International Immunology

International Immunology Advance Access originally published online on August 28, 2006
International Immunology 2006 18(10):1405-1411; doi:10.1093/intimm/dxl082

This Article Abstract FREE Full Text (PDF) All Versions of this Article: 18/10/1405    most recent dxl082v1 Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Add to My Personal Archive Download to citation manager Search for citing articles in: ISI Web of Science (11) Request Permissions Google Scholar Articles by Sato, S. Articles by Akira, S. Search for Related Content PubMed PubMed Citation Articles by Sato, S. Articles by Akira, S. Social Bookmarking          What's this?

© The Japanese Society for Immunology. 2006. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org

TAK1 is indispensable for development of T cells and prevention of colitis by the generation of regulatory T cells

Shintaro Sato¹, Hideki Sanjo², Tohru Tsujimura³, Jun Ninomiya-Tsuji⁴, Masahiro Yamamoto², Taro Kawai¹, Osamu Takeuchi^1,2 and Shizuo Akira^1,2

¹ Akira Innate Immunity Project, Exploratory Research for Advanced Technology, Japan Science and Technology Agency, Osaka University, 3-1 Yamada-oka, Suita, Osaka 565-0871, Japan
² Department of Host Defense, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamada-oka, Suita, Osaka 565-0871, Japan
³ Department of Pathology, Hyogo College of Medicine, 1-1 Mukogawa-cho, Nishinomiya, Hyogo 663-8501, Japan
⁴ Department of Environmental and Molecular Toxicology, North Carolina State University, Raleigh, NC 27695-7633, USA

Correspondence to: S. Akira; E-mail: sakira{at}biken.osaka-u.ac.jp

	Abstract

Top Abstract Introduction Methods Results and Discussion References

 
Transforming growth factor (TGF)-ß-activating kinase 1 (TAK1) is critical for Toll-like receptor- and tumor necrosis factor-mediated cellular responses. In B cells, TAK1 is essential for the activation of mitogen-activated protein kinases (MAPKs), but not nuclear factor-B (NF-B), in antigen receptor signaling. In this study, we generate T cell-specific TAK1-deficient (Lck^Cre/+Tak1^flox/flox) mice and show that TAK1 is indispensable for the maintenance of peripheral CD4 and CD8 T cells. In thymocytes, TAK1 is essential for TCR-mediated activation of both NF-B and MAPKs. Additionally, Lck^Cre/+Tak1^flox/flox mice developed colitis as they aged. In these mice, accumulations of activated/memory T cells as well as B cells were observed. Development of regulatory T (Treg) cells in thymus was abrogated in Lck^Cre/+Tak1^flox/flox mice, suggesting that the loss of Treg cells is the cause of the disease. Together, the results show that TAK1, by controlling the generation of central Treg cells, is important for preventing spontaneously developing colitis.

Keywords: IBD, regulatory T cells, signal transduction, TCR

	Introduction

Top Abstract Introduction Methods Results and Discussion References

 
Transforming growth factor (TGF)-ß-activating kinase 1 (TAK1) is a serine/threonine kinase that belongs to the mitogen-activated protein kinase kinase kinase family and plays crucial roles in signaling pathways initiated by IL-1ß receptor (IL-1R), tumor necrosis factor- receptor (TNFR) and Toll-like receptors (TLRs) (1, 2). IL-1ß/TLR ligands and TNF stimulations lead to the rapid recruitment of TAK1 to TNFR-associated factor (TRAF)-6 and TRAF2, respectively, and TAK1 phosphorylates IB kinase (IKK)-ß and MKK6 (3). Activated IKK complex in turn phosphorylates IB to initiate its degradation, and freed nuclear factor-B (NF-B) translocates into the nucleus and induces the expression of genes involved in inflammation (4). On the other hand, phosphorylation of MKK6 leads to the activation of mitogen-activated protein kinases (MAPKs) including c-Jun N-terminal kinase (JNK) and p38 (5, 6). TAK1-deficient cells fail to activate NF-B and MAPK in response to these ligands, indicating that TAK1 is essential for the activation of signaling pathways in response to these stimuli (1, 2).

In addition to receptors belonging to the IL-1R/TLR and TNF superfamily, TAK1 is also involved in antigen receptor signaling in B lymphocytes (1). B cell receptor (BCR) stimulation triggers the rapid activation of protein kinase C (PKC)-ß, and it transduces signals via a complex composed of CARD11, Bcl10 and MALT1, leading to the activation of the IKK complex (7). Although TAK1-deficient B cells develop normally, except for B-1 B cells, BCR-mediated proliferation is severely impaired in TAK1-deficient B cells (1). Interestingly, TAK1 is recruited to the Bcl10 complex and controls the activation of JNK, but not NF-B, in BCR signaling (1).

TCR signaling utilizes mechanisms similar to those of BCR. The importance of NF-B in T cell development and TCR signaling has been extensively studied. TCR signaling also activates the IKK complex via a complex of Bcl10, CARD11 and MALT1 downstream of PKC (7). In contrast to B cells, it has been shown that in vitro RNA interference-mediated knockdown of TAK1 in Jurkat cells results in diminished NF-B activation in TCR signaling (8). However, the role of TAK1 in T cells in vivo has yet to be clarified.

In the present study, we show that TAK1 plays indispensable roles in T cell development and the maintenance of tolerance by establishing and using T cell-specific TAK1-deficient (Lck^Cre/+Tak1^flox/flox) mice. In contrast to BCR signaling, TCR-mediated NF-B activation was dependent on TAK1. Surprisingly, T cell-specific TAK1-deficient mice developed severe colitis as they aged. Peripheral T cells isolated from old Lck^Cre/+Tak1^flox/flox mice showed an activated phenotype, which could account for the observed pathological phenotypes. Furthermore, thymocytes and peripheral T cells from young Lck^Cre/+Tak1^flox/flox mice did not contain the population of regulatory T (Treg) cells. Our study uncovers novel physiological roles for TAK1 in T cell regulation and functioning.

	Methods

Top Abstract Introduction Methods Results and Discussion References

 
Generation of T cell-specific TAK1-deficient mice
Tak1^flox/flox mice were generated as previously described (1). Lck-Cre mice (9) were bred with Tak1^flox/flox mice to generate mice carrying Lck-cre and floxed Tak1 (Lck^Cre/+Tak1^flox/+) genes. These mice were then mated with Tak1^flox/flox mice. Offspring carrying Lck-cre and floxed Tak1 genes (Lck^Cre/+Tak1^flox/+ or Lck^Cre/+Tak1^flox/flox) were used for analysis.

Southern blot analysis
Southern blot analysis for the deletion of floxed Tak1 allele was performed as described previously (1).

Immunoblot analysis
Cells were lysed in a lysis buffer containing 0.5% Nonidet-P40, 150 mM NaCl, 20 mM Tris–Cl (pH 7.5), 1 mM EDTA and a protease inhibitor cocktail (Complete; Roche Diagnostics, Indianapolis, IN, USA). Lysates were separated by SDS-PAGE and transferred onto polyvinylidene difluoride membrane. The membrane was blotted with the indicated antibodies, and visualized with an enhanced chemiluminescence system (Perkin Elmer Life Sciences, Boston, MA, USA). Anti-TAK1 were as described previously (1). Anti-phospho-JNK, phospho-ERK and phospho-IB were purchased from Cell Signaling Technology (Beverly, MA, USA). Anti-JNK, ERK and IB were from SantaCruz (Santa Cruz, CA, USA).

Flow cytometry
Single-cell suspensions were prepared from thymus, spleen and lymph node. Cells were stained with FITC-, PE-, PerCP- or allophycoerythrin (APC)-conjugated antibodies (PharMingen), and then analyzed on a FACSCalibur (Becton Dickinson, Lincoln Park, NJ, USA). For Foxp3 staining, we used an APC-conjugated anti-Foxp3 and Foxp3 staining buffer set (eBioscience, San Diego, CA, USA), and followed the manufacturer's protocols.

Proliferation assay
Thymocytes (2 x 10⁵) were cultured in 96-well plates for 36 h with the indicated concentration of anti-CD3 with or without 2 µg ml^–1 of anti-CD28 (PharMingen). One microcurie of [³H]-thymidine ([³H]TdR) was pulsed for the last 12 h, and then [³H]TdR uptake was measured in a ß-scintillation counter (Packard).

Cell viability
Thymocytes were treated with the indicated concentration of recombinant mouse TNF (R&D Systems, Minneapolis, MN, USA) for 24 h. Cell viability was examined using Annexin V–Cy3 (BioVision, Mountain View, CA, USA) and FACSCalibur.

Electrophoretic mobility shift assay
Thymocytes (2 x 10⁷) were treated with stimuli for the indicated periods. Nuclear extracts were purified from cells and incubated with a specific probe for the NF-B DNA-binding site, electrophoresed and then visualized by autoradiography as described previously (1).

Histopathological analysis
Paraffin-embedded colon samples were sectioned and stained with hematoxylin and eosin. Transverse sections are shown in the figures. Inflammation was graded semi-quantitatively from 0 (no change) to 5 (most severe colitis) for each region of the colon (cecum, and ascending, transverse, and descending colon, and rectum). The summation of the score for each segment of the colon provides a total disease score per mouse (0–25). The scoring was performed in a blinded manner by two independent investigators.

Statistical analysis
Data are expressed as the mean ± SD. P-values were determined using the Student's t-test.

	Results and Discussion

Top Abstract Introduction Methods Results and Discussion References

 
TAK1 is required for T cell maturation and homeostasis
To investigate the functional role of TAK1 in T cells, we generated T cell-specific TAK1-deficient mice by mating floxed Tak1 (Tak1^flox/flox) mice with transgenic mice expressing Cre under the control of the proximal Lck promoter (9). Lck^Cre/+Tak1^flox/+ and Lck^Cre/+Tak1^flox/flox mice were born at the expected Mendelian ratios, and young mice (4- to 12-weeks old) showed no gross defects in growth and survival in specific pathogen-free (SPF) conditions. Southern blot analysis showed almost complete Cre-mediated deletion of Tak1 in thymocytes from Lck^Cre/+Tak1^flox/flox mice (Fig. 1A). We also checked the deletion of TAK1 by immunoblotting using lysate prepared from thymocytes. Although Cre-mediated deletion led to the production of a truncated/mutated TAK1 lacking the ability to activate NF-B and AP-1 (1), intact TAK1 disappeared in Lck^Cre/+Tak1^flox/flox thymocytes (Fig. 1B).

View larger version (45K): [in this window] [in a new window] [Download PowerPoint slide]   Fig. 1 T cell development in LckCre/+Tak1flox/flox mice. (A) Southern blot analysis of genomic DNA from thymocytes for deletion of the floxed Tak1 allele. (B) Immunoblot analysis of thymocytes from LckCre/+Tak1flox/+ and LckCre/+Tak1flox/flox mice. Cells were lysed and immunoprecipitated with anti-TAK1. Then immunoprecipitants were subjected to SDS-PAGE and immunoblotted with the anti-TAK1. H.C., heavy chain of antibody. (C–F) Flow cytometric analysis of T cell development in the thymus (C and D), spleen (E) and lymph node (F) from LckCre/+Tak1flox/+ and LckCre/+Tak1flox/flox mice. Cells from 8-week-old mice were stained with the indicated antibodies. Percentages of cells within each quadrant are shown (C, E and F). The panels represent histograms of control cells (black line) and LckCre/+Tak1flox/flox cells (filled line) (D). Results are representative of four different experiments.

 
First, we analyzed whether T cell-specific TAK1 deficiency affected T cell development. Flow cytometric analysis of thymocyte populations in Lck^Cre/+Tak1^flox/flox mice revealed that single-positive (SP) cells, especially CD8 SP thymocytes, were markedly reduced (Fig. 1C and Table 1), although the population of CD4⁺CD8⁺ double-positive (DP) thymocytes was not altered between the control and Lck^Cre/+Tak1^flox/flox mice. The double-negative (DN) compartment was further analyzed for the expression of CD44 and CD25, which can define four subsets of DN thymocytes (10). There were no differences in the DN1 to DN4 subsets between control Lck^Cre/+Tak1^flox/+ and Lck^Cre/+Tak1^flox/flox mice (Fig. 1C). It has been reported that mature CD8 SP cells progressively down-regulate HSA (or CD24) surface expression when they go through their final maturation steps in the thymus (11). In both CD8 SP and CD4 SP cells in Lck^Cre/+Tak1^flox/flox mice, we found a severe reduction of HSA^low mature cells, whereas the number of HSA^high immature cells was not affected (Fig. 1D). These results suggest that, in Lck^Cre/+Tak1^flox/flox mice, SP thymocytes progressively disappear during their final maturation steps in the thymus. An analysis of splenocytes and lymph node cells showed that peripheral T cells were remarkably reduced in Lck^Cre/+Tak1^flox/flox mice (Fig. 1E and F and Table 1). Taken together, these data indicate that TAK1 is indispensable for the development of peripheral T cells.

View this table: [in this window] [in a new window]   Table 1 Number of T and B cell subsets in 8-week-old control and LckCre/+Tak1flox/flox mice

 
TAK1 is essential for TCR-mediated proliferation and survival after TNF stimulation
Since there were few peripheral T cells in Lck^Cre/+Tak1^flox/flox mice, we examined T cell responses against TCR stimulation using thymocytes. As shown in Fig. 2(A), thymocytes prepared from Lck^Cre/+Tak1^flox/flox mice poorly proliferated in response to activation of anti-CD3 compared with control thymocytes, even in the presence of the co-stimulatory anti-CD28. We have previously shown that TAK1 is essential for the survival of fibroblasts in response to TNF stimulation by controlling NF-B (1). We also investigated the viability of thymocytes in response to TNF. While thymocytes prepared from Lck^Cre/+Tak1^flox/+ mice were viable after TNF stimulation, this stimulation induced cell death in thymocytes from Lck^Cre/+Tak1^flox/flox mice in a dose-dependent manner (Fig. 2B), indicating that anti-, but not pro-, apoptotic signal induced by TNF is diminished in Lck^Cre/+Tak1^flox/flox mice. Taken together, these results indicate that TAK1 is required for TCR- and TNF-mediated cellular responses in T cells.

View larger version (14K): [in this window] [in a new window] [Download PowerPoint slide]   Fig. 2 Impaired responses to TCR cross-linking and TNF in LckCre/+Tak1flox/flox thymocytes. (A) Thymocytes from LckCre/+Tak1flox/+ or LckCre/+Tak1flox/flox mice were treated with the indicated concentration of anti-CD3 in the presence or absence of 2 µg ml–1 anti-CD28 for 24 h. Proliferation was examined by [3H]TdR incorporation. Values are mean ± SD of triplicate cultures representative of three experiments. (B) Control and LckCre/+Tak1flox/flox thymocytes were treated with the indicated concentration of TNF for 24 h. Viability of the cells was determined by Annexin V–Cy3 staining. Three independent experiments were performed in triplicate. Values are shown as the mean ± SD percentage of viable cells after 24 h.

 
TCR-mediated activation of NF-B and MAPKs is abrogated in Lck^Cre/+Tak1^flox/flox thymocytes
We next examined the activation of signaling molecules. With TCR cross-linking or phorbol ester/calcium ionophore (PMA/iono) stimulation, JNK and ERK were activated in thymocytes prepared from control mice (Fig. 3A). In sharp contrast, activation of JNK, but not ERK, was abrogated in TAK1-deficient thymocytes. Then we examined activation of NF-B in response to TCR or PMA/iono stimulation. TCR-mediated IB phosphorylation or PMA/iono-induced IB degradation was severely impaired in TAK1-deficient thymocytes (Fig. 3B). In accord with this, the induction of DNA–NF-B binding activity was also impaired in TAK1-deficient thymocytes compared with control cells (Fig. 3C). These results indicate that TAK1 is indispensable for the activation of both NF-B and JNK in TCR signaling.

View larger version (68K): [in this window] [in a new window] [Download PowerPoint slide]   Fig. 3 Impaired activation of JNK and NF-B in TAK1-deficient thymocytes. (A and B) MAPK and IB activation in thymocytes. Thymocytes from LckCre/+Tak1flox/+ or LckCre/+Tak1flox/flox mice were untreated or treated with 10 µg ml–1 anti-CD3 plus 2 µg ml–1 anti-CD28 (left panels) or with 50 ng ml–1 each of PMA and ionomycine (PMA + iono, right panels) for the indicated periods. Whole-cell lysates were prepared and subjected to immunoblot analysis with the indicated antibodies. (C) NF-B activation in thymocytes. Control and LckCre/+Tak1flox/flox thymocytes were untreated or treated with anti-CD3/anti-CD28 (left panel) or with PMA/iono (right panel) for the indicated periods, and then nuclear extracts were prepared. NF-B DNA-binding activity was determined by electrophoretic mobility shift assay. Results are representative of four different experiments.

 
Inflammatory bowel diseases in Lck^Cre/+Tak1^flox/flox mice
Lck^Cre/+Tak1^flox/flox mice appeared normal and healthy in the SPF facility until 3 months of age; however, we found that most Lck^Cre/+Tak1^flox/flox mice began showing wasting and severe diarrhea from 4 to 6 months of age. These old Lck^Cre/+Tak1^flox/flox mice also showed stiff colon, rectal prolapse and enlarged spleen and lymph nodes compared with littermate control mice. Upon histological examination of tissues from these mice, we observed prominent ulceration, loss of goblet cells in the epithelial layer and mononuclear cell infiltration of the colon (Fig. 4A and B). The small intestine was never affected (data not shown), and the littermate control mice showed no or only a few symptoms and histological change in their intestine.

View larger version (34K): [in this window] [in a new window] [Download PowerPoint slide]   Fig. 4 IBDs in old LckCre/+Tak1flox/flox mice. (A) Histopathologic analysis of colon from a 6-month-old LckCre/+Tak1flox/+ (a) or an LckCre/+Tak1flox/flox mice (b). The lumen of the colon is at the top of each representative photomicrograph. Hematoxylin–eosin staining is shown at x100. (B) The colitis scores shown for individual mice at 6 months of age were total scores for individual sections as described in Methods. (C) Flow cytometric analysis for the expression of CD44 and CD62L on CD3+ T cell prepared from 40- or 4-week-old mice. Cells were stained with the indicated antibodies. Percentages of cells within each quadrant are shown. Results are representative of four different experiments. (D) Southern blot analysis of genomic DNA from purified splenic CD3+ T cells and B220+ B cells to check the deletion of floxed Tak1 allele.

 
Along with splenomegaly, total cell numbers of splenocytes were increased in old Lck^Cre/+Tak1^flox/flox mice, and the percentage of myeloid lineage cells such as macrophages and neutrophils was up-regulated (data not shown). In addition, the percentage of peripheral T cells in old Lck^Cre/+Tak1^flox/flox mice seemed to be slightly higher than that in young Lck^Cre/+Tak1^flox/flox mice (data not shown). Flow cytometric analysis of the expression of CD44 and CD62L revealed that there were almost no CD62L^highCD44^low naive T cells remaining in the spleen and lymph nodes of old Lck^Cre/+Tak1^flox/flox mice (Fig. 4C). Given that >90% of the peripheral T cells of Lck^Cre/+Tak1^flox/flox mice were CD44^high, most of these cells were considered as activated/memory T cells. To assess the Cre-mediated deletion of Tak1 in peripheral T cells of old mice, CD3⁺ cells or B220⁺ cells were sorted from spleen of control and Lck^Cre/+Tak1^flox/flox mice. Southern blot analysis showed that there was no allele in T cells from old Lck^Cre/+Tak1^flox/flox mice, but there was in old Lck^Cre/+Tak1^flox/+ mice (Fig. 4D), indicating that the remained and/or up-regulated peripheral T cells were ‘leaked’ cells, which had had escaped from Cre-mediated deletion. These results suggest that TAK1 plays a critical role in preventing the development of colitis by controlling homeostasis of T cells.

Role of Treg cells in the development of colitis in Lck^Cre/+Tak1^flox/flox mice
The development of autoimmune bowel disease is sometimes caused by the lack of suppression in immune responses. For instance, mice deficient in IL-2, IL-2Rß or TGF-ß1 and transgenic mice expressing dominant-negative TGFßRII spontaneously develop colitis with massive enlargement of the spleen and lymph nodes because of defects in the maintenance of Treg cells (12–19). These cells have been shown to play a central role in the maintenance of immune tolerance and the termination of immune responses (20–22). A subset of Treg cells develops intrathymically under the control of the specific transcription factor, Foxp3 (20–22). To investigate the role of TAK1 in the development of Treg cells, we checked the population of CD4⁺CD25⁺Foxp3⁺ T cells in Lck^Cre/+Tak1^flox/flox mice. Thymocytes and splenocytes were prepared from 6-week-old Lck^Cre/+Tak1^flox/+ and Lck^Cre/+Tak1^flox/flox mice, stained with appropriate antibodies and then analyzed by flow cytometry. While Treg cells substantially existed in both thymus and spleen of control mice, these cells were not detectable in both organs of Lck^Cre/+Tak1^flox/flox mice (Fig. 5A). Furthermore, the expression level of CD45RB in CD4 SP thymocytes of Lck^Cre/+Tak1^flox/flox mice was similar to that of control mice (Fig. 5B). These results suggest that TAK1 is indispensable for development of thymocytes to Treg precursor cells, but not to CD4⁺CD45RB^high cells, which are capable of inducing colitis when injected in SCID mice (23).

View larger version (18K): [in this window] [in a new window] [Download PowerPoint slide]   Fig. 5 Impaired development of Treg cells in LckCre/+Tak1flox/flox mice. Flow cytometric analysis of the expression of CD25, Foxp3 (A) and CD45RB (B) by CD4 SP thymocytes or CD4+ splenocytes prepared from 6-week-old mice. Cells were stained with the indicated antibodies as described in Methods. Percentages of cells within each quadrant are shown. Results are representative of four different experiments.

 
In summary, we showed that TAK1 is critical for the maintenance of mature T cells, and also for the prevention of spontaneous colitis. Furthermore, TAK1-deficient thymocytes showed impaired TCR-mediated responses, activation of NF-B and MAPKs. The mice also showed severe defects in the maintenance of mature CD4 SP and CD8 SP thymocytes and of peripheral T cells. Nevertheless, the numbers of DN and DP thymocytes were not altered in Lck^Cre/+Tak1^flox/flox mice. It is assumed that Lck^Cre/+-mediated deletion is initiated at the onset of CD44 in earlier DN stages of thymocyte development, suggesting that TAK1 is not critical for the development and maintenance of DP cells.

Involvement of TAK1 in the maintenance of SP thymocytes and peripheral T cells reminds us of the role of the components of the IKK complex in T cells. For instance, it is reported that T cell-specific deletion of NEMO/IKK- or the expression of IKK-ß without kinase activity results in the diminished generation of peripheral CD4⁺ and CD8⁺ T cells (24). These results suggest that TAK1 regulates appropriate T cell development by activating the NF-B pathway. Whereas IKK activity is also essential for the maintenance of mature B cells, we have shown that B cells do not require TAK1 for their development and maintenance, except for peritoneal B-1 B cells (1). These results indicate that the requirement for TAK1 in the activation of IKK complex is determined in a cell type-specific fashion.

Unexpectedly, T cell-specific TAK1-deficient mice spontaneously developed colitis as they aged. Although TAK1-deficient thymocytes were hyporeactive to TCR stimulation, old T cell-specific TAK1-deficient mice showed splenomegaly, lymphadenopathy with full of B cells and a high percentage of activated/memory T cells. Further analysis of thymocytes revealed that intrathymic development of CD4⁺CD25⁺Foxp3⁺ Treg cells was totally dependent on TAK1. Since it has been reported that the expression of Foxp3 in CD4 SP thymocytes is mostly limited to HSA^low population (25), it may be possible that the defect in the development of Treg cells correlates with blockade in the maturation of SP thymocytes. Although cytokines such as IL-2 and TGF-ß1 are critical for the maintenance of Treg cells, they were shown to be dispensable for the thymic development of natural Treg cells (26). The mechanisms for thymic development of natural Treg cells are not currently well understood. It has been shown that Bcl10 and PKC are critical for the development of thymic Treg cells (27), suggesting that TCR-signaling pathway leading to the activation of NF-B is critical for natural Treg cells development.

Although Lck^Cre/+Tak1^flox/flox mice also showed a defect in the maintenance of naive peripheral T cells, thymic SP cells appeared and the defect was more profound in the development of natural Treg cells. Therefore, we assume that the balance between effector and Treg cells is perturbed in the absence of TAK1, and that it is responsible for the spontaneously developed colitis. However, further studies are required to clarify the precise mechanisms for the development of colitis under conditions of TAK1 deficiency.

During the preparation of the manuscript, the generation of Cd4-Cre:Tak1^flox/flox mice, in which TAK1 is deleted in T cells, was reported (28). In that study, development of any diseases was not reported, although the T cell maintenance was similarly defective. The difference may be because of the difference in the promoters regulating Cre expression. The Lck promoter induces expression of the gene early in T cell development, compared with the Cd4 promoter, which starts to express DP stage of thymocytes. Our mouse model appears quite useful for investigating the mechanisms of autoimmune colitis. Future studies should clarify the detailed mechanisms of how TAK1 controls the development of various subsets of T cells and maintains immune tolerance.

	Acknowledgements

 
We thank J. Takeda (Osaka University, Suita, Japan) for providing Lck-Cre mice, K. Nakamura for cell sorting, A. Shibano, M. Shiokawa, Y. Fujiwara, A. Miyabe and N. Kitagaki for technical assistance and M. Hashimoto for secretarial assistance. This work was supported by grants from Special Coordination Funds, the Ministry of Education, Culture, Sports, Science and Technology.

	Abbreviations

 

APC, allophycoerythrin

BCR, B cell receptor

DN, double negative

DP, double positive

[3H]TdR, [3H]-thymidine

IKK, IB kinase

JNK, c-Jun N-terminal kinase

MAPK, mitogen-activated protein kinase

NF-B, nuclear factor-B

PKC, protein kinase C

PMA/iono, phorbol ester/calcium ionophore

SP, single positive

SPF, specific pathogen free

TAK1, transforming growth factor-ß-activating kinase 1

TGF, transforming growth factor

TLR, Toll-like receptor

TNF, tumor necrosis factor

TRAF, TNFR-associated factor

Treg, regulatory T

	Notes

 
Transmitting editor: K. Takatsu

Received 3 July 2006, accepted 24 July 2006.

	References

Top Abstract Introduction Methods Results and Discussion References

 

1. Sato S, Sanjo H, Takeda K, et al. (2005) Essential function for the kinase TAK1 in innate and adaptive immune responses. Nat. Immunol. 6:1087.[CrossRef][Web of Science][Medline]
2. Shim JH, Xiao C, Paschal AE, et al. (2005) TAK1, but not TAB1 or TAB2, plays an essential role in multiple signaling pathways in vivo. Genes Dev. 19:2668.[Abstract/Free Full Text]
3. Ninomiya-Tsuji J, Kishimoto K, Hiyama A, Inoue J, Cao Z, Matsumoto K. (1999) The kinase TAK1 can activate the NIK-I kappaB as well as the MAP kinase cascade in the IL-1 signalling pathway. Nature 398:252.[CrossRef][Medline]
4. Ghosh S and Karin M. (2002) Missing pieces in the NF-kappaB puzzle. Cell 109:S81.
5. Raingeaud J, Whitmarsh AJ, Barrett T, Derijard B, Davis RJ. (1996) MKK3- and MKK6-regulated gene expression is mediated by the p38 mitogen-activated protein kinase signal transduction pathway. Mol. Cell. Biol. 16:1247.[Abstract]
6. Davis RJ. (2000) Signal transduction by the JNK group of MAP kinases. Cell 103:239.[CrossRef][Web of Science][Medline]
7. Thome M. (2004) CARMA1, BCL-10 and MALT1 in lymphocyte development and activation. Nat. Rev. Immunol. 4:348.[CrossRef][Web of Science][Medline]
8. Sun L, Deng L, Ea CK, Xia ZP, Chen ZJ. (2004) The TRAF6 ubiquitin ligase and TAK1 kinase mediate IKK activation by BCL10 and MALT1 in T lymphocytes. Mol. Cell. 14:289.[CrossRef][Web of Science][Medline]
9. Takahama Y, Ohishi K, Tokoro Y, et al. (1998) Functional competence of T cells in the absence of glycosylphosphatidylinositol-anchored proteins caused by T cell-specific disruption of the Pig-a gene. Eur. J. Immunol. 28:2159.[CrossRef][Web of Science][Medline]
10. Godfrey DI, Kennedy J, Suda T, Zlotnik A. (1993) A developmental pathway involving four phenotypically and functionally distinct subsets of CD3-CD4-CD8- triple-negative adult mouse thymocytes defined by CD44 and CD25 expression. J. Immunol. 150:4244.[Abstract]
11. Tian T, Zhang J, Gao L, Qian XP, Chen WF. (2005) Heterogeneity within medullary-type TCRalphabeta(+)CD3(+)CD4(–)CD8(+) thymocytes in normal mouse thymus. Int. Immunol. 13:313.
12. Willerford DM, Chen J, Ferry JA, Davidson L, Ma A, Alt FW. (1995) Interleukin-2 receptor alpha chain regulates the size and content of the peripheral lymphoid compartment. Immunity 3:521.[CrossRef][Web of Science][Medline]
13. Schorle H, Holtschke T, Hunig T, Schimpl A, Horak I. (1991) Development and function of T cells in mice rendered interleukin-2 deficient by gene targeting. Nature 352:621.[CrossRef][Medline]
14. Sadlack B, Merz H, Schorle H, Schimpl A, Feller AC, Horak I. (1993) Ulcerative colitis-like disease in mice with a disrupted interleukin-2 gene. Cell 75:253.[CrossRef][Web of Science][Medline]
15. Nelson BH. (2004) IL-2, regulatory T cells, and tolerance. J. Immunol. 172:3983.[Abstract/Free Full Text]
16. Shull MM, Ormsby I, Kier AB, et al. (1992) Targeted disruption of the mouse transforming growth factor-beta1 gene results in multifocal inflammatory disease. Nature 359:693.[CrossRef][Medline]
17. Kulkarni AB, Huh CG, Becker D, et al. (1993) Transforming growth factor beta 1 null mutation in mice causes excessive inflammatory response and early death. Proc. Natl Acad. Sci. USA 90:770.[Abstract/Free Full Text]
18. Gorelik L and Flavell RA. (2000) Abrogation of TGF beta signaling in T cells leads to spontaneous T cell differentiation and autoimmune disease. Immunity 12:171.[CrossRef][Web of Science][Medline]
19. Aoki CA, Borchers AT, Li M, et al. (2005) Transforming growth factor beta (TGF-ß) and autoimmunity. Autoimmun. Rev. 4:450.[CrossRef][Web of Science][Medline]
20. Schwartz RH. (2005) Natural regulatory T cells and self-tolerance. Nat. Immunol. 6:327.[CrossRef][Web of Science][Medline]
21. Fontenot JD and Rudensky AY. (2005) A well adapted regulatory contrivance: regulatory T cell development and the forkhead family transcription factor Foxp3. Nat. Immunol. 6:331.[CrossRef][Web of Science][Medline]
22. Sakaguchi S. (2005) Naturally arising Foxp3-expressing CD25+CD4+ regulatory T cells in immunological tolerance to self and non-self. Nat. Immunol. 6:345.[CrossRef][Web of Science][Medline]
23. Powrie F, Leach MW, Mauze S, Menon S, Caddle LB, Coffman RL. (1994) Inhibition of Th1 responses prevents inflammatory bowel disease in scid mice reconstituted with CD45RB^high CD4⁺ T cells. Immunity 1:553.[CrossRef][Web of Science][Medline]
24. Schmidt-Supprian M, Courtois G, Tian J, et al. (2003) Mature T cells depend on signaling through the IKK complex. Immunity 19:377.[CrossRef][Web of Science][Medline]
25. Fontenot JD, Rasmussen JP, Williams LM, Dooley JL, Farr AG, Rudensky AY. (2005) Regulatory T cell lineage specification by the forkhead transcription factor foxp3. Immunity 22:329.[CrossRef][Web of Science][Medline]
26. Weaver CT, Harrington LE, Mangan PR, Gavrieli M, Murphy KM. (2006) Th17: an effector CD4 T cell lineage with regulatory T cell ties. Immunity 24:677.[CrossRef][Medline]
27. Schmidt-Supprian M, Tian J, Grant EP, et al. (2004) Differential dependence of CD4⁺CD25⁺ regulatory and natural killer-like T cells on signals leading to NF-kappaB activation. Proc. Natl Acad. Sci. USA 101:4566.[Abstract/Free Full Text]
28. Wan YY, Chi H, Xie M, Schneider MD, Flavell RA. (2006) The kinase TAK1 integrates antigen and cytokine receptor signaling for T cell development, survival and function. Nat. Immunol. 7:851.[CrossRef][Web of Science][Medline]

CiteULike    Connotea    Del.icio.us    What's this?

This article has been cited by other articles:

				R. D. Schulteis, H. Chu, X. Dai, Y. Chen, B. Edwards, D. Haribhai, C. B. Williams, S. Malarkannan, M. J. Hessner, S. Glisic-Milosavljevic, et al. Impaired survival of peripheral T cells, disrupted NK/NKT cell development, and liver failure in mice lacking Gimap5 Blood, December 15, 2008; 112(13): 4905 - 4914. [Abstract] [Full Text] [PDF]

				R. R. McCully and J. L. Pomerantz The Protein Kinase C-Responsive Inhibitory Domain of CARD11 Functions in NF-{kappa}B Activation To Regulate the Association of Multiple Signaling Cofactors That Differentially Depend on Bcl10 and MALT1 for Association Mol. Cell. Biol., September 15, 2008; 28(18): 5668 - 5686. [Abstract] [Full Text] [PDF]

				R. Kajino-Sakamoto, M. Inagaki, E. Lippert, S. Akira, S. Robine, K. Matsumoto, C. Jobin, and J. Ninomiya-Tsuji Enterocyte-Derived TAK1 Signaling Prevents Epithelium Apoptosis and the Development of Ileitis and Colitis J. Immunol., July 15, 2008; 181(2): 1143 - 1152. [Abstract] [Full Text] [PDF]

				M. Tang, X. Wei, Y. Guo, P. Breslin, S. Zhang, S. Zhang, W. Wei, Z. Xia, M. Diaz, S. Akira, et al. TAK1 is required for the survival of hematopoietic cells and hepatocytes in mice J. Exp. Med., July 7, 2008; 205(7): 1611 - 1619. [Abstract] [Full Text] [PDF]

				S. Suzuki, P. Singhirunnusorn, A. Mori, S. Yamaoka, I. Kitajima, I. Saiki, and H. Sakurai Constitutive Activation of TAK1 by HTLV-1 Tax-dependent Overexpression of TAB2 Induces Activation of JNK-ATF2 but Not IKK-NF-{kappa}B J. Biol. Chem., August 31, 2007; 282(35): 25177 - 25181. [Abstract] [Full Text] [PDF]

				W. W. Reiley, W. Jin, A. J. Lee, A. Wright, X. Wu, E. F. Tewalt, T. O. Leonard, C. C. Norbury, L. Fitzpatrick, M. Zhang, et al. Deubiquitinating enzyme CYLD negatively regulates the ubiquitin-dependent kinase Tak1 and prevents abnormal T cell responses J. Exp. Med., June 11, 2007; 204(6): 1475 - 1485. [Abstract] [Full Text] [PDF]

				K. A. Alford, S. Glennie, B. R. Turrell, L. Rawlinson, J. Saklatvala, and J. L. E. Dean Heat Shock Protein 27 Functions in Inflammatory Gene Expression and Transforming Growth Factor-beta-activated Kinase-1 (TAK1)-mediated Signaling J. Biol. Chem., March 2, 2007; 282(9): 6232 - 6241. [Abstract] [Full Text] [PDF]

This Article Abstract FREE Full Text (PDF) All Versions of this Article: 18/10/1405    most recent dxl082v1 Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Add to My Personal Archive Download to citation manager Search for citing articles in: ISI Web of Science (11) Request Permissions Google Scholar Articles by Sato, S. Articles by Akira, S. Search for Related Content PubMed PubMed Citation Articles by Sato, S. Articles by Akira, S. Social Bookmarking          What's this?

Online ISSN 1460-2377 - Print ISSN 0953-8178

Copyright © 2009 Japanese Society for Immunology

Oxford Journals Oxford University Press

• Site Map
• Privacy Policy
• Frequently Asked Questions

Other Oxford University Press sites: Oxford University Press Oxford Journals China Oxford Journals Japan American National Biography Booksellers' Information Service Children's Fiction and Poetry Children's Reference Corporate & Special Sales Dictionaries Dictionary of National Biography Digital Reference English Language Teaching Higher Education Textbooks Humanities International Education Unit Law Medicine Music Online Products Oxford English Dictionary Reference Rights and Permissions Science School Books Social Sciences Very Short Introductions World's Classics