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

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.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

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.

References

  1. 1

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

  2. 2

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

  3. 3

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

  4. 4

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

  5. 5

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

  6. 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).

  7. 7

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

  8. 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).

  9. 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).

  10. 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).

  11. 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).

  12. 12

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

  13. 13

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

  14. 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).

  15. 15

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

  16. 16

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

  17. 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).

  18. 18

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

  19. 19

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

  20. 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).

  21. 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).

  22. 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).

  23. 23

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

  24. 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).

  25. 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).

  26. 26

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

  27. 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).

  28. 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).

  29. 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).

  30. 30

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

  31. 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).

  32. 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).

  33. 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).

  34. 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).

  35. 35

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

  36. 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).

  37. 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).

  38. 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).

  39. 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).

  40. 40

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

  41. 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).

  42. 42

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

  43. 43

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

  44. 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).

  45. 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).

  46. 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).

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

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

Further reading