem cell ,Cell Division ,Research Paper ,Developmental Biology - Abstract
Caspases are fundamental components of the mammalian apoptotic machinery, but the precise contribution of individual caspases is controversial. CPP32 (caspase 3) is a prototypical caspase that becomes activated during apoptosis. In this study, we took a comprehensive approach to examining the role of CPP32 in apoptosis using mice, embryonic stem (ES) cells, and mouse embryonic fibroblasts (MEFs) deficient for CPP32. CPP32(ex3-/-) mice have reduced viability and, consistent with an earlier report, display defective neuronal apoptosis and neurological defects. Inactivation of CPP32 dramatically reduces apoptosis in diverse settings, including activation-induced cell death (AICD) of peripheral T cells, as well as chemotherapy-induced apoptosis of oncogenically transformed CPP32(-/-) MEFs. As well, the requirement for CPP32 can be remarkably stimulus-dependent: In ES cells, CPP32 is necessary for efficient apoptosis following UV- but not gamma-irradiation. Conversely, the same stimulus can show a tissue-specific dependence on CPP32: Hence, TNFalpha treatment induces normal levels of apoptosis in CPP32 deficient thymocytes, but defective apoptosis in oncogenically transformed MEFs. Finally, in some settings, CPP32 is required for certain apoptotic events but not others: Select CPP32(ex3-/-) cell types undergoing cell death are incapable of chromatin condensation and DNA degradation, but display other hallmarks of apoptosis. Together, these results indicate that CPP32 is an essential component in apoptotic events that is remarkably system- and stimulus-dependent. Consequently, drugs that inhibit CPP32 may preferentially disrupt specific forms of cell death.
- Published
- 1998
38. Lymphocyte-derived ACh regulates local innate but not adaptive immunity
- Author
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Colin Reardon, Tak W. Mak, Kevin J. Tracey, Mauricio Rosas-Ballina, Michael W. Tusche, Peder S. Olofsson, Anne Brüstle, Dirk Brenner, and Gordon S. Duncan
- Subjects
CD4-Positive T-Lymphocytes ,medicine.medical_specialty ,Lymphoid Tissue ,Neuroimmunomodulation ,Recombinant Fusion Proteins ,Lymphocyte ,Population ,Mice, Transgenic ,Adaptive Immunity ,Biology ,Choline O-Acetyltransferase ,Mice ,Immune system ,Pregnancy ,Immunity ,Internal medicine ,medicine ,Animals ,Lymphocytes ,education ,Mice, Knockout ,B-Lymphocytes ,education.field_of_study ,Multidisciplinary ,Innate immune system ,Macrophages ,Toll-Like Receptors ,Dendritic Cells ,Biological Sciences ,Acquired immune system ,Choline acetyltransferase ,Acetylcholine ,Immunity, Innate ,Receptors, Neurotransmitter ,Cell biology ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Endocrinology ,Myeloid Differentiation Factor 88 ,Metagenome ,Female ,Cell activation ,Signal Transduction - Abstract
Appropriate control of immune responses is a critical determinant of health. Here, we show that choline acetyltransferase (ChAT) is expressed and ACh is produced by B cells and other immune cells that have an impact on innate immunity. ChAT expression occurs in mucosal-associated lymph tissue, subsequent to microbial colonization, and is reduced by antibiotic treatment. MyD88-dependent Toll-like receptor up-regulates ChAT in a transient manner. Unlike the previously described CD4 + T-cell population that is stimulated by norepinephrine to release ACh, ChAT + B cells release ACh after stimulation with sulfated cholecystokinin but not norepinephrine. ACh-producing B-cells reduce peritoneal neutrophil recruitment during sterile endotoxemia independent of the vagus nerve, without affecting innate immune cell activation. Endothelial cells treated with ACh in vitro reduced endothelial cell adhesion molecule expression in a muscarinic receptor-dependent manner. Despite this ability, ChAT + B cells were unable to suppress effector T-cell function in vivo. Therefore, ACh produced by lymphocytes has specific functions, with ChAT + B cells controlling the local recruitment of neutrophils.
- Published
- 2013
39. The transcription factor interferon regulatory factor-1 is essential for natural killer cell function in vivo
- Author
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Gordon S. Duncan, Toshifumi Matsuyama, David Kägi, H W Mittrücker, and Tak W. Mak
- Subjects
Cytotoxicity, Immunologic ,Cell Transplantation ,Immunology ,Lymphocytic Choriomeningitis ,Lymphocyte Activation ,Mice ,Interleukin 21 ,Interferon ,MHC class I ,medicine ,Animals ,Immunology and Allergy ,Cytotoxic T cell ,Lectins, C-Type ,Antigens ,Lymphokine-activated killer cell ,biology ,H-2 Antigens ,Proteins ,Articles ,Interferon-beta ,Neoplasms, Experimental ,Phosphoproteins ,Interleukin-12 ,Mice, Mutant Strains ,DNA-Binding Proteins ,Killer Cells, Natural ,Mice, Inbred C57BL ,IRF1 ,Antigens, Surface ,Cancer research ,biology.protein ,Interleukin 12 ,Interleukin-2 ,NK Cell Lectin-Like Receptor Subfamily B ,Interferon Regulatory Factor-1 ,Transcription Factors ,medicine.drug - Abstract
The activation of natural killer (NK) cells, cytotoxic lymphocytes capable of major histocompatibility complex (MHC)-unrestricted killing and early antiviral defense, is temporally related to the increased interferon (IFN)-alpha/beta production that is seen in the viral infection of mice. Type I IFN (IFN-alpha/beta) are expressed in many cell types early after primary viral infection and have been shown to mediate resistance against a variety of viruses. In this study, the role of the transcriptional activator IFN regulatory factor-1 (IRF-1) in murine NK cell activity was assessed. IRF-1-deficient mice displayed a normal frequency of NK marker-positive cells, but exhibited greatly reduced NK cell-mediated cytotoxicity after both virus infection and stimulation with the IFN inducer polyinosinic:polycytidilic acid in vivo. In vitro, cytolytic activity in IRF-1-deficient NK cells remained defective after stimulation with IFN-beta, IL-2, and IL-12. IRF-1-deficient mice were unable to eliminate syngeneic MHC class I-negative tumor cells in vivo, and had a reduced ability to reject parental semi-allogeneic donor cells from the circulation. Thus, IRF-1 is essential for the induction of NK cell-mediated cytotoxicity and for the in vivo effector functions that are mediated by this activity.
- Published
- 1996
40. LFA-1-deficient mice show normal CTL responses to virus but fail to reject immunogenic tumor
- Author
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Tak W. Mak, Gordon S. Duncan, Martin F. Bachmann, Rudolf Schmits, Arda Shahinian, Andrew Wakeham, D. M. Baker, G. Shumaker, Pamela S. Ohashi, D. D. Hickstein, A. Van Der Heiden, Thomas M. Kündig, and John J.L. Simard
- Subjects
Integrins ,Lymphocyte ,Molecular Sequence Data ,Immunology ,Priming (immunology) ,chemical and pharmacologic phenomena ,CD18 ,Biology ,Vesicular stomatitis Indiana virus ,Mice ,Immune system ,Antigen ,medicine ,Animals ,Lymphocytic choriomeningitis virus ,Immunology and Allergy ,Cytotoxic T cell ,Lymphocyte function-associated antigen 1 ,Base Sequence ,Homozygote ,hemic and immune systems ,Articles ,Lymphocyte Function-Associated Antigen-1 ,Mice, Mutant Strains ,Mice, Inbred C57BL ,CTL ,medicine.anatomical_structure ,Mutagenesis ,Virus Diseases ,CD18 Antigens ,T-Lymphocytes, Cytotoxic - Abstract
The leukocyte integrin LFA-1 (CD11a/CD18) plays an important role in lymphocyte recirculation and homotypic interactions. Leukocytes from mice lacking CD11a displayed defects in in vitro homotypic aggregation, in proliferation in mixed lymphocyte reactions, and in response to mitogen. Mutant mice mounted normal cytotoxic T cell (CTL) responses against systemic LCMV and VSV infections and showed normal ex vivo CTL function. However, LFA-1-deficient mice did not reject immunogenic tumors grafted into footpads and did not demonstrate priming response against tumor-specific antigen. Thus CD11a deficiency causes a selective defect in induction of peripheral immune responses whereas responses to systemic infection are normal.
- Published
- 1996
41. iRhom2 Regulation of TACE Controls TNF-Mediated Protection Against Listeria and Responses to LPS
- Author
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Rama Khokha, Dieter Häussinger, Tak W. Mak, Philipp A. Lang, Carl P. Blobel, Kazuhito Ohishi, Koichi Hamada, Thorsten Maretzky, Pamela S. Ohashi, David R. McIlwain, Sathish Kumar Maney, Haifeng C. Xu, Gordon S. Duncan, Thorsten Berger, Aditya Murthy, Annick Itie-Youten, Andrew Wakeham, and Karl S. Lang
- Subjects
Lipopolysaccharides ,Lipopolysaccharide ,Cell ,Medizin ,Biology ,ADAM17 Protein ,Article ,Sepsis ,chemistry.chemical_compound ,Inactive Rhomboid Protein 2 ,Immunity ,medicine ,Animals ,Humans ,Listeriosis ,Multidisciplinary ,Innate immune system ,Septic shock ,Tumor Necrosis Factor-alpha ,medicine.disease ,Shock, Septic ,Immunity, Innate ,ADAM Proteins ,medicine.anatomical_structure ,chemistry ,Immunology ,Tumor necrosis factor alpha ,Carrier Proteins ,Signal Transduction - Abstract
TACE Trafficking The cytokine tumor necrosis factor (TNF) is a major driver of inflammation and contributes to the immune pathology seen in a variety of diseases, including inflammatory bowel disease, rheumatoid arthritis, and sepsis. Soluble TNF is produced by cleavage of its ectodomain by the ADAM family metalloprotease, TNFα-converting enzyme (TACE). However, the molecular regulation of TACE is not understood (see the Perspective by Lichtenthaler ). Adrain et al. (p. 225 ) and McIlwain et al. (p. 229 ) now show that the rhomboid family member iRhom2 interacts with TACE in macrophages and is required for its proper intracellular trafficking and activation. In the absence of iRhom2, TACE was not released from the endoplasmic reticulum, and active protease did not reach the cell surface. Because of an inability to produce TNF, iRhom2-deficient mice were more resistant to lipopolysaccharide-induced septic shock but could not adequately control a Listeria monocytogenes infection.
- Published
- 2012
42. The interaction between caveolin-1 and Rho-GTPases promotes metastasis by controlling the expression of alpha5-integrin and the activation of Src, Ras and Erk
- Author
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Tak W. Mak, M. Quintela-Fandino, Amber Ablack, Michael W. Tusche, Alessandro Rufini, E Arpaia, Pamela S. Ohashi, Anne Brüstle, Heiko Blaser, John D. Lewis, Gordon S. Duncan, C K Fleming, Hon S. Leong, Jennifer Silvester, Shruti Nambiar, and Evan F. Lind
- Subjects
MAPK/ERK pathway ,rho GTP-Binding Proteins ,Cancer Research ,RHOA ,Caveolin 1 ,RhoC ,Chick Embryo ,Integrin alpha5 ,Mice, SCID ,Metastasis ,Mice ,0302 clinical medicine ,Cell Movement ,Neoplasm Metastasis ,Phosphorylation ,Extracellular Signal-Regulated MAP Kinases ,0303 health sciences ,biology ,Cell migration ,3. Good health ,Cell biology ,src-Family Kinases ,rhoC GTP-Binding Protein ,030220 oncology & carcinogenesis ,RNA Interference ,Original Article ,Proto-oncogene tyrosine-protein kinase Src ,Protein Binding ,Src ,caveolin-1 ,Immunoblotting ,Molecular Sequence Data ,RAC1 ,03 medical and health sciences ,Cell Line, Tumor ,intravital microscopy ,Genetics ,medicine ,Animals ,cancer ,Amino Acid Sequence ,Rho-GTPase ,Molecular Biology ,030304 developmental biology ,Sequence Homology, Amino Acid ,Neoplasms, Experimental ,medicine.disease ,Enzyme Activation ,Mice, Inbred C57BL ,Crk-Associated Substrate Protein ,alpha5-integrin ,Cancer cell ,biology.protein ,Cancer research ,ras Proteins - Abstract
Proteins containing a caveolin-binding domain (CBD), such as the Rho-GTPases, can interact with caveolin-1 (Cav1) through its caveolin scaffold domain. Rho-GTPases are important regulators of p130(Cas), which is crucial for both normal cell migration and Src kinase-mediated metastasis of cancer cells. However, although Rho-GTPases (particularly RhoC) and Cav1 have been linked to cancer progression and metastasis, the underlying molecular mechanisms are largely unknown. To investigate the function of Cav1-Rho-GTPase interaction in metastasis, we disrupted Cav1-Rho-GTPase binding in melanoma and mammary epithelial tumor cells by overexpressing CBD, and examined the loss-of-function of RhoC in metastatic cancer cells. Cancer cells overexpressing CBD or lacking RhoC had reduced p130(Cas) phosphorylation and Rac1 activation, resulting in an inhibition of migration and invasion in vitro. The activity of Src and the activation of its downstream targets FAK, Pyk2, Ras and extracellular signal-regulated kinase (Erk)1/2 were also impaired. A reduction in α5-integrin expression, which is required for binding to fibronectin and thus cell migration and survival, was observed in CBD-expressing cells and cells lacking RhoC. As a result of these defects, CBD-expressing melanoma cells had a reduced ability to metastasize in recipient mice, and impaired extravasation and survival in secondary sites in chicken embryos. Our data indicate that interaction between Cav1 and Rho-GTPases (most likely RhoC but not RhoA) promotes metastasis by stimulating α5-integrin expression and regulating the Src-dependent activation of p130(Cas)/Rac1, FAK/Pyk2 and Ras/Erk1/2 signaling cascades.
- Published
- 2011
43. Correction for Su et al., 14-3-3σ regulates B-cell homeostasis through stabilization of FOXO1
- Author
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Kazuo Yamamoto, Andrew Wakeham, Tak W. Mak, Philipp A. Lang, Gordon S. Duncan, Pamela S. Ohashi, Zhenyue Hao, Hiroki Yoshida, Kiichi Murakami, Heiko Hermeking, Ashley Young, Wen-Jye Lin, Atsushi Hirao, Yu-Wen Su, and Bert Vogelstein
- Subjects
Multidisciplinary ,B cell homeostasis ,FOXO1 ,Biology ,Corrections ,Cell biology - Published
- 2011
44. CARD6 is interferon inducible but not involved in nucleotide-binding oligomerization domain protein signaling leading to NF-kappaB activation
- Author
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Tak W. Mak, Almut Dufner, Alisha R. Elford, Håkan Hall, Pamela S. Ohashi, Gordon S. Duncan, and Andrew Wakeham
- Subjects
Heterozygote ,Microinjections ,Protein domain ,Mice, Transgenic ,Nod ,Biology ,Lymphocytic choriomeningitis ,Mice ,Downregulation and upregulation ,Interferon ,medicine ,Animals ,Molecular Biology ,Alleles ,Cells, Cultured ,Crosses, Genetic ,Embryonic Stem Cells ,Adaptor Proteins, Signal Transducing ,Mice, Knockout ,Recombination, Genetic ,NF-kappa B ,Cell Biology ,Articles ,Fibroblasts ,medicine.disease ,NFKB1 ,Embryo, Mammalian ,Molecular biology ,Recombinant Proteins ,Clone Cells ,Protein Structure, Tertiary ,CARD Signaling Adaptor Proteins ,Enzyme Activation ,Mice, Inbred C57BL ,Nod Signaling Adaptor Proteins ,Blastocyst ,Electroporation ,Mutation ,Interferons ,Signal transduction ,medicine.drug ,Signal Transduction - Abstract
We have previously reported the cloning and characterization of CARD6, a caspase recruitment domain (CARD)-containing protein that is structurally related to the interferon (IFN)-inducible GTPases. CARD6 associates with microtubules and with receptor-interacting protein 2 (RIP2). RIP2 mediates NF-kappaB activation induced by the intracellular nucleotide-binding oligomerization domain (NOD) receptors that sense bacterial peptidoglycan. Here we report that the expression of CARD6 and RIP2 in bone marrow-derived macrophages is rapidly induced by beta IFN and gamma IFN. This IFN-induced upregulation of CARD6 is suppressed by lipopolysaccharide (LPS), in contrast to LPS's enhancement of IFN-induced RIP2 upregulation. We generated CARD6-deficient (CARD6(-/-)) mice and carried out extensive analyses of signaling pathways mediating innate and adaptive immune responses, including the NOD pathways, but did not detect any abnormalities. Moreover, CARD6(-/-) mice were just as susceptible as wild-type mice to infection by Salmonella enterica serovar Typhimurium, Listeria monocytogenes, Candida albicans, lymphocytic choriomeningitis virus, or mouse adenovirus type 1. Thus, although structural and in vitro analyses strongly suggest an important role for CARD6 in immune defense, the physiological function of CARD6 remains obscure.
- Published
- 2007
45. Tissue inhibitor of metalloproteinase 3 regulates TNF-dependent systemic inflammation
- Author
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David S, Smookler, Fazilat F, Mohammed, Zamaneh, Kassiri, Gordon S, Duncan, Tak W, Mak, and Rama, Khokha
- Subjects
Inflammation ,Lipopolysaccharides ,Mice, Inbred C57BL ,Mice, Knockout ,Tissue Inhibitor of Metalloproteinase-3 ,Kinetics ,Mice ,Tumor Necrosis Factor-alpha ,Macrophages ,Animals ,Cells, Cultured ,Immunity, Innate ,Receptors, Tumor Necrosis Factor - Abstract
Host response to infectious agents must be rapid and powerful. One mechanism is the release of presynthesized membrane-bound TNF. TNF shedding is mediated by TNF-alpha converting enzyme, which is selectively inhibited by the tissue inhibitor of metalloproteinase 3 (TIMP3). We show that loss of TIMP3 impacts innate immunity by dysregulating cleavage of TNF and its receptors. Cultured timp3-/- macrophages release more TNF in response to LPS than wild-type macrophages. In timp3-/- mice, LPS causes serum levels of TNF and its receptors to rise more rapidly and remain higher compared with wild-type mice. The altered kinetics of ligand and receptor shedding enhances TNF signaling in timp3-/- mice, indicated by elevated serum IL-6. Physiologically, timp3-/- mice are more susceptible to LPS-induced mortality. Ablation of the TNF receptor gene p55 (Tnfrsf1a) or treatment with a synthetic metalloproteinase inhibitor rescues timp3-/- mice. Thus, TIMP3 is essential for normal innate immune function.
- Published
- 2006
46. RhoC is dispensable for embryogenesis and tumor initiation but essential for metastasis
- Author
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Rama Khokha, Annick You-Ten, Andrew Wakeham, Otto Sanchez-Sweatman, Gordon S. Duncan, Anne Hakem, and Tak W. Mak
- Subjects
rho GTP-Binding Proteins ,Stress fiber ,Cell Survival ,T-Lymphocytes ,RhoC ,Embryonic Development ,Mice, Transgenic ,GTPase ,Tumor initiation ,Lymphocyte Activation ,Metastasis ,Research Communications ,Mice ,Cell Movement ,Pregnancy ,Genetics ,medicine ,Animals ,Neoplasm Metastasis ,Mice, Knockout ,B-Lymphocytes ,biology ,Melanoma ,Cell migration ,Neoplasms, Experimental ,medicine.disease ,Mice, Inbred C57BL ,Cell Transformation, Neoplastic ,rhoC GTP-Binding Protein ,Immunology ,Cancer research ,biology.protein ,ras Proteins ,Female ,RhoC GTP-Binding Protein ,Developmental Biology - Abstract
The Rho proteins are Ras-related guanosine triphosphatases (GTPases) that function in cytoskeletal reorganization, cell migration, and stress fiber and focal adhesion formation. Overexpression of RhoC enhances the ability of melanoma cells to exit the blood and colonize the lungs. However, in vivo confirmation of RhoC's role in metastasis has awaited a RhoC-deficient mouse model. Here we report the generation of RhoC-deficient mice and show that RhoC is dispensable for embryonic and post-natal development. We demonstrate that loss of RhoC does not affect tumor development but decreases tumor cell motility and metastatic cell survival leading to a drastic inhibition of metastasis.
- Published
- 2005
47. Specific ablation of the apoptotic functions of cytochrome C reveals a differential requirement for cytochrome C and Apaf-1 in apoptosis
- Author
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Zhenyue Hao, Yingju Jang, Hitoshi Okada, Xiaodong Wang, Andrew Wakeham, Andrew E. H. Elia, Wen Chen Yeh, Gordon S. Duncan, Thomas Calzascia, Pamela S. Ohashi, Annick You-Ten, Tak W. Mak, Chia Che Chang, and Min Fang
- Subjects
Transgene ,T-Lymphocytes ,Mutant ,Caspase 1 ,Apoptosis ,Mice, Transgenic ,Biology ,Lymphocyte Activation ,Nervous System Malformations ,General Biochemistry, Genetics and Molecular Biology ,Electron Transport ,Mice ,Lymphocyte homeostasis ,Animals ,APAF1 ,Mice, Knockout ,Enzyme Precursors ,Biochemistry, Genetics and Molecular Biology(all) ,Cytochrome c ,Cytochromes c ,Proteins ,Mice, Mutant Strains ,Cell biology ,Enzyme Activation ,Apoptotic Protease-Activating Factor 1 ,Gamma Rays ,Caspases ,Immunology ,biology.protein ,Apoptosome - Abstract
SummaryAs components of the apoptosome, a caspase-activating complex, cytochrome c (Cyt c) and Apaf-1 are thought to play critical roles during apoptosis. Due to the obligate function of Cyt c in electron transport, its requirement for apoptosis in animals has been difficult to establish. We generated “knockin” mice expressing a mutant Cyt c (KA allele), which retains normal electron transfer function but fails to activate Apaf-1. Most KA/KA mice displayed embryonic or perinatal lethality caused by defects in the central nervous system, and surviving mice exhibited impaired lymphocyte homeostasis. Although fibroblasts from the KA/KA mice were resistant to apoptosis, their thymocytes were markedly more sensitive to death stimuli than Apaf-1−/− thymocytes. Upon treatment with γ irradiation, procaspases were efficiently activated in apoptotic KA/KA thymocytes, but Apaf-1 oligomerization was not observed. These studies indicate the existence of a Cyt c- and apoptosome-independent but Apaf-1-dependent mechanism(s) for caspase activation.
- Published
- 2004
48. The B7 family member B7-H3 preferentially down-regulates T helper type 1-mediated immune responses
- Author
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Stephen Chung, Woong-Kyung Suh, Edward M. Bertram, Tania H. Watts, Tom Horan, Joan da Costa, Mark R. Bray, Tak W. Mak, Suzanne Plyte, Beata U. Gajewska, Andrew E. H. Elia, Pauline Campbell, Pamela S. Ohashi, Matthew A. Gronski, Jacob Bukczynski, Sudha Arya, Kevin Gaida, Hitoshi Okada, Gordon S. Duncan, Steven Kiyoshi Yoshinaga, Manel Jordana, Andrew Wakeham, Annick Itie, and Wojciech Dawicki
- Subjects
B7 Antigens ,Encephalomyelitis, Autoimmune, Experimental ,T cell ,Immunology ,Receptors, Antigen, T-Cell ,Down-Regulation ,Biology ,Lymphocyte Activation ,Vesicular stomatitis Indiana virus ,Interleukin 21 ,Interferon-gamma ,Mice ,Immune system ,Antigen ,medicine ,Immunology and Allergy ,Animals ,Lymphocytic choriomeningitis virus ,Interleukin 4 ,Autoantibodies ,Mice, Knockout ,Experimental autoimmune encephalomyelitis ,T-cell receptor ,Dendritic Cells ,Th1 Cells ,medicine.disease ,Flow Cytometry ,Orthomyxoviridae ,Molecular biology ,Mice, Inbred C57BL ,medicine.anatomical_structure ,biology.protein ,B7-1 Antigen ,Interleukin-4 ,Antibody - Abstract
We investigated the in vivo function of the B7 family member B7-H3 (also known as B7RP-2) by gene targeting. B7-H3 inhibited T cell proliferation mediated by antibody to T cell receptor or allogeneic antigen-presenting cells. B7-H3-deficient mice developed more severe airway inflammation than did wild-type mice in conditions in which T helper cells differentiated toward type 1 (T(H)1) rather than type 2 (T(H)2). B7-H3 expression was consistently enhanced by interferon-gamma but suppressed by interleukin 4 in dendritic cells. B7-H3-deficient mice developed experimental autoimmune encephalomyelitis several days earlier than their wild-type littermates, and accumulated higher concentrations of autoantibodies to DNA. Thus, B7-H3 is a negative regulator that preferentially affects T(H)1 responses.
- Published
- 2003
49. Costimulation through the inducible costimulator ligand is essential for both T helper and B cell functions in T cell-dependent B cell responses
- Author
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Beata U. Gajewska, Matthew A. Gronski, Manel Jordana, Alexandra Ho, Arda Shahinian, Melania Pintilie, Bernhard Odermatt, Andrew Wakeham, John S Whorisky, Tak W. Mak, Friedhelm Bladt, Louis-Martin Boucher, Urs Eriksson, Anna Tafuri, Denis Bouchard, Tony Pawson, Steve K. Yoshinaga, Gordon S. Duncan, and Pamela S. Ohashi
- Subjects
Antigens, Differentiation, T-Lymphocyte ,Adoptive cell transfer ,T cell ,Immunology ,Lymphocyte Cooperation ,Biology ,Affinity maturation ,Inducible T-Cell Co-Stimulator Protein ,Inducible T-Cell Co-Stimulator Ligand ,Mice ,medicine ,Immunology and Allergy ,Animals ,B cell ,B-Lymphocytes ,T follicular helper cell differentiation ,Proteins ,T-Lymphocytes, Helper-Inducer ,Ligand (biochemistry) ,Cell biology ,ICOS LIGAND ,medicine.anatomical_structure ,Immunoglobulin G ,Gene Deletion - Abstract
Costimulation through the inducible costimulator (ICOS) and its ligand (ICOSL) is essential for T cell-dependent B cell responses, but the cellular and temporal dynamics underlying its in vivo effects are poorly defined. Here we have shown that Icosl(-/-) and Icos(-/-) mice had similar phenotypes and that ICOS-ICOSL costimulation modulated the early but not late phases of IgG1 affinity maturation. Exploiting the adoptive transfer of T or B cells from primed Icosl(-/-) mice, we provided genetic evidence that costimulation through ICOSL was essential for primary but not secondary helper T cell responses and for the control of both T and B cell activities, resulting in T cell-dependent IgG1 production.
- Published
- 2002
50. Chk2 is a tumor suppressor that regulates apoptosis in both an ataxia telangiectasia mutated (ATM)-dependent and an ATM-independent manner
- Author
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Jorge A. Wong, Stephen J. Elledge, Scott W. Lowe, Elisa de Stanchina, Robert G. Bristow, Andrew J. Elia, Tak W. Mak, Penny A. Jeggo, Takashi Sakai, Alison Cheung, Toshio Suda, Gordon S. Duncan, Atsushi Hirao, Pierre Marie Girard, Hitoshi Okada, Andrew Wakeham, and Talin Sarkissian
- Subjects
CD4-Positive T-Lymphocytes ,Male ,Cell cycle checkpoint ,Time Factors ,Apoptosis ,Cell Cycle Proteins ,Ataxia Telangiectasia Mutated Proteins ,CD8-Positive T-Lymphocytes ,medicine.disease_cause ,environment and public health ,Mice ,Tissue Distribution ,Phosphorylation ,Checkpoint Kinase 2 ,Cell Growth and Development ,Mice, Knockout ,Alanine ,Cell Cycle ,Cell cycle ,DNA-Binding Proteins ,Female ,biological phenomena, cell phenomena, and immunity ,Protein Binding ,animal structures ,9,10-Dimethyl-1,2-benzanthracene ,Thymus Gland ,Biology ,Protein Serine-Threonine Kinases ,Models, Biological ,medicine ,Animals ,Molecular Biology ,Models, Genetic ,Tumor Suppressor Proteins ,Dose-Response Relationship, Radiation ,Cell Biology ,Neoplasms, Experimental ,medicine.disease ,Genes, p53 ,enzymes and coenzymes (carbohydrates) ,Ataxia-telangiectasia ,Mutation ,Cancer research ,Carcinogens ,Tumor Suppressor Protein p53 ,Carcinogenesis ,Ataxia telangiectasia and Rad3 related ,Protein Kinases - Abstract
In response to ionizing radiation (IR), the tumor suppressor p53 is stabilized and promotes either cell cycle arrest or apoptosis. Chk2 activated by IR contributes to this stabilization, possibly by direct phosphorylation. Like p53, Chk2 is mutated in patients with Li-Fraumeni syndrome. Since the ataxia telangiectasia mutated (ATM) gene is required for IR-induced activation of Chk2, it has been assumed that ATM and Chk2 act in a linear pathway leading to p53 activation. To clarify the role of Chk2 in tumorigenesis, we generated gene-targeted Chk2-deficient mice. Unlike ATM(-/-) and p53(-/-) mice, Chk2(-/-) mice do not spontaneously develop tumors, although Chk2 does suppress 7,12-dimethylbenzanthracene-induced skin tumors. Tissues from Chk2(-/-) mice, including those from the thymus, central nervous system, fibroblasts, epidermis, and hair follicles, show significant defects in IR-induced apoptosis or impaired G(1)/S arrest. Quantitative comparison of the G(1)/S checkpoint, apoptosis, and expression of p53 proteins in Chk2(-/-) versus ATM(-/-) thymocytes suggested that Chk2 can regulate p53-dependent apoptosis in an ATM-independent manner. IR-induced apoptosis was restored in Chk2(-/-) thymocytes by reintroduction of the wild-type Chk2 gene but not by a Chk2 gene in which the sites phosphorylated by ATM and ataxia telangiectasia and rad3(+) related (ATR) were mutated to alanine. ATR may thus selectively contribute to p53-mediated apoptosis. These data indicate that distinct pathways regulate the activation of p53 leading to cell cycle arrest or apoptosis.
- Published
- 2002
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