Korean J Hematol.  2008 Dec;43(4):219-231. 10.5045/kjh.2008.43.4.219.

Enhancement of Graft-versus-leukemia Effects by Mesenchymal Stem Cells in Mixed Chimerisim after a Murine Non-myeloablative Hematopoietic Stem Cell Transplantation

• Lim JY
• Kim BG
• Moon SK
• Min CK

• KMID: 1485080
• DOI: http://doi.org/10.5045/kjh.2008.43.4.219

Abstract

BACKGROUND: Mesenchymal stem cells (MSCs) may be useful for reducing graft-versus-host disease (GVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). The GVHD and a graft-versus-leukemia (GVL) effect are inversely related. We therefore wanted to determine whether MSCs can preserve the GVL effect following experimental allo-HSCT.
METHODS
After non-myeloablative allogeneic hematopoietic stem cell transplantation (NM-HSCT) using C57BL/6 (H-2b)-->B6D2F1 (H-2b/d), some mice received donor lymphocyte infusion (DLI) for induction of GVL effects by virtue of complete chimerism (CC), while the other groups did not receive DLI with persistence of mixed chimerism (MC). All mice were inoculated subcutaneously with P815 tumor cells and were intravenously treated with either donor MSCs or diluents.
RESULTS
Between the DLI-treated groups with CC, tumor-related deaths and tumor growths were comparable irrespective to the infusion of MSCs. On the contrary, among mice without DLI which showed MC, the administration of MSCs significantly delayed tumor-related deaths compared to those without the administration of MSCs (50-day percent survival, 54.5% vs. 18.1%, P=0.0225). In the MC animals, tumor growth seemed to be more delayed in the mice injected with MSCs than in the controls (P=0.09). Donor MSCs injection was associated with increased donor effector/memory CD62L- T cells in MC but not in CC.
CONCLUSION
In spite of the observed immunosuppressive effects of donor MSCs, our results indicate that the GVL effects were not influenced by the injection of MSCs but that under a given condition such as MC, the injection of donor MSCs could result in enhanced GVL effects.

Keyword

Mesenchymal stem cell; Graft-versus-host disease; Graft-versus-leukemia effect; Allogeneic hematopoietic stem cell transplantation

MeSH Terms

Animals
Chimerism
Graft vs Host Disease
Hematopoietic Stem Cell Transplantation
Hematopoietic Stem Cells
Humans
Lymphocytes
Mesenchymal Stromal Cells
Mice
T-Lymphocytes
Tissue Donors
Virtues

Figure

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  Fig. 1 Immunophenotype of murine MSCs. Donor MSCs were stained with surface antibodies and analyzed by FACS.

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  Fig. 2 Responding B6 splenocytes (5×105 cells/well) were incubated for 4 days with mitomicin-treated B6 (Syngeneic) or F1 splenocytes (Allogeneic) in the presence of 1×105 MSCs or diluent alone (Control). Addition of MSCs inhibited T cell proliferative response compared to the controls (∗P=0.005).

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  Fig. 3 The percentage of H-2d negative donor cells in peripheral blood following donor lymphocyte infusion.

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  Fig. 4 Effect of MSCs on survival according to the chimeric status. Sublethally irradiated (400cGy) B6D2F1 (H-2b/d) mice were reconstituted with 107 C57BL/6 (H-2b) bone marrow cells. All mice were found to be a state of mixed chimeric (MC). At 14 days after NM-HSCT, group C and D mice received DLI at a dose of 20×106 spleen cells from donor mice for induction of GVL effects by virtue of complete chimerism (CC), while groups A and B did not receive DLI with persistence of MC. At the same, all mice were inoculated subcutaneously with 1×106 P815 cells and then the recipients were intravenously treated on days 15, 18 and 21 with either donor MSCs (group B and D, 5×105/day) or diluents (group A and C). Kalan-Meier survival curves demonstrate a difference between the MC and CC groups (group A vs. B, ∗P=0.0225; group C vs. D, P=0.371).

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  Fig. 5 Flow cytometry analysis of donor and recipient CD62L− memory/effector CD4+ or CD8+ T cells in peripheral blood obtained 21 days after the injection of MSCs. The percentage of circulating donor cells was determined by calculating the number of cells negative for H-2d. Each graph represents one of two similar experiments. (A), Group A; (B), Group B; (C), Group C; (D), Group D.

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  Fig. 6 Tissue distribution of MSCs injected into the recipients with MC (group B). In addition to the tumor tissue, immunohistochemical analysis of lungs, liver, and spleen sections was used to visualize the prelabeled MSCs by their PKH+ red fluorescence (magnification: ×100). Mice in Group A did not receive the prelabeled MSCs.

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