Progenitor cell hyperplasia with rare development of myeloid leukemia in interleukin 11 bone marrow chimeras.

Abstract

Post 5-fluorouracil-treated murine bone marrow cells infected with a recombinant retrovirus (murine stem cell virus-interleukin 11 [MSCV-IL-11]) bearing a human IL-11 gene were transplanted into lethally irradiated syngeneic mice. Analysis of proviral integration sites in DNA prepared from hematopoietic tissues and purified cell populations of long-term reconstituted primary and secondary recipients demonstrated polyclonal engraftment by multipotential stem cells. High levels (100-1,500 U/ml) of IL-11 were detected in the plasma of the MSCV-IL-11 mice. Systemic effects of chronic IL-11 exposure included loss of body fat, thymus atrophy, some alterations in plasma protein levels, frequent inflammation of the eyelids, and often a hyperactive state. A sustained rise in peripheral platelet levels (approximately 1.5-fold) was seen throughout the observation period (4-17 wk). No changes were observed in the total number of circulating leukocytes in the majority of the transplanted animals (including 10 primary and 18 secondary recipients) despite a > 20-fold elevation in myeloid progenitor cell content in the spleen. The exceptions were members of one transplant pedigree which presented with myeloid leukemia during the secondary transplant phase. A clonal origin of the disease was determined, with significant expansion of the MSCV-IL-11-marked clone having occurred in the spleen of the primary host. Culturing of leukemic spleen cells from a quaternary recipient led to the establishment of a permanent cell line (denoted PGMD1). IL-11-producing PGMD1 myeloid leukemic cells are dependent on IL-3 for continuous growth in vitro and they differentiate into granulocytes and macrophages in response to granulocyte/macrophage colony-stimulating factor. The inability of autogenously produced IL-11 to support autonomous growth of PGMD1 cells argues against a mechanism of transformation involving a classical autocrine loop.

The DN2 Myeloid-T (DN2mt) Progenitor is a Target Cell for Leukemic Transformation by the TLX1 Oncogene

“…The murine IL-3-dependent bone marrow DN2mt progenitor [17,21,28], PGMD1 [36] and M-NSF-60 (ATCC No. CRL-1838; American Type Culture Collection, Manassas, VA) cell lines were routinely maintained in Iscove’s Modified Dulbecco’s Medium (IMDM; Mediatech Inc., Herndon, VA) containing 4 mM L-glutamine, 50 IU/ml penicillin, 50 μg/ml streptomycin, 10% heat-inactivated fetal bovine serum (FBS, Cambrex BioScience Walkersville, Inc., Walkersville, MD), 10% conditioned medium from X630-rlL3 cells (a source of recombinant IL-3) plus/minus 0.75 mg/ml geneticin (Invitrogen Corp., Carlsbad, CA) as applicable.…”

FDA licensure of neumega® to prevent severe chemotherapy‐induced thrombocytopenia

“…NEUMEGAO has been shown to increase blood platelet counts when administered in vivo as a single agent to nonmyelosuppressed rodents, dogs and nonhuman primates. Moreover, thrombocytosis and hematopoietic progenitor cell hyperplasia were observed in lethally irradiated mice transplanted with mouse bone marrow cells that had been retrovirally transfected with the human (h) IL-11 gene [48].…”

“…NEUMEGA@ has had either no effect or minor and inconsistent effects on the growth of myeloma cells [52,53] and did not reverse steroid-induced growth arrest in three human myeloma cell lines [54]. NEUMEGAO has no direct effect on acute myelogenous leukemia (AML) cells but has been shown to augment the proliferation of at least some AML cells stimulated by other cytokines [55, 561. In the study by Huwley et al [48], cells from one secondarily transplanted pedigree showed a leukemic phenotype, and a transformed cell line expressing the IL-11 gene was produced from one quarternarily transplanted animal [48]. This cell line was dependent on LL-3 for continuous growth in vitro, and the cells differentiated into mature leukocytes in vitro in response to GM-CSF treatment.…”

The myeloma-associated oncogene fibroblast growth factor receptor 3 is transforming in hematopoietic cells

“…The transplantation of BM cells was performed as previously described. 19,20 In brief, BM was flushed from the hind limbs of donor mice that had been injected 4 days previously with 150 mg/kg 5-flurorouracil (Roche, Laval, QB) in phosphate-buffered saline (PBS). Following erythrocyte lysis, nucleated cells were cultured in 100-mm dishes at a density of 5 ϫ 10 5 cells/mL in prestimulation medium (Iscoves modified Dulbecco medium [IMDM]; Life Technologies), 10% fetal calf serum (FCS; Life Technologies), 1% penicillin/streptomycin (Life Technologies), 2 mM glutamine (Life Technologies), 50 M 2-mercaptoethanol (2-ME) supplemented with 10% conditioned medium from X630-rIL3 cells, 21 10% conditioned medium from Sp2/mIL6 cells, 22 and 2% to 5% conditioned medium from Sp2/soluble Flt3 ligand (sFLt3L) for 2 days.…”