Establishment of a Murine Graft-versus-Myeloma Model Using Allogeneic Stem Cell Transplantation

Abstract

BackgroundMultiple myeloma (MM) is a malignant plasma cell disorder with poor long-term survival and high recurrence rates. Despite evidence of graft-versus-myeloma (GvM) effects, the use of allogeneic hematopoietic stem cell transplantation (allo-SCT) remains controversial in MM. In the current study, we investigated the anti-myeloma effects of allo-SCT from B10.D2 mice into MHC-matched myeloma-bearing Balb/cJ mice, with concomitant development of chronic graft-versus-host disease (GvHD).Methods and resultsBalb/cJ mice were injected intravenously with luciferase-transfected MOPC315.BM cells, and received an allogeneic (B10.D2 donor) or autologous (Balb/cJ donor) transplant 30 days later. We observed a GvM effect in 94% of the allogeneic transplanted mice, as the luciferase signal completely disappeared after transplantation, whereas all the autologous transplanted mice showed myeloma progression. Lower serum paraprotein levels and lower myeloma infiltration in bone marrow and spleen in the allogeneic setting confirmed the observed GvM effect. In addition, the treated mice also displayed chronic GvHD symptoms. In vivo and in vitro data suggested the involvement of effector memory CD4 and CD8 T cells associated with the GvM response. The essential role of CD8 T cells was demonstrated in vivo where CD8 T-cell depletion of the graft resulted in reduced GvM effects. Finally, TCR Vβ spectratyping analysis identified Vβ families within CD4 and CD8 T cells, which were associated with both GvM effects and GvHD, whereas other Vβ families within CD4 T cells were associated exclusively with either GvM or GvHD responses.ConclusionsWe successfully established an immunocompetent murine model of graft-versus-myeloma. This is the first murine GvM model using immunocompetent mice that develop MM which closely resembles human MM disease and that are treated after disease establishment with an allo-SCT. Importantly, using TCR Vβ spectratyping, we also demonstrated the presence of GvM unique responses potentially associated with the curative capacity of this immunotherapeutic approach.

Granulocytic myeloid-derived suppressor cells promote angiogenesis in the context of multiple myeloma

“…injections) and monitoring of MOPC315.BM tumor development by bioluminescence were performed as previously described [31, 32]. For 5TGM1 solid tumor growth, 5TGM1 cells were suspended in phosphate buffered saline (PBS) containing 1 mg/ml of matrigel and were injected subcutaneously (2.5×10 5 cells/200μl/mouse) into the right flank of C57BL/KaLwRij mice.…”

Limited Impact of Imatinib in a Murine Model of Sclerodermatous Chronic Graft-versus-Host Disease

“…Further, these data are consistent with recent observations by Binsfeld et al . in a murine model of graft-versus-myeloma effect consisting in the transplantation of BALB/cJ mice, previously injected with luciferase-transfected MOPC315.BM myeloma cells, with bone marrow cells and splenocytes from B10.D2 mice [33]. …”

“…Briefly, recipient Balb/cJ mice (H-2 d ) were lethally irradiated with 7 Gy total body irradiation using a 137 Cs irradiator (GammaCell 40, Nordion, Ontario, Canada). Six hours later, recipients were injected with 200 μL of the prepared cell suspension (2x10 6 spleen cells + 1x10 6 bone marrow cells for the moderate scl-cGVHD model [32] or 70x10 6 spleen cells + 10x10 6 bone marrow cells for the severe scl-cGVHD model [31, 33]) from B10.D2 (H-2 d ) mice. Recipient mice were monitored daily, while recipient body weights and GVHD scores (see next section) were recorded every 3 days.…”

“…Scl-cGvHD severity was assessed using a clinical scoring system as previously reported [31, 33]. Briefly, ear-tagged animals were individually scored 3 times/week for five parameters: weight loss (grade 1, loss between 10–20%; grade 2, loss > 20%), posture (1, kyphosis only at rest; 2, severe kyphosis when the animal moves), activity (1, moderate activity impairment; 2, no move unless stimulated), skin (1, erythema or scaling tail; 2, lesions on the body surface), and hair loss (1, loss > 1 cm 2 ; 2 loss > 2 cm 2 ).…”

Ex Vivo Oncolytic Virotherapy with Myxoma Virus Arms Multiple Allogeneic Bone Marrow Transplant Leukocytes to Enhance Graft versus Tumor

“…Thus, we tested the potential for ex vivo MYXV virotherapy to eliminate pre-seeded residual disease in an immunocompetent murine model of MM by exploiting tagged murine MOPC315.BM cells 22, 23. MOPC315.BM cells are a murine plasmacytoma cell line that can engraft into syngeneic immunocompetent BALB/c mice and cause a progressive disease highly recapitulative of human MM, including BM invasion, osteolysis, and paralysis 22, 23. We report that, in stark contrast to all human MM tested, this murine MOPC315.BM cell line is resistant to direct infection with MYXV because of a defect in virus binding to these cells.…”