Induction of natural killer T cell–dependent alloreactivity by administration of granulocyte colony–stimulating factor after bone marrow transplantation

Article metrics


Granulocyte colony–stimulating factor (G-CSF) is often used to hasten neutrophil recovery after allogeneic bone marrow transplantation (BMT), but the clinical and immunological consequences evoked remain unclear1. We examined the effect of G-CSF administration after transplantation in mouse models and found that exposure to either standard G-CSF or pegylated-G-CSF soon after BMT substantially increased graft-versus-host disease (GVHD). This effect was dependent on total body irradiation (TBI) rendering host dendritic cells (DCs) responsive to G-CSF by upregulating their expression of the G-CSF receptor. Stimulation of host DCs by G-CSF subsequently unleashed a cascade of events characterized by donor natural killer T cell (NKT cell) activation, interferon-γ secretion and CD40-dependent amplification of donor cytotoxic T lymphocyte function during the effector phase of GVHD. Crucially, the detrimental effects of G-CSF were only present when it was administered after TBI conditioning and at a time when residual host antigen presenting cells were still present, perhaps explaining the conflicting and somewhat controversial clinical studies from the large European and North American BMT registries2,3. These data have major implications for the use of G-CSF in disease states where NKT cell activation may have effects on outcome.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1: Administration of G-CSF or peg–G-CSF following bone marrow transplantation increases GVHD.
Figure 2: Administration of G-CSF after BMT activates donor NKT cells and enhances GVHD in an NKT cell− and CD8+ T cell–dependent manner.
Figure 3: G-CSF treatment after BMT increases NKT cell–dependent GVHD via effects on residual host tissue.
Figure 4: Conditioning with TBI, but not chemotherapy, enhances G-CSFR expression on host DCs and exacerbates GVHD after G-CSF administration.


  1. 1

    Morris, E.S., MacDonald, K.P. & Hill, G.R. Stem cell mobilization with G-CSF analogs: a rational approach to separate GVHD and GVL? Blood 107, 3430–3435 (2006).

  2. 2

    Ringden, O. et al. Treatment with granulocyte colony–stimulating factor after allogeneic bone marrow transplantation for acute leukemia increases the risk of graft-versus-host disease and death: a study from the Acute Leukemia Working Party of the European Group for Blood and Marrow Transplantation. J. Clin. Oncol. 22, 416–423 (2004).

  3. 3

    Khoury, H.J. et al. Impact of posttransplantation G-CSF on outcomes of allogeneic hematopoietic stem cell transplantation. Blood 107, 1712–1716 (2006).

  4. 4

    Klumpp, T.R., Mangan, K.F., Goldberg, S.L., Pearlman, E.S. & Macdonald, J.S. Granulocyte colony–stimulating factor accelerates neutrophil engraftment following peripheral-blood stem-cell transplantation: a prospective, randomized trial. J. Clin. Oncol. 13, 1323–1327 (1995).

  5. 5

    McQuaker, I.G. et al. Low-dose filgrastim significantly enhances neutrophil recovery following autologous peripheral-blood stem-cell transplantation in patients with lymphoproliferative disorders: evidence for clinical and economic benefit. J. Clin. Oncol. 15, 451–457 (1997).

  6. 6

    Pagliuca, A., Carrington, P.A., Pettengell, R., Tule, S. & Keidan, J. Guidelines on the use of colony-stimulating factors in haematological malignancies. Br. J. Haematol. 123, 22–33 (2003).

  7. 7

    Morris, E.S. et al. NKT cell–dependent leukemia eradication following stem cell mobilization with potent G-CSF analogs. J. Clin. Invest. 115, 3093–3103 (2005).

  8. 8

    Patterson, S. et al. Human invariant NKT cells display alloreactivity instructed by invariant TCR-CD1d interaction and killer Ig receptors. J. Immunol. 181, 3268–3276 (2008).

  9. 9

    Liu, F., Wu, H.Y., Wesselschmidt, R., Kornaga, T. & Link, D.C. Impaired production and increased apoptosis of neutrophils in granulocyte colony–stimulating factor receptor–deficient mice. Immunity 5, 491–501 (1996).

  10. 10

    Shlomchik, W.D. et al. Prevention of graft versus host disease by inactivation of host antigen-presenting cells. Science 285, 412–415 (1999).

  11. 11

    Ochando, J.C. et al. Alloantigen-presenting plasmacytoid dendritic cells mediate tolerance to vascularized grafts. Nat. Immunol. 7, 652–662 (2006).

  12. 12

    Merad, M. et al. Depletion of host Langerhans cells before transplantation of donor alloreactive T cells prevents skin graft-versus-host disease. Nat. Med. 10, 510–517 (2004).

  13. 13

    Hill, G.R. & Ferrara, J.L.M. The primacy of the gastrointestinal tract as a target organ of graft-versus-host disease: Rationale for the use of cytokine shields in allogeneic bone marrow transplantation. Blood 95, 2754–2759 (2000).

  14. 14

    Morecki, S. et al. Effect of KRN7000 on induced graft-vs-host disease. Exp. Hematol. 32, 630–637 (2004).

  15. 15

    Hashimoto, D. et al. Stimulation of host NKT cells by synthetic glycolipid regulates acute graft-versus-host disease by inducing TH2 polarization of donor T cells. J. Immunol. 174, 551–556 (2005).

  16. 16

    Zeng, D. et al. Bone marrow NK1.1 and NK1.1+ T cells reciprocally regulate acute graft versus host disease. J. Exp. Med. 189, 1073–1081 (1999).

  17. 17

    Pillai, A.B., George, T.I., Dutt, S., Teo, P. & Strober, S. Host NKT cells can prevent graft-versus-host disease and permit graft antitumor activity after bone marrow transplantation. J. Immunol. 178, 6242–6251 (2007).

  18. 18

    Matte, C.C. et al. Donor APCs are required for maximal GVHD but not for GVL. Nat. Med. 10, 987–992 (2004).

  19. 19

    MacDonald, K.P. et al. Donor pretreatment with progenipoietin-1 is superior to G-CSF in preventing graft-versus-host disease after allogeneic stem cell transplantation. Blood 101, 2033–2042 (2003).

  20. 20

    Morris, E.S. et al. Donor treatment with pegylated G-CSF augments the generation of IL-10 producing regulatory T cells and promotes transplant tolerance. Blood 103, 3573–3581 (2004).

  21. 21

    Burman, A.C. et al. IFNγ differentially controls the development of idiopathic pneumonia syndrome and GVHD of the gastrointestinal tract. Blood 110, 1064–1072 (2007).

  22. 22

    Rowe, V. et al. Host B cells produce IL-10 following TBI and attenuate acute GVHD after allogeneic bone marrow transplantation. Blood 108, 2485–2492 (2006).

  23. 23

    Westerhof, G.R. et al. Comparison of different busulfan analogues for depletion of hematopoietic stem cells and promotion of donor-type chimerism in murine bone marrow transplant recipients. Cancer Res. 60, 5470–5478 (2000).

  24. 24

    Jung, S. et al. In vivo depletion of CD11c+ dendritic cells abrogates priming of CD8+ T cells by exogenous cell-associated antigens. Immunity 17, 211–220 (2002).

  25. 25

    Hill, G.R. et al. Total body irradiation and acute graft versus host disease. The role of gastrointestinal damage and inflammatory cytokines. Blood 90, 3204–3213 (1997).

  26. 26

    Cooke, K.R. et al. Tumor necrosis factor-α neutralization reduces lung injury after experimental allogeneic bone marrow transplantation. Transplantation 70, 272–279 (2000).

  27. 27

    Banovic, T. et al. TGF-β in allogeneic stem cell transplantation: friend or foe? Blood 106, 2206–2214 (2005).

  28. 28

    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 (2000).

  29. 29

    Matsuda, J.L. et al. Tracking the response of natural killer T cells to a glycolipid antigen using CD1d tetramers. J. Exp. Med. 192, 741–754 (2000).

  30. 30

    Hammond, K.J. et al. CD1d-restricted NKT cells: an interstrain comparison. J. Immunol. 167, 1164–1173 (2001).

Download references


We thank D.C. Link (Washington University) and A.W. Roberts (The Walter and Eliza Hall Institute, Melbourne) for the Csf3r−/− mice and D. Pellicci (University of Melbourne) for the generation of CD1d tetramers used in this study. The TEa mice were supplied by J. Bromberg (Mount Sinai School of Medicine, New York). K.P.A.M. and C.R.E. are National Health and Medical Research Council (NHMRC) R.D. Wright Fellows. D.I.G. and M.J.S. are NHMRC Research Fellows. G.R.H. is a NHMRC Practitioner Fellow.

Author information

E.S.M. designed and performed experiments and contributed to the manuscript. K.P.A.M., R.D.K., H.M.M., T.B., A.L.J.D., V.R., Y.A.W., N.C.R., A.C.B. and K.A.M. performed experiments. C.R.E. contributed reagents and data interpretation. D.I.G. and M.J.S. contributed vital reagents and experimental design and contributed to the manuscript. G.R.H. designed the study and wrote the manuscript.

Correspondence to Geoffrey R Hill.

Ethics declarations

Competing interests

The bone marrow transplantation laboratory has previously received funding from Amgen USA.

Supplementary information

Supplementary Text and Figures

Supplementary Figs. 1 and 2 (PDF 223 kb)

Rights and permissions

Reprints and Permissions

About this article

Further reading