Journal of Molecular Medicine

, 86:999 | Cite as

In vivo dendritic cell depletion reduces breeding efficiency, affecting implantation and early placental development in mice

  • Gesa Krey
  • Pierre Frank
  • Valerie Shaikly
  • Gabriela Barrientos
  • Rosalia Cordo-Russo
  • Frauke Ringel
  • Petra Moschansky
  • Igor V. Chernukhin
  • Metodi Metodiev
  • Nelson Fernández
  • Burghard F. Klapp
  • Petra C. Arck
  • Sandra M. Blois
Original Article


Implantation of mammalian embryos into their mother’s uterus ensures optimal nourishment and protection throughout development. Complex molecular interactions characterize the implantation process, and an optimal synchronization of the components of this embryo-maternal dialogue is crucial for a successful reproductive outcome. In the present study, we investigated the role of dendritic cells (DC) during implantation process using a transgenic mouse system (DTRtg) that allows transient depletion of CD11c+ cells in vivo through administration of diphtheria toxin. We observed that DC depletion impairs the implantation process, resulting in a reduced breeding efficiency. Furthermore, the maturity of uterine natural killer cells at dendritic cell knockout (DCKO) implantation sites was affected as well; as demonstrated by decreased perforin expression and reduced numbers of periodic-acid-Schiff (PAS)-positive cells. This was accompanied by disarrangements in decidual vascular development. In the present study, we were also able to identify a novel DC-dependent protein, phosphatidylinositol transfer protein β (PITPβ), involved in implantation and trophoblast development using a proteomic approach. Indeed, DCKO mice exhibited substantial anomalies in placental development, including hypocellularity of the spongiotrophoblast and labyrinthine layers and reduced numbers of trophoblast giant cells. Giant cells also down-regulated their expression of two characteristic markers of trophoblast differentiation, placental lactogen 1 and proliferin. In view of these findings, dendritic cells emerge as possible modulators in the orchestration of events leading to the establishment and maintenance of pregnancy.


Dendritic cells Implantation Natural killer cells Placentation 



This work was supported by research grants from the Charité (AF-2007-011) to S.M.B. G.B. and R.C-R. received a scholarship from the German Student Exchange Program (Deutscher Akademischer Austauschdienst). S.M.B is a fellow of the Habilitation program at the Charité, University Medicine Berlin. P.C.A. and S.M.B. are part of the EMBIC Network of Excellence, co-financed by the European Commission throughout the FP6 framework program Life Science, Genomics and Biotechnology for Health.

Author contributions

G.K. performed all the experiments and contributed to manuscript writing; V.S. designed and performed proteomics experiments. P.F., G.B., and P.M. performed histological analysis and animal experiments; R.C-R. and F.R. assisted with the PCR experiments; I.C. and M.M. assisted with MALDI-MS experiments; B.F.K and N.F provided advice, P.C.A. contributed with reagents and S.M.B. supervised the work, designed the experiments and wrote the manuscript.


  1. 1.
    Aluvihare VR, Kallikourdis M, Betz AG (2005) Tolerance, suppression and the fetal allograft. J Mol Med 83.2:88–96 (Feb)CrossRefGoogle Scholar
  2. 2.
    Carson DD, Bagchi I, Dey SK, Enders AC, Fazleabas AT, Lessey BA, Yoshinaga K (2000) Embryo implantation. Dev Biol 223.2:217–237 (Jul 15)CrossRefGoogle Scholar
  3. 3.
    Norwitz ER, Schust DJ, Fisher SJ (2001) Implantation and the survival of early pregnancy. N Engl J Med 345.13:1400–1408 (Nov 8)CrossRefGoogle Scholar
  4. 4.
    Lydon JP, DeMayo FJ, Funk CR, Mani SK, Hughes AR, Montgomery CA Jr, Shyamala G, Conneely OM, O'Malley BW (1995) Mice lacking progesterone receptor exhibit pleiotropic reproductive abnormalities. Genes Dev 9.18:2266–2278 (Sep 15)CrossRefGoogle Scholar
  5. 5.
    Moffett-King A (2002) Natural killer cells and pregnancy. Nat Rev Immunol 2.9:656–663 (Sep)CrossRefGoogle Scholar
  6. 6.
    Croy BA, Esadeg S, Chantakru S, van den Heuvel M, Paffaro VA, He H, Black GP, Ashkar AA, Kiso Y, Zhang J (2003) Update on pathways regulating the activation of uterine natural killer cells, their interactions with decidual spiral arteries and homing of their precursors to the uterus. J Reprod Immunol 59.2:175–191 (Aug)CrossRefGoogle Scholar
  7. 7.
    Verma S, Hiby SE, Loke YW, King A (2000) Human decidual natural killer cells express the receptor for and respond to the cytokine interleukin 15. Biol Reprod 62.4:959–968 (Apr)CrossRefGoogle Scholar
  8. 8.
    Shinozaki M, Hirahashi J, Lebedeva T, Liew FY, Salant DJ, Maron R, Kelley VR (2002) IL-15, a survival factor for kidney epithelial cells, counteracts apoptosis and inflammation during nephritis. J Clin Invest 109.7:951–960 (Apr)Google Scholar
  9. 9.
    Waldmann TA, Dubois S, Tagaya Y (2001) Contrasting roles of IL-2 and IL-15 in the life and death of lymphocytes: implications for immunotherapy. Immunity 14.2:105–110 (Feb)Google Scholar
  10. 10.
    Ain R, Canham LN, Soares MJ (2003) Gestation stage-dependent intrauterine trophoblast cell invasion in the rat and mouse: novel endocrine phenotype and regulation. Dev Biol 260.1:176–190 (Aug 1)CrossRefGoogle Scholar
  11. 11.
    Ashkar AA, Di Santo JP, Croy BA (2000) Interferon gamma contributes to initiation of uterine vascular modification, decidual integrity, and uterine natural killer cell maturation during normal murine pregnancy. J Exp Med 192.2:259–270 (Jul 17)CrossRefGoogle Scholar
  12. 12.
    Croy BA, He H, Esadeg S, Wei Q, McCartney D, Zhang J, Borzychowski A, Ashkar AA, Black GP, Evans SS, Chantakru S, van den Heuvel M, Paffaro VA Jr, Yamada AT (2003) Uterine natural killer cells: insights into their cellular and molecular biology from mouse modelling. Reproduction 126.2:149–160 (Aug)CrossRefGoogle Scholar
  13. 13.
    Guimond MJ, Luross JA, Wang B, Terhorst C, Danial S, Croy BA (1997) Absence of natural killer cells during murine pregnancy is associated with reproductive compromise in TgE26 mice. Biol Reprod 56.1:169–179 (Jan)CrossRefGoogle Scholar
  14. 14.
    Blois SM, Alba Soto CD, Tometten M, Klapp BF, Margni RA, Arck PC (2004) Lineage, maturity, and phenotype of uterine murine dendritic cells throughout gestation indicate a protective role in maintaining pregnancy. Biol Reprod 70.4:1018–1023 (Apr)Google Scholar
  15. 15.
    Haimovici F, Anderson DJ (1993) Cytokines and growth factors in implantation. Microsc Res Tech 25.3:201–207 (Jun 15)CrossRefGoogle Scholar
  16. 16.
    Blois SM, Ilarregui JM, Tometten M, Garcia M, Orsal AS, Cordo-Russo R, Toscano MA, Bianco GA, Kobelt P, Handjiski B, Tirado I, Markert UR, Klapp BF, Poirier F, Szekeres-Bartho J, Rabinovich GA, Arck PC (2007) A pivotal role for galectin-1 in fetomaternal tolerance. Nat Med 13(12):1450–1457 (Dec)PubMedCrossRefGoogle Scholar
  17. 17.
    Laskarin G, Kämmerer U, Rukavina D, Thomson AW, Fernandez N, Blois SM (2007) Antigen-presenting cells and materno-fetal tolerance: an emerging role for dendritic cells. Am J Reprod Immunol 58(3):255–67 (Sep)PubMedCrossRefGoogle Scholar
  18. 18.
    Ferlazzo G, Pack M, Thomas D, Paludan C, Schmid D, Strowig T, Bougras G, Muller WA, Moretta L, Munz C (2004) Distinct roles of IL-12 and IL-15 in human natural killer cell activation by dendritic cells from secondary lymphoid organs. Proc Natl Acad Sci U S A 101.47:16606–16611 (Nov 23)CrossRefGoogle Scholar
  19. 19.
    Yu Y, Hagihara M, Ando K, Gansuvd B, Matsuzawa H, Tsuchiya T, Ueda Y, Inoue H, Hotta T, Kato S (2001) Enhancement of human cord blood CD34+ cell derived NK cell cytotoxicity by dendritic cells. J Immunol 166:1590–1600PubMedGoogle Scholar
  20. 20.
    Piccioli D, Sbrana S, Melandri E, Valiante N (2002) Contact-dependent stimulation and inhibition of dendritic cells by natural killer cells. J Exp Med 195:335–341PubMedCrossRefGoogle Scholar
  21. 21.
    Blois SM, Barrientos G, Garcia MG, Orsal AS, Tometten M, Cordo-Russo R, Klapp BF, Santoni A, Fernandez N, Terness P, Arck PC (2008) Interaction between dendritic cells and natural killer cells during pregnancy in mice. J Mol Med (doi: 10.1007/s00109-008-0342-2)
  22. 22.
    Jung S, Unutmaz D, Wong P, Sano G, De los Santos K, Sparwasser T, Wu S, Vuthoori S, Ko K, Zavala F, Pamer EG, Littman DR, Lang RA (2002) In vivo depletion of CD11c(+) dendritic cells abrogates priming of CD8(+) T cells by exogenous cell-associated antigens. Immunity 17.2:211–220 (Aug)CrossRefGoogle Scholar
  23. 23.
    Probst HC, Tschannen K, Odermatt B, Schwendener R, Zinkernagel RM, Van Den Broek M (2005) Histological analysis of CD11c-DTR/GFP mice after in vivo depletion of dendritic cells. Clin Exp Immunol 141.3:398–404 (Sep)CrossRefGoogle Scholar
  24. 24.
    Kammerer U, Eggert AO, Kapp M, McLellan AD, Geijtenbeek TB, Dietl J, van Kooyk Y, Kampgen E (2003) Unique appearance of proliferating antigen-presenting cells expressing DC-SIGN (CD209) in the decidua of early human pregnancy. Am J Pathol 162.3:887–896 (Mar)Google Scholar
  25. 25.
    Ye W, Zheng LM, Young JD, Liu CC (1996) The involvement of interleukin (IL)-15 in regulating the differentiation of granulated metrial gland cells in mouse pregnant uterus. J Exp Med 184.6:2405–2410 (Dec 1)CrossRefGoogle Scholar
  26. 26.
    Wang H, Dey S (2006) Roadmap to embryo implantation: clues from mouse models. Nat Rev Genet 7.3:185–199CrossRefGoogle Scholar
  27. 27.
    Yang Z, Tschopp O, Hemmings-Mieszczak M, Feng J, Brodbeck D, Perentes E, Hemmings B (2003) Protein kinase B alpha/Akt1 regulates placental development and fetal growth. J Biol Chem 278:32124–32131PubMedCrossRefGoogle Scholar
  28. 28.
    Riley J, Moley K (2006) Glucose utilization and the PI3-K-pathway: mechanisms for cell survival in preimplantation embryos. Reproduction 131:823–835PubMedCrossRefGoogle Scholar
  29. 29.
    Qiu Q, Yang M, Tsang B, Gruslin A (2004) EGF-induced trophoblast secretion of MMP-9 and TIMP-1 involves activation of both PI3K and MAPK signalling pathways. Reproduction 128:355–363PubMedCrossRefGoogle Scholar
  30. 30.
    Kamei T, Jones S, Chapman B, McGonigle K, Dai G, Soares M (2002) The phosphatidylinositol 3-Kinase/Akt signaling pathway modulates the endocrine differentiation of trophoblast cells. Mol Endocrinol 16(7):1469–1481PubMedCrossRefGoogle Scholar
  31. 31.
    Faria TN, Soares MJ (1991) Trophoblast cell differentiation: establishment, characterization, and modulation of a rat trophoblast cell line expressing members of the placental prolactin family. Endocrinology 129.6:2895–2906 (Dec)CrossRefGoogle Scholar
  32. 32.
    Blois SM, Kammerer U, Soto CA, Tometten MC, Shaikly V, Barrientos G, Jurd R, Rukavina D, Thomson AW, Klapp BF, Fernandez N, Arck PC (2007) Dendritic cells: key to fetal tolerance? Biol Reprod 77.4:590–598 (Oct)CrossRefGoogle Scholar
  33. 33.
    Parr MB, Parr EL (1989) Immunohistochemical investigation of secretory component and immunoglobulin A in the genital tract of the female rat. J Reprod Fertil 85.1:105–113 (Jan)Google Scholar
  34. 34.
    Ashkar AA, Black GP, Wei Q, He H, Liang L, Head JR, Croy BA (2003) Assessment of requirements for IL-15 and IFN regulatory factors in uterine NK cell differentiation and function during pregnancy. J Immunol 171.6:2937–2944 (Sep 15)Google Scholar
  35. 35.
    Barber EM, Pollard JW (2003) The uterine NK cell population requires IL-15 but these cells are not required for pregnancy nor the resolution of a Listeria monocytogenes infection. J Immunol 171.1:37–46 (Jul 1)Google Scholar
  36. 36.
    Lucas M, Schachterle W, Oberle K, Aichele P, Diefenbach A (2007) Dendritic cells prime natural killer cells by trans-presenting interleukin 15. Immunity 26.4:503–517 (Apr)CrossRefGoogle Scholar
  37. 37.
    Burnett TG, Hunt JS (2000) Nitric oxide synthase-2 and expression of perforin in uterine NK cells. J Immunol 164.10:5245–5250 (May 15)Google Scholar
  38. 38.
    Croy BA, Luross JA, Guimond MJ, Hunt JS (1996) Uterine natural killer cells: insights into lineage relationships and functions from studies of pregnancies in mutant and transgenic mice. Nat Immunol 15.1:22–33Google Scholar
  39. 39.
    Croy BA, Ashkar AA, Foster RA, DiSanto JP, Magram J, Carson D, Gendler SJ, Grusby MJ, Wagner N, Muller W, Guimond MJ (1997) Histological studies of gene-ablated mice support important functional roles for natural killer cells in the uterus during pregnancy. J Reprod Immunol 35.2:111–133 (Nov 15)CrossRefGoogle Scholar
  40. 40.
    Wang C, Tanaka T, Nakamura H, Umesaki N, Hirai K, Ishiko O, Ogita S, Kaneda K (2003) Granulated metrial gland cells in the murine uterus: localization, kinetics, and the functional role in angiogenesis during pregnancy. Microsc Res Tech 60.4:420–429 (Mar 1)CrossRefGoogle Scholar
  41. 41.
    Croy BA, Ashkar AA, Minhas K, Greenwood JD (2000) Can murine uterine natural killer cells give insights into the pathogenesis of preeclampsia? J Soc Gynecol Investig 7.1:12–20 (Jan–Feb)CrossRefGoogle Scholar
  42. 42.
    Cockcroft S, Carvou N (2007) Biochemical and biological functions of class I phosphatidylinositol transfer proteins. Biochim Biophys Acta 1771.6:677–691 (Jun)Google Scholar
  43. 43.
    Lee KF, Kwok KL, Chung MK, Lee YL, Chow JF, Yeung WS (2005) Phospholipid transfer protein (PLTP) mRNA expression is stimulated by developing embryos in the oviduct. J Cell Biochem 95.4:740–749 (Jul 1)CrossRefGoogle Scholar
  44. 44.
    Cunningham E, Thomas G, Ball A, Hiles I, Cockcroft S (1995) Phosphatidylinositol transfer protein dictates the rate of inositol trisphosphate production by promoting the synthesis of PIP2. Curr Biol 5.7:775–783CrossRefGoogle Scholar
  45. 45.
    Martin T (1995) Intracellular signalling. New directions for phosphatidylinositol transfer. Curr Biol 5.9:990–992CrossRefGoogle Scholar
  46. 46.
    Panaretou C, Domin J, Cockcroft S, Waterfield M (1997) Characterization of p150, an adaptor protein for the human phosphatidylinositol (PtdIns) 3-kinase. Substrate presentation by phosphatidylinositol transfer protein to the p150.Ptdins 3-kinase complex. J Biol Chem 272.4:2477–2485Google Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Gesa Krey
    • 1
  • Pierre Frank
    • 1
  • Valerie Shaikly
    • 2
  • Gabriela Barrientos
    • 1
  • Rosalia Cordo-Russo
    • 1
  • Frauke Ringel
    • 1
  • Petra Moschansky
    • 1
  • Igor V. Chernukhin
    • 2
  • Metodi Metodiev
    • 2
  • Nelson Fernández
    • 2
  • Burghard F. Klapp
    • 1
  • Petra C. Arck
    • 1
  • Sandra M. Blois
    • 1
  1. 1.Charité Centrum 12 für Innere Medizin und Dermatologie, Reproductive Immunology Research GroupUniversity Medicine of BerlinBerlinGermany
  2. 2.Department of Biological SciencesUniversity of EssexColchesterUK

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