CD69-null mice protected from arthritis induced with anti-type II collagen antibodies

Kaoru Murata1,2, Masamichi Inami1, Akihiro Hasegawa1, Shuichi Kubo1, Motoko Kimura1, Masakatsu Yamashita1,4, Hiroyuki Hosokawa1, Tomokazu Nagao7, Kazuo Suzuki7, Kahoko Hashimoto5, Hiroshi Shinkai2, Haruhiko Koseki3, Masaru Taniguchi1,6, Steven F. Ziegler8 and Toshinori Nakayama1

Departments of 1 Molecular Immunology and Medical Immunology, 2 Clinical Biology of Extracellular Matrix and 3 Molecular Embryology, Graduate School of Medicine, Chiba University, Chiba 260-8670, Japan 4 PRESTO, Japan Science and Technology Corp., 5 Laboratory for Dendritic Cell Immunobiology and 6 Laboratory for Immune Regulation, RIKEN Research Center for Allergy and Immunology, Japan 7 Biodefense Laboratory, Department of Bioactive Molecule, National Institute of Infectious Diseases, Tokyo 162-8640, Japan 8 Immunology Program, Benaroya Research Institute at Virginia Mason, Seattle, WA 98101, USA

Correspondence to: T. Nakayama; E-mail: nakayama{at}med.m.chiba-u.ac.jp
Transmitting editor: M. Miyasaka


    Abstract
 Top
 Abstract
 Introduction
 Methods
 Results
 Discussion
 References
 
CD69, known as an early activation marker antigen on T and B cells, is also expressed on platelets and activated neutrophils, suggesting certain roles in inflammatory diseases. In order to address the role of CD69 in the pathogenesis of arthritis, we established CD69-null mice. CD69-null mice displayed a markedly attenuated arthritic inflammatory response when injected with anti-type II collagen antibodies. Cell transfer experiments with neutrophils, but not T cells or spleen cells, from wild-type mice into CD69-null mice restored the induction of arthritis. These results indicate a critical role for CD69 in neutrophil function in arthritis induction during the effector phase. Thus, CD69 would be a possible therapeutic target for arthritis in human patients.

Keywords: IL-6, neutrophil, rheumatoid arthritis


    Introduction
 Top
 Abstract
 Introduction
 Methods
 Results
 Discussion
 References
 
Collagen-induced arthritis (CIA) is an experimental model of rheumatoid arthritis (RA), although there are several differences in the features of inflammation (1,2). In this model, antibodies against type II collagen are critical for the pathogenesis of arthritis (35). Treatment with a mixture of four monoclonal anti-type II collagen mAb and lipopolysaccharide (LPS) reproducibly induced arthritis in various strains of mice (6). This model system has been used for investigating underlying cellular and molecular mechanisms during the effector phase of CIA (7,8).

CD69 is a type II membrane protein expressed as a homodimer of heavily glycosylated subunits (9). Both T and B cells begin to express CD69 within a few hours after stimulation, and it is known as an early activation marker antigen of lymphocytes (10). Regulatory roles for CD69 in TCR-mediated signaling events have also been suggested (10). Freshly prepared thymocytes undergoing selection events express CD69 (11,12) and regulatory roles for CD69 expression in T cell development in the thymus were suggested (13,14). A mild effect on B cell development was also reported (15). Constitutive expression of CD69 was noted on platelets (10), and activated neutrophils and eosinophils express CD69 on their cell surface; however, the role of CD69 in immune responses and inflammation in the periphery has not been well clarified.

An analysis of infiltrating inflammatory cells in the joints of RA patients suggested the involvement of CD69 in the disease. Co-culture of T cells and monocytic cells from the RA patients resulted in tumor necrosis factor (TNF)-{alpha} production, which was inhibited by pre-treatment of T cells with CD69-specific antibodies (16). These data suggested a role for CD69 on T cells in the pathogenesis of RA. To extend these studies, we established CD69-null mice to assess the role of CD69 in inflammation leading to arthritis. We found that the induction of arthritis with anti-type II collagen mAb was inhibited in the CD69-null mice, suggesting a crucial role for CD69 in the pathogenesis of CIA, particularly during the effector phase.


    Methods
 Top
 Abstract
 Introduction
 Methods
 Results
 Discussion
 References
 
Establishment of CD69-null mice
Genomic clones encoding the mouse CD69 gene were isolated from a C57BL/6 mouse genomic library (17). To introduce null mutations into the CD69 locus, we generated a replacement vector to remove the first exon of the CD69 gene encompassing the initiation codon (Supplementary Data available at International Immunology online, Supplementary Fig. 1). CD69-null (CD69-KO) mice with a 129/Ola background were established and backcrossed 11 times to the BALB/c background. The null expression of CD69 molecules on activated T cells was confirmed by flow cytometry (Supplementary Fig. 2). T (Supplementary Fig. 3) and B (data not shown) cell development appeared to be unaffected. All mice used in this study were maintained under specific pathogen-free conditions. Animal care was in accordance with the guidelines of Chiba University.

Induction and scoring of arthritis
Arthritis is induced by an arthritogenic mAb mixture purchased from Immuno-Biological Laboratories (Gunma, Japan) as described (6,7). Eight-week-old CD69-KO female mice were injected i.v. with 2 mg of anti-type II collagen mAb. Two days later, LPS (50 µg) was i.p. injected. Swelling of the hind paws was examined every other day thereafter. The severity of the arthritis was graded on a clinical score of 0–3: 0, normal; 1, slight swelling and/or erythema; 2, pronounced edematous swelling; 3, joint rigidity. Each limb was graded, giving a maximum score of 12.

Cytokine and chemokine expression in the joint (hind paw)
Ten days after the initial mAb injection, hind paws were homogenized as described (7) and total RNA was isolated using TRIzol (Life Technologies, Gaithersburg, MD) (18). The PCR primer sequences were as follows: mouse IL-1ß, 5'-CTGAAAGCTCTCCACCTC-3' (sense) and 5'-GGTGCTGAT

Histology
Hind legs were removed, fixed in 4% paraformaldehyde/PBS, decalcified in 10% EDTA, dehydrated in 50–100% ethanol, propanol and xylene, and embedded in paraffin as described (7,19). Then, the samples were sectioned and stained with hematoxylin & eosin, and examined for pathological changes under a light microscope at x80 and x160 magnification.

In situ hybridization
In situ hybridization was performed as described (20). Knee joints decalcified in 10% EDTA were processed for paraffin embedding. Riboprobes were prepared using T3 and T7 RNA polymerase using a cDNA template encoding the CD69 coding region (9).

Neutrophil preparation and administration
Wild-type BALB/c mice were injected i.p. with 2 ml of 4% thioglycolate (Merck, Darmstadt, Germany) and peritoneal neutrophils were recovered 4 h later by collecting peritoneal lavage with 5 ml of saline (21). More than 80% of the cells were neutrophils with Gr-1 staining (data not shown). Ten million neutrophils were injected i.v. into CD69-KO mice on day 0 and 2.

Statistical analysis
The Fisher’s PLSD test was used to evaluate the statistical significance of the data presented.


    Results
 Top
 Abstract
 Introduction
 Methods
 Results
 Discussion
 References
 
Arthritis induced by anti-type II collagen antibodies in CD69-KO mice
The goal of this study was to evaluate the role of CD69 in the pathogenesis of arthritis. Wild-type and CD69-KO mice on a BALB/c background were treated with anti-type II collagen antibodies on day 0 and with LPS on day 2. The severity of the arthritis was scored and is depicted in Fig. 1(A). The first sign of arthritis was detected on day 4 and the arthritis was established in the control wild-type mice around day 10, as reported previously (7). The severity of it was clearly reduced in CD69-KO mice and essentially no rigidity was detected. Figure 1(B) shows macroscopic views of a representative arthritis joint 10 days after anti-type II collagen antibody injection. The massive swelling observed in the joints of wild-type mice treated with anti-type II collagen antibodies and LPS (Fig. 1B, b) was not observed in those of CD69-KO mice (Fig. 1B, d). Figure 1(C) shows representative results of a histological analysis of the joints with arthritis. A large number of neutrophils had infiltrated around the joint, and the structure of the synovial membrane and bone were destroyed in wild-type mice (Fig. 1C, b). More than 95% of the infiltrated cells were neutrophils by morphological criteria (Fig. 1C, e) and a low percentage of macrophages was detected. As expected, the levels of neutrophil infiltration were dramatically diminished in the joints of CD69-KO mice (Fig. 1C, d).





View larger version (227K):
[in this window]
[in a new window]
 
Fig. 1. Arthritis induced by anti-type II collagen antibodies in CD69-KO mice. Wild-type and CD69-KO mice (four mice each) with a BALB/c background were treated with anti-type II collagen antibodies on day 0 and LPS on day 2. Total three independent experiments were performed with similar results. (A) The severity of the arthritis was scored and depicted. *Statistically significant (between wild-type and CD69-KO group with treatment, P < 0.01). (B) A representative picture of a hind paw in each group (a: wild-type, no treatment; b: wild-type, antibody treatment; c: CD69-KO, no treatment; and d: CD69-KO, antibody treatment). (C) Histological analysis of arthritic joints with hematoxylin & eosin staining. The joints were prepared on day 10 (a: wild-type, no treatment; b and e: wild-type, antibody treatment; c: CD69-KO, no treatment; d: CD69-KO, antibody treatment; a–d: x80 and e: x160).

 
Expression of cytokines and chemokines in the arthritic hind paws of CD69-KO mice
The levels of certain cytokines (IL-1ß and IL-6) and chemokines (MIP-1{alpha} and MCP-1) appeared to be highly correlated with the severity of arthritis induced with anti-type II collagen antibodies and LPS (7). Thus, the transcriptional expression levels of these cytokines and chemokines in the arthritic joints of CD69-KO mice were examined by semi-quantitative RT-PCR analysis. As shown in Fig. 2(A), the increase observed in mRNA levels of IL-1ß, IL-6 and MCP-1 were all decreased in the CD69-KO groups, suggesting that the severity of the arthritis was considerably low. The decrease was detected even on day 10. The expression of TNF-{alpha} was not detected in either group, and that of MIP-1{alpha} was detected, but not changed, after treatment with anti-type II collagen mAb and LPS (data not shown).



View larger version (63K):
[in this window]
[in a new window]
 
Fig. 2. Expression of cytokines and chemokines in the arthritic hind paws of CD69-KO mice. (A) The transcriptional expression levels of IL-1ß, IL-6, MCP-1 and GAPDH in the arthritic joints of CD69-KO mice 4 and 10 days after antibody injection were examined by semi-quantitative RT-PCR analysis. Four mice per group were used and representative data are shown. Densitometric units of the band of the highest concentration were measured and normalized with the corresponding GAPDH band. The numbers from left to right are as follows: IL-1ß (day 4): 0.02, 1.00, 0.07 and 0.03; IL-1ß (day 10): 0.06, 1.00, 0.04 and 0.04; IL-6 (day 4): 0.38, 1.00, 0.07 and 0.05; IL-6 (day 10): 0.08, 1.00, 0.06 and 0.07; MCP-1 (day 4): 0.41, 1.00, 0.15 and 0.11; MCP-1 (day 10): 0.75, 1.00, 0.14 and 0.21. (B) The transcriptional expression levels of CXCL1, CXCL4, CXCL7, MCP-2, KC and GAPDH in the arthritic joints of CD69-KO mice 4 days after antibody injection were examined by semi-quantitative RT-PCR analysis. The densitometric values measured as in (A) are as follows: CXCL1 (day 4): 0.03, 1.00, 0.02 and 0.02; CXCL1 (day 10): 0.27, 1.00, 0.18 and 0.11; CXCL4 (day 4): 0.91, 1.00, 0.85 and 0.90; CXCL4 (day 10): 0.97, 1.00, 0.99 and 1.01; CXCL7 (day 4): 0.66, 1.00, 0.73 and 0.79; CXCL7 (day 10): 0.87, 1.00, 0.88 and 0.70; MCP-2 (day 4): 1.15, 1.00, 0.99 and 0.94; MCP-2 (day 10): 0.52, 1.00, 0.49 and 0.40; KC (day 4): 1.03, 1.00, 1.12 and 1.06; and KC (day 10): 0.93, 1.00, 1.00 and 0.97.

 
Next, chemokines involved in neutrophil migration (CXCL1, CXCL4, CXCL7, MCP-2 and KC) were examined (Fig. 2B). The transcription of CXCL1 was induced after treatment with anti-type II collagen antibodies and LPS, and the expression levels were significantly lower in CD69-KO mice on day 4. The difference was also detected on day 10, but the expression of CXCL1 was decreased in both wild-type and CD69-KO mice. Significant levels of transcription of other genes were detected in the normal joint (no treatment) and no induction was observed after the treatment on day 4. Also no difference between wild-type and CD69-KO mice was observed. On day 10, the expression levels of MCP-2 in CD69-KO mice with treatment were significantly lower than those of wild-type mice. The levels of CXCL4, CXCL7 and KC mRNA were not significantly different between CD69-KO and wild-type mice. These results suggest the possibility that the induction of CXCL1 is involved in neutrophil migration in the arthritic joint. Also, decreased neutrophil migration in CD69-KO mice would be due, at least in part, to the decreased CXCL1 expression in the joint. In addition, the involvement of MCP-1 and MCP-2 was suggested.

Expression of CD69 molecules on infiltrating neutrophils in the arthritic joints
Next, we examined the expression of CD69 in the infiltrating neutrophils in the joints of wild-type mice injected with the anti-type II collagen mAb. In situ hybridization was used to determine CD69 mRNA expression in infiltrating neutrophils. As shown in Fig. 3(a), significant expression of CD69 mRNA was detected in the infiltrating neutrophils (arrow) in the arthritic joints of anti-type II collagen-treated mice. No significant expression of CD69 mRNA was observed in the synovial membrane in the joints of untreated mice (Fig. 3b).



View larger version (140K):
[in this window]
[in a new window]
 
Fig. 3. Expression of CD69 in infiltrating neutrophils in the arthritic joint. The expression of CD69 in the infiltrating neutrophils in the joints was assessed by in situ hybridization. The joints were prepared on day 10 (a: wild-type, antibody treatment; b: wild-type, no treatment). Neutrophils were determined by morphological criteria. Arrows indicate representative CD69 mRNA+ neutrophils.

 
Transfer of CD69-expressing neutrophils into CD69-null mice reconstituted the anti-type II collagen-mediated arthritic response
The results thus far suggested that CD69 molecules on neutrophils are critical for the arthritis induced by anti-type II collagen mAb and LPS. In order to examine this hypothesis, 1 x 107 neutrophils prepared from wild-type BALB/c mice by thioglycolate treatment were injected i.v. into CD69-KO mice on the same day as antibody injection and also on day 2. The clinical scores of wild-type and CD69-KO mice after neutrophil cell transfer were indistinguishable, and were significantly higher than those of CD69-KO mice at all time points tested (Fig. 4). The restored induction of arthritis was not observed by cell transfer of large numbers of T cells (1 x 108) or whole-spleen cells (1 x 108) from wild-type mice (data not shown). These results suggest that CD69 expression on neutrophils is crucial for the induction of the arthritis.



View larger version (24K):
[in this window]
[in a new window]
 
Fig. 4. Arthritis induced by anti-type II collagen mAb was restored by cell transfer of wild-type neutrophils into CD69-KO mice. The arthritis was induced as in Fig. 1. Ten million neutrophils prepared from wild-type BALB/c mice by thioglycolate treatment were injected i.v. into CD69-KO mice on days 0 and 2. The clinical scores of the wild-type and CD69-KO mice with or without neutrophil cell transfer are shown. *Statistically significant (between neutrophil transferred and non-transferred groups in CD69-KO mice, P < 0.01). **Statistically significant (between neutrophil transferred and non-transferred groups in CD69-KO mice, P < 0.05).

 

    Discussion
 Top
 Abstract
 Introduction
 Methods
 Results
 Discussion
 References
 
Clinical and biological manifestations of rheumatoid arthritis are heterogeneous, reflecting a multitude of causative factors and the presence of complex pathophysiological mechanisms (22). CIA has been used to dissect the underlying complex pathogenesis of rheumatoid arthritis, although there are several differences in the feature of inflammation (1,2). Here, we demonstrate the involvement of CD69 in the pathogenesis of arthritis using newly established CD69-KO mice and an experimental mouse arthritis model induced with anti-type II collagen antibodies. Moreover, CD69 expression on neutrophils was suggested to be critical for the induction of arthritis. Although CD69 is highly expressed on activated T and B cells, the induction of arthritis with anti-type II collagen mAb and LPS is independent of T and B cells (7), and thus we expected that CD69 expressed on other cell types could be important. Neutrophils are known to be important in the murine model of arthritis (23), and massive infiltration was observed in our model (Fig. 1C). Wild-type neutrophils (two injections, 1 x 107) could restore the arthritic induction almost completely in CD69-KO mice (Fig. 4), but transfer of large numbers (two injections, 1 x 108) of wild-type T cells or spleen cells into CD69-KO mice showed no restoration of arthritis.

The induction of pro-inflammatory cytokines such as IL-1ß and IL-6 in the arthritic joints was significantly lower in CD69-KO groups (Fig. 2A). This may reflect the decreased severity of arthritis in CD69-KO mice. In contrast, diminished expression of CXCL1, MCP-1 and MCP-2 may contribute to the decreased migration of neutrophils into the joint. However, it is not known how CD69 deficiency resulted in the diminished induction of these chemokine mRNAs in the joints of CD69-KO mice at this time.

Our data demonstrates a CD69-dependent role for neutrophils in the effector phase of anti-type II collagen mAb-induced arthritis. As for the mechanism behind the involvement of CD69 molecules in the induction of arthritis, particularly in the effector phase, one possibility is that CD69 molecules are involved in the migration and interaction of neutrophils with antibody-bound collagen-bearing tissues in the joint (24) through putative CD69 ligands. CD69 is a C-type lectin family glycoprotein and a role in cell adhesion was suggested (9,13). Another possibility is that the expression of CD69 molecules on neutrophils is required for their inflammatory response in the joints of affected mice. A role for CD69 as a co-receptor in cellular responses has been shown in T cells (10). In fact, similar to T and B cells, the cell surface expression of CD69 on neutrophils was significantly enhanced after activation with phorbol myristate acetate and ionomycin (K. Murata et al., unpublished observation). It has not been clarified whether CD69 can be induced by other stimulants such as LPS, autoantibodies and/or inflammatory cytokines in vivo. Also, it is of great interest to assess the role of CD69 on neutrophils in various innate immune responses, autoimmune responses and inflammation. Further studies on the role of CD69 in inflammation await characterization of its ligands.

In summary, the expression of CD69 is found to control the development of arthritis induced with anti-type II collagen mAb. This suggests a crucial role for CD69 in the pathogenesis of CIA, particularly in the effector phase, and that the specific blockade of the CD69 interaction may have therapeutic value in the arthritis.


    Acknowledgements
 
The authors are grateful to Drs Daisuke Kitamura, Masato Kubo, Hidetada Kawana and Kenichiro Seino for helpful comments on the establishment of CD69-null mice, pathological diagnosis and neutrophil preparation. The authors also thank Ms Kaoru Sugaya for excellent technical assistance and Ms Nina Maeshima for comments during the preparation of the manuscript. This work was supported by grants from the Ministry of Education, Culture, Sports, Science and Technology (Japan) (Grants-in-Aid for: Scientific Research, Priority Areas Research 13218016 and 12051203; Scientific Research B 14370107; and Special Coordination Funds for Promoting Science and Technology), the Ministry of Health, Labor and Welfare (Japan), the Program for Promotion of Fundamental Studies in Health Science of the Organization for Pharmaceutical Safety and Research (Japan), and the Human Frontier Science Program Research Grant (RG00168/2000-M206). S. F. Z. is partially funded by NIH grant AI.


    Abbreviations
 
CD69-KO—CD69-null

CIA—collagen-induced arthritis

LPS—lipopolysaccharide

TNF—tumor necrosis factor

RA—rheumatoid arthritis


    References
 Top
 Abstract
 Introduction
 Methods
 Results
 Discussion
 References
 

  1. Myers, L. K., Rosloniec, E. F., Cremer, M. A. and Kang, A. H. 1997. Collagen-induced arthritis, an animal model of autoimmunity. Life Sci. 61:1861.[CrossRef][ISI][Medline]
  2. Holmdahl, R., Bockermann, R., Backlund, J. and Yamada, H. 2002. The molecular pathogenesis of collagen-induced arthritis in mice—a model for rheumatoid arthritis. Ageing Res. Rev. 1:135.[CrossRef][ISI][Medline]
  3. Hirano, T. 2002. Revival of the autoantibody model in rheumatoid arthritis. Nat. Immunol. 3:342.[CrossRef][ISI][Medline]
  4. Matsumoto, I., Maccioni, M., Lee, D. M., Maurice, M., Simmons, B., Brenner, M., Mathis, D. and Benoist, C. 2002. How antibodies to a ubiquitous cytoplasmic enzyme may provoke joint-specific autoimmune disease. Nat. Immunol. 3:360.[CrossRef][ISI][Medline]
  5. Wipke, B. T., Wang, Z., Kim, J., McCarthy, T. J. and Allen, P. M. 2002. Dynamic visualization of a joint-specific autoimmune response through positron emission tomography. Nat. Immunol. 3:366.[CrossRef][ISI][Medline]
  6. Terato, K., Hasty, K. A., Reife, R. A., Cremer, M. A., Kang, A. H. and Stuart, J. M. 1992. Induction of arthritis with monoclonal antibodies to collagen. J. Immunol. 148:2103.[Abstract/Free Full Text]
  7. Kagari, T., Doi, H. and Shimozato, T. 2002. The importance of IL-1ß and TNF-{alpha}, and the noninvolvement of IL-6, in the development of monoclonal antibody-induced arthritis. J. Immunol. 169:1459.[Abstract/Free Full Text]
  8. Johansson, A. C., Hansson, A. S., Nandakumar, K. S., Backlund, J. and Holmdahl, R. 2001. IL-10-deficient B10.Q mice develop more severe collagen-induced arthritis, but are protected from arthritis induced with anti-type II collagen antibodies. J. Immunol. 167:3505.[Abstract/Free Full Text]
  9. Ziegler, S. F., Ramsdell, F., Hjerrild, K. A., Armitage, R. J., Grabstein, K. H., Hennen, K. B., Farrah, T., Fanslow, W. C., Shevach, E. M. and Alderson, M. R. 1993. Molecular characterization of the early activation antigen CD69: a type II membrane glycoprotein related to a family of natural killer cell activation antigens. Eur. J. Immunol. 23:1643.[ISI][Medline]
  10. Testi, R., D‘Ambrosio, D., De Maria, R. and Santoni, A. 1994. The CD69 receptor: a multipurpose cell-surface trigger for hematopoietic cells. Immunol. Today 15:479.[CrossRef][ISI][Medline]
  11. Yamashita, I., Nagata, T., Tada, T. and Nakayama, T. 1993. CD69 cell surface expression identifies developing thymocytes which audition for T cell antigen receptor-mediated positive selection. Int. Immunol. 5:1139.[Abstract]
  12. Bendelac, A., Matzinger, P., Seder, R. A., Paul, W. E. and Schwartz, R. H. 1992. Activation events during thymic selection. J. Exp. Med. 175:731.[Abstract]
  13. Nakayama, T., Kasprowicz, D. J., Yamashita, M., Schubert, L. A., Gillard, G., Kimura, M., Didierlaurent, A., Koseki, H. and Ziegler, S. F. 2002. The generation of mature, single-positive thymocytes in vivo is dysregulated by CD69 blockade or overexpression. J. Immunol. 168:87.[Abstract/Free Full Text]
  14. Feng, C., Woodside, K. J., Vance, B. A., El-Khoury, D., Canelles, M., Lee, J., Gress, R., Fowlkes, B. J., Shores, E. W. and Love, P. E. 2002. A potential role for CD69 in thymocyte emigration. Int. Immunol. 14:535.[Abstract/Free Full Text]
  15. Lauzurica, P., Sancho, D., Torres, M., Albella, B., Marazuela, M., Merino, T., Bueren, J. A., Martinez, A. C. and Sanchez-Madrid, F. 2000. Phenotypic and functional characteristics of hematopoietic cell lineages in CD69-deficient mice. Blood 95:2312.[Abstract/Free Full Text]
  16. McInnes, I. B., Leung, B. P., Sturrock, R. D., Field, M. and Liew, F. Y. 1997. Interleukin-15 mediates T cell-dependent regulation of tumor necrosis factor-{alpha} production in rheumatoid arthritis. Nat. Med. 3:189.[ISI][Medline]
  17. Ziegler, S. F., Levin, S. D., Johnson, L., Copeland, N. G., Gilbert, D. J., Jenkins, N. A., Baker, E., Sutherland, G. R., Feldhaus, A. L. and Ramsdell, F. 1994. The mouse CD69 gene. Structure, expression, and mapping to the NK gene complex. J. Immunol. 152:1228.[Abstract/Free Full Text]
  18. Kimura, M., Koseki, Y., Yamashita, M., Watanabe, N., Shimizu, C., Katsumoto, T., Kitamura, T., Taniguchi, M., Koseki, H. and Nakayama, T. 2001. Regulation of Th2 cell differentiation by mel-18, a mammalian polycomb group gene. Immunity 15:275.[ISI][Medline]
  19. Shibata, Y., Kamata, T., Kimura, M., Yamashita, M., Wang, C. R., Murata, K., Miyazaki, M., Taniguchi, M., Watanabe, N. and Nakayama, T. 2002. Ras activation in T cells determines the development of antigen-induced airway hyperresponsiveness and eosinophilic inflammation. J. Immunol. 169:2134.[Abstract/Free Full Text]
  20. Hoshino, M., Sone, M., Fukata, M., Kuroda, S., Kaibuchi, K., Nabeshima, Y. and Hama, C. 1999. Identification of the stef gene that encodes a novel guanine nucleotide exchange factor specific for Rac1. J. Biol. Chem. 274:17837.[Abstract/Free Full Text]
  21. Ajuebor, M. N., Das, A. M., Virag, L., Flower, R. J., Szabo, C. and Perretti, M. 1999. Role of resident peritoneal macrophages and mast cells in chemokine production and neutrophil migration in acute inflammation: evidence for an inhibitory loop involving endogenous IL-10. J. Immunol. 162:1685.[Abstract/Free Full Text]
  22. Choy, E. H. and Panayi, G. S. 2001. Cytokine pathways and joint inflammation in rheumatoid arthritis. N. Engl. J. Med. 344:907.[Free Full Text]
  23. Wipke, B. T. and Allen, P. M. 2001. Essential role of neutrophils in the initiation and progression of a murine model of rheumatoid arthritis. J. Immunol. 167:1601.[Abstract/Free Full Text]
  24. Birner, U., Issekutz, T. B., Walter, U. and Issekutz, A. C. 2000. The role of {alpha}4 and LFA-1 integrins in selectin-independent monocyte and neutrophil migration to joints of rats with adjuvant arthritis. Int. Immunol. 12:141.[Abstract/Free Full Text]




This Article
Abstract
FREE Full Text (PDF)
Supplementary data
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 (8)
Request Permissions
Google Scholar
Articles by Murata, K.
Articles by Nakayama, T.
PubMed
PubMed Citation
Articles by Murata, K.
Articles by Nakayama, T.