Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Crucial functions of the Rap1 effector molecule RAPL in lymphocyte and dendritic cell trafficking

Nature Immunology volume 5, pages 1045–1051 (2004)Cite this article

Abstract

Immunosurveillance requires the coordinated regulation of chemokines and adhesion molecules to guide immune cell migration. However, the critical molecule for governing the high trafficking capability of immune cells is not clear. Here we show that the effector molecule RAPL is indispensable in the integrin-mediated adhesion and migration of lymphocytes and dendritic cells. RAPL deficiency caused defective chemokine-triggered lymphocyte adhesion and migration to secondary lymphoid organs, resulting in atrophic lymphoid follicles and deficient marginal zone B cells, concomitant with increased immature B cells in the blood. Furthermore, splenic dendritic cells were diminished and defective in adhesion. After being activated with inflammatory stimuli, skin and splenic dendritic cells failed to migrate into either the draining lymph nodes or the white pulp of the spleen. Thus, RAPL is a crucial immune cell trafficking regulator essential for immunosurveillance.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Impairment of chemokine-stimulated lymphocyte adhesion.
Figure 2: Defective lymphocyte trafficking to secondary lymphoid organs.
Figure 3: Abnormal trafficking of thymocytes and immature B cells and deficient marginal zone B cells.
Figure 4: Impairment of DC adhesion and migration.
Figure 5: Impaired DC trafficking from skin to draining lymph node.

Similar content being viewed by others

References

  1. Steinman, R.M. The dendritic cell system and its role in immunogenicity. Annu. Rev. Immunol. 9, 271–296 (1991).

    Article  CAS  Google Scholar 

  2. Steinman, R.M., Pack, M. & Inaba, K. Dendritic cells in the T-cell areas of lymphoid organs. Immunol. Rev. 156, 25–37 (1997).

    Article  CAS  Google Scholar 

  3. Ardavin, C. Origin, precursors and differentiation of mouse dendritic cells. Nat. Rev. Immunol. 3, 582–590 (2003).

    Article  CAS  Google Scholar 

  4. Springer, T.A. Traffic signals on endothelium for lymphocyte recirculation and leukocyte emigration. Annu. Rev. Physiol. 57, 827–872 (1995).

    Article  CAS  Google Scholar 

  5. Butcher, E.C. & Picker, L.J. Lymphocyte homing and homeostasis. Science 272, 60–66 (1996).

    Article  CAS  Google Scholar 

  6. Katagiri, K. et al. Rap1 is a potent activation signal for leukocyte function-associated antigen 1 distinct from protein kinase C and phosphatidylinositol-3-kinase. Mol. Cell. Biol. 20, 1956–1969 (2000).

    Article  CAS  Google Scholar 

  7. Reedquist, K.A. et al. The small GTPase, Rap1, mediates CD31-induced integrin adhesion. J. Cell Biol. 148, 1151–1158 (2000).

    Article  CAS  Google Scholar 

  8. Katagiri, K., Hattori, M., Minato, N. & Kinashi, T. Rap1 functions as a key regulator of T-cell and antigen-presenting cell interactions and modulates T-cell responses. Mol. Cell. Biol. 22, 1001–1015 (2002).

    Article  CAS  Google Scholar 

  9. Shimonaka, M. et al. Rap1 translates chemokine signals to integrin activation, cell polarization, and motility across vascular endothelium under flow. J. Cell Biol. 161, 417–427 (2003).

    Article  CAS  Google Scholar 

  10. Katagiri, K., Maeda, A., Shimonaka, M. & Kinashi, T. RAPL, a Rap1-binding molecule that mediates Rap1-induced adhesion through spatial regulation of LFA-1. Nat. Immunol. 4, 741–748 (2003).

    Article  CAS  Google Scholar 

  11. Kinashi, T. & Katagiri, K. Regulation of lymphocyte adhesion and migration by the small GTPase Rap1 and its effector molecule, RAPL. Immunol. Lett. 93, 1–5 (2004).

    Article  CAS  Google Scholar 

  12. Dustin, M.L., Bivona, T.G. & Philips, M.R. Membranes as messengers in T cell adhesion signaling. Nat. Immunol. 5, 363–372 (2004).

    Article  CAS  Google Scholar 

  13. Chaffin, K.E. & Perlmutter, R.M. A pertussis toxin-sensitive process controls thymocyte emigration. Eur. J. Immunol. 21, 2565–2573 (1991).

    Article  CAS  Google Scholar 

  14. Ueno, T. et al. Role for CCR7 ligands in the emigration of newly generated T lymphocytes from the neonatal thymus. Immunity 16, 205–218 (2002).

    Article  CAS  Google Scholar 

  15. Fukui, Y. et al. Haematopoietic cell-specific CDM family protein DOCK2 is essential for lymphocyte migration. Nature 412, 826–831 (2001).

    Article  CAS  Google Scholar 

  16. Matloubian, M. et al. Lymphocyte egress from thymus and peripheral lymphoid organs is dependent on S1P receptor 1. Nature 427, 355–360 (2004).

    Article  CAS  Google Scholar 

  17. Loder, F. et al. B cell development in the spleen takes place in discrete steps and is determined by the quality of B cell receptor-derived signals. J. Exp. Med. 190, 75–89 (1999).

    Article  CAS  Google Scholar 

  18. Martin, F. & Kearney, J.F. Marginal-zone B cells. Nat. Rev. Immunol. 2, 323–335 (2002).

    Article  CAS  Google Scholar 

  19. Lu, T.T. & Cyster, J.G. Integrin-mediated long-term B cell retention in the splenic marginal zone. Science 297, 409–412 (2002).

    Article  CAS  Google Scholar 

  20. Kitajima, T., Ariizumi, K., Bergstresser, P.R. & Takashima, A. A novel mechanism of glucocorticoid-induced immune suppression: the inhibition of T cell-mediated terminal maturation of a murine dendritic cell line. J. Clin. Invest. 98, 142–147 (1996).

    Article  CAS  Google Scholar 

  21. Pribila, J.T., Itano, A.A., Mueller, K.L. & Shimizu, Y. The α1β1 and αEβ7 integrins define a subset of dendritic cells in peripheral lymph nodes with unique adhesive and antigen uptake properties. J. Immunol. 172, 282–291 (2004).

    Article  CAS  Google Scholar 

  22. Vremec, D., Pooley, J., Hochrein, H., Wu, L. & Shortman, K. CD4 and CD8 expression by dendritic cell subtypes in mouse thymus and spleen. J. Immunol. 164, 2978–2986 (2000).

    Article  CAS  Google Scholar 

  23. De Smedt, T. et al. Regulation of dendritic cell numbers and maturation by lipopolysaccharide in vivo. J. Exp. Med. 184, 1413–1424 (1996).

    Article  CAS  Google Scholar 

  24. Macatonia, S.E., Knight, S.C., Edwards, A.J., Griffiths, S. & Fryer, P. Localization of antigen on lymph node dendritic cells after exposure to the contact sensitizer fluorescein isothiocyanate. Functional and morphological studies. J. Exp. Med. 166, 1654–1667 (1987).

    Article  CAS  Google Scholar 

  25. Gunn, M.D. et al. Mice lacking expression of secondary lymphoid organ chemokine have defects in lymphocyte homing and dendritic cell localization. J. Exp. Med. 189, 451–460 (1999).

    Article  CAS  Google Scholar 

  26. Forster, R. et al. CCR7 coordinates the primary immune response by establishing functional microenvironments in secondary lymphoid organs. Cell 99, 23–33 (1999).

    Article  CAS  Google Scholar 

  27. Price, A.A., Cumberbatch, M., Kimber, I. & Ager, A. Alpha 6 integrins are required for Langerhans cell migration from the epidermis. J. Exp. Med. 186, 1725–1735 (1997).

    Article  CAS  Google Scholar 

  28. Xu, H. et al. The role of ICAM-1 molecule in the migration of Langerhans cells in the skin and regional lymph node. Eur. J. Immunol. 31, 3085–3093 (2001).

    Article  CAS  Google Scholar 

  29. Schneeberger, E.E., Vu, Q., LeBlanc, B.W. & Doerschuk, C.M. The accumulation of dendritic cells in the lung is impaired in CD18−/− but not in ICAM-1−/− mutant mice. J. Immunol. 164, 2472–2478 (2000).

    Article  CAS  Google Scholar 

  30. Berlin-Rufenach, C. et al. Lymphocyte migration in lymphocyte function-associated antigen (LFA)-1-deficient mice. J. Exp. Med. 189, 1467–1478 (1999).

    Article  CAS  Google Scholar 

  31. Stein, J.V. et al. The CC chemokine thymus-derived chemotactic agent 4 (TCA-4, secondary lymphoid tissue chemokine, 6Ckine, exodus-2) triggers lymphocyte function-associated antigen 1-mediated arrest of rolling T lymphocytes in peripheral lymph node high endothelial venules. J. Exp. Med. 191, 61–76 (2000).

    Article  CAS  Google Scholar 

  32. Warnock, R.A. et al. The role of chemokines in the microenvironmental control of T versus B cell arrest in Peyer's patch high endothelial venules. J. Exp. Med. 191, 77–88 (2000).

    Article  CAS  Google Scholar 

  33. Lo, C.G., Lu, T.T. & Cyster, J.G. Integrin-dependence of lymphocyte entry into the splenic white pulp. J. Exp. Med. 197, 353–361 (2003).

    Article  CAS  Google Scholar 

  34. Cinamon, G. et al. Sphingosine 1-phosphate receptor 1 promotes B cell localization in the splenic marginal zone. Nat. Immunol. 5, 713–720 (2004).

    Article  CAS  Google Scholar 

  35. Pillai, S. The chosen few? Positive selection and the generation of naive B lymphocytes. Immunity 10, 493–502 (1999).

    Article  CAS  Google Scholar 

  36. Petrie, H.T. Cell migration and the control of post-natal T-cell lymphopoiesis in the thymus. Nat. Rev. Immunol. 3, 859–866 (2003).

    Article  CAS  Google Scholar 

  37. Sanui, T. et al. DOCK2 is essential for antigen-induced translocation of TCR and lipid rafts, but not PKC-θ and LFA-1, in T cells. Immunity 19, 119–129 (2003).

    Article  CAS  Google Scholar 

  38. Amsen, D., Kruisbeek, A., Bos, J.L. & Reedquist, K. Activation of the Ras-related GTPase Rap1 by thymocyte TCR engagement and during selection. Eur. J. Immunol. 30, 2832–2841 (2000).

    Article  CAS  Google Scholar 

  39. Sebzda, E., Bracke, M., Tugal, T., Hogg, N. & Cantrell, D.A. Rap1A positively regulates T cells via integrin activation rather than inhibiting lymphocyte signaling. Nat. Immunol. 3, 251–258 (2002).

    Article  CAS  Google Scholar 

  40. Kamath, A.T. et al. The development, maturation, and turnover rate of mouse spleen dendritic cell populations. J. Immunol. 165, 6762–6770 (2000).

    Article  CAS  Google Scholar 

  41. Leenen, P.J. et al. Heterogeneity of mouse spleen dendritic cells: in vivo phagocytic activity, expression of macrophage markers, and subpopulation turnover. J. Immunol. 160, 2166–2173 (1998).

    CAS  PubMed  Google Scholar 

  42. Imhof, B.A. & Aurrand-Lions, M. Adhesion mechanisms regulating the migration of monocytes. Nat. Rev. Immunol. 4, 432–444 (2004).

    Article  CAS  Google Scholar 

  43. Yagi, T. et al. A novel ES cell line, TT2, with high germline-differentiating potency. Anal. Biochem. 214, 70–76 (1993).

    Article  CAS  Google Scholar 

  44. Tachibana, M. et al. G9a histone methyltransferase plays a dominant role in euchromatic histone H3 lysine 9 methylation and is essential for early embryogenesis. Genes. Dev. 16, 1779–1791 (2002).

    Article  CAS  Google Scholar 

  45. Inaba, K. et al. Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with granulocyte/macrophage colony-stimulating factor. J. Exp. Med. 176, 1693–1702 (1992).

    Article  CAS  Google Scholar 

  46. Martin-Fontecha, A. et al. Regulation of dendritic cell migration to the draining lymph node: impact on T lymphocyte traffic and priming. J. Exp. Med. 198, 615–621 (2003).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank A. Takashima (University of Texas, Dallas, Texas) for the XS52 cell line; Y. Fukui (Kyusyu University, Fukuoka, Japan) for discussions; and M. Imai and M. Hirata for technical assistance. Supported by the Ministry of Education, Sciences, Sports, and Cultures and Cell Science Research Foundation (16043224 and 14370112 to T.K. and 13140202 and 16390116 to K.I.).

Author information

Authors and Affiliations

  1. Department of Molecular Immunology and Allergy, Graduate School of Medicine, Kyoto University, Kyoto, 606-8501, Japan

    Koko Katagiri, Noriko Ohnishi & Tatsuo Kinashi

  2. Department of Dermatology, Graduate School of Medicine, Kyoto University, Kyoto, 606-8507, Japan

    Kenji Kabashima

  3. Department of Animal Development and Physiology, Division of Systemic Life Science, Laboratory of Immunobiology, Graduate School of Biostudies, Graduate School of Science, Kyoto University, Kyoto, 606-8502, Japan

    Tomonori Iyoda & Kayo Inaba

  4. Center for Animal Resources and Development, Kumamoto University, Kumamoto, 860-0811, Japan

    Naoki Takeda

  5. Experimental Research Center for Infectious Diseases, Institute for Virus Research, Kyoto University, Kyoto, 606-8507, Japan

    Yoichi Shinkai

Authors
  1. Koko Katagiri
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Noriko Ohnishi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kenji Kabashima
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Tomonori Iyoda
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Naoki Takeda
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yoichi Shinkai
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Kayo Inaba
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Tatsuo Kinashi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Tatsuo Kinashi.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Targeted disruption of the Rassf5 gene by homologous recombination. (PDF 916 kb)

Supplementary Fig. 2

Expression of LFA-1, CCR7, and CXCR4 in lymphocytes. (PDF 192 kb)

Supplementary Fig. 3

Adhesion and transmission of lymphocytes under shear flow. (PDF 164 kb)

Supplementary Video 1

Transmigration of lymphocytes infected with control adenovirus through endothelial monolayers without CCL21. (MOV 530 kb)

LN T cells from Balb/c mice transgenic for the adenovirus receptor were incubated with control adenovirus and cultured for 2 days before under-flow adhesion assays. Adenovirus-infected cells express GFP as a marker.

Time-lapse images of control adenovirus-infected lymphocytes were collected every 10 seconds, and the video was created at 10 frames/second using QuickTime Pro (Apple Computer Inc., Cupertino, CA).

Supplementary Video 2

Transmigration of lymphocytes infected with control adenovirus through endothelial monolayers with CCL21. (MOV 599 kb)

LN T cells from Balb/c mice transgenic for the adenovirus receptor were incubated with control adenovirus and cultured for 2 days before under-flow adhesion assays. Adenovirus-infected cells express GFP as a marker.

Time-lapse images of control adenovirus-infected lymphocytes stimulated with CCL21 were collected every 10 seconds, and the video was created at 10 frames/second using QuickTime Pro (Apple Computer Inc., Cupertino, CA).

Supplementary Video 3

Transmigration of lymphocytes infected with RAPL adenovirus through endothelial monolayers in the absence of CCL21 (MOV 764 kb)

LN T cells from Balb/c mice transgenic for the adenovirus receptor were incubated with RAPL-expressing adenovirus and cultured for 2 days before under-flow adhesion assays. Adenovirus-infected cells express GFP as a marker.

Time-lapse images of RAPL adenovirus-infected lymphocytes were collected every 10 seconds, and the video was created at 10 frames/second using QuickTime Pro (Apple Computer Inc., Cupertino, CA).

Rights and permissions

Reprints and permissions

About this article

Cite this article

Katagiri, K., Ohnishi, N., Kabashima, K. et al. Crucial functions of the Rap1 effector molecule RAPL in lymphocyte and dendritic cell trafficking. Nat Immunol 5, 1045–1051 (2004). https://doi.org/10.1038/ni1111

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ni1111

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing