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Limited engraftment of low-risk myelodysplastic syndrome cells in NOD/SCID gamma-C chain knockout mice

Leukemia volume 24, pages 1662–1664 (2010)Cite this article

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The development of increasingly immunocompromised mice has allowed for improved engraftment of human tissue in xenograft hosts. NOD/SCID IL2Rgamma-c null mice (NSG) represent the latest iterative attempt to limit both the innate and acquired immune function in mice, thus preventing immune surveillance during xenografting. NSG mice have severe T- and B-cell impairment, as well as a complete absence of natural killer-cells.1, 2 Carroll and colleagues3 have recently reported in Leukemia the engraftment of human acute myeloid leukemia cells in the bone marrow of NSG mice, suggesting that NSG mice may be more permissive to human malignant myeloid cell engraftment than the previous xenograft hosts. To date, there has been only limited success in establishing a robust xenograft model of low-risk myelodysplastic syndromes (MDS) despite the use of a variety of immune deficient and transgenic mice. In an attempt to improve on these models, we tested whether the increased deficiency in natural killer cell function in NSG mice might improve the myeloid cell engraftment of MDS samples with low myeloblast counts and provide a useful pre-clinical model of low-risk MDS.4, 5, 6, 7

We attempted to improve engraftment of human myeloid cells by injecting whole bone marrow cells intra-tibial (to abrogate potential trafficking/homing defects), by T-cell depleting-bone marrow samples (to limit the number of T cells that may contribute to graft versus host disease) and by injecting CD34+ purified cells. Four out of five animals that received an intra-tibial injection of bone marrow cells died unexpectedly 40–56 days after injection (MDS samples 5, 6). The remaining intra-tibial injected mouse from MDS sample 6 had 0.12% peripheral blood human engraftment at 6 weeks (100% T-cells), suggesting that graft-versus-host disease may have contributed to the death of the other intra-tibial injected mice. Next, we retro-orbital injected T-cell depleted-bone marrow cells (>2 log reduction in T cells) from MDS samples 5 and 6. Both samples had 0.2% human cell engraftment in the spleen, but no engraftment in the peripheral blood or bone marrow. Surprisingly, engrafted human cells were predominantly CD3+ (70–80%), suggesting that T-cell expansion in NSG mice is robust even when less than 900 T cells were injected. Finally, we injected CD34+ purified cells (autoMACS, Miltenyi, Auburn, CA, USA) from normal or MDS bone marrow cells. One mouse receiving normal CD34+ bone marrow cells had 0.17% bone marrow engraftment, which consisted predominantly of B cells (95%). CD34+ purified cells from three MDS samples (5, 6, 8) were injected retro-orbital, and only MDS sample 8 (13% myeloblasts) had human cell engraftment in the bone marrow (0.16, 17.7% of engrafted cells were CD33+ following injection of 2 × 106 cells) (Figures 1a and b). To ensure that the viability of CD34+ injected cells was not compromised during the purification process; we assessed their growth in normoxic methylcellulose culture (H4544, Stem Cell Technologies, Vancouver, BC, Canada). CD34+ purified cells from the normal bone marrow and MDS sample 8 produced 52 and 14 CFU per 1,000 cells plated, respectively. Interphase fluorescence in situ hybridization analysis from unfractionated and CD34+ purified methylcellulose colonies from MDS sample 8 detected trisomy 8 in 190/200 and 188/200 cells, respectively, indicating that the CD34+ purified cells were clonal. In contrast, CD34+ cells from MDS samples 5 and 6 did not produce methylcellulose colonies, which may reflect the variable growth of MDS cells in normoxic vs hypoxic (3% oxygen) conditions. Collectively, these results indicate that intra-tibial injection, T-cell depletion or CD34+ purification do not significantly improve the engraftment of low-blast count MDS samples in NSG mice.

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Acknowledgements

We thank Tim Ley, Tim Graubert, Dan Link and Gerry Linette for helpful scientific discussions and reagents. This work was supported by National Institutes of Health Grant HL082973, and the Howard Hughes Medical Institute Physician-Scientist Early Career Award (MJW).

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Authors and Affiliations

  1. Department of Medicine, Washington University School of Medicine, St Louis, MO, USA

    M G Martin, J S Welch, G L Uy, T A Fehniger & M J Walter

  2. Division of Oncology, Washington University School of Medicine, St Louis, MO, USA

    M G Martin, J S Welch, G L Uy, T A Fehniger & M J Walter

  3. Siteman Cancer Center, Washington University School of Medicine, St Louis, MO, USA

    G L Uy, T A Fehniger, S Kulkarni & M J Walter

  4. Department of Pathology and Immunology, Washington University School of Medicine, St Louis, MO, USA

    S Kulkarni & E J Duncavage

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Correspondence to M J Walter.

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Martin, M., Welch, J., Uy, G. et al. Limited engraftment of low-risk myelodysplastic syndrome cells in NOD/SCID gamma-C chain knockout mice. Leukemia 24, 1662–1664 (2010). https://doi.org/10.1038/leu.2010.156

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