152 results on '"Osami Kanagawa"'
Search Results
2. Vav links the T cell antigen receptor to the actin cytoskeleton and T cell activation independently of intrinsic Guanine nucleotide exchange activity.
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Ana V Miletic, Daniel B Graham, Kumiko Sakata-Sogawa, Michio Hiroshima, Michael J Hamann, Saso Cemerski, Tracie Kloeppel, Daniel D Billadeau, Osami Kanagawa, Makio Tokunaga, and Wojciech Swat
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Medicine ,Science - Abstract
BACKGROUND:T cell receptor (TCR) engagement leads to formation of signaling microclusters and induction of rapid and dynamic changes in the actin cytoskeleton, although the exact mechanism by which the TCR initiates actin polymerization is incompletely understood. The Vav family of guanine nucleotide exchange factors (GEF) has been implicated in generation of TCR signals and immune synapse formation, however, it is currently not known if Vav's GEF activity is required in T cell activation by the TCR in general, and in actin polymerization downstream of the TCR in particular. METHODOLOGY/PRINCIPAL FINDINGS:Here, we report that Vav1 assembles into signaling microclusters at TCR contact sites and is critical for TCR-initiated actin polymerization. Surprisingly, Vav1 functions in TCR signaling and Ca(++) mobilization via a mechanism that does not appear to strictly depend on the intrinsic GEF activity. CONCLUSIONS/SIGNIFICANCE:We propose here a model in which Vav functions primarily as a tyrosine phosphorylated linker-protein for TCR activation of T cells. Our results indicate that, contrary to expectations based on previously published studies including from our own laboratory, pharmacological inhibition of Vav1's intrinsic GEF activity may not be an effective strategy for T cell-directed immunosuppressive therapy.
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- 2009
- Full Text
- View/download PDF
3. Construction of an open-access database that integrates cross-reference information from the transcriptome and proteome of immune cells.
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Atsushi Hijikata, Hiroshi Kitamura, Yayoi Kimura, Ryo Yokoyama, Yuichi Aiba, Yanyuan Bao, Shigeharu Fujita, Koji Hase, Shohei Hori, Yasuyuki Ishii, Osami Kanagawa, Hiroshi Kawamoto, Kazuya Kawano, Haruhiko Koseki, Masato Kubo, Ai Kurita-Miki, Tomohiro Kurosaki, Kyoko Masuda, Mitsumasa Nakata, Keisuke Oboki, Hiroshi Ohno, Mariko Okamoto, Yoshimichi Okayama, Jiyang O-Wang, Hirohisa Saito, Takashi Saito, Machie Sakuma, Katsuaki Sato, Kaori Sato, Ken-ichiro Seino, Ruka Setoguchi, Yuki Tamura, Masato Tanaka, Masaru Taniguchi, Ichiro Taniuchi, Annabelle Teng, Takeshi Watanabe, Hiroshi Watarai, Sho Yamasaki, and Osamu Ohara
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- 2007
- Full Text
- View/download PDF
4. Massive clonal expansion of polycytotoxic skin and blood CD8
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Axel Patrice, Villani, Aurore, Rozieres, Benoît, Bensaid, Klara Kristin, Eriksson, Amandine, Mosnier, Floriane, Albert, Virginie, Mutez, Océane, Brassard, Tugba, Baysal, Mathilde, Tardieu, Omran, Allatif, Floriane, Fusil, Thibault, Andrieu, Denis, Jullien, Valérie, Dubois, Catherine, Giannoli, Henri, Gruffat, Marc, Pallardy, François-Loïc, Cosset, Audrey, Nosbaum, Osami, Kanagawa, Janet L, Maryanski, Daniel, Yerly, Jean-François, Nicolas, and Marc, Vocanson
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integumentary system ,musculoskeletal, neural, and ocular physiology ,Immunology ,Receptors, Antigen, T-Cell ,SciAdv r-articles ,macromolecular substances ,CD8-Positive T-Lymphocytes ,Clone Cells ,Immunophenotyping ,nervous system ,Stevens-Johnson Syndrome ,Humans ,Prospective Studies ,Research Articles ,Research Article - Abstract
This study highlights the key role of polycytotoxic CD8+ T cells in the severity of toxic epidermal necrolysis., Toxic epidermal necrolysis (TEN) is a life-threatening cutaneous adverse drug reaction. To better understand why skin symptoms are so severe, we conducted a prospective immunophenotyping study on skin and blood. Mass cytometry results confirmed that effector memory polycytotoxic CD8+ T cells (CTLs) are the main leucocytes in TEN blisters at the acute phase. Deep T cell receptor (TCR) repertoire sequencing identified massive expansion of unique CDR3 clonotypes in blister cells. The same clones were highly expanded in patient’s blood, and the degree of their expansion showed significant correlation with disease severity. By transducing α and β chains of the expanded clonotypes into a TCR-defective cell line, we confirmed that those cells were drug specific. Collectively, these results suggest that the relative clonal expansion and phenotype of skin-recruited CTLs condition the clinical presentation of cutaneous adverse drug reactions.
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- 2020
5. Nitric oxide production in islets from nonobese diabetic mice: aminoguanidine-sensitive and -resistant stages in the immunological diabetic process
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Corbett, John A., Mikhael, Anwar, Jun Shimizu, Frederick, Katherine, Misko, Thomas P., McDaniel, Michael L., Osami Kanagawa, and Unanue, Emil R.
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Nitric oxide -- Analysis ,Islands of Langerhans -- Physiological aspects ,Aminoguanidine -- Analysis ,Immunological research -- Observations ,Science and technology - Abstract
Production of nitric oxide (NO) in nonobese diabetic islets of diabetic female mice is attributed to immunological diabetogenic events. Pharmacological function of the drug aminoguanidine, an inactivator of NO synthase during the diabetic process, is investigated. Inhibitory role of aminoguanidine is a function of inhibitor levels and quantity of spleen cells transported during the diabetic process. The action of the drug leads to a decline in the formation of NO, and postpones the onset of diabetes by three to eight days.
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- 1993
6. Involvement of Receptor Activator of Nuclear Factor-κB Ligand (RANKL)-induced Incomplete Cytokinesis in the Polyploidization of Osteoclasts
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Michio Tomura, Masaru Ishii, Yongwon Choi, Noriko Takegahara, Hyunsoo Kim, Atsushi Miyawaki, Asako Sakaue-Sawano, Hiroki Mizuno, and Osami Kanagawa
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cell division ,Male ,0301 basic medicine ,Cell division ,Osteoclasts ,Osteolysis ,Biochemistry ,Cell Fusion ,0302 clinical medicine ,Osteogenesis ,Phosphorylation ,Cells, Cultured ,Cell fusion ,biology ,imaging ,food and beverages ,pathological conditions, signs and symptoms ,Cell cycle ,Cell biology ,medicine.anatomical_structure ,RANKL ,030220 oncology & carcinogenesis ,osteoclast ,musculoskeletal diseases ,Recombinant Fusion Proteins ,Bone Marrow Cells ,Mice, Transgenic ,Polyploidy ,03 medical and health sciences ,Multinucleate ,Osteoclast ,Quinoxalines ,medicine ,Animals ,Molecular Biology ,Crosses, Genetic ,Myeloid Progenitor Cells ,Cytokinesis ,Cell Nucleus ,incomplete cytokinesis ,Cell growth ,flow cytometry ,RANK Ligand ,fungi ,Cell Biology ,Luminescent Proteins ,cell proliferation ,030104 developmental biology ,biology.protein ,Cancer research ,Benzimidazoles ,Protein Processing, Post-Translational ,Proto-Oncogene Proteins c-akt ,Biomarkers - Abstract
Osteoclasts are specialized polyploid cells that resorb bone. Upon stimulation with receptor activator of nuclear factor-κB ligand (RANKL), myeloid precursors commit to becoming polyploid, largely via cell fusion. Polyploidization of osteoclasts is necessary for their bone-resorbing activity, but the mechanisms by which polyploidization is controlled remain to be determined. Here, we demonstrated that in addition to cell fusion, incomplete cytokinesis also plays a role in osteoclast polyploidization. In in vitro cultured osteoclasts derived from mice expressing the fluorescent ubiquitin-based cell cycle indicator (Fucci), RANKL induced polyploidy by incomplete cytokinesis as well as cell fusion. Polyploid cells generated by incomplete cytokinesis had the potential to subsequently undergo cell fusion. Nuclear polyploidy was also observed in osteoclasts in vivo, suggesting the involvement of incomplete cytokinesis in physiological polyploidization. Furthermore, RANKL-induced incomplete cytokinesis was reduced by inhibition of Akt, resulting in impaired multinucleated osteoclast formation. Taken together, these results reveal that RANKL-induced incomplete cytokinesis contributes to polyploidization of osteoclasts via Akt activation.
- Published
- 2016
7. Recirculating Memory T Cells Are a Unique Subset of CD4+ T Cells with a Distinct Phenotype and Migratory Pattern
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Andrew D. Luster, Osami Kanagawa, Shannon K. Bromley, Sha Yan, and Michio Tomura
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CD4-Positive T-Lymphocytes ,Interleukin 2 ,Receptors, CCR7 ,Pathology ,medicine.medical_specialty ,Lymphoid Tissue ,Ultraviolet Rays ,CD40 Ligand ,Immunology ,Population ,Receptors, Lymphocyte Homing ,Mice, Transgenic ,Biology ,Article ,Immunophenotyping ,Mice ,Antigen ,Antigens, CD ,Cell Movement ,T-Lymphocyte Subsets ,medicine ,Animals ,Immunology and Allergy ,Cytotoxic T cell ,Cell Lineage ,education ,Skin ,education.field_of_study ,CD40 ,integumentary system ,hemic and immune systems ,Molecular biology ,Mice, Inbred C57BL ,Luminescent Proteins ,Lymphatic system ,medicine.anatomical_structure ,Microscopy, Fluorescence ,Radiation Chimera ,biology.protein ,Interleukin-2 ,Lymph Nodes ,Immunologic Memory ,Memory T cell ,medicine.drug - Abstract
Several populations of memory T cells have been described that differ in their migration and function. In this study, we have identified a unique subset of memory T cells, which we have named recirculating memory T cells (TRCM). By exposing Kaede transgenic mouse skin to violet light, we tracked the fate of cutaneous T cells. One population of memory CD4+ T cells remained in the skin. A second population migrated from the skin into draining lymph nodes (LNs) in a CCR7-dependent manner. These migrating CD4+ T cells expressed a novel cell surface phenotype (CCR7int/+CD62LintCD69−CD103+/− E-selectin ligands+) that is distinct from memory T cell subsets described to date. Unlike memory T cell subsets that remain resident within tissues long-term, or that migrate either exclusively between lymphoid tissues or into peripheral nonlymphoid sites, CD4+ TRCM migrate from the skin into draining LNs. From the draining LNs, CD4+ TRCM reenter into the circulation, distal LNs, and sites of non-specific cutaneous inflammation. In addition, CD4+ TRCM upregulated CD40L and secreted IL-2 following polyclonal stimulation. Our results identify a novel subset of recirculating memory CD4+ T cells equipped to deliver help to both distal lymphoid and cutaneous tissues.
- Published
- 2013
8. Nuclear transferred embryonic stem cells for analysis of B1 B-lymphocyte development
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Osami Kanagawa, Mitsuyo Takase, Sidonia Fagarasan, Ryuji Iida, Mikako Maruya, Satoshi Nishigami, Atsushi Miyawaki, Teruhiko Wakayama, and Asako Sakaue-Sawano
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Nuclear Transfer Techniques ,Lymphocyte ,Cellular differentiation ,Immunology ,Receptors, Antigen, B-Cell ,Mice, Transgenic ,Cell Line ,Immunoglobulin kappa-Chains ,Mice ,Immunoglobulin lambda-Chains ,Antigen ,medicine ,Animals ,Immunology and Allergy ,Cell Lineage ,Gene Rearrangement, B-Lymphocyte ,Peritoneal Cavity ,Embryonic Stem Cells ,B-Lymphocytes ,Mice, Inbred BALB C ,CD40 ,biology ,Cell Differentiation ,General Medicine ,Embryonic stem cell ,Immunity, Innate ,Cell biology ,Mice, Inbred C57BL ,B-1 cell ,medicine.anatomical_structure ,Mice, Inbred DBA ,Cell culture ,biology.protein ,Immunoglobulin Heavy Chains ,Clone (B-cell biology) - Abstract
The transfer of nuclei of fully differentiated cells into enucleated oocytes is a well-recognized method for the generation of embryonic stem (ES) cells. Here, we demonstrate that nuclear transferred ES (NT-ES) cells can be established with high efficiency using innate-like B lymphocytes as donor cells. We established two mouse lines carrying rearranged immunoglobulin heavy and light chains using NT-ES cells containing nuclei from peritoneal cavity B1 cells. Analysis of B1 clone lines revealed that the B1-cell generation critically depends on the interaction between antigen (possibly self-antigen) and surface immunoglobulin, while the B1-cell maintenance requires the peritoneal environment. The B1-cell expansion takes place in spleen, and is held in check by competitor B2 cells. The results indicate that the NT-ES method could replace the transgenic or knock-in mouse approaches currently used to study the biology of cells that undergo somatic rearrangements of their antigen receptor genes.
- Published
- 2012
9. Activated regulatory T cells are the major T cell type emigrating from the skin during a cutaneous immune response in mice
- Author
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Gyohei Egawa, Jason G. Cyster, Herman Waldmann, Tetsuya Honda, Hideaki Tanizaki, Yoshiki Tokura, Osami Kanagawa, Yoshiki Miyachi, Kenji Kabashima, Takeshi Watanabe, Atsushi Otsuka, Michio Tomura, and Shohei Hori
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integumentary system ,Transgene ,T cell ,FOXP3 ,hemic and immune systems ,chemical and pharmacologic phenomena ,Endogeny ,General Medicine ,Biology ,Immune system ,medicine.anatomical_structure ,Downregulation and upregulation ,In vivo ,Immunology ,medicine ,Lymph - Abstract
Tregs play an important role in protecting the skin from autoimmune attack. However, the extent of Treg trafficking between the skin and draining lymph nodes (DLNs) is unknown. We set out to investigate this using mice engineered to express the photoconvertible fluorescence protein Kaede, which changes from green to red when exposed to violet light. By exposing the skin of Kaede-transgenic mice to violet light, we were able to label T cells in the periphery under physiological conditions with Kaede-red and demonstrated that both memory phenotype CD4+Foxp3- non-Tregs and CD4+Foxp3+ Tregs migrated from the skin to DLNs in the steady state. During cutaneous immune responses, Tregs constituted the major emigrants and inhibited immune responses more robustly than did LN-resident Tregs. We consistently observed that cutaneous immune responses were prolonged by depletion of endogenous Tregs in vivo. In addition, the circulating Tregs specifically included activated CD25hi Tregs that demonstrated a strong inhibitory function. Together, our results suggest that Tregs in circulation infiltrate the periphery, traffic to DLNs, and then recirculate back to the skin, contributing to the downregulation of cutaneous immune responses.
- Published
- 2016
10. Conjoint 3rd Australasian Wound & Tissue Repair Society and 9th Australasian Society for Dermatology Research Conference
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Osami Kanagawa, Penny E. Lovat, A. Anfosso, D. S. Hill, Nikolas K. Haass, and Ichiko Kinjyo
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Pathology ,medicine.medical_specialty ,business.industry ,Melanoma ,Endoplasmic reticulum ,medicine ,Unfolded protein response ,Cancer research ,Surgery ,Dermatology ,Cell cycle ,medicine.disease ,business - Published
- 2012
11. Vitamin A-dependent transcriptional activation of the nuclear factor of activated T cells c1 (NFATc1) is critical for the development and survival of B1 cells
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Osami Kanagawa, Teruhiko Wakayama, Keiichiro Suzuki, Sidonia Fagarasan, Mikako Maruya, Hanae Fujimoto, and Michio Miyajima
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Transcriptional Activation ,Transcription, Genetic ,Receptors, Retinoic Acid ,T-Lymphocytes ,CD1 ,Mice, Transgenic ,Cell Separation ,Mice ,Animals ,Vitamin A ,Antigen-presenting cell ,Cell Proliferation ,Interleukin 3 ,B-Lymphocytes ,Multidisciplinary ,CD40 ,NFATC Transcription Factors ,integumentary system ,biology ,ZAP70 ,Biological Sciences ,Flow Cytometry ,Natural killer T cell ,Molecular biology ,B-1 cell ,Gene Expression Regulation ,biology.protein ,Interleukin 12 ,Female - Abstract
B1 cells represent a distinct subset of B cells that produce most of the natural serum IgM and much of the gut IgA and function as an important component of early immune responses to pathogens. The development of B1 cells depends on the nuclear factor of activated T cells c1 (NFATc1), a transcription factor abundantly expressed by B1 cells but not by conventional B2 cells. However, the factors that regulate the expression of NFATc1 in B1 cells remain unknown. Here we show that a vitamin A-deficient diet results in reduction of NFATc1 expression in B1 cells and almost complete loss of the B1 cell compartment. As a consequence, vitamin A-deficient mice have reduced serum IgM and are unable to mount T cell-independent antibody responses against bacterial antigens. We demonstrate that injection of all-trans retinoic acid induces the expression of NFATc1, particularly from the constitutive P2 promoter, and leads to the increase of the B1 cells. Thus, the retinoic acid-dependent pathway is critical for regulating NFATc1 expression and for maintenance of the natural memory B cell compartment.
- Published
- 2010
12. No Requirement of Trans Presentations of IL-15 for Human CD8 T Cell Proliferation
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Hidemi Uchiyama, Shaun K. Olsen, Mitsuyo Takase, Osami Kanagawa, and Naruhisa Ota
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T cell ,Immunology ,Antigen presentation ,CD1 ,Mice, Transgenic ,Cell Separation ,CD8-Positive T-Lymphocytes ,Biology ,Lymphocyte Activation ,Polymerase Chain Reaction ,Mice ,Interleukin 21 ,Interleukin-15 Receptor alpha Subunit ,medicine ,Animals ,Humans ,Immunology and Allergy ,Cytotoxic T cell ,IL-2 receptor ,Antigen-presenting cell ,Cell Proliferation ,Interleukin-15 ,ZAP70 ,Flow Cytometry ,Cell biology ,medicine.anatomical_structure - Abstract
The trans presentation of IL-15 by cells expressing the specific high-affinity receptor α-chain (IL-15Rα) to cells expressing the signaling receptor β-chain and γ-chain is essential for the generation and maintenance of CD8 memory T cells, NK cells, and NKT cells in an in vivo mouse system. We have also demonstrated in vitro that cell-surface IL-15Rα on cells expressing all the receptor components present IL-15 to receptor β-chain/γ-chain coexpressed on the same cell surface (cis presentation). However, although mouse CD8 T cells express all the IL-15R components, they show no evidence of cis presentation. In this study, we demonstrate that increased expression of mouse IL-15Rα in mouse CD8 T cells by retrovirus-mediated gene transfer changes the ability of the T cell to use cis presentation on the cell surface, indicating that cis presentation requires high expression of mouse IL-15Rα on the cell surface. Using cell lines expressing human or mouse receptors, we demonstrate that cis presentation occurs more efficiently in the human receptor–ligand combination than in that of the mouse system. Moreover, we found that primary human CD8 T cells do not require trans presentation of human IL-15 in vitro. These findings raise the possibility that the maintenance and generation of memory CD8 T cells are achieved via distinct mechanisms in humans and mice. Therefore, careful study of the human immune system, rather than extrapolation from the murine model, is necessary to achieve more complete understanding of memory CD8 T cell development in humans.
- Published
- 2010
13. Regulatory T cells (PP-045)
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E. Tasaki, Y. Okuno, E. M. Bertram, H. Isaka, S. Delbauve, T. Nakashima, K. A. Newell, J. C. Antvorskov, H. Fan, S. Kawashima, G. Ying, D. Horwitz, T. Thomas, M. Feuerer, Akihiko Yoshimura, C. Elly, K. Wang, T. Kanzaki, K. Buschard, T. P. Arstila, T. Huenig, F. Mair, N. Perdue, J. Sprent, L. Lu, L. Akahira, Y. Yoshida, H. Feng, W. Wang, Howard L. Weiner, Thorsten Buch, T. Laurinolli, D. Lim, F. X. Qin, X. Cao, J. Bártová, Y. m. Cao, I. R. van Driel, S. Hong, C. Tsao, L. M. Kastner, E. M. Ross, J. Wang, T. Honda, S. Park, Y. Chung, S. Schwele, R. Haque, B. Li, T. Yoshikawa, Matthew C. Cook, B. H. Hahn, M. G. Netea, Aleksandar Bulog, I. Iwamoto, J. Liu, P. Fundova, M. Kibata, Y. Lee, C. C. Goodnow, F. Conti, J. S. LeMasurier, Jeffrey A. Bluestone, J. Kie, K. Soejima, N. Erfani, X. R. Zhou, R. J. Steptoe, I. Kim, M. A. Fernandez, C. S. Constantinescu, S. Bauchiero, X. Jin, D. S. Y. Tan, Holly A. Bolton, S. Hu, L. R. Wedderburn, V. Flamand, Z. Liu, K. Sugimura, S. Kunkel, J. M. Rolland, Takatoku Oida, T. Ho, Tobias Bopp, L. Zheng, Y. Li, S. Shen, K. Michishita, M. Shin, R. Germain, Y. Tang, S. Klein-Hessling, Q. h. Wang, X. P. Peng, S. Nakamura, Daniel J. Campbell, Irma Joosten, X. Jiang, M. Braitch, Z. Yao, Masahide Hamaguchi, V. Pancre, H. Lee, Y. Zheng, Y. Cui, W. Müller, R. White, H. Yue, Ai Harashima, J. Perheentupa, M. Tomura, A. T. Endharti, S. Gattenloehner, Kenji Kabashima, M. Goldman, Z. Sarraf, Mihoko Shibuya, T. Sasaki, Y. Kanno, M. Yamamoto, J. Lee, G. Y. Zhou, A. M. Fischer, Y. Kwon, Christophe Benoist, Z. Chen, M. Nakatsugawa, Y. Liu, Y. Park, K. Yamamoto, Dirk H. Busch, C. S. Constantinecsu, T. Hamamoto, A. Carpentier, Y. Zhou, Hans-Dieter Volk, Tomohisa Okamura, J. Tsukada, K. Webster, E. Rabellino, I. Debock, F. Lin, Vladimir Mićović, T. Wakayama, Josef Bodor, H. Bae, G. Metzner, A. Kwan, Ari Waisman, C. Pecli, C. Wu, J. Song, S. Imoto, S. Han, H. Tanizaki, M. Mariotti-Ferrandiz, J. Paik, S. M. Salonen, K. Kawahata, B. Adams, M. H. Nyirenda, C. L. Hardy, K. Isobe, L. M. Lu, J. R. Killebrew, O. Morales, D. P. Funda, A. R. Kendal, A. McNally, R. A. DePinho, Ryuji Iida, N. Sasaki, S. Kim, N. Yang, L. H. Rossi, G. Brestrich, L. Hsu, N. Wada, C. Lu, C. B. Schmidt-Weber, V. Seyfert-Margolis, A. J. van der ven, T. Nakatsura, V. Nikolaev, H. Collins, A. R. Kitching, Carla Jones, B. Yen, Z. J. Jiao, C. Chan, T. Nakagaki, M. Takeuchi, Hirofumi Shoda, G. Lee, G. Wang, T. V. Hogan, M. Plebanski, F. Stenard, F. Liu, A. Suto, T. Otani, K. Tsuji-Takayama, D. E. Furst, Y. Takasaki, Tim Sparwasser, A. Tanaka, M. Asakawa, J. H. Louie, D. Brand, D. W. Dwyer, S. Nakano, A. Okamoto, M. Imamura, L. Chi, C. Zhu, J. Zhu, P. Roubal, S. Dharancy, M. Dohi, E. Kekäläinen, S. I. Alexander, H. Kojima, Dong Han, Q. Wang, N. Delhem, K. Takahashi, D. M. Heery, J. Sun, N. Sakemura, B. Gran, T. Toraya, A. Feng, X. Shen, C. A. Garcia Santana, H. Itoh, S. Zheng, N. MacDonald, U. Behn, A. Facciabene, B. Chiang, W. Kastenmuller, M. W. Leung, Shuji Sumitomo, X. Qu, S. Huang, R. Molinaro, A. Kokešová, M. Bozza, S. Z. Josefowicz, K. Bourcier, D. Bourges, K. Hirose, S. Rahimifar, F. Yamasaki, K. Nistala, E. Serfling, E. Schmitt, Friederike Berberich-Siebelt, Y. Wang, G. Coukos, F. Gross, I. Drexler, L. Yang, T. Gogishvili, S. Kagami, Y. Harada, H. Nakajima, E. Market, Y. Chen, A. Sledzinska, Lin Tian, Amy L. Putnam, F. Cunha, Anthony E.J. Dubois, M. I. Greene, S. Kamei, J. J. Lafaille, D. Kolodin, P. A. Gleeson, Ranjeny Thomas, W. Bolton, Xiao-Sen Li, I. Osawa, C. Miroux, Y. Kang, J. Drbohlav, A. Chaudhry, C. F. Benjamim, C. Kang, A. Ghaderi, G. Zhang, R. P. Singh, M. Hong, S. R. Holdsworth, L. Sanvito, L. Wang, J. j. Wu, T. Kobata, M. Kuo, B. Kim, M. Michels, A. I. Kokaji, Z. Li, W. Shi, J. Yao, X. Xia, W. Chang, S. Choi, R. Nakagawa, Y. y. Pan, M. Vaeth, C. Canetti, F. Lei, P. Hsu, J. Alves-Filho, S. Hori, A. Šalaková, B. L. Wang, J. Wilkinson, P. Zdziarski, M. Fairhurst, Keishi Fujio, X. Zheng, H. Suzuki, R. K. Dinesh, Shimon Sakaguchi, Z. Jiang, S. Hemmers, B Fazekas de St Groth, M. Zelman-Femiak, H. Moncrieffe, L. Han, J. Park, M. Suzuki, S. Eaton, D. Sakurai, T. Inoue, F. Lhommé, S. Nishimoto, Ines Mrakovčić-Šutić, R. Mohamud, Matthias Klein, S. D'Costa, R. E. O'Hehir, N. Morishima, J. Cha, M. Schmueck, M. Uchiyama, K. Imi, H. Hase, E. Boleslowski, A. Liston, C. Park, P. Reinke, C. Shen, H. Wakashin, D. A. Horwitz, Ohkura Naganari, Herman Waldmann, A. Okochi, A. Khoo, I. Kubajewska, J. Seoh, A. Roemhild, Alexander Y. Rudensky, Diane Mathis, J. A. Altin, C. Liu, M. Lohse, T. Shimada, B. Yang, W. Liu, L. Turka, M. Kweon, G. Gasteiger, Hans J. P. M. Koenen, Y. Kim, S. Okuda, Burkhard Becher, and Osami Kanagawa
- Subjects
Immunology ,Immunology and Allergy ,General Medicine - Published
- 2010
14. Epidermal γδ T cells sense precancerous cellular dysregulation and initiate immune responses
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Nobumichi Hozumi, Sachiko Hirose, Osami Kanagawa, Eri Shimura, Freddy Radtke, Daniel Metzger, Naoko Nakano, and Pierre Chambon
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Keratinocytes ,Receptors, CCR7 ,Chemokine ,Skin Neoplasms ,T-Lymphocytes ,T cell ,Immunology ,Biology ,Mice ,Immune system ,Antigen ,Antibody Specificity ,Cell Movement ,T-Lymphocyte Subsets ,medicine ,Animals ,Immunology and Allergy ,Receptor, Notch1 ,Lymph node ,Tissue homeostasis ,Mice, Knockout ,B-Lymphocytes ,integumentary system ,Receptors, Antigen, T-Cell, gamma-delta ,General Medicine ,T lymphocyte ,beta-Galactosidase ,Acquired immune system ,Cell biology ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Immunoglobulin G ,biology.protein ,Lymph Nodes ,Epidermis ,Precancerous Conditions - Abstract
Hyperplasia associated with a loss of tissue homeostasis can induce DNA replication stress, leading to precancerous dysregulation. Epidermal {gamma}{delta} T cells reside in the primary barrier that protects against diverse environmental insults; however, the functions of these T cells in tissue surveillance are not completely understood. In mice with inducible Notch1 inactivation in keratinocytes that causes epidermal hyperplasia, epidermal {gamma}{delta} T cells sensed stressed keratinocytes and migrated into the cutaneous draining lymph nodes. Simultaneous induction of β-galactosidase (β-Gal) as a putative antigen expressed in the process of precancerous dysregulation and Notch1 ablation in the epidermis resulted in elevated β-Gal-specific IgG2a production. Epidermal {gamma}{delta} T cells were found to have the capacity to express chemokine (C-C motif) receptor 7 and migrate into the lymph nodes. Cutaneous draining lymph node cells in Notch1-inactivated mice expressed high levels of IFN-{gamma} upon anti-CD3 plus anti-CD28 stimulation. Furthermore, induced expression of β-Gal in mice that lacked epidermal {gamma}{delta} T cells failed to induce anti-β-Gal IgG. These results suggest that epidermal {gamma}{delta} T cells play an essential role in the initiation process of epidermal antigen-specific humoral immune responses and demonstrate the importance of epidermal {gamma}{delta} T cells in sensing precancerous dysregulation and activating adaptive immunity.
- Published
- 2010
15. Critical role of IFN-γ in CFA-mediated protection of NOD mice from diabetes development
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Yoshiko Mori, Tetsuro Kodaka, Takako Kato, Osami Kanagawa, and Edith M. Kanagawa
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CD4-Positive T-Lymphocytes ,medicine.medical_specialty ,Adoptive cell transfer ,medicine.medical_treatment ,Freund's Adjuvant ,Immunology ,Spleen ,Nod ,Biology ,medicine.disease_cause ,T-Lymphocytes, Regulatory ,Autoimmunity ,Interferon-gamma ,Mice ,Adjuvants, Immunologic ,Mice, Inbred NOD ,Internal medicine ,medicine ,Animals ,Immunology and Allergy ,Receptor ,Receptors, Interferon ,NOD mice ,Mice, Knockout ,General Medicine ,Adoptive Transfer ,In vitro ,Diabetes Mellitus, Type 1 ,Endocrinology ,Cytokine ,medicine.anatomical_structure - Abstract
IFN-gamma signaling-deficient non-obese diabetic (NOD) mice develop diabetes with similar kinetics to those of wild-type NOD mice. However, the immunization of IFN-gamma signaling-deficient NOD mice with CFA failed to induce long-term protection, whereas wild-type NOD mice receiving CFA remained diabetes-free. CFA also failed to protect IFN-gamma receptor-deficient (IFN-gammaR(-/-)) NOD mice from the autoimmune rejection of transplanted islets, as it does in diabetic NOD mice, and from disease transfer by spleen cells from diabetic NOD mice. These data clearly show that the pro-inflammatory cytokine IFN-gamma is necessary for the CFA-mediated protection of NOD mice from diabetes. There is no difference in the T(h)1/T(h)17 balance between IFN-gammaR(-/-) NOD and wild-type NOD mice. There is also no difference in the total numbers and percentages of regulatory T (Treg) cells in the lymph node CD4(+) T-cell populations between IFN-gammaR(-/-) NOD and wild-type NOD mice. However, pathogenic T cells lacking IFN-gammaR are resistant to the suppressive effect of Treg cells, both in vivo and in vitro. Therefore, it is likely that CFA-mediated protection against diabetes development depends on a change in the balance between Treg cells and pathogenic T cells, and IFN-gamma signaling seems to control the susceptibility of pathogenic T cells to the inhibitory activity of Treg cells.
- Published
- 2009
16. Suppression of induced pluripotent stem cell generation by the p53–p21 pathway
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Keisuke Okita, Tomoko Ichisaka, Shinya Yamanaka, Kazutoshi Takahashi, Hyenjong Hong, Masato Nakagawa, Takashi Aoi, and Osami Kanagawa
- Subjects
Cell signaling ,Multidisciplinary ,SOX2 ,KLF4 ,Somatic cell ,Cellular differentiation ,Transfection ,Biology ,Induced pluripotent stem cell ,Embryonic stem cell ,Molecular biology ,Article - Abstract
Induced pluripotent stem (iPS) cells can be generated from somatic cells by the introduction of Oct3/4 (also known as Pou5f1), Sox2, Klf4 and c-Myc, in mouse and in human. The efficiency of this process, however, is low. Pluripotency can be induced without c-Myc, but with even lower efficiency. A p53 (also known as TP53 in humans and Trp53 in mice) short-interfering RNA (siRNA) was recently shown to promote human iPS cell generation, but the specificity and mechanisms remain to be determined. Here we report that up to 10% of transduced mouse embryonic fibroblasts lacking p53 became iPS cells, even without the Myc retrovirus. The p53 deletion also promoted the induction of integration-free mouse iPS cells with plasmid transfection. Furthermore, in the p53-null background, iPS cells were generated from terminally differentiated T lymphocytes. The suppression of p53 also increased the efficiency of human iPS cell generation. DNA microarray analyses identified 34 p53-regulated genes that are common in mouse and human fibroblasts. Functional analyses of these genes demonstrate that the p53-p21 pathway serves as a barrier not only in tumorigenicity, but also in iPS cell generation.
- Published
- 2009
17. Preferential Generation of Follicular B Helper T Cells from Foxp3 + T Cells in Gut Peyer's Patches
- Author
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Shohei Hori, Keiichiro Suzuki, Osami Kanagawa, Tasuku Honjo, Sidonia Fagarasan, Masayuki Tsuji, Noriko Komatsu, and Shimpei Kawamoto
- Subjects
Adoptive cell transfer ,Multidisciplinary ,CD40 ,biology ,T cell ,Follicular B helper T cells ,FOXP3 ,Germinal center ,T lymphocyte ,Cell biology ,Peyer Patch ,medicine.anatomical_structure ,Immunology ,biology.protein ,medicine - Abstract
Most of the immunoglobulin A (IgA) in the gut is generated by B cells in the germinal centers of Peyer's patches through a process that requires the presence of CD4 + follicular B helper T(T FH ) cells. The nature of these T FH cells in Peyer's patches has been elusive. Here, we demonstrate that suppressive Foxp3 + CD4 + T cells can differentiate into T FH cells in mouse Peyer's patches. The conversion of Foxp3 + T cells into T FH cells requires the loss of Foxp3 expression and subsequent interaction with B cells. Thus, environmental cues present in gut Peyer's patches promote the selective differentiation of distinct helper T cell subsets, such as T FH cells.
- Published
- 2009
18. Production of healthy cloned mice from bodies frozen at −20°C for 16 years
- Author
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Takamasa Iwaki, Teruhiko Wakayama, Sayaka Wakayama, Osami Kanagawa, Takafusa Hikichi, Hiroshi Ohta, and Eiji Mizutani
- Subjects
Cryopreservation ,Cloning ,Nuclear Transfer Techniques ,Multidisciplinary ,Cloning, Organism ,Brain ,Embryo ,Extinct species ,Biological Sciences ,Biology ,Molecular biology ,Embryonic stem cell ,Embryo Culture Techniques ,Mice ,Embryo Culture Technique ,Animals ,Reprogramming ,Embryonic Stem Cells - Abstract
Cloning animals by nuclear transfer provides an opportunity to preserve endangered mammalian species. However, it has been suggested that the “resurrection” of frozen extinct species (such as the woolly mammoth) is impracticable, as no live cells are available, and the genomic material that remains is inevitably degraded. Here we report production of cloned mice from bodies kept frozen at −20 °C for up to 16 years without any cryoprotection. As all of the cells were ruptured after thawing, we used a modified cloning method and examined nuclei from several organs for use in nuclear transfer attempts. Using brain nuclei as nuclear donors, we established embryonic stem cell lines from the cloned embryos. Healthy cloned mice were then produced from these nuclear transferred embryonic stem cells by serial nuclear transfer. Thus, nuclear transfer techniques could be used to “resurrect” animals or maintain valuable genomic stocks from tissues frozen for prolonged periods without any cryopreservation.
- Published
- 2008
19. Selective impairment of Fc RI-mediated allergic reaction in Gads-deficient mice
- Author
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Mitsuyo Takase-Utsugi, Machie Sakuma, Sho Yamasaki, Osami Kanagawa, Eri Ishikawa, Takashi Saito, and Keigo Nishida
- Subjects
endocrine system ,endocrine system diseases ,medicine.medical_treatment ,Immunology ,Immunoglobulin E ,Mice ,Immune system ,In vivo ,Hypersensitivity ,medicine ,Animals ,Immunology and Allergy ,Calcium Signaling ,Mast Cells ,B cell ,Adaptor Proteins, Signal Transducing ,Immunosuppression Therapy ,Mice, Knockout ,biology ,Receptors, IgE ,Chemistry ,Passive Cutaneous Anaphylaxis ,Toll-Like Receptors ,Degranulation ,nutritional and metabolic diseases ,General Medicine ,Mast cell ,Immunity, Innate ,Haematopoiesis ,Immunity, Active ,Cytokine ,medicine.anatomical_structure ,biology.protein - Abstract
Gads is a Grb2-like adaptor protein expressed in hematopoietic cells. We demonstrated that mast cells from Gads(-/-) mice have selective functional defects. Bone marrow-derived mast cells from Gads(-/-) mice failed to induce Ca(2+) mobilization, degranulation and cytokine production upon cross-linking of FcepsilonRI. In vivo passive cutaneous anaphylaxis was also greatly impaired in Gads(-/-) mice. In contrast, Gads was dispensable for Toll-like receptor-mediated cytokine production in mast cells. Accordingly, mast cell-dependent resistance to acute peritoneal bacterial infection is not reduced in Gads(-/-) mice in vivo. Moreover, mature T and B cell responses and antibody production upon immunization were apparently normal in Gads(-/-) mice. Thus, inhibition of Gads in vivo would suppress the IgE-mediated allergic reaction with minimum adverse effects on both innate and acquired immune responses, and Gads could be an ideal target for the control of allergic responses.
- Published
- 2008
20. Monitoring cellular movement in vivo with photoconvertible fluorescence protein 'Kaede' transgenic mice
- Author
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Yoshihiro Miwa, Osami Kanagawa, Satoshi Karasawa, Junko Tanaka, Atsushi Miyawaki, Michio Tomura, and Naoki Yoshida
- Subjects
Genetically modified mouse ,Multidisciplinary ,Lymphocyte ,Fluorescent Antibody Technique ,Mice, Transgenic ,Cell migration ,Biological Sciences ,Biology ,Flow Cytometry ,Cell biology ,Luminescent Proteins ,Mice ,Lymphatic system ,medicine.anatomical_structure ,Immune system ,Cell Movement ,In vivo ,Immune System ,medicine ,Animals ,Humans ,Kaede ,CD8 ,HeLa Cells - Abstract
Kaede is a photoconvertible fluorescence protein that changes from green to red upon exposure to violet light. The photoconversion of intracellular Kaede has no effect on cellular function. Using transgenic mice expressing the Kaede protein, we demonstrated that movement of cells with the photoconverted Kaede protein could be monitored from lymphoid organs to other tissues as well as from skin to the draining lymph node. Analysis of the kinetics of cellular movement revealed that each subset of cells in the lymph node, such as CD4 + T, CD8 + T, B, and dendritic cells, has a distinct migration pattern in vivo . Thus, the Kaede transgenic mouse system would be an ideal tool to monitor precise cellular movement in vivo at different stages of immune response to pathogens as well as in autoimmune diseases.
- Published
- 2008
21. Protection of IFN-γ signaling-deficient NOD mice from diabetes by cyclophosphamide
- Author
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Tetsuro Kodaka, Shohei Hori, Osami Kanagawa, Takako Kato, Yoshiko Mori, and Edith M. Kanagawa
- Subjects
Genetically modified mouse ,Cyclophosphamide ,Regulatory T cell ,Immunology ,Nod ,Biology ,T-Lymphocytes, Regulatory ,Diabetes Mellitus, Experimental ,Interferon-gamma ,Islets of Langerhans ,Mice ,Mice, Inbred NOD ,Diabetes mellitus ,medicine ,Animals ,Immunology and Allergy ,Beta (finance) ,NOD mice ,Mice, Knockout ,Pancreatic islets ,General Medicine ,medicine.disease ,Diabetes Mellitus, Type 1 ,medicine.anatomical_structure ,Immunosuppressive Agents ,medicine.drug - Abstract
Non-obese diabetic (NOD) mice that are genetically deficient in either IFN-gamma or beta chain of the IFN-gammaR develop diabetes with similar kinetics to wild-type NOD mice. In the current study, we demonstrated that treatment of IFN-gamma signaling-deficient NOD mice with cyclophosphamide (CY) not only fails to induce acute diabetes but also confers permanent protection from diabetes. Protection was mediated by the preferential generation of regulatory T cells (Treg cells) that are capable of suppressing the diabetogenic process, with no change in the total number and function of Treg cells. Moreover, CY treatment of IFN-gamma signaling-deficient NOD mice reversed the ongoing pathogenic process and eliminated cellular infiltrates of pancreatic islets. While these results have been derived using a genetically modified mouse model of diabetes, they indicate that knowledge of host genetic factors and environmental factors influencing the development of Type I diabetes mellitus may provide a rational approach to develop a means to reverse the development of Type I diabetes in human.
- Published
- 2008
22. Crystal Structure of the Interleukin-15·Interleukin-15 Receptor α Complex
- Author
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Hidemi Uchiyama, Mikako Shirouzu, Shaun K. Olsen, Osami Kanagawa, Shigeyuki Yokoyama, Kazutaka Murayama, Mutsuko Kukimoto-Niino, Takaho Terada, Mitsutoshi Toyama, Naruhisa Ota, and S. Kishishita
- Subjects
Sushi domain ,Molecular model ,Stereochemistry ,Interleukin ,Cell Biology ,Biology ,Acquired immune system ,Biochemistry ,Epitope ,Cell biology ,Interleukin-15 receptor ,Protein structure ,Interleukin 15 ,Molecular Biology - Abstract
Interleukin (IL)-15 is a pleiotropic cytokine that plays a pivotal role in both innate and adaptive immunity. IL-15 is unique among cytokines due to its participation in a trans signaling mechanism in which IL-15 receptorα (IL-15Rα) from one subset of cells presents IL-15 to neighboring IL-2Rβ/γc-expressing cells. Here we present the crystal structure of IL-15 in complex with the sushi domain of IL-15Rα. The structure reveals that theα receptor-binding epitope of IL-15 adopts a unique conformation, which, together with amino acid substitutions, permits specific interactions with IL-15Rα that account for the exceptionally high affinity of the IL-15·IL-15Rα complex. Interestingly, analysis of the topology of IL-15 and IL-15Rα at the IL-15·IL-15Rα interface suggests that IL-15 should be capable of participating in a cis signaling mechanism similar to that of the related cytokine IL-2. Indeed, we present biochemical data demonstrating that IL-15 is capable of efficiently signaling in cis through IL-15Rα and IL-2Rβ/γc expressed on the surface of a single cell. Based on our data we propose that cis presentation of IL-15 may be important in certain biological contexts and that flexibility of IL-15Rα permits IL-15 and its three receptor components to be assembled identically at the ligand-receptor interface whether IL-15 is presented in cis or trans. Finally, we have gained insights into IL-15·IL-15Rα·IL-2Rβ·γc quaternary complex assembly through the use of molecular modeling.
- Published
- 2007
23. Real-time tracking of cell cycle progression during CD8+ effector and memory T-cell differentiation
- Author
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Michio Tomura, Philip D. Hodgkin, Paulus Mrass, Atsushi Doi, Cameron J. Wellard, Osami Kanagawa, Sioh Yang Tan, Wolfgang Weninger, Asako Sakaue-Sawano, Ichiko Kinjyo, Atsushi Miyawaki, Jim S. Qin, William Ritchie, and Lois L. Cavanagh
- Subjects
Transgene ,Cellular differentiation ,General Physics and Astronomy ,Mice, Transgenic ,Editorials: Cell Cycle Features ,CD8-Positive T-Lymphocytes ,Biology ,Bioinformatics ,Article ,General Biochemistry, Genetics and Molecular Biology ,Immune system ,Genes, Reporter ,Memory cell ,Animals ,Multidisciplinary ,Effector ,Gene Expression Profiling ,Cell Cycle ,Cell Differentiation ,General Chemistry ,Cell cycle ,Phenotype ,3. Good health ,Cell biology ,Mice, Inbred C57BL ,Transcriptome ,CD8 - Abstract
The precise pathways of memory T-cell differentiation are incompletely understood. Here we exploit transgenic mice expressing fluorescent cell cycle indicators to longitudinally track the division dynamics of individual CD8+ T cells. During influenza virus infection in vivo, naive T cells enter a CD62Lintermediate state of fast proliferation, which continues for at least nine generations. At the peak of the anti-viral immune response, a subpopulation of these cells markedly reduces their cycling speed and acquires a CD62Lhi central memory cell phenotype. Construction of T-cell family division trees in vitro reveals two patterns of proliferation dynamics. While cells initially divide rapidly with moderate stochastic variations of cycling times after each generation, a slow-cycling subpopulation displaying a CD62Lhi memory phenotype appears after eight divisions. Phenotype and cell cycle duration are inherited by the progeny of slow cyclers. We propose that memory precursors cell-intrinsically modulate their proliferative activity to diversify differentiation pathways., CD8+ memory T cells appear during infection via a process of selection and differentiation that remains poorly understood. Using a fluorescent indicator of cell cycle progression, Kinjyo et al. show that slow-cycling memory precursors are derived from fast-cycling-activated T cells in influenza-infected mice.
- Published
- 2015
24. Antibody repertoire diversification through VH gene replacement in mice cloned from an IgA plasma cell
- Author
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Teruhiko Wakayama, Federica Mainoldi, Martina P. Bach, Mikako Maruya, Hassan Jumaa, Osami Kanagawa, Satoshi Kishigami, Rashmi Kumar, Sidonia Fagarasan, and Stefano Casola
- Subjects
Immunoglobulin A ,Multidisciplinary ,biology ,V(D)J recombination ,Gene rearrangement ,Plasma cell ,Molecular biology ,medicine.anatomical_structure ,Antibody Repertoire ,biology.protein ,medicine ,Immunoglobulin heavy chain ,Antibody ,Gene - Abstract
In mammals, VDJ recombination is responsible for the establishment of a highly diversified preimmune antibody repertoire. Acquisition of a functional Ig heavy (H) chain variable (V) gene rearrangement is thought to prevent further recombination at the IgH locus. Here, we describe VHQ52(NT); Vκgr32(NT) Ig monoclonal mice reprogrammed from the nucleus of an intestinal IgA(+) plasma cell. In VHQ52(NT) mice, IgA replaced IgM to drive early B-cell development and peripheral B-cell maturation. In VHQ52(NT) animals, over 20% of mature B cells disrupted the single productive, nonautoimmune IgH rearrangement through VH replacement and exchanged it with a highly diversified pool of IgH specificities. VH replacement occurred in early pro-B cells, was independent of pre-B-cell receptor signaling, and involved predominantly one adjacent VH germ-line gene. VH replacement was also identified in 5% of peripheral B cells of mice inheriting a different productive VH rearrangement expressed in the form of an IgM H chain. In summary, editing of a productive IgH rearrangement through VH replacement can account for up to 20% of the IgH repertoire expressed by mature B cells.
- Published
- 2015
25. NF-κB–inducing kinase controls lymphocyte and osteoclast activities in inflammatory arthritis
- Author
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Deborah V. Novack, Osami Kanagawa, Kunihiko Aya, Muhammad Alhawagri, Hideki Kitaura, and Amanda Hagen-Stapleton
- Subjects
Adoptive cell transfer ,DNA, Complementary ,Inflammatory arthritis ,Lymphocyte ,Antigen presentation ,Osteoclasts ,Arthritis ,Mice, Transgenic ,Inflammation ,Protein Serine-Threonine Kinases ,Biology ,medicine.disease_cause ,Autoimmunity ,Mice ,Immune system ,Mice, Inbred NOD ,medicine ,Animals ,Lymphocytes ,Mice, Knockout ,Base Sequence ,NF-kappa B ,General Medicine ,medicine.disease ,Adoptive Transfer ,Arthritis, Experimental ,DNA-Binding Proteins ,medicine.anatomical_structure ,Immunology ,medicine.symptom ,Research Article - Abstract
NF-(kappa)B is an important component of both autoimmunity and bone destruction in RA. NF-(kappa)B-inducing kinase (NIK) is a key mediator of the alternative arm of the NF-(kappa)B pathway, which is characterized by the nuclear translocation of RelB/p52 complexes. Mice lacking functional NIK have no peripheral lymph nodes, defective B and T cells, and impaired receptor activator of NF-kappaB ligand-stimulated osteoclastogenesis. We investigated the role of NIK in murine models of inflammatory arthritis using Nik-/- mice. The serum transfer arthritis model is initiated by preformed antibodies and required only intact neutrophil and complement systems in recipients. While Nik-/- mice had inflammation equivalent to that of Nik+/+ controls, they showed significantly less periarticular osteoclastogenesis and less bone erosion. In contrast, Nik-/- mice were completely resistant to antigen-induced arthritis (AIA), which requires intact antigen presentation and lymphocyte function but not lymph nodes. Additionally, transfer of Nik+/+ splenocytes or T cells to Rag2-/- mice conferred susceptibility to AIA, while transfer of Nik-/- cells did not. Nik-/- mice were also resistant to a genetic, spontaneous form of arthritis, generated in mice expressing both the KRN T cell receptor and H-2. Thus, NIK is important in the immune and bone-destructive components of inflammatory arthritis and represents a possible therapeutic target for these diseases.
- Published
- 2005
26. Impact of early expression of TCRα chain on thymocyte development
- Author
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Osami Kanagawa and Ching-Yu Huang
- Subjects
CD4-Positive T-Lymphocytes ,Receptors, Antigen, T-Cell, alpha-beta ,CD3 ,Transgene ,Immunology ,Mice, Transgenic ,chemical and pharmacologic phenomena ,CD8-Positive T-Lymphocytes ,Gene Rearrangement, T-Lymphocyte ,Mice ,Animals ,Immunology and Allergy ,Gene Rearrangement, beta-Chain T-Cell Antigen Receptor ,Receptor ,Mice, Knockout ,Regulation of gene expression ,Membrane Glycoproteins ,biology ,T-cell receptor ,hemic and immune systems ,Gene rearrangement ,Flow Cytometry ,Specific Pathogen-Free Organisms ,Cell biology ,Mice, Inbred C57BL ,Thymocyte ,biology.protein ,Gene Rearrangement, alpha-Chain T-Cell Antigen Receptor ,CD8 ,Signal Transduction - Abstract
CD4(-)CD8(-) thymocytes expressing a transgenic T cell receptor (TCR) alpha chain have decreased capacity to give rise to CD4(+)CD8(+) thymocytes when compared with wild-type thymocytes. This inefficient CD4(-)CD8(-) to CD4(+)CD8(+) maturation is mediated by the transgenic TCR alpha chain pairing with endogenous TCR beta chain but not with endogenous TCR gamma chain. Comparison between TCR alpha chain-transgenic mice with or without a functional pre-TCR alpha (pT alpha ) chain reveals that the formation of transgenic alpha/endogenous beta TCR on CD4(-)CD8(-) thymocytes inhibits the formation of pre-TCR, but at the same time mediates CD4(-)CD8(-) to CD4(+)CD8(+) maturation in the absence of pre-TCR, albeit inefficiently. These results indicate that alpha beta TCR and pre-TCR provide different signals for thymocyte development. They also suggest that the precise regulation of the sequential rearrangements of TCR beta and alpha loci and the cellular expansion induced by the pre-TCR may both be evolved to ensure the efficient generation of mature alpha beta T cells.
- Published
- 2004
27. Regulation of autoimmune diabetes by non-islet-specific T cells— a role for the glucocorticoid-induced TNF receptor
- Author
-
Shimon Sakaguchi, Jonathan D. Katz, Jun Shimizu, Anish Suri, Osami Kanagawa, and Emil R. Unanue
- Subjects
Adoptive cell transfer ,medicine.medical_specialty ,T-Lymphocytes ,Immunology ,Mice, Transgenic ,Mice, SCID ,medicine.disease_cause ,Receptors, Tumor Necrosis Factor ,Autoimmunity ,Islets of Langerhans ,Mice ,Interleukin 21 ,Mice, Inbred NOD ,Internal medicine ,medicine ,Animals ,Immunology and Allergy ,Cytotoxic T cell ,IL-2 receptor ,Mice, Inbred BALB C ,business.industry ,ZAP70 ,Adoptive Transfer ,Diabetes Mellitus, Type 1 ,Endocrinology ,Cytokines ,Cytokine secretion ,business ,Glucocorticoid ,medicine.drug - Abstract
Diabetogenic BDC2.5 CD4 T cells induce diabetes when injected into NOD.scid mice. However, when co-transferred with the OVA-specific DO11.10 CD4 T cells, BDC2.5 T cells failed to cause diabetes. This inhibition depended upon the stimulation of DO11.10 T cells only with soluble OVA, which skewed their differentiation to a Th2-type pattern of cytokine secretion in vivo. However, in vivo neutralization of IL-4, IL-10 or TGF-beta using monoclonal antibodies did not prevent the inhibition whereas treatment with an antibody against the glucocorticoid-induced TNF receptor abrogated the protection from disease. In the protected mice, the diabetogenic T cells could be isolated from their spleens and shown to transfer diabetes when injected into new NOD.scid recipients. Thus, the inhibition took place without the physical or functional elimination of the diabetogenic T cells.
- Published
- 2004
28. Paradoxical Dampening of Anti-Islet Self-Reactivity but Promotion of Diabetes by OX40 Ligand
- Author
-
Arlene H. Sharpe, Andy I. Chen, Christophe Benoist, Diane Mathis, Osami Kanagawa, Laurent Poirot, Elzbieta Hyatt, Natalia Martin-Orozco, and Zhibin Chen
- Subjects
CD4-Positive T-Lymphocytes ,medicine.medical_specialty ,T cell ,Immunology ,Cell ,Priming (immunology) ,Mice, Transgenic ,OX40 Ligand ,Biology ,Ligands ,Lymphocyte Activation ,Autoantigens ,Immunophenotyping ,Islets of Langerhans ,Mice ,Mice, Inbred NOD ,T-Lymphocyte Subsets ,Internal medicine ,Immune Tolerance ,medicine ,Splenocyte ,Animals ,Immunology and Allergy ,education ,Mice, Knockout ,geography ,education.field_of_study ,Membrane Glycoproteins ,geography.geographical_feature_category ,Cell growth ,Receptors, Interleukin-2 ,Islet ,Adoptive Transfer ,Tumor Necrosis Factor Receptor Superfamily, Member 7 ,OX40 ligand ,Diabetes Mellitus, Type 1 ,medicine.anatomical_structure ,Endocrinology ,Tumor Necrosis Factors ,Disease Progression ,T cell migration - Abstract
Costimulatory signals received by diabetogenic T cells during priming by or upon secondary encounter with autoantigen are decisive in determining the outcome of autoimmune attack. The OX40-OX40 ligand (OX40L) costimulatory pathway is known to influence T cell responses, prompting us to examine its role in autoimmune diabetes. A null allele at OX40L completely prevented diabetes development in nonobese diabetic mice and strongly reduced its incidence in a TCR transgenic model (BDC2.5). However, somewhat paradoxically, the initial activation of T cells responsive to islet β cell Ag was slightly faster and more efficient in the absence of OX40L, with an increased degree of cell proliferation and survival in the deficient hosts. Activated T cell migration into and retention within the islets was also slightly accelerated. When challenged in vitro, splenocytes from BDC2.5.OX40Lo/o mice showed no altered reactivity to exogenously added peptide, no bias to the Th1 or Th2 phenotype, and no alteration in T cell survival. Thus, the OX40/OX40L axis has the paradoxical effect of dampening the early activation and migration of autoimmune T cells, but sustains the long-term progression to autoimmune destruction.
- Published
- 2003
29. Interferon-producing Cells Fail to Induce Proliferation of Naive T Cells but Can Promote Expansion and T Helper 1 Differentiation of Antigen-experienced Unpolarized T Cells
- Author
-
Osami Kanagawa, Andrew Pekosz, Emil R. Unanue, Marina Cella, Ravi K. Veeraswamy, Anne Krug, and Marco Colonna
- Subjects
Male ,T-Lymphocytes ,T cell ,Immunology ,Antigen presentation ,Antigen-Presenting Cells ,Biology ,Lymphocyte Activation ,Article ,Mice ,Interleukin 21 ,Leukocytes ,medicine ,Animals ,Immunology and Allergy ,Cytotoxic T cell ,IL-2 receptor ,Antigen-presenting cell ,Cell Polarity ,CD28 ,Cell Differentiation ,Dendritic Cells ,Th1 Cells ,Natural killer T cell ,Interleukin-12 ,Cell biology ,medicine.anatomical_structure ,Interferon Type I ,Interleukin-2 ,interferon-producing cells ,unpolarized T cells - Abstract
Interferon-producing cells (IPCs) secrete high levels of type I interferon in response to certain viruses. The lack of lineage markers, the expression of major histocompatibility complex (MHC) class II and the capacity to stimulate allogeneic T cells have led these cells to be classified as a subset of dendritic cells (DCs), called plasmacytoid DCs (PDCs). However, the role of IPCs/PDCs in initiating primary immune responses remains elusive. Here we examined the antigen presenting capacity of murine IPCs in antigen specific systems. While CD8α+ and CD11b+ DCs induced logarithmic expansion of naive CD4 and CD8 T cells, without conferring T helper commitment at a first encounter, primary IPCs lacked the ability to stimulate naive T cells. However, when antigen-experienced, nonpolarized T cells expanded by classical DC subsets, were restimulated by IPCs, they proliferated and produced high amounts of IFN-γ. These data indicate that IPCs can effectively stimulate preactivated or memory-type T cells and exert an immune-regulatory role. They also suggest that expansion of naive T cells and acquisition of effector function during antigen-specific T cell responses may involve different antigen-presenting cell (APC) types. Independent and coordinated control of T cell proliferation and differentiation would provide the immune system with greater flexibility in regulating immune responses.
- Published
- 2003
30. Caractérisation phénotypique des leucocytes cutanés à la phase aiguë d’un syndrome de Lyell chez un patient VIH+ au stade SIDA
- Author
-
B. Ben Said, Osami Kanagawa, Floriane Albert, Marc Vocanson, J.-F. Nicolas, and Axel-Patrice Villani
- Subjects
Dermatology - Abstract
Introduction La necrolyse epidermique toxique (NET) ou syndrome de Lyell, est une reaction cutanee medicamenteuse severe au cours de laquelle on observe un infiltrat leucocytaire CD4+ et CD8+. Les lymphocytes T (LT) CD8+ sont consideres comme les effecteurs de cette reaction mais les LT CD4+ pourraient egalement jouer un role dans la presentation de l’antigene. Nous rapportons ici un cas particulier de NET chez un patient sideen, avec un taux residuel tres faible de LT CD4+, qui illustre bien le role majeur et suffisant des LT CD8+ au cours de ces reactions. Observation Un homme de 35 ans consultait pour l’apparition brutale d’une dermatose bulleuse extensive febrile associee a une atteinte muqueuse severe avec signe de Nikolski. L’interrogatoire trouvait un diagnostic recent de SIDA (charge virale 650 000 copies/mL) decouvert fortuitement sur une pneumocystose pour laquelle le patient etait traite depuis 10 jours par trimethoprime-sulfomethoxazole seul. L’histologie confirmait le diagnostic clinique de syndrome de Lyell avec une necrolyse epidermique massive. A l’acme de la reaction, le decollement cutane atteignait 100 % de la surface corporelle (SCORE of Toxci Epidermal Necrolysis [SCORTEN] – calcule a 3). Le phenotypage lymphocytaire montrait un deficit marque des LT CD4+ a 66 elements/mm 3 avec une augmentation compensatrice des LT CD8+ circulants a 1648 elements/mm 3 . L’analyse en cytometrie de masse des leucocytes infiltrant le tissu cutane (leucocytes extraits du liquide de bulle a j2) montrait une presence quasi exclusive de LT CD8+ (86 % des leucocytes infiltrant le tissu cutane) pour moins de 1 % de LT CD4+ (dont 84 % de LT regulateurs, soit 0,83 % des lymphocytes TCRab+ infiltrant la peau). Les cellules natural killer etaient egalement faiblement representees (6,76 % des cellules infiltrant le tissu cutane). L’analyse specifique des marqueurs de cytotoxicite mettait en evidence une predominance de LT CD8+ co-exprimant la granulysine, le granzyme B, le granzyme A, TWEAK, la perforine et a un moindre degre le CD226 et le CD253. Les autres marqueurs de cytotoxicite CD107a, CD137 et CD178 (FasL) etaient peu ou pas exprimes. L’analyse phenotypique montrait par ailleurs que ces LT CD8+ infiltrant le tissu cutane etaient CD27+ et CD45RA−, suggerant un profil de cellules effectrices memoires. Discussion Notre patient a developpe une NET extremement severe (decollement 100 %) et ce malgre un SIDA avance (donc des LT CD8+ moins fonctionnels) et en l’absence de LT CD4+. Cela demontre bien que les LT CD8+ sont les principales cellules effectrices de la NET et qu’elles peuvent etre activees meme en l’absence de LT CD4+, confirmant ce qui est decrit dans les modeles animaux d’hypersensibilite retardee aux chimiques. Conclusion Notre cas illustre bien l’importance des LT CD8+ cytotoxiques polyfonctionnels effecteurs memoires au decours de la NET, et ce meme en l’absence de LT CD4+.
- Published
- 2015
31. The Mechanism Underlying T Cell Help for Induction of an Antigen-Specific In Vivo Humoral Immune Response to Intact Streptococcus pneumoniae Is Dependent on the Type of Antigen
- Author
-
Osami Kanagawa, Clifford M. Snapper, Ching-Liang Chu, Richard J. Riese, Abdul Qadir Khan, Yi Shen, Zheng-Qi Wu, and Harold A. Chapman
- Subjects
Male ,Phosphorylcholine ,T-Lymphocytes ,T cell ,Immunology ,Receptors, Antigen, T-Cell ,Mice ,Immune system ,Bacterial Proteins ,Antigen ,medicine ,Animals ,Immunology and Allergy ,Cytotoxic T cell ,Antigen-presenting cell ,biology ,T-cell receptor ,Germinal center ,Antibodies, Bacterial ,Cathepsins ,Molecular biology ,Streptococcus pneumoniae ,medicine.anatomical_structure ,Immunoglobulin G ,B7-1 Antigen ,biology.protein ,Antibody - Abstract
Little is known concerning the role of T cells in regulating an anti-polysaccharide Ig response to an intact pathogen. We previously reported that the in vivo Ig responses to Streptococcus pneumoniae (strain R36A), specific for pneumococcal surface protein A (PspA) and for the phosphorylcholine (PC) determinant of C-polysaccharide, were both dependent on TCR-αβ+ T cells and B7-dependent costimulation, although only PspA-specific memory was generated. In this report, we show that the T cell help underlying these two Ag-specific Ig responses is distinct. Using H-Y-specific T cell transgenic mice made “nonleaky” by crossing with mice genetically deficient for TCR-α, we demonstrate that the T cell help for the anti-PC, in contrast to the anti-PspA, response is TCR-nonspecific and occurs normally in the absence of germinal center formation, although it is still dependent on B7-dependent costimulation. Consistent with these data, we demonstrate, using cathepsin S−/− mice, that although the anti-PC response is largely dependent on CD4+ T cells, there is a reduced (or lack of) dependence, relative to the anti-PspA response, on the generation of new peptide-MHC class II complexes. In this regard, the T cell help for an optimal anti-PC response is delivered more rapidly than that required for an optimal anti-PspA response. Collectively, these data demonstrate a novel accelerated TCR-nonspecific B7-dependent form of T cell help for augmenting a polysaccharide-specific Ig response to an intact bacterium without the generation of memory.
- Published
- 2002
32. Superantigen-Induced TCR α Locus Secondary Rearrangement: Role in Tolerance Induction
- Author
-
Osami Kanagawa, Ching-Yu Huang, Gillian E. Wu, and Rachel Golub
- Subjects
Receptors, Antigen, T-Cell, alpha-beta ,T-Lymphocytes ,Immunology ,Down-Regulation ,Cytochrome c Group ,Mice, Transgenic ,chemical and pharmacologic phenomena ,Locus (genetics) ,Endogeny ,Biology ,Lymphocyte Activation ,Mice ,chemistry.chemical_compound ,In vivo ,hemic and lymphatic diseases ,Immune Tolerance ,Superantigen ,Animals ,Immunology and Allergy ,Antigens ,Columbidae ,Gene ,Mice, Knockout ,Superantigens ,T-cell receptor ,Cell Differentiation ,hemic and immune systems ,Molecular biology ,biological factors ,Mice, Inbred C57BL ,Tolerance induction ,chemistry ,Injections, Intravenous ,Mice, Inbred CBA ,Immunization ,Gene Rearrangement, alpha-Chain T-Cell Antigen Receptor ,Genes, T-Cell Receptor alpha ,Spleen ,DNA - Abstract
Immunization with superantigen in vivo induces transient activation of superantigen-specific T cells, followed by a superantigen-nonresponsive state. In this study, using a TCR α knock-in mouse in which the knock-in α-chain can be replaced with endogenous α-chain through secondary rearrangement, we show that immunization of superantigen changes the TCR α-chain expression on peripheral superantigen-specific T cells, induces expression of recombination-activating genes, and generates DNA double-strand breaks at the TCR α-chain locus. These results suggest that viral superantigens are capable of inducing peripheral TCR revision. Our findings thus provide a new perspective on pathogen-immune system interaction.
- Published
- 2002
33. In APCs, the Autologous Peptides Selected by the Diabetogenic I-Ag7 Molecule Are Unique and Determined by the Amino Acid Changes in the P9 Pocket
- Author
-
Michael L. Gross, Anish Suri, Osami Kanagawa, Emil R. Unanue, Koen van der Drift, and Ilan Vidavsky
- Subjects
Molecular Sequence Data ,Immunology ,Mutant ,Antigen-Presenting Cells ,Sequence (biology) ,Major histocompatibility complex ,Autoantigens ,Epitope ,Tacrolimus Binding Proteins ,Mice ,Residue (chemistry) ,Antigens, CD ,Tumor Cells, Cultured ,Animals ,Immunology and Allergy ,Molecule ,Amino Acid Sequence ,chemistry.chemical_classification ,Antigen Presentation ,Membrane Glycoproteins ,biology ,Histocompatibility Antigens Class II ,Lysosome-Associated Membrane Glycoproteins ,Adenovirus E2 Proteins ,Peptide Fragments ,Amino acid ,Diabetes Mellitus, Type 1 ,Amino Acid Substitution ,chemistry ,Biochemistry ,biology.protein ,Muramidase ,Lysosomes ,Protein Binding - Abstract
We demonstrate in this study the great degree of specificity in peptides selected by a class II MHC molecule during processing. In this specific case of the diabetogenic I-Ag7 molecule, the P9 pocket of I-Ag7 plays a critical role in determining the final outcome of epitope selection, a conclusion that is important in interpreting the role of this molecule in autoimmunity. Specifically, we examined the display of naturally processed peptides from APCs expressing either I-Ag7 molecules or a mutant I-Ag7 molecule in which the β57Ser residue was changed to an Asp residue. Using mass spectrometry analysis, we identified over 50 naturally processed peptides selected by I-Ag7-expressing APCs. Many peptides were selected as families with a core sequence and variable flanks. Peptides selected by I-Ag7 were unusually rich in the presence of acidic residues toward their C termini. Many peptides contained short sequences of two to three acidic residues. In binding analysis, we determined the core sequences of many peptides and the interaction of the acidic residues with the P9 pocket. However, different sets of peptides were isolated from APCs bearing a modified I-Ag7 molecule. These peptides did not favor acidic residues toward the carboxyl terminus.
- Published
- 2002
34. Tracking and quantification of dendritic cell migration and antigen trafficking between the skin and lymph nodes
- Author
-
Osami Kanagawa, Akihiro Hata, Kouji Matsushima, Takeshi Watanabe, Francis H. W. Shand, Hidekazu Tsutsui, Satoshi Matsuoka, Kenji Kabashima, Ryoyo Ikebuchi, Michio Tomura, Kayo Inaba, Atsushi Miyawaki, Satoshi Ueha, and Yasutaka Nakanishi
- Subjects
Receptors, CCR7 ,Langerin ,Light ,Endogeny ,chemical and pharmacologic phenomena ,Mice, Transgenic ,Article ,Mice ,Antigen ,In vivo ,Antigens, CD ,Cell Movement ,Fluorescent protein ,Animals ,Immune homeostasis ,Gene Knock-In Techniques ,Dendritic cell migration ,Skin ,Mice, Knockout ,Multidisciplinary ,biology ,integumentary system ,Proteins ,hemic and immune systems ,Dendritic Cells ,Cell biology ,Mice, Inbred C57BL ,Microscopy, Fluorescence ,Langerhans Cells ,Immunology ,biology.protein ,Dermatitis, Irritant ,Lymph ,Lymph Nodes ,Integrin alpha Chains - Abstract
Skin-derived dendritic cells (DCs) play a crucial role in the maintenance of immune homeostasis due to their role in antigen trafficking from the skin to the draining lymph nodes (dLNs). To quantify the spatiotemporal regulation of skin-derived DCs in vivo, we generated knock-in mice expressing the photoconvertible fluorescent protein KikGR. By exposing the skin or dLN of these mice to violet light, we were able to label and track the migration and turnover of endogenous skin-derived DCs. Langerhans cells and CD103(+)DCs, including Langerin(+)CD103(+)dermal DCs (DDCs), remained in the dLN for 4-4.5 days after migration from the skin, while CD103(-)DDCs persisted for only two days. Application of a skin irritant (chemical stress) induced a transient >10-fold increase in CD103(-)DDC migration from the skin to the dLN. Tape stripping (mechanical injury) induced a long-lasting four-fold increase in CD103(-)DDC migration to the dLN and accelerated the trafficking of exogenous protein antigens by these cells. Both stresses increased the turnover of CD103(-)DDCs within the dLN, causing these cells to die within one day of arrival. Therefore, CD103(-)DDCs act as sentinels against skin invasion that respond with increased cellular migration and antigen trafficking from the skin to the dLNs.
- Published
- 2014
35. Vα gene replacement in a TCRα knock-in mouse
- Author
-
Rachel Golub, Osami Kanagawa, Gillian E. Wu, and Ching-Yu Huang
- Subjects
medicine.anatomical_structure ,Immune system ,Gene replacement ,Transgene ,T cell ,Immunology ,T-cell receptor ,medicine ,Immunology and Allergy ,Locus (genetics) ,Biology ,Knock in mouse ,Molecular biology - Abstract
Using a TCRα chain knock-in mouse, we demonstrate that V-gene replacement can operate in the T cell receptor α locus. Functional TCRα chain transcripts generated by Vα-gene replacement at the site of the Vα-embedded heptamer were identified in splenic T cells. This finding shows that Vα-gene replacement can likely be used to shape the peripheral T cell repertoire.The conservation of the embedded heptamer in most Vα segments adds support to the notion that V-gene replacement is a mechanism maintained to diversify the immune system and that argues that itis common to B and T cells.
- Published
- 2001
36. Cutting Edge: A Single MHC Anchor Residue Alters the Conformation of a Peptide-MHC Complex Inducing T Cells That Survive Negative Selection
- Author
-
Daniel A. Peterson, Osami Kanagawa, Emil R. Unanue, and Richard J. DiPaolo
- Subjects
Genetically modified mouse ,Cell Survival ,Macromolecular Substances ,Protein Conformation ,Molecular Sequence Data ,Immunology ,Mice, Transgenic ,chemical and pharmacologic phenomena ,Peptide ,Lymphocyte Activation ,medicine.disease_cause ,Major histocompatibility complex ,Autoimmunity ,Mice ,Negative selection ,T-Lymphocyte Subsets ,Immune Tolerance ,medicine ,Animals ,Point Mutation ,Immunology and Allergy ,Amino Acid Sequence ,chemistry.chemical_classification ,Antigen Presentation ,Aspartic Acid ,Alanine ,biology ,Point mutation ,Histocompatibility Antigens Class II ,Cell Differentiation ,hemic and immune systems ,Molecular biology ,Peptide Fragments ,Amino acid ,Mice, Inbred C57BL ,chemistry ,biology.protein ,Muramidase ,Peptide/MHC Complex - Abstract
We generated transgenic mice that expressed hen egg-white lysozyme (HEL) under a class II MHC promoter. The A7 line expressed HEL with a point mutation in the Asp52 residue, the main anchor amino acid responsible for the selection of the chemically dominant family of peptides (52–60) by I-Ak molecules. Mice expressing HEL with Ala52 were completely unresponsive when immunized with the same protein, i.e., HEL A52. However, the same mice immunized with wild-type HEL elicited T cells that recognized a conformation of the 52–61 core sequence uniquely different between Asp52 and Ala52 containing peptides. Importantly, some T cells also recognized the HEL A52 peptide given exogenously but not the same peptide processed from HEL A52 protein. Thus, a core MHC anchor residue influences markedly the specificity of the T cells. We discuss the relevance of these findings to autoimmunity and vaccination with altered peptides.
- Published
- 2001
37. An Instructive Component in T Helper Cell Type 2 (Th2) Development Mediated by Gata-3
- Author
-
Wenjun Ouyang, J. David Farrar, Andreas Radbruch, Kenneth M. Murphy, Max Löhning, Osami Kanagawa, and Mario Assenmacher
- Subjects
Cellular differentiation ,Immunology ,Population ,T lymphocytes ,instruction ,Cell Separation ,GATA3 Transcription Factor ,Biology ,Cell fate determination ,GATA-3 ,Transfection ,Cell Line ,Immunophenotyping ,03 medical and health sciences ,Interferon-gamma ,Mice ,0302 clinical medicine ,Th2 Cells ,medicine ,cytokine ,Immunology and Allergy ,Animals ,stochastic ,Progenitor cell ,education ,Interleukin 4 ,030304 developmental biology ,Genetics ,0303 health sciences ,education.field_of_study ,Stochastic Processes ,Stem Cells ,Brief Definitive Report ,Cell Differentiation ,T helper cell ,Th1 Cells ,Cell biology ,Clone Cells ,DNA-Binding Proteins ,medicine.anatomical_structure ,Retroviridae ,Trans-Activators ,Cytokines ,Cytokine secretion ,Interleukin-4 ,Stem cell ,STAT6 Transcription Factor ,Cell Division ,030215 immunology - Abstract
Although interleukin (IL)-12 and IL-4 polarize naive CD4+ T cells toward T helper cell type 1 (Th1) or Th2 phenotypes, it is not known whether cytokines instruct the developmental fate in uncommitted progenitors or select for outgrowth of cells that have stochastically committed to a particular fate. To distinguish these instructive and selective models, we used surface affinity matrix technology to isolate committed progenitors based on cytokine secretion phenotype and developed retroviral-based tagging approaches to directly monitor individual progenitor fate decisions at the clonal and population levels. We observe IL-4–dependent redirection of phenotype in cells that have already committed to a non–IL-4–producing fate, inconsistent with predictions of the selective model. Further, retroviral tagging of naive progenitors with the Th2-specific transcription factor GATA-3 provided direct evidence for instructive differentiation, and no evidence for the selective outgrowth of cells committed to either the Th1 or Th2 fate. These data would seem to exclude selection as an exclusive mechanism in Th1/Th2 differentiation, and support an instructive model of cytokine-driven transcriptional programming of cell fate decisions.
- Published
- 2001
38. Congenital Nephrotic Syndrome in Mice Lacking CD2-Associated Protein
- Author
-
Vladimir Karpitskii, Osami Kanagawa, Jeffrey H. Miner, Andrey S. Shaw, Neng-Yao Shih, Ancho Nguyen, Jun Li, and Michael L. Dustin
- Subjects
medicine.medical_specialty ,Nephrotic Syndrome ,Recombinant Fusion Proteins ,T-Lymphocytes ,Kidney Glomerulus ,Podocyte foot ,Lymphocyte Activation ,Cell junction ,Basement Membrane ,Nephrin ,Mice ,Internal medicine ,medicine ,Animals ,Congenital nephrotic syndrome ,Adaptor Proteins, Signal Transducing ,Mice, Knockout ,Extracellular Matrix Proteins ,Multidisciplinary ,biology ,Mesangial cell ,Membrane Proteins ,Proteins ,Epithelial Cells ,Glomerulonephritis ,medicine.disease ,Glomerular Mesangium ,Cytoskeletal Proteins ,Microscopy, Electron ,Intercellular Junctions ,Endocrinology ,biology.protein ,Podocin ,Slit diaphragm - Abstract
CD2-associated protein (CD2AP) is an 80-kilodalton protein that is critical for stabilizing contacts between T cells and antigen-presenting cells. In CD2AP-deficient mice, immune function was compromised, but the mice died at 6 to 7 weeks of age from renal failure. In the kidney, CD2AP was expressed primarily in glomerular epithelial cells. Knockout mice exhibited defects in epithelial cell foot processes, accompanied by mesangial cell hyperplasia and extracellular matrix deposition. Supporting a role for CD2AP in the specialized cell junction known as the slit diaphragm, CD2AP associated with nephrin, the primary component of the slit diaphragm.
- Published
- 1999
39. Quantitative Analysis of the T Cell Repertoire that Escapes Negative Selection
- Author
-
Richard J. DiPaolo, Emil R. Unanue, Osami Kanagawa, and Daniel A. Peterson
- Subjects
Genetically modified mouse ,Transgene ,Molecular Sequence Data ,Immunology ,Clonal Deletion ,Mice, Transgenic ,chemical and pharmacologic phenomena ,Peptide ,Biology ,medicine.disease_cause ,Epitope ,Autoimmunity ,Epitopes ,Mice ,Negative selection ,chemistry.chemical_compound ,T-Lymphocyte Subsets ,Immune Tolerance ,medicine ,Animals ,Immunology and Allergy ,Amino Acid Sequence ,Lymphocyte Count ,chemistry.chemical_classification ,Antigen Presentation ,Histocompatibility Antigens Class II ,hemic and immune systems ,Molecular biology ,Peptide Fragments ,Infectious Diseases ,Immunization ,chemistry ,Muramidase ,Lysozyme ,Chickens - Abstract
Mice expressing hen egg-white lysozyme (HEL) as a transgene are unresponsive to immunization with the HEL protein. Profound tolerance was found even in situations where the amounts of I-A k -peptide complexes was 100 or less per APC. Among the few T cells that escaped tolerance, we did not observe differential responses to the different HEL epitopes, perhaps because of the very high sensitivity of the negative selection process. The same HEL transgenic mice that did not respond to HEL responded to immunization with the 46–61 peptide of HEL. These peptide-specific T cells that escaped negative selection belonged to a set that reacted with a particular conformer of the HEL peptide-I-A k (type B). The presence of type B reactive T cells should be considered in autoimmunity.
- Published
- 1999
40. Cutting Edge: Negative Selection of Immature Thymocytes by a Few Peptide-MHC Complexes: Differential Sensitivity of Immature and Mature T Cells
- Author
-
Daniel A. Peterson, Richard J. DiPaolo, Osami Kanagawa, and Emil R. Unanue
- Subjects
Immunology ,Immunology and Allergy - Abstract
We quantitated the number of peptide-class II MHC complexes required to affect the deletion or activation of 3A9 TCR transgenic thymocytes. Deletion of immature double positive thymocytes was very sensitive, taking place with approximately three peptide-MHC complexes per APC. However, the activation of mature CD4+ thymocytes required 100-fold more complexes per APC. Therefore, a “biochemical margin of safety” exists at the level of the APC. To be activated, autoreactive T cells in peripheral lymphoid tissues require a relatively high level of peptide-MHC complexes.
- Published
- 1999
41. TCR-Independent Pathways Mediate the Effects of Antigen Dose and Altered Peptide Ligands on Th Cell Polarization
- Author
-
Arash Grakoui, David L. Donermeyer, Osami Kanagawa, Kenneth M. Murphy, and Paul M. Allen
- Subjects
Immunology ,Immunology and Allergy - Abstract
We examined the role of the peptide/MHC ligand in CD4+ T cell differentiation into Th1 or Th2 cells using a TCR αβ transgenic mouse specific for hemoglobin (Hb)(64-76)/I-Ek. We identified two altered peptide ligands of Hb(64-76) that retain strong agonist activity but, at a given dose, induce cytokine patterns distinct from the Hb(64-76) peptide. The ability of these peptides to produce distinct cytokine patterns at identical doses is not due to an intrinsic qualitative property. Each peptide can induce Th2 cytokines at low concentrations and Th1 cytokines at high concentrations and has a unique range of concentrations at which these distinct effects occur. The pattern of cytokines produced from limiting dilution of naive T cells demonstrated that the potential to develop an individual Th1 or Th2 cell is stochastic, independent of Ag dose. We propose that the basis for the observed effects on the Th1/Th2 balance shown by the altered peptide ligands and the amount of Ag dose involves the modification of soluble factors in bulk cultures that are the driving force that polarize the population to either a Th1 or Th2 phenotype.
- Published
- 1999
42. Cutting Edge: Thymic Positive Selection and Peripheral Activation of Islet Antigen-Specific T Cells: Separation of Two Diabetogenic Steps by an I-Ag7 Class II MHC β-Chain Mutant
- Author
-
Osami Kanagawa, Barbara A. Vaupel, Guan Xu, Emil R. Unanue, and Jonathan D. Katz
- Subjects
Immunology ,Immunology and Allergy - Abstract
The diabetes-susceptible class II MHC genes (in human and mouse) share unique nonaspartic acid residues at position 57 of the class II β-chain. Transgenic expression of a mutant I-Ag7, substituting histidine and serine at position 56 and 57 of β-chain with proline and aspartic acid (I-Ag7PD), respectively, inhibits diabetes development in the nonobese diabetic mouse model. Here, we demonstrate that immature thymocytes expressing a diabetogenic islet Ag-specific transgenic TCR are positively selected by I-Ag7PD class II MHC to give rise to mature CD4+ T cells. However, splenic APCs expressing the same I-Ag7PD fail to present pancreatic islet Ag to mature T cells bearing this diabetogenic TCR. These results indicate that nonaspartic acid residues at position 57 of class II MHC β-chain is important for diabetogenic CD4+ T cell activation in the periphery but is not essential for the formation of a diabetogenic T cell repertoire in the thymus.
- Published
- 1998
43. Rapid deletion of rearranged T cell antigen receptor (TCR) Vα-Jα segment by secondary rearrangement in the thymus: Role of continuous rearrangement of TCR α chain gene and positive selection in the T cell repertoire formation
- Author
-
Ching-Yu Huang, Osami Kanagawa, and Fanping Wang
- Subjects
T-Lymphocytes ,T cell ,CD3 ,Cellular differentiation ,Gene Expression ,Mice, Transgenic ,chemical and pharmacologic phenomena ,Mice ,medicine ,Animals ,Sequence Deletion ,Mice, Knockout ,Multidisciplinary ,biology ,T-cell receptor ,Cell Differentiation ,hemic and immune systems ,Gene rearrangement ,Biological Sciences ,Molecular biology ,Thymocyte ,medicine.anatomical_structure ,biology.protein ,Gene Rearrangement, alpha-Chain T-Cell Antigen Receptor ,Alpha chain ,CD8 - Abstract
A rearranged T cell receptor (TCR) Vα and Jα gene from a cytochrome c- specific T cell hybridoma was introduced into the genomic Jα region. The introduced TCR α chain gene is expressed in a majority of CD3 positive and CD4 CD8 double-negative immature thymocytes. However, only a few percent of the double-positive and single-positive thymocytes express this TCR α chain. This decrease is caused by a rearrangement of TCR α chain locus, which deletes the introduced TCR gene. Analysis of the mice carrying the introduced TCR α chain and the transgenic TCR β chain from the original cytochrome c- specific T cell hybridoma revealed that positive selection efficiently rescues double-positive thymocytes from the loss of the introduced TCR α chain gene. In the mice with negatively selecting conditions, T cells expressing the introduced TCR αβ chains were deleted at the double-positive stage. However, a large number of thymocytes escape negative selection by using an endogenous TCR α chain created by secondary rearrangement maintaining normal thymocyte development. These results suggest that secondary rearrangements of the TCR α chain gene play an important role in the formation of the T cell repertoire.
- Published
- 1998
44. Induction of Endogenous Mammary Tumor Virus in Lymphocytes Infected with Murine Acquired Immunodeficiency Syndrome Virus
- Author
-
Rafick-Pierre Sekaly, Louise Doyon, Osami Kanagawa, Barbara A. Vaupel, and Shinyo Gayama
- Subjects
Lymphoma, B-Cell ,viruses ,Molecular Sequence Data ,Immunology ,chemical and pharmacologic phenomena ,Biology ,Virus ,Mice ,Open Reading Frames ,Retrovirus ,Murine Acquired Immunodeficiency Syndrome ,Mammary tumor virus ,parasitic diseases ,Tumor Cells, Cultured ,medicine ,Superantigen ,Animals ,Amino Acid Sequence ,Lymphocytes ,RNA, Messenger ,B cell ,Immunodeficiency ,Superantigens ,medicine.disease ,biology.organism_classification ,Genes, gag ,Virology ,Open reading frame ,medicine.anatomical_structure ,Mammary Tumor Virus, Mouse ,Cell culture ,Spleen - Abstract
Mice infected with murine acquired immunodeficiency syndrome (MAIDS) virus developed lymphoadenopathy and profound immunodeficiency. Concomitantly the expression of endogenous mammary tumor virus (MTV) mRNA increased significantly, especially for the 1.7-kb 3' open reading frame (ORF) mRNA encoding MTV superantigen. B cell lines that are established from MAIDS mice and exhibit superantigen activity also express a high level of 1.7-kb endogenous MTV and mRNA. Infection of a B cell tumor line in vitro with retrovirus containing the cloned MAIDS virus gene induced superantigen activity and this cell line also expressed the 1.7-kb superantigen coding MTV 3' ORF mRNA. These results strongly suggest a link between MAIDS virus infection and the induction of endogenous superantigen activity. This may play an important role in the pathogenesis of the MAIDS virus.
- Published
- 1998
45. Autoreactivity of T cells from nonobese diabetic mice: An I-Ag7-dependent reaction
- Author
-
Osami Kanagawa, Steven M. Martin, Barbara A. Vaupel, Emil R. Unanue, and Eugenio Carrasco-Marín
- Subjects
Ovalbumin ,T-Lymphocytes ,T cell ,Autoimmunity ,Biology ,Major histocompatibility complex ,Mice ,Antigen ,Mice, Inbred NOD ,medicine ,Animals ,Cytotoxic T cell ,Antigen-presenting cell ,NOD mice ,Mice, Inbred BALB C ,Multidisciplinary ,Histocompatibility Antigens Class II ,Biological Sciences ,Natural killer T cell ,Cell biology ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Immunology ,biology.protein ,Lymph Nodes ,CD8 - Abstract
Mice bearing the I-Ag7class II major histocompatibility complex molecules contain a high number of spontaneous autoreactive T cells, as estimated by limiting-dilution assays. We found this autoreactivity in various strains that bear the I-Ag7molecule, such as the nonobese diabetic (NOD) mouse strain, which spontaneously develops autoimmune diabetes. However, NOD mice strains that do not express the I-Ag7molecule, but instead express I-Ab, do not have a high incidence of autoreactive T cells. About 15% of the autoreactive T cells also recognize the I-Ag7molecule expressed in the T2 line, which is defective in the processing of protein antigens. We interpret this to mean that some of the T cells may interact with class II molecules that are either devoid of peptides or contain a limited peptide content. We also find a high component of autoreactivity among antigen-specific T cell clones. These T cell clones proliferate specifically to protein antigens but also have a high level of reactivity to antigen-presenting cells not pulsed with antigen. Thus, the library of T cell receptors in NOD mice is skewed to autoreactivity, which we speculate is based on the weak peptide-binding properties of I-Ag7molecules.
- Published
- 1998
46. Differences Between Responses of Naive and Activated T Cells to Anergy Induction
- Author
-
Robert J. Hayashi, Dennis Y. Loh, Osami Kanagawa, and Fanping Wang
- Subjects
Immunology ,Immunology and Allergy - Abstract
T cell unresponsiveness to Ag stimulation can be induced by several means. The precise mechanism by which this process occurs remains poorly understood. Preincubating T cells with either EDCI-fixed APC or ionomycin is a proven means of inducing T cell anergy with reduced IL-2 production in response to Ag stimulation. Using T cells from mice expressing the TCR transgene DO11.10, which is specific for a peptide (323–339) derived from hen egg OVA, we demonstrate that naive cells obtained directly from the host are resistant to the anergy induction by either fixed APC or ionomycin. TCR transgenic mice also deficient in the recombination-activating gene-2 (RAG-2−/−), preventing the formation of T cells with endogenous TCRs, were immunized with OVA, and in vivo activated T cells with low expression of CD62 were isolated. These primed cells possess the same sensitivity to ionomycin-induced anergy as in vitro activated cell lines. This unresponsive state most profoundly affects Ag-induced IL-2 production, with IFN-γ and IL-3 affected to a lesser degree and no effect observed on IL-4 production. Thus, T cells in vivo can be distinguished phenotypically by their susceptibility to anergic stimuli. Anergy so induced affects selected T cell functions.
- Published
- 1998
47. ChronicListeriaInfection in SCID Mice: Requirements for the Carrier State and the Dual Role of T Cells in Transferring Protection or Suppression
- Author
-
Vatsala Bhardwaj, Osami Kanagawa, Paul E. Swanson, and Emil R. Unanue
- Subjects
Immunology ,Immunology and Allergy - Abstract
Listeriosis in mice with the SCID mutation results in a chronic infection. The chronic infection is characterized by abundant granulomas and neutrophil infiltrates. Both lesions were particularly noticeable in the liver. In the liver, about 95% are granulomas with 5% microabscesses involving intrahepatic infection. The majority of Listeria resided in membrane-bound vacuolar structures of the macrophages and not in the cytosol. Three manipulations resulted in alterations in the equilibrium between granulomas and liver microabscesses, with massive transfer of the infection to the hepatocyte and dissolution of the granulomas: depletion of neutrophils and neutralization of IFN-γ and TNF-α. We did not find a role for IL-12, IL-10, or nitric oxide. Adoptive transfer studies showed a decisive role for both CD4+ and CD8+ T cells for an effective immune response, i.e., clearance of bacteria, granuloma formation with lymphocytes, and disappearance of microabscess. Clearance of Listeria was induced by transfer of CD8+ T cells from mice with targeted disruption of the IFN-γ structural gene (IfgTM1KO), even in the presence of neutralizing mAb to IFN-γ. In marked contrast, transfer of CD4+ T cells from IfgTM1KO mice exacerbated the infection in the chronically infected SCID mice, resulting in increased mortality with dissolution of the granulomas and severe hepatic infection with neutrophil infiltration. Thus, these data indicate that both IFN-γ-dependent and -independent mechanisms are operative in the context of a chronic listerial infection.
- Published
- 1998
48. The role of I-Ag7 beta chain in peptide binding and antigen recognition by T cells
- Author
-
Osami Kanagawa, Emil R. Unanue, and Jun Shimizu
- Subjects
CD74 ,T-Lymphocytes ,T cell ,Immunology ,Peptide binding ,Biology ,Lymphocyte Activation ,Major histocompatibility complex ,Islets of Langerhans ,Mice ,Mice, Inbred NOD ,MHC class I ,medicine ,Animals ,Immunology and Allergy ,Antigens ,Hybridomas ,Antigen processing ,Histocompatibility Antigens Class II ,General Medicine ,MHC restriction ,Molecular biology ,Clone Cells ,Cell biology ,Diabetes Mellitus, Type 1 ,medicine.anatomical_structure ,biology.protein ,Immunization ,Peptides ,Protein Binding ,Peptide/MHC Complex - Abstract
We examine here how the beta chain of the class II MHC molecule I-Ag7 influences T cell recognition. Three sets of T cell clones were identified. The first set recognizes peptides bound to I-Ag7, I-Ad and I-Ag7 mutant in which the allele-specific residues His and Ser at position 56 and 57 were changed to the Pro at residue 56 and to non-polymorphic Asp at residue 57. The second set responds to the antigen presented only by I-Ag7 and does not recognize the peptides bound to the other class II molecules. The third set is also specific for I-Ag7 as a result of the poor binding of the peptide to I-Ad and the mutant I-Ag7. These results indicate that positions 56 and 57 of the I-Ag7 class II MHC beta chain play a role in both T cell recognition of the MHC-peptide complex and peptide binding to MHC. These two different functions may be involved in I-Ag7-restricted beta cell antigen recognition by diabetogenic T cell clones.
- Published
- 1997
49. Optimizing fluorescence excitation and detection for intravital two-photon microscopy
- Author
-
Dan, Suan, Henry R, Hampton, Michio, Tomura, Osami, Kanagawa, Tatyana, Chtanova, and Tri Giang, Phan
- Subjects
Mice ,Microscopy, Confocal ,Microscopy, Fluorescence ,Cell Tracking ,Calibration ,Green Fluorescent Proteins ,Animals ,Mice, Transgenic ,Lymph Nodes ,Signal-To-Noise Ratio ,Software - Abstract
Commercial two-photon microscope systems incorporating turnkey ultrafast lasers have made the technology more user-friendly and accessible to nonspecialized biology laboratories. This has been accompanied by the development of an exciting range of new fluorescent proteins and dyes such as near-infra-red fluorescent proteins and optical highlighters. However, the two-photon absorption properties of these fluorescent molecules are not widely available and cannot be reliably predicted from their single photon absorption spectra. Furthermore, the spectral characteristics of fluorescent proteins in vivo can be affected by the local environment and light scattering by deep tissue and can vary greatly from one laboratory to the next. Here, we describe a simple protocol for determining the two-photon excitation peaks of fluorescent reporters that can be tailored to the relevant tissue samples to suit the imaging goals of individual biological laboratories.
- Published
- 2013
50. Contrasting quiescent G0 phase with mitotic cell cycling in the mouse immune system
- Author
-
Osami Kanagawa, Akihiro Hata, Michio Tomura, Kenji Ohtawa, Atsushi Miyawaki, Asako Sakaue-Sawano, Mitsuyo Takase-Utsugi, and Yoshiko Mori
- Subjects
Male ,Genetically modified mouse ,Cell division ,Hematopoietic System ,Cellular differentiation ,Transgene ,lcsh:Medicine ,Mice, Transgenic ,Biology ,Resting Phase, Cell Cycle ,Flow cytometry ,Mice ,Immune system ,medicine ,Animals ,lcsh:Science ,Cells, Cultured ,Multidisciplinary ,medicine.diagnostic_test ,Cell Cycle ,lcsh:R ,G1 Phase ,Cell cycle ,Flow Cytometry ,Cell biology ,Haematopoiesis ,Immune System ,Female ,lcsh:Q ,Cell Division ,Research Article - Abstract
A transgenic mouse line expressing Fucci (fluorescent ubiquitination-based cell-cycle indicator) probes allows us to monitor the cell cycle in the hematopoietic system. Two populations with high and low intensities of Fucci signals for Cdt1(30/120) accumulation were identified by FACS analysis, and these correspond to quiescent G0 and cycling G1 cells, respectively. We observed the transition of immune cells between quiescent and proliferative phases in lymphoid organs during differentiation and immune responses.
- Published
- 2013
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