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PubMed1975

Human bone marrow lymphocytes. I. Distribution of lymphocyte subpopulations in the bone marrow of normal individuals.

This study was undertaken to determine the proportions and in vitro immune capacities of lymphocyte populations in the bone marrows of normal humans. Relatively pure mononuclear cell suspensions were obtained from bone marrow aspirates by linear sucrose gradient centrifugations. Simultaneous peripheral blood and bone marrow specimens from each individual were assayed for lymphocyte surface markers and mitogen responsiveness. Maximal possible contamination of bone marrow aspirates by peripheral blood was determined by performing aspirates on individuals who had received 51chromium-labeled autologous erythrocytes. Rhymus-derived (T) lymphocytes, as determined by the sheep red blood cell (E) rosette assay, comprised 8.6-(plus or minus 1.6)% of the total bone marrow lymphocyte pool. Bone marrow-derived (B) lymphocytes, as determined by the presence of a complement receptor, made up 15.4-(plus or minus 1.9)% of the lymphocyte pool whereas 74.6 (plus or minus 2.4)% of mononuclear cells lacked easily detectable surface markers. These findings could not be explained by contamination with peripheral blood lymphocytes since contamination was corrected for in the calculations. Lymphocyte-enriched suspensions of bone marrow cells responded to stimulation with phytohemagglutinin, concanalin A, and particularly pokeweed mitogen. In vitro incubations of bone marrow and peripheral blood lymphocytes with tritiated thymidine followed by determinations of E and erythrocyte antibody complement (EAC) rosettes were performed. Simultaneous rosetteradioautographs demonstrated that the proliferative potential of bone marrow B lymphocytes was greater than peripheral blood B lymphocytes (P less than 0.01). On the other hand, the proliferative potential of bone marrow T lymphocytes was the same as that of peripheral blood T lymphocytes. These findings demonstrate that in addition to containing B lymphocytes the normal bone marrow contains a small fraction of T lymphocytes similar to the mature T lymphocyte pool found in the peripheral blood. These T cells most probably enter the bone marrow parenchyma as part of the normal recirculating lymphocyte pool.

Adult↗
PubMed2004

Replacement of recipient stromal/mesenchymal cells after bone marrow transplantation using bone fragments and cultured osteoblast-like cells.

Abstract We present our experience on treatment of three children with potentially fatal diseases using a unique protocol for non-myeloablative bone marrow transplantation. The protocol was designed to promote engraftment of bone marrow stromal/mesenchymal cells (SC/MSCs) based on the knowledge from preclinical models over the last three decades. Accordingly, our protocol is the first to test the use of bone fragments as an ideal vehicle to transplant such cells residing in the bone core. Because of the paucity of knowledge for optimum transplantation of SC/MSCs in humans, we used a multifaceted approach and implanted bone fragments both intraperitoneally and directly into bone on day 0 of BMT. We also infused cultured donor osteoblast-like cells intravenously post-BMT. We were able to achieve high levels of stroma cell engraftment as defined by molecular analyses of bone biopsy specimens.

Bone Marrow Transplantation↗
PubMed1980

Functional study and detection of HLA-D products on fractionated human bone marrow cells.

Bone marrow cells from nine normal human volunteers obtained from the Iliac crest, were used in this work for antigen determination and functional studies. The bone marrow aspirated cells were sequentially separated: elimination of erythrocyte, granulocytes and monocytes achieved by Ficoll-Isopaque centrifugation followed by plastic adherence. Purified bone marrow cells were finally separated by size using velocity sedimentation. The slow sedimenting small cells were shown to be mainly T lymphocytes, probably of blood origin. The medium sized bone marrow cells were shown to contain myeloid precursors (CFu-c). Large immature cells were in cycle actively synthesizing DNA molecules. HLA-D and HLA-DR detections on the fractionated cells were performed using three techniques: fluorescence with specific anti HLA-DR allo and xeno antisera; primed lymphocyte typing (PLT) with anti HLA-DR monospecific in vitro primed lymphocytes and detection of the HLA-D stimulating product using the bone marrow fractionated cells as stimulators in a mixed leukocyte culture. Concordant results were obtained with the three techniques. Lymphocytes in the bone marrow express HLA-D products a peripheral lymphocytes. Bone marrow fractions depleted of lymphocytes and monocytes also contain approximately 20% of cells expressing HLA-D products The meaning of the expression of HLA-D products on immature precursors non-lymphoid cells is discussed.

Bone Marrow Cells↗
PubMed1998

No neuronal regulation of murine bone marrow function.

Bone marrow is innervated by efferent (sympathetic) and afferent nerves, but it is not clear whether these nerves affect cell formation or release in any significant way. To elucidate this problem, we studied mice neonatally sympathectomized with 6-hydroxydopamine and adult mice in which one hind limb was surgically denervated. Progenitor and transit cell numbers and proliferative activity were estimated in bone marrow, blood, and spleen. In addition, we performed unilateral electrical stimulation of nerve fibers to tibial marrow and applied a cell mobilizing stimulus (bleeding, granulocyte colony-stimulating factor injection, or intraperitoneal injection of a chemotactic substance) to investigate cell egress from the marrow. Blood flow to hindleg bone marrow was assessed with the radioactive microsphere technique. Except for a smaller bone marrow cell population and lower body weight in neonatally sympathectomized mice, we found no clear indications that bone marrow innervation influenced cell production. Also, the innervation did not detectably affect cell release from the marrow. Electrical stimulation of hind limb nerves did not change the blood flow to the marrow, whereas it markedly decreased blood flow to the overlying muscle. We therefore conclude that no obvious function can be ascribed to tibial marrow innervation in the mouse.

Animals↗
PubMed1988

Talc granulomatosis in the rat: the relationship between osteoblast insufficiency and adjacent bone marrow hyperplasia.

Bone loss in talc granulomatosis is paralleled by hyperplasia of bone marrow in the rat. To test the hypothetical relation between those two phenomena, bone matrix osteoinduction was employed as a model in which bone formation and bone marrow appearance are separated in time. Implantation of demineralized bone matrix to normal rats was followed by three talc injections (one weekly), starting 1 week after matrix implantation. Implants of demineralized bone and [3H]thymidine-labeled tibial metaphyses from talc-injected and normal rats were analyzed histologically and evaluated for alkaline and tartrate-resistant acid phosphatase activity on days 7, 14, 21, and 30 after matrix implantation. Analysis of tibial autoradiographs showed a marked growth arrest and bone marrow hyperplasia in talc-injected rats 7 days after first talc injection. Alkaline phosphatase activity in homogenates of bone implants was low in talc-injected rats on day 14 after implantation. Moreover, histology of the bone implants showed numerical and functional inhibition of osteoblasts on the same day, causing marked growth delay. Bone marrow appeared as late as day 21 after bone matrix implantation. We conclude that hyperplasia of the adjacent bone marrow is not the cause of bone loss in talc granulomatosis, but rather its compensatory consequence.

Animals↗
PubMed1991

Supernatant cell counts from long-term bone marrow culture correlate with the speed of engraftment following autologous bone marrow transplantation.

This study evaluates the relationship between bone marrow growth in a long-term bone marrow culture (LTBMC) system and speed of engraftment of the same marrow following autologous bone marrow transplantation (ABMT). Bone marrow from 21 patients transplanted with unmanipulated, non-cryopreserved autologous marrow was cultured. Samples from 21 normal donors were cultured to establish the normal supernatant cell count range. Supernatant counts from LTBMCs established from marrow taken from patients at the time of bone marrow harvest were compared with the time to neutrophil and platelet engraftment. Supernatant counts, particularly after 1 week in culture, showed close correlation with time to neutrophil and platelet engraftment following ABMT (r = 0.733, p less than 0.01; r = 0.735, p less than 0.01 respectively). Where supernatant cell counts were within the normal range rapid engraftment was predicted (neutrophils greater than 0.5 x 10(9)/l within 21 days, platelets greater than 50 x 10(9)/l within 28 days) and if supernatant counts were below this range, engraftment was predicted to be delayed. After 1 week in culture, the speed of neutrophil and platelet engraftment were correctly predicted in 19 and 18 cases respectively. Preliminary data suggest that LTBMC of marrow obtained 2-6 weeks before harvesting provides similar data, thus allowing the opportunity to intervene, for example with growth factors, in selected patients.

Adolescent↗
PubMed1984

Pulmonary bone-marrow embolisation after unfiltered autologous bone-marrow transplantation.

A 15-year-old girl had Ewing's sarcoma of the right scapula 1978 and was treated with local radiotherapy and adjuvant chemotherapy. She remained in complete remission until September 1982 when she developed a single pulmonary metastasis in the right lower lobe. She received a high dose of Melphalan twice, always followed by autologous bone-marrow transplantation. A segmental right lower lobectomy revealed haematopoietic bone-marrow tissue emboli within the alveolar capillaries and foreign body granulomatous lesions in the whole surgical specimen. No residual Ewing's sarcoma growth was seen.

Adolescent↗
PubMed1989

Elimination of malignant clonogenic breast cancer cells from human bone marrow.

Autologous bone marrow transplantation is a promising approach to the treatment of breast cancer but is at present limited to patients without bone marrow metastases. To eliminate malignant clonogenic breast cancer cells from normal human bone marrow, immunomagnetic separation has been combined with chemoseparation using 4-hydroperoxycyclophosphamide. Breast cancer cell lines have been mixed with a 10-fold excess of irradiated human bone marrow from normal donors. Mixtures have been incubated with a combination of five different monoclonal antibodies which bind to epithelial cell surface antigens of Mr 42,000, 55,000, 72,000, 200,000, and greater than 200,000. Antiglobulin coated microspheres which contained magnetite were added, and tumor cells were trapped in a magnetic field. Elimination of tumor cells from the decanted marrow was measured in a limiting dilution assay. Two treatments with antibody and microspheres permitted elimination of 2-4 logs of clonogenic breast cancer cells, depending upon the cell line studied. Similar treatment of nonirradiated normal marrow failed to affect levels of colony forming units-granulocyte-macrophage significantly. Use of immunomagnetic purging in combination with 4-hydroperoxycyclophosphamide eliminated up to 5 logs of tumor cells but reduced the recovery of colony forming units-granulocyte-macrophage. If prompt engraftment is observed following reinfusion of similarly treated marrow in phase I trials, these techniques should permit extension of autologous bone marrow transplantation to a larger population of breast cancer patients.

Antibodies, Monoclonal↗
PubMed1979

Bone marrow transplantation.

Bone marrow transplantation can be considered in any disease state resulting in the malfunction or absence of part or all bone marrow elements. Diseases such as aplastic anemia, leukemia, and immunodeficiency disease are being treated with bone marrow transplantation. As with any organ transplant, graft rejection is a possibility. In bone marrow transplantation, there is the additional, unique problem of graft versus host disease. In order to prevent or minimize graft rejection, the immunocompetence of the recipient and the degree of disparity between donor and recipient at the major histocompatibility complex (MHC) loci are considered. The results of bone marrow transplantation are variable, and the mortality rate is still relatively high. However, progress is being made, and in many instances, normal bone marrow function can be restored in patients with whom other treatment has failed.

Anemia, Aplastic↗
PubMed1997

Bone-marrow MR imaging before and after autologous marrow transplantation in lymphoma patients without known bone-marrow involvement.

PURPOSE: To study lumbar bone marrow by means of MR imaging before and after bone-marrow transplantation in lymphoma patients. Particular emphasis was paid to heterogeneity and to focal manifestations, i.e. appearances that could simulate tumor. MATERIAL AND METHODS: Twenty-two patients who were disease-free for a minimum of 30 months after transplantation were studied in 107 MR examinations. Two radiologists visually evaluated coronal T1-weighted and short inversion time inversion-recovery (STIR) images. RESULTS: T1-weighted images demonstrated a more heterogeneous marrow after transplantation than before it. Sharply defined focal low signal intensity areas appeared on this sequence in 5 (23%) of the 22 patients at between 21 and 60 weeks after transplantation. The mean age of these 5 patients was 48.4 years (range 42-54 years). The difference in age between these 5 patients and the remaining 17 patients, who had a mean age of 33.4 years (range 14-51 years), was statistically significant (p < 0.01, Student's t-test, 2-sided test). CONCLUSION: Sharply defined focal low signal intensity areas may be seen on T1-weighted images of bone marrow in patients who are in complete remission after transplantation, particularly in those aged over 40-45 years.

Adolescent↗
PubMed2004

Race, education, and knowledge of bone marrow registry: indicators of willingness to donate bone marrow among African Americans and Caucasians.

BACKGROUND: As bone marrow transplantation (BMT) increases, the availability of suitable donors becomes critical, especially for African Americans, who require a large donor pool to find a suitable match. Previous studies indicated willingness to donate marrow may be a barrier for achieving a large donor pool. METHODS: We conducted a random-sample, statewide telephone survey of 421 Caucasians and 408 African Americans in South Carolina to determine if racial differences in willingness to donate bone marrow exist. We assessed a general level of willingness, asking, "Will you be willing to be a marrow donor?" We assessed an additional level of willingness, asking, "Are you willing to be contacted about bone marrow donation?" RESULTS: We detected no racial differences in general willingness to donate (Caucasians 34%, African Americans 32%, P=.52), although there was a difference in willingness to be contacted to sign-up for the registry (Caucasians 18.3%, African Americans 11%, P=.003). African Americans were more aware that better matches occur within the same race (P <.0001). Caucasians were more knowledgeable about the registry (P <.0001). Younger, more highly educated respondents indicated a greater willingness to be donors. In both races, fear of pain was the most common reason for unwillingness to donate, and it was significantly higher in African Americans. CONCLUSION: Our study suggests reported lack of general willingness does not explain the racial disparities in BMT. Many who expressed willingness to donate were not willing to be contacted to sign up for the registry, especially African Americans. Education and adequate pain control may improve minority recruitment.

Adult↗
PubMed2000

Kinetics of bone marrow repopulation in lethally irradiated rats after transplantation of vascularized bone marrow in syngeneic hind limb.

Grafting of the recipient with bone marrow cell suspension provides only few stromal cells and no autologous environment for cooperation between hemopoietic and stromal cells. Vascularized bone marrow grafts provide the recipient with bone marrow hemopoietic and stromal cells in their natural spatial relationship. It is expected that such a graft will resume its function soon after transplantation and almost immediately repopulate bone marrow cavities of the irradiated recipient as well as supply the sites of cytopoiesis with the necessary growth factors. The aim of the study was to investigate the process of repopulation of bone marrow cavities and lymphoid organs of irradiated recipient by bone marrow cells from syngeneic hind limb transplant. Lewis rats received total body irradiation of 8 Gy followed by orthotopic transplantation of syngeneic limb or i.v. infusion of equivalent amount of bone marrow cells. In control experiments the hind limb was shielded with lead plate during total body irradiation. Ten days after irradiation and hind limb transplantation the yield of nucleated cells from tibia was reaching values of normal animals. In rats receiving i.v. bone marrow cell infusion it was 40% of control values and in rats repopulated from shielded own hind limb it was 60% of controls. In all groups a higher percentage of early and immediate normoblasts and a reduced pool of juvenile and segmented neutrophils was observed. Thirty days after irradiation and repopulation procedures all parameters were returning to normal levels in each group. The results indicate that bone marrow cell transplantation in hind limb graft is highly effective in lethally irradiated animals in reconstituting bone marrow.

Animals↗
PubMed1988

Pattern of response of megakaryocyte colony-stimulating activity in the serum of patients undergoing bone marrow transplantation.

Bone marrow transplantation has become an accepted form of therapy for several malignant, hematologic, and genetic disorders. Platelet recovery is delayed after bone marrow transplantation. To better understand the mechanisms involved in platelet recovery we studied 23 patients undergoing bone marrow transplantation for the presence of megakaryocyte colony-stimulating activity (Mk-CSA) in their serum. Shortly after beginning the pretransplant preparative regimen the Mk-CSA level in the serum of these patients increased. This increase was transient, and the level returned to baseline, to later increase again. The second increase in Mk-CSA level occurred during the second week after bone marrow transplantation at the time of hematopoietic recovery. Most patients who failed to engraft did not show a rise in Mk-CSA during the second week after transplantation. All patients showing engraftment had an Mk-CSA rise during the second week after transplantation. The difference between these two groups was highly significant (p = 0.0007). The biphasic response of Mk-CSA after bone marrow transplantation is similar to the response seen in a rat model after lethal irradiation. We postulate that the first elevation in Mk-CSA is due to tissue injury and nonspecific response whereas the second elevation of Mk-CSA is a physiologic response to marrow aplasia and associated with effective bone marrow engraftment.

Adolescent↗
PubMed1991

Pediatric bone marrow transplantation.

Bone marrow transplantation is a high-visibility, high-technology discipline with a growing list of potentially curative applications in neoplastic, hematologic, immunologic, and genetic diseases of children. The clinical problems experienced by children who have received bone marrow grafts involve pediatricians in both general and subspecialty practice and require a working knowledge of the applied immunobiology of bone marrow transplantation. As the transplantation procedure has evolved from a research tool to an established therapeutic modality, collaboration between basic scientists and clinicians has led to a greater understanding of the pathophysiology of the immunobiologic events that occur after allogeneic bone marrow transplantation and, thus, to improvements in the clinical care of transplant recipients. As its applications in pediatric medicine grow, bone marrow transplantation presents additional challenges for the future: expansion of the allogeneic donor pool, use of more effective or aggressive preparative regimens and autologous bone marrow-purging strategies, more effective prevention and treatment of graft-versus-host disease, and development of gene-replacement therapy by infusion of modified autologous bone marrow cells.

Bone Marrow Transplantation↗