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PubMed1990

Radioimmune imaging of bone marrow in patients with suspected bone metastases from primary breast cancer.

Radioimmune imaging of bone marrow was performed by technetium-99m- (99mTc) labeled antigranulocyte monoclonal antibody BW 250/183 (AGMoAb) scans in 32 patients with suspected bone metastases from primary breast cancer. AGMoAb scans showed bone marrow defects in 25/32 (78%) patients; bone invasion was subsequently confirmed in 23 (72%) patients. Conventional bone scans performed within the same week detected bone metastases in 17/32 (53%) patients (p less than 0.001). AGMoAb scans detected more sites indicating metastatic disease than bone scans in 12 of these 17 patients (71%). All patients with bone metastases in the axial skeleton had bone marrow defects at least at the sites of bone metastases. Of 15 patients with normal, or indicative of, benign disease bone scans, 8 patients (53%) presented with bone marrow defects in the AGMoAb scans. Bone invasion was confirmed in six of them. AGMoAb bone marrow scans provide a method for the early detection of bone metastatic invasion in patients with breast cancer and suspected bone metastases.

Antibodies, Monoclonal↗
PubMed1982

Ratio of blood and marrow-derived cells in bone marrow transplantation.

The content of the cell suspensions used in bone marrow transplantation can vary by at least twelvefold in the ratio of bone marrow-derived to blood-derived cells. The ratio is somewhat higher in young donors but is similar for male and female donors and is not closely correlated with the incidence of graft v host reactions.

Adolescent↗
PubMed1985

Effects of monoclonal antibody and complement treatment of human marrow on hematopoiesis in continuous bone marrow culture.

Long-term bone marrow cultures were established from single-cell suspensions of human bone marrow that had been treated with monoclonal antibodies and complement. Each treated cell suspension was evaluated for production of hematopoietic stem cells over 20 weeks. Treatment with antibody to HLA-DR (Ia), B1, J2, or J5 did not remove adherent cells including those differentiating to adipocytes in 17-hydroxycorticosteroid. In contrast, treatment with monoclonal antibody directed against human beta 2-microglobulin reduced adipocyte numbers by 100-fold and reduced the total adherent cell density over 70%. Cumulative total nonadherent cell and granulocyte-macrophage colony-forming units (GM-CFUc) production over 20 weeks was not significantly altered by one cycle of anti-Ia plus complement or up to three cycles of treatment with complement and anti-J2, -J5, or -B1. However, one cycle of treatment with anti-beta 2-micro-globulin depressed production of both GM-CFUc and nonadherent cells by over 100-fold compared to other treatment groups. While one cycle of treatment of anti-Ia and complement killed all detectable cells forming CFU-erythroid-granulocyte-megakaryocyte-macrophage, blast-forming units (erythroid), and GM-CFUc, GM cluster-forming cells survived. Treatment of marrow with three cycles of anti-Ia and complement removed all detectable GM colony- and GM cluster-forming cells; however, this marrow produced fewer cumulative Ia-positive GM-CFUc. Long-term bone marrow cultures may prove to be an interesting system for in vitro analysis of the effects of new immunotherapeutic agents including other monoclonal antibodies prior to clinical use.

Adipose Tissue↗
PubMed1999

A comparative study of blood and bone marrow cultures in cadaveric bone donation.

The results of blood cultures taken from cadaveric bone donors were compared with bone marrow and also swab cultures of the procured grafts. In eight of the 95 donors evaluated, pathogenic micro-organisms were detected in the blood sample. In two, identical micro-organisms were cultured from the blood and bone marrow sample whilst the swab cultures were negative. Considering the low sensitivity of the swab culture, the organisms detected in the blood culture were likely to have spread haematogenously and considered to be present in the explanted grafts. Bacteriological screening of bone donors is extremely important since the transmission of micro-organisms via an allograft can lead to serious complications in the recipient. Positive blood cultures provide important information on the presence of pathogenic micro-organisms in grafts obtained from cadaveric bone donors and are therefore essential in deciding whether a graft is bacteriologically acceptable for transplantation.

Blood↗
PubMed1999

Bone marrow pre-B-1 (Bomb1): a quantitative trait locus inducing bone marrow pre-B-cell expansion in lymphoma-prone SL/Kh mice.

Abnormalities of regulatory genes in early B-cell development often lead to lymphomagenesis. Our previous study showed that there is an abnormal transient expansion of bone marrow (BM) pre-B cells in lymphoma-prone SL/Kh strain mice. Such expansion is a genetic property of SL/Kh stem cells rather than BM microenvironments. Using the percentage of BP1+ B220+ pre-B cells in total BM lymphoid cells as a quantitative parameter, we studied the genetic control of BM pre-B cells in 159 F2 offspring of crosses between SL/Kh and NFS/N mice and 334 back-crosses to SL/Kh mice. A highly significant quantitative trait locus was identified on the distal segment of chromosome 3, showing logarithm of odds scores of 22.7 in the F2 cohort and 10.7 in back-cross mice. This quantitative trait locus, named bone marrow pre-B-1, colocalized with lymphoid enhancer factor-1, which encodes a high mobility group DNA-binding protein that is expressed in T and pre-B cells.

Animals↗
PubMed1995

Prospective randomised double-blind trial of the in vivo use of recombinant human erythropoietin in bone marrow transplantation from HLA-identical sibling donors. The Australian Bone Marrow Transplant Study Group.

Ninety one patients between the ages of 17 and 58 years undergoing histocompatible allogeneic transplants from sibling donors were entered into a double-blind randomised trial to evaluate the effect of human erythropoietin (rhu EPO) at a dose of 300 units per kg/day given thrice weekly by intravenous injection on erythropoiesis and on erythrocyte and platelet transfusion requirements. Dose was ceased when the haemoglobin exceeded 12g/dL and recommenced if the haemoglobin fell below 12 g/dL, at 150 units/kg/day. If the haemoglobin exceeded 12 g/dL on a further occasion, the dose of rhu EPO was not given. Patients received two units of erythrocytes when the haemoglobin dropped below 8.5g/dL and received platelet transfusions when the count dropped below 20 x 10(9)/L. Univariate analysis revealed a significantly higher reticulocyte count, haemoglobin concentration and bone marrow erythropoiesis after day 14 in the group receiving rhu EPO but this was not reflected in decreased erythrocyte transfusion (7 +/- 5 in controls versus 6 +/- 5 in rhu EPO group) or in platelet transfusions (11 +/- 7 in controls versus 11 +/- 9 in rhu EPO group). Hospitalisation in each group was the same (29 +/- 8 in the control group and 28 +/- 8 in the rhu EPO group). However, in the multivariate analysis, the administration of rhu EPO was associated with an 18% reduction in erythrocyte transfusion requirement when other variables were taken into account. No side-effects due to rhu EPO were detected in this study.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed2004

Correlation of bone marrow lipid water content with bone mineral density on the lumbar spine.

STUDY DESIGN: Prospective study. OBJECTIVES: To assess the proton MR spectroscopy (1H MRS) of vertebral bone marrow and correlate the lipid water ratio (LWR) and spectral line width (LW) with bone mineral density (BMD) in female subjects. SUMMARY OF BACKGROUND DATA: The mechanism of bone marrow fat accumulation and bone mineral content is poorly understood. Proton MR spectroscopy was used to demonstrate the lipid and water spectra in the bone marrow. We try to assess the possible interaction between the bone marrow lipid content, aging, and BMD. METHODS: Proton MRS and BMD of the lumbar spine were performed in 52 female subjects (mean age, 58 years; SD, 10 years). They were 13 premenopausal and 39 postmenopausal women. The BMD (g/cm2) was measured using dual energy radiograph absorptiometry at the lumbar spine. Single voxel 1H MRS was measured at L3 vertebral body by stimulated echo-acquisition mode (STEAM) sequence and demonstrated two major peaks (lipid and water). Comparisons of the differences between the two subgroups were made. Pearson's correlation was also calculated to explore the association of 1H MRS measurements with age and BMD. Partial correlation was further conducted when controlling the variable such as age or BMD. RESULTS: BMD and LWR had statistically significant difference between the pre- and postmenopausal subgroups (P < 0.001), while lipid LW had a borderline difference and water LW had no difference. LWR was positively correlated with age (r = 0.52 and P < 0.0001) and negatively correlated with BMD (r = -0.40 and P = 0.003) for all the subjects. Lipid LW was negatively correlated with age (r = -0.32 and P = 0.0197) and positively correlated with BMD (r = 0.67 and P < 0.0001). When controlling for BMD effect, only LWR is statistically correlated with age (partial r = 0.39, P = 0.0045), while only the lipid LW is statistically correlated with BMD when controlling for age (partial r = 0.63, P < 0.0001). None of the correlations between water LW and age or BMD was significant. In the subgroups, only the lipid LW is significantly correlated with BMD (r = 0.78, P = 0.0016 in premenopausal women; r = 0.62, P < 0.0001 in postmenopausal women). CONCLUSION: The LWR had a positive correlation with the age, while the lipid LW had a positive correlation with BMD, even after controlling the age factor. The bone marrow lipid water content and metabolism acted as important roles in the internal environment of bone and influenced bone mineralization.

Absorptiometry, Photon↗
PubMed1998

Bone marrow cell development and trabecular bone dynamics after ovariectomy in ddy mice.

To clarify the relationship between the sequential changes of trabecular bone turnover and bone marrow cell development in ovariectomized (ovx) mice, bilateral tibiae of 8-week-old ddy mice were obtained. Histomorphometric analyses of the trabecular bone of the proximal tibia of ovx mice revealed increases in the bone formation rate and the osteoclast surface for the first 28 days postovariectomy. The trabecular bone volume showed a rapid decrease for the first 28 days and a steady state for the subsequent 14 days. In bone marrow cell culture experiments, the numbers of total and nonadherent bone marrow cells per tibia obtained from the ovx mice increased. The formation of osteogenic nodules and osteoclast-like multinucleated cells in the marrow cultures obtained from ovx limbs showed a significant increase on days 14 and 28 and returned to the sham-operated level by day 42. The numbers of colony forming units (fibroblastic) and colony forming units (granulocytes and macrophages) that developed from the marrow cells did not differ between the ovx and sham limbs at any time during the study period. Fluorescence-activated cell-sorter analysis revealed no population changes in the cell development of macrophages. These results demonstrate that there are two stages in the development of osteopenia after ovx. During the first 28 days after ovx, the ovariectomy enhances the developmental process from bone marrow stromal cells to osteoblasts and the terminal differentiation from osteoclast precursors to mature osteoclasts. The trabecular bone turnover also increases. In the subsequent 14 days, the changes in the osteogenic and osteoclastogenic potentials of the bone marrow cells are alleviated and the trabecular bone dynamics are in a steady state. The changes in bone marrow cell development are closely associated with those at the trabecular bone surface.

Animals↗
PubMed1993

High-dose weekly intravenous immunoglobulin to prevent infections in patients undergoing autologous bone marrow transplantation or severe myelosuppressive therapy. A study of the American Bone Marrow Transplant Group.

OBJECTIVE: To determine whether intravenous immunoglobulin (IVIG) prevents severe infections during autologous bone marrow transplantation or equivalent high-dose myelosuppressive therapy. DESIGN: Randomized, stratified, nonblinded study. SETTING: Three tertiary care university hospitals. PATIENTS: One hundred seventy patients entered the study; 82 received IVIG and 88 were untreated controls. The study groups were similar for parameters capable of influencing the likelihood of infection. INTERVENTIONS: Intravenous immunoglobulin was given weekly at a dose of 500 mg/kg body weight from the initiation of cytotoxic therapy to the resolution of neutropenia. MEASUREMENTS: The development of bloodstream or other clinically proven infection, platelet use, and the development of alloimmunity to platelet transfusion. RESULTS: Clinical infection, bacteremia, and fungemia occurred in 43%, 35%, and 6% of the IVIG-treated patients and in 44%, 34%, and 9% of the control patients. Gram-positive bacteremia and gram-negative bacteremia occurred in 28% and 11% of the IVIG group and in 23% and 13% of the control group. Death due to infection occurred in 4.9% of IVIG recipients and in 2.3% of controls. None of these observations was statistically significant (P > 0.2). Survival to hospital discharge was achieved in 86.6% of the IVIG group and in 96.6% of the control group. The survival difference (10%; 95% CI, 1.7% to 18.3%; P = 0.02) was due to a higher incidence of regimen-related toxic death in the IVIG-treated group. CONCLUSIONS: The use of IVIG did not prevent infection. Fewer deaths occurred among controls due to a higher incidence of fatal hepatic veno-occlusive disease in patients receiving IVIG.

Adolescent↗
PubMed1993

Studies on the bone marrow cells from an excised pipe bone.

Samples of rib bone were collected and preserved under various conditions. The effect of preserving a piece of bone on the yield of hematopoietic stem cells was analyzed. Bone marrow cells were collected by means of flushing the bone marrow cavity with medium. The initial number of nucleated bone marrow cells collected from the donor's excised rib ranged from 0.8 x 10(8) to 2.3 x 10(8) per 1 cm of rib with an average of 1.21 x 10(8) (n = 7). In addition, the initial number of mononuclear bone marrow cells ranged from 1.1 x 10(7) to 4.8 x 10(7) per 1 cm of rib with an average of 2.47 x 10(7) (n = 7). The percentage loss of mononuclear bone marrow cells from rib preserved in medium for 48 hours at 4 degrees C, was less than 30%. A colony-forming assay by the methylcellulose method, complemented with r-G-CSF, r-GM-CSF, r-IL3, r-IL6, and r-erythropoietin, was done serially using bone marrow buffy-coat cells. These cells were obtained from the pieces of rib that were stored under various conditions. The number of CFU-GM and BFU-E that proliferated in this culture was 41.1 +/- 5.5 and 15.2 +/- 2.7 per dish. If the bone was preserved at 4 degrees C for 48 hours following excision, the recovery of CFU-GM and BFU-E from the bone marrow cells was greater than 50%. The effect of cryopreserving bone marrow mononuclear cells on the yield of hematopoietic cells was also analyzed. The percentage loss of proliferated hematopoietic cells by cryopreservation was less than 30% for bone marrow cells. Bone marrow cells were collected from pieces of rib obtained from cadaveric donors by means of the same method. The average number of nucleated bone marrow cells and MNCs was 1.26 x 10(7) and 2.30 x 10(7) per 1 cm of rib, respectively. The average number of CFU-GM and BFU-E that was detected by the colony-forming assay was 33.4 +/- 5.35 and 11.7 +/- 1.75 per dish, respectively. The percentage loss of proliferated hematopoietic cells by cryopreservation was less than 40% for bone marrow cells from cadaveric donors. These data, which reveal that hematopoietic bone marrow stem cells could be collected from a piece of pipe bone that was without circulation, suggest that it is possible to use pipe bone from cadaveric donors as a source of hematopoietic stem cells for bone marrow transplants.(ABSTRACT TRUNCATED AT 400 WORDS)

Aged↗
PubMed1993

Preclinical studies for adoptive immunotherapy in bone marrow transplantation. Generation of anti-CD3 activated cytotoxic T cells from normal donors and autologous bone marrow transplant candidates.

In order to obtain T cells for adoptive immunotherapy after autologous bone marrow transplantation (ABMT) for patients with resistant hematological malignancies, a culture system was developed for growing T cells and inducing non-MHC-restricted cytotoxicity using anti-CD3 monoclonal antibody (OKT3) activation. In this investigation, we show that (1) peripheral blood lymphocytes or bone marrow mononuclear cells (BMMNC) from normal donors and cancer patients can be activated with OKT3 and grown in interleukin-2; (2) normal BMMNC activated with OKT3/IL-2 exhibited non-MHC-restricted cytotoxicity and surface markers comparable to that exhibited by normal PBL activated with OKT3/IL-2; (3) both proliferation and cytotoxic functions were IL-2-dependent; (4) PBL activated with OKT3/IL-2 after cryogenic storage grew and killed comparable to PBL activated with OKT3/IL-2 prior to cryopreservation; (5) OKT3/IL-2-activated PBL and BMMNC obtained from 5 patients with non-Hodgkin's lymphomas (NHL) and 1 patient with acute myelogenous leukemia (AML) increased cell numbers 41-75-fold in 2 weeks of culture; 5 of 6 patients with NHL or AML had PBL and BMMNC that exhibited cytotoxic activity; and (6) contaminating leukemic cells did not overgrow in OKT3/IL-2-activated cultures and could no longer be detected on cytospin specimens after 3 weeks of culture. These data show that T cells in PBL or BMMNC from ABMT candidates can be activated with OKT3/IL-2 for adoptive immunotherapy in combination with ABMT.

Antibodies↗
PubMed1992

Over-expression of c-src or v-src in bone marrow stromal cells stimulates hematopoiesis in long-term bone marrow culture.

The S17 murine stromal cell line was infected with retroviral vectors encoding the v-src and c-src oncogenes and cells expressing high levels of either pp60v-src or pp60c-src were isolated. Long-term bone marrow cultures (LTBMCs) established with these different stromal cell lines showed that progenitor cells proliferated to a greater extent in cultures with stromal cells that over-expressed either c-src or v-src. An increase in the number of granulocytes, monocytes, and colony-forming units granulocyte-macrophage (CFU-GM) in the nonadherent cell population of LTBMCs prepared with S17/v-src or S17/c-src stromal cells was observed. Conditioned media from the S17/v-src and S17/src stromal cell lines stimulated the formation of CFU-GM in the absence of additional hematopoietic cell growth factors. Conditioned media from S17/v-src and S17/c-src stimulated proliferation of the granulocyte-macrophage colony-stimulating factor (GM-CSF)-responsive cell line FDCP-1 and this stimulation was inhibited by neutralizing antisera to murine GM-CSF. An increase in the concentration of GM-CSF was confirmed by enzyme-linked immunosorbent assay. No secretion of interleukin-1 alpha (IL-1 alpha) or tumor necrosis factor-alpha was detected by any of the stromal cell lines. There was no increase in the secretion of either CSF-1 or IL-6 by either S17/v-src or S17/c-src. The addition of 1 micrograms/mL monoclonal anti-GM-CSF antibody to LTBMCs caused a decrease in the number of nonadherent cells in cultures established with each of the different stromal cell lines. Northern blot analysis showed no difference in the level of GM-CSF RNA among the different stromal cell lines. These studies suggest that the increased proliferation of hematopoietic progenitor cells in LTBMCs with S17/v-src or S17/c-src cells may result from a posttranscriptional event that elevates production of GM-CSF by the S17/c-src and S17/v-src stromal cells.

3T3 Cells↗
PubMed1988

Effects of T cell depletion in radiation bone marrow chimeras. II. Requirement for allogeneic T cells in the reconstituting bone marrow inoculum for subsequent resistance to breaking of tolerance.

The ability of normal recipient-type lymphocytes to break tolerance in long-term allogenic radiation chimeras has been investigated. Reconstitution of lethally irradiated mice with a mixture of syngeneic and allogeneic T cell-depleted (TCD) bone marrow (BM) has previously been shown to lead to mixed chimerism and permanent, specific tolerance to donor and host alloantigen (3-5). If allogeneic T cells are not depleted from the reconstituting inoculum, complete allogeneic chimerism results; however, no clinical evidence for GVHD is observed, presumably due to the protective effect provided by syngeneic TCD BM. This model has now been used to study the effects of allogenic T cells administered in reconstituting BM inocula on stability of long-term tolerance. We have attempted to break tolerance in long-term chimeras originally reconstituted with TCD or non-TCD BM by challenging them with inocula containing normal, nontolerant recipient strain lymphocytes. tolerance was broken with remarkable ease in recipients of mixed marrow inocula in which both original BM components were TCD. In contrast, tolerance in chimeras originally reconstituted with non-TCD allogeneic BM was not affected by such inocula. Susceptibility to loss of chimerism and tolerance was not related to initial levels of chimerism per se, but rather to T cell depletion of allogeneic BM, since chimeras reconstituted with TCD allogeneic BM alone (mean level of allogeneic chimerism 98%) were as susceptible as mixed chimeras to the tolerance-breaking effects of such inocula. The possible contribution of GVH reactivity to this resistance was investigated using an F1 into parent strain combination. In these animals, the use of non-TCD F1 BM inocula for reconstitution did not lead to resistance to the tolerance-breaking effects of recipient strain splenocytes. These results suggest that the ability of T cells in allogeneic BM inocula to confer resistance to late graft failure may be related to their graft-vs.-host reactivity, even in situations in which they do not cause clinical GVHD. These findings may have relevance to the mechanism whereby T cell depletion of allogeneic BM leads to an increased incidence of late graft failure in clinical BM transplantation situations.

Animals↗
PubMed1989

Cryopreservation of marrow, purging and autologous bone marrow transplantation in childhood.

Since 1984 bone marrow from 42 children with acute lymphoblastic leukaemia, non Hodgkin's lymphoma and neuroblastoma was cryopreserved. In 5 cases (c-ALL, NHL and B type) the marrow was purged by using a cocktail of three monoclonal antibodies (VIL A1, VIB C5, VIB-E3). Up to now 13 children (ALL/10, neuroblastoma/3) were autografted (one of them after purging) after supralethal chemoradiotherapy. Except one child with early death all patients had engraftment: a level of 1.0.10(9)/l leukocytes was reached at days 10-33 (median, 19); platelet level over 60.10(9)/l at days 32-60 (median, 41). 2 children died on treatment related complications, one on infection after full haematological restitution, 2 patients alive with relapse, 8/13 alive in CCR and well.

Antibodies, Monoclonal↗