Thermal Sensitivity and Thermally Enhanced Radiosensitivity of Murine Bone Marrow Granulocyte-Macrophage Colony-Forming Units(CFU-GM).

Experiments presented here were aimed at the description of hematopoiesis repair and in vivo homing of transplanted separated CD117+B220–bone marrow cells after whole-body lethal irradiation at LD 9Gy. ROSA 26 mice were used as donors of marrow cells for transplantation [B6;129S/Gt (ROSA)26Sor] and were tagged with lacZ gene, and F2 hybrid mice [B6129SF2/J] were used as recipients of bone marrow transplanted cells. Hematopoiesis repair was provided by transplantation, both suspension of whole bone marrow cells (5x106) and isolated CD117+B220–cells (5x104). Mice survived up to thirty days after irradiation. We demonstrated that transplantation of suspension of whole bone marrow cells led to faster recovery of CFU-GM (Granulocyte-macrophage colony forming units) in bone marrow and in the spleen too. It is not clear what the share of residential and transplanted cells is in the repair process. Our results demonstrate that sufficient hematopoietic repair occurs after transplantation of CD117+B220–(lacZ+) in lethally irradiated mice, and the difference in CFU-GM numbers in the bone marrow and spleen found on day 8 posttransplant has no influence on the survival of lethally irradiated mice (30 days follow-up).

Download Full-text

Enhanced survival but reduced engraftment in murine recipients of recombinant granulocyte/macrophage colony-stimulating factor following transplantation of T-cell-depleted histoincompatible bone marrow

Blood ◽

10.1182/blood.v72.4.1148.1148 ◽

1988 ◽

Vol 72 (4) ◽

pp. 1148-1154 ◽

Cited By ~ 15

Author(s):

BR Blazar ◽

MB Widmer ◽

CC Soderling ◽

S Gillis ◽

DA Vallera

Keyword(s):

Bone Marrow ◽

T Cell ◽

Hematological Parameters ◽

Donor Cell ◽

Colony Stimulating Factor ◽

Murine Bone Marrow ◽

Macrophage Colony Stimulating Factor ◽

Macrophage Colony ◽

Stimulating Factor

Abstract In vivo administration of murine recombinant granulocyte/macrophage colony stimulating factor (rGM-CSF) was evaluated for effects on survival and engraftment in an allogeneic murine bone marrow transplantation (BMT) model involving T-cell depletion of donor marrow. The model provides a high incidence of graft failure/rejection. Recipients of continuous subcutaneous infusions of rGM-CSF had a significant survival advantage when compared with untreated controls. However, a significantly lower incidence of donor cell engraftment was noted. Hematological parameters were not substantially affected. When rGM-CSF was administered intraperitoneally (IP), twice daily injections closely approximated the effects of continuous infusion on survival. Single IP injections were without significant effects on survival or engraftment. These results demonstrate that prolonged frequent in vivo exposure to rGM-CSF can significantly improve survival but significantly decreases donor cell repopulation in recipients of T-cell- depleted histoincompatible marrow grafts.

Download Full-text

The Effects of Mustine Hydrochloride on the Colony Forming Units of Murine Bone Marrow

Acta Haematologica ◽

10.1159/000208586 ◽

1971 ◽

Vol 46 (5) ◽

pp. 271-274 ◽

Cited By ~ 3

Author(s):

J.G. Sharp ◽

Brynmor Thomas

Keyword(s):

Bone Marrow ◽

Murine Bone Marrow ◽

Colony Forming Units

Download Full-text

Hematopoietic regulatory factors produced in long-term murine bone marrow cultures and the effect of in vitro irradiation

Blood ◽

10.1182/blood.v64.2.516.bloodjournal642516 ◽

1984 ◽

Vol 64 (2) ◽

pp. 516-525 ◽

Cited By ~ 1

Author(s):

RJ Gualtieri ◽

RK Shadduck ◽

DG Baker ◽

PJ Quesenberry

Keyword(s):

Bone Marrow ◽

Stromal Cells ◽

Factor Vii ◽

Murine Bone Marrow ◽

L Cell ◽

Bone Marrow Cultures ◽

Macrophage Colony ◽

Marrow Cultures

The nature of hematopoietic regulatory factors elaborated by the adherent (stromal) cells of long-term murine bone marrow cultures and the effect of in vitro stromal irradiation (XRT) on the production of these factors was investigated. Using an in situ stromal assay employing a double layer of semisolid agar, it was possible to demonstrate stromal elaboration of at least two colony-stimulating activities, ie, granulocyte/macrophage colony-stimulating activity (G/M- CSA) and megakaryocyte colony-stimulating activity (Meg-CSA). Exposure of the stroma to XRT resulted in dose-dependent elevations of both activities that correlated inversely with total myeloid cell mass as determined by concurrent reductions in total supernatant cell recoveries from irradiated cultures. Mixture experiments that combined control and irradiated stroma revealed that the hematopoietically active control stroma could block detection of XRT-related G/M-CSA elevations. These data implicate a local negative feedback mechanism in the regulation of hematopoiesis. Antiserum directed against purified L cell colony-stimulating factor (CSF) reduced granulocyte/macrophage colony formation in the target layer but did not effect the increased Meg-CSA. While a radioimmunoassay for L-cell type CSF was unable to detect significant differences in concentrated media from control and irradiated cultures, bioassays of these media revealed XRT-related G/M- CSA elevations. These results indicate that the G/M-CSA elaborated in these cultures is immunologically distinct from the Meg-CSA produced, and although distinct from L cell CSF, the G/M-CSA is crossreactive with the L cell CSF antiserum. Morphologic, histochemical, and factor VII antigen immunofluorescent studies were performed on the stromal cell population responsible for production of these stimulatory activities. In addition to “fat” cells, the stromal cells remaining after XRT were composed of two predominant cell populations. These included a major population of acid phosphatase and nonspecific esterase-positive macrophage-like cells and a minor population of factor VII antigen negative epithelioid cells.

Download Full-text

Two phases of engraftment established by serial bone marrow transplantation in mice

Blood ◽

10.1182/blood.v73.2.397.bloodjournal732397 ◽

1989 ◽

Vol 73 (2) ◽

pp. 397-401 ◽

Cited By ~ 1

Author(s):

RJ Jones ◽

P Celano ◽

SJ Sharkis ◽

LL Sensenbrenner

Keyword(s):

Bone Marrow ◽

Bone Marrow Transplantation ◽

Marrow Transplantation ◽

Time Interval ◽

Serial Transfer ◽

Colony Forming Units ◽

Macrophage Colony ◽

Two Phases ◽

Serial Transplantation

Serially transplanted bone marrow eventually fails to reconstitute lethally irradiated mice. The reasons for this loss of repopulating ability are unknown. We showed that serial bone marrow transplantation changed the ratio of hematopoietic progenitors in bone marrow. The numbers of granulocyte-macrophage colony-forming units (CFU-GM) in the bone marrow did not change with serial transplantation. Spleen CFU (CFU- S) numbers decreased with serial transfer but remained at levels which should be associated with engraftment, even on the transfers which were unsuccessful. The CFU-S, therefore, did not appear to be the cells responsible for long-term hematopoietic repopulation. The number of successful serial transfers was dependent on the size of the grafts, and prolonging the time interval between transfers reestablished the ability of the serially transplanted marrow to reconstitute lethally irradiated recipients. Serial bone marrow transplantation dissociated two phases of engraftment. The first unsustained phase was maintained with repeated serial transfer and appeared to be produced by committed progenitors. The second sustained phase was eventually lost with repeated serial transfer and was apparently due to the pluripotent stem cell.

Download Full-text

FcγRII (CD32) Is Linked to Apoptotic Pathways in Murine Granulocyte Precursors and Mature Eosinophils

Blood ◽

10.1182/blood.v90.3.1267.1267_1267_1274 ◽

1997 ◽

Vol 90 (3) ◽

pp. 1267-1274 ◽

Cited By ~ 1

Author(s):

Belen de Andrés ◽

Allen L. Mueller ◽

Arthur Blum ◽

Joel Weinstock ◽

Sjef Verbeek ◽

...

Keyword(s):

Bone Marrow ◽

Bone Marrow Cells ◽

Tissue Injury ◽

Chromatin Condensation ◽

Annexin V ◽

Murine Bone Marrow ◽

Gm Csf ◽

Bone Marrow Cultures ◽

Macrophage Colony ◽

Apoptotic Pathways

Murine granulocytes and precursors express low-affinity IgG Fc receptors (FcγR). We investigated the effects of FcγR ligation on the development of eosinophils in cultures of normal murine bone marrow. Eosinophilopoiesis was induced by culture of bone marrow cells in the presence of cytokines (granulocyte-macrophage colony-stimulating factor [GM-CSF], interleukin-3 [IL-3], and IL-5). Addition to the cultures of 2.4G2, a rat monoclonal antibody (mAb) that reacts with FcγRII (CD32) and FcγRIII (CD16), induced granulocyte apoptosis within 24 hours. Granulocytes in cultures that contained 2.4G2 showed chromatin condensation, binding of Annexin-V, and fas induction, and by electron microscopy, apoptosis was most commonly observed in cells of the eosinophil lineage. Since murine granulocytes can express both FcγRII (CD32) and FcγRIII (CD16), we investigated the effect of 2.4G2 on cultures of bone marrow obtained from FcγRIII (CD16) gene–disrupted mice and found that the apoptosis induced with 2.4G2 was CD16-independent. Studies with bone marrow cultures from B6MLR-lpr/lpr and C3H/HEJ-gld/gld mice established that the FcγRII (CD32)-triggered apoptosis was fas-fasL–dependent. When mature eosinophils isolated from hepatic granulomas of Schistosoma mansoni–infected mice were cultured in cytokines in the presence of 2.4G2, the eosinophils underwent apoptosis within 24 hours. These findings identify a previously unknown linkage between FcγR on eosinophils and fas-mediated apoptosis, a connection that could be relevant to mechanisms by which eosinophils mediate tissue injury and antibody-dependent cellular cytotoxicity reactions.

Download Full-text

Forssman glycosphingolipid as an immunohistochemical marker for mouse stromal macrophages in hematopoietic foci

Blood ◽

10.1182/blood.v72.1.42.42 ◽

1988 ◽

Vol 72 (1) ◽

pp. 42-48 ◽

Cited By ~ 22

Author(s):

Y Sadahira ◽

M Mori ◽

M Awai ◽

S Watarai ◽

T Yasuda

Keyword(s):

Bone Marrow ◽

Hematopoietic Cells ◽

Hematopoietic Tissue ◽

Electron Microscopic ◽

Circulating Cells ◽

Immunohistochemical Marker ◽

Colony Forming Units ◽

Macrophage Colony ◽

Splenic Red Pulp ◽

Red Pulp

Abstract The immunohistochemical distribution of Forssman glycosphingolipid (GSL) in mouse hematopoietic tissue was examined by using light and electron microscopic immunoperoxidase methods with a highly specific rabbit anti-Forssman GSL antibody. Bone marrow, splenic red pulp, and thymic macrophages, which are closely associated with hematopoietic cells, were stained by the antibody, whereas hematopoietic cells, circulating cells, alveolar macrophages, Kupffer cells, peritoneal resident macrophages, and macrophages derived from granulocyte- macrophage colony-forming units cultured in the presence of L-cell- conditioned medium were not stained. In addition, thymic cortical epithelial cells, the framework of reticular cells of the cortical and medullary regions of the mesenteric lymph node and periarterial lymphoid sheath of the spleen, and some vessels of the tissues examined were also stained. After phlebotomy, the fine cytoplasmic processes of Forssman-positive splenic red pulp macrophages were distributed extensively throughout the erythroid colonies. On the other hand, after hypertransfusion, these macrophages retracted their processes, became round, and tended to aggregate. These results indicate that Forssman GSL can be used as an immunohistochemical marker for stromal macrophages in the hematopoietic foci of the bone marrow and splenic red pulp.

Download Full-text