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PubMed2003

[Bone marrow suppression].

Bone marrow suppression is one of the most popular side effects of cancer chemotherapy. Bone marrow suppression includes neutropenia, anemia and thrombocytopenia. Clinical application of cytokine is useful in the treatment of bone marrow suppression due to cancer chemotherapy. Granulocyte colony-stimulating factor(G-CSF) is useful in the treatment of neutropenia. Erythropoietin(EPO) is useful is the treatment of anemia. However, EPO is not widely used in Japan yet. Clinical application of varions cytokines will bring forth much improvement in the treatment of bone marrow suppression due to cancer chemotherapy.

Anemia↗
PubMed2001

Topics in bone marrow biopsy pathology: role of marrow topography in myelodysplastic syndromes and evaluation of post-treatment and post-bone marrow transplant biopsies.

The role of bone marrow biopsy is expanding. Bone marrow biopsy is considered a "gold standard" for assessing cellularity and infiltrative process. More recently, biopsies are increasingly used for immunohistochemical stains and molecular studies such as in situ hybridization and laser microdissection, further augmenting morphologic interpretation. Biopsies are playing a greater role in early diagnosis of myelodysplastic syndromes (MDS). Both primary and secondary MDS are increasing (5% to 12%), and can present a significant challenge in early diagnosis. Present MDS diagnostic criteria are based purely on marrow aspirate smears, peripheral blood smears, and ancillary studies, and may not be adequate in some instances. With the current understanding of marrow topography and hematopoietic microenvironment, bone marrow biopsies are greatly useful in early diagnosis and grading of MDS. Marrow biopsies also are helpful in evaluation of postchemotherapy and post-bone marrow transplant patients. Assessment of topographic alteration and certain corroborating immunohistochemical and molecular studies can only be performed on bone marrow biopsies, and these ancillary studies will contribute significantly to the understanding of hematopoietic disorders. Bone marrow biopsy, in conjunction with other modalities such as flow cytometry, will play a significant role in diagnostic hematopathology. Ann Diagn Pathol 5:110-120, 2001.

Biopsy↗
PubMed1991

The role of immunoglobulin in bone marrow transplantation.

Bone marrow transplantation has proven its value as a therapeutic approach to a variety of human diseases, primarily the hematopoietic malignancies. The major clinical problems preventing successful outcome of marrow transplant have been defined and therapeutic approaches to preventing or treating these complications have led to increased long-term disease survival. Passively administered antibody given as intravenous immunoglobulin has been studied as a therapeutic modality following bone marrow transplantation. Studies have demonstrated the efficacy of immunoglobulin in reducing bacterial infections in the posttransplant period; reducing severe CMV infections in allogeneic marrow transplant recipients who are seronegative for the virus and susceptible to primary infection; reducing mortality from CMV pneumonia in combination with ganciclovir; and reducing acute graft-versus-host disease following allogeneic BMT. In many cases alternate therapeutic strategies offer comparable or greater efficacy (eg, selective CMV-negative blood products for CMV seronegative allogeneic BMT recipients receiving bone marrow from a seronegative donor). However, it is clear that intravenous immunoglobulin has a place in the therapeutic armamentarium of bone marrow transplantation. Future controlled clinical trials are necessary to establish its exact role and to define which preparations and dose schedules provide the greatest therapeutic benefits.

Acute Disease↗
PubMed1987

Prolongation of graft survival in ALS-treated mice by donor-specific bone marrow. Density gradient fractionation of the active bone marrow cells.

Antilymphocyte serum (ALS)-treated recipient mice that are infused with donor bone marrow cells have been shown to enjoy prolonged skin graft survival. In the present experiments, discontinuous density gradient centrifugation in Percoll was used to prepare fractions of these cells. Two of four fractions had good graft-prolonging ability. In experiments in which the density of the gradients was varied, we were able to estimate the density of a major portion of the cells active in graft prolongation as being in a range of 1.061 to 1.066 g/ml, and to produce one fraction with superior graft-prolonging activity. This fraction constituted only 10% of the recovered cells, was enriched for small-to-medium lymphocytes, Ia+, Thy-1+, and IgM+ cells, and contained cells bearing a marker known to be present on active cells, Fc gamma R. Additionally, this fraction contained cells capable of suppressing the mixed lymphocyte reaction, although two denser fractions, of poor-to-good graft-prolonging ability, also shared this ability. The effectiveness of this method in bringing about a high-capacity, rapid isolation of cells that can augment graft survival in immunosuppressed recipients makes it well suited for future clinical application.

Animals↗
PubMed1975

[99mTc-sulfur-rhenium colloid and 111In-indium citrate in bone-marrow scintigraphy].

Bone marrow scinitigraphy using 111In-Indium-citrate and 99mTc-sulfur rhenium colloid was compared with each other in 6 male and 3 female patients. Our results in all patients were in favour of 111In-citrate which caused a better delineation of morphological details than 99mTc-sulfur rhenium colloid did. In the first days after intravenous administration of 111In-citrate, radioindium accumulated markedly in the pudendal region, this finding being more distinct in male patients than in female ones. On account of these results a considerable radiation dose has to be assumed in bone marrow scinitigraphy using 111In-citrate as long as the absorbed dose from 111In has not been estimated under consideration of the radioindium accumulation in the pudendal region. 111In-citrate should be applied in bone marrow scanning only exceptionally and 99mTc-sulfur rhenium colloid be preferred in the routine diagnostics of bone marrow.

Adult↗
PubMed1992

Study of possible correlations between in vitro growth of bone marrow stromal and hematopoietic precursor cells early after allogeneic bone marrow transplantation.

Growth characteristics of stromal cells, assessed as adherent cells in long-term bone marrow cultures, and of hematopoietic progenitor cells, prior to and shortly after allogeneic bone marrow transplantation (BMT), were investigated, more specifically with regard to their possible correlations. The main constituent cells of the bone marrow stroma, that is, endothelial cells, reticular cells/fibroblasts, and monocytes/macrophages, showed an as yet inexplicable increased growth in samples taken from recipients prior to BMT, as compared with the growth in samples from their healthy donors and those taken after BMT. In the third week after BMT the in vitro outgrowth of hematopoietic precursors was severely depressed, but cell numbers in the adherent layer were normal. No relationship between in vitro growth of hematopoietic precursor cells and stromal cells was observed at that time. Probably the precursor cells growing in vitro are committed progenitor cells, relatively independent of stromal influences. In the eight week after grafting, endothelial cell outgrowth in vitro was highly correlated with granulocyte-macrophage colony-forming unit (CFU-GM) colony formation and to a lesser extent with mixed-lineage colony-forming unit (CFU-Mix) colony formation. This may indicate the reappearance of cytokine-mediated influences or the reappearance of a direct interaction, for example, by cell-cell contact between stromal cells and hematopoietic progenitor cells at that time.

Bone Marrow↗
PubMed1976

Bone marrow transplantation from donors with aplastic anemia. A report from the ACS/NIH bone marrow transplant registry.

Bone marrow transplantation from HLA-matched allogeneic donors was used to treat two series of patients with severe aplastic anemia. No significant differences were detected in all comparisons made between the Registry series (38 patients) and the Seattle series (24 patients), and pooled data from the two series were used in the analyses. Currently, 55% of the patients who received bone marrow transplants within three months of diagnosis are alive, but only 13% of the patients who received bone marrow transplants more than nine months after diagnosis are alive. The difference between the two groups was significant (P less than .02). Patients less than 21 years of age had a significantly higher survival rate than those patients who were 21 years or older at the time of transplantation (P less than .02). Survival rates were significantly higher for patients who had received 15 or fewer pretransplant transfusions than those who received more (P less than .05).

Adolescent↗
PubMed2001

Reconstitution of the CD45RO(+) and CD20(+) lymphoid marrow population following allogeneic bone marrow transplantation for Ph(+) CML.

Following bone marrow transplantation (BMT) investigations on the recovery of the B and T lymphocyte populations have focused on the peripheral blood and only marginally regard the bone marrow. An immunohistochemical and morphometric study was performed on 352 trephine biopsies derived from 123 patients with chronic myelogenous leukemia (CML) at standardized endpoints before and after allogeneic BMT and compared to a control group. The purpose of this investigation was to quantify the B-CD20(+) and T-CD45RO(+) lymphocyte subsets and to determine possible relationships with the occurrence of acute and chronic GVHD. Moreover, we studied the dynamics of lymphocyte repopulation in the post-transplant period, correlations with the total peripheral lymphocyte count and differences associated with sibling vs alternate HLA-compatible (unmanipulated) marrow grafts. Morphometric analysis revealed a very fast regeneration of CD45RO(+) and CD20(+) marrow lymphocytes in the first 2 weeks following BMT. In less than 2 months, in most patients, the post-transplant quantity of lymphocytes was comparable to that of the normal bone marrow. This finding was opposed to the profound depression of the absolute lymphocyte count in the peripheral blood. No relevant relationships could be calculated between engraftment status and the lymphocyte repopulation in the bone marrow. On the other hand, significant correlations were calculable between the development of (chronic and acute) GVHD including severity with the number of CD45RO(+) lymphocytes. In non-related graft constellations a more frequent evolution of acute grade III + IV GVHD was detectable. This complication was accompanied by an increased quantity of CD45RO(+) lymphocytes in the marrow.

Adolescent↗
PubMed1989

Potential therapeutic role of cisplatinum in autologous bone marrow transplantation: in vitro eradication of neuroblastoma cells from bone marrow.

Cisplatinum may prove to be a valuable agent for the elimination of diseased cells in the bone marrow of patients with neuroblastoma. In this study, we measured the efficacy of cisplatinum on human neuroblastoma cell lines and on normal human bone marrow progenitors, GM-CFC and CFU-F. Data indicate that the therapeutic index of cisplatinum is high. We set up an experimental model consisting of a mixture of human bone marrow and human neuroblastoma cells in order to confirm these preliminary results in purging conditions. Results indicate that cisplatinum exhibits a high and specific tumoricidal property and appears to be valid in bone marrow purging.

Bone Marrow↗
PubMed2001

Bone marrow aspiration.

Bone marrow aspiration biopsies are carried out principally to permit cytological assessment but also for immunophenotypic, cytogenetic, molecular genetic, and other specialised investigations. Often, a trephine biopsy is carried out as part of the same procedure. Bone marrow aspirations should be carried out by trained individuals who are aware of the indications, contraindications, and hazards of the procedure. They should follow a standard operating procedure. The operator should have made an adequate assessment of clinical and haematological features to ensure both that appropriate indications exist and that all relevant tests are performed. For the patient's comfort and safety, the posterior iliac crest is generally the preferred site of aspiration. Films of aspirated marrow and, when appropriate, films of crushed particles should be made and labelled. Once thoroughly dry, films should be fixed and stained. As a minimum, a Romanowsky stain and a Perls' stain are required. A cover slip should be applied. The bone marrow films should be assessed and reported in a systematic manner so that nothing of importance is overlooked, using a low power, then intermediate, then high power objective. A differential count should be performed. An interpretation of the findings, in the light of the clinical and haematological features, should be given. The report should be signed or computer authorised, using a secure password, and issued in a timely manner.

Biopsy, Needle↗
PubMed1999

Correction of bone marrow failure in dyskeratosis congenita by bone marrow transplantation.

Dyskeratosis congenita is recognized by its dermal lesions and constitutional aplastic anemia in some cases. We report successful allogeneic bone marrow transplantation in two siblings with this disease from their sister, and their long term follow-up. We used reduced doses of cyclophosphamide and busulfan for conditioning instead of total body irradiation. Also, we report late adverse effects of transplantation which are not distinguishable from the natural course of disease.

Adolescent↗
PubMed1983

The influence of colcemid concentration on mitotic index and length of bone marrow chromosomes.

Bone marrow cells from 25 patients with different hematological disorders were cultured overnight and then exposed to colcemid, varying both concentration and time of exposure, in order to obtain optimal results. PHA stimulated lymphocytes were treated in a similar manner for comparison. Results showed that with concentrations at or below 0.4 microgram/ml, the mitotic index of PHA stimulated lymphocytes was reduced and the metaphase spreads were of poor quality, but spreads of bone marrow chromosomes were not adversely affected at even much lower concentrations. Furthermore, both chromosome length and mitotic index were found to increase with lower concentrations. These observations, together with results obtained by comparing mitotic indices after varying exposure times, suggest that bone marrow cells may be inhibited from passing through G2 by high concentrations of colcemid. Thus by lowering the concentration of the colcemid for bone marrow cultures, we have been able to increase greatly the yield of bandable metaphases.

Bone Marrow↗
PubMed2001

[Bone Marrow Cells Activated by Autologous Dendritic Cells Purges Bone Marrow from Patients with Chronic Myelogenous Leukemia]

In order to investigate the effect of autologous dendritic cells (DCs) activating bone marrow cells and purging bone marrow from chronic myelogenous leukemia (CML) patients, DCs were separated by negative selection system of human cells from bone marrow mononuclear cells (BMMNCs) of 2 CML patients in hematological remission and harvested after 3 days of culture in IMDM containing autologous plasma, rhGM-CSF and rhTNFalpha at 37 degrees C, 5% CO(2) humidified atmosphere. BMMNCs from the patients were also used to set up long-term culture (LTC) system in T-25 plastic flasks. The LTCs included three groups, i.e., control, addition of rhIL-2, and co-culture with autologous DCs. Half of non-adherent cells were collected, counted and assayed for CFU-GM weekly. Then, equivalent volume of fresh medium was replaced to maintain the culture. The culture was discontinued if the non-adherent cells count was less than 2 x 10(5). Adherent cells were collected for CFU-GM assay and flow cytometry for CD34 and P210. The colonies originating from the adherent cells were picked up under the inverted microscope. RNA was extracted, and BCR/ABL measured by nested reverse transcription polymerase chain reaction (RT-PCR). The results showed that the CFU-GM yields of non-adherent cells declined after 1 to 2 weeks co-cultured with autologous DCs, and it paralleled with group with rhIL-2. P210(+) cell percentage was also decreased. From the third week on, however, the decrease of CFU-GM yields slowed down, while CFU-GM in the system with rhIL-2 continued to fall. In system co-cultured with autologous DCs, the adherent cells contained the least percentagcs of CD34(+) cells and P210(+) cells percentage. However, the expression of BCR/ABL in CFU-GM colonies derieved from the adherent cells of DCs co-cultured had no significant difference with those from the culture without DCs. Our results suggest that co-culture of marrow cells with autologous DCs could significantly diminish the leukemic progenitors cells including both mature and primitive progenitor cells. Autologous dendritic cells might be used for ex vivo purging of CML marrow.

Journal Article↗
PubMed2000

Effect of interleukin-3, stem cell factor and granulocyte-macrophage colony-stimulating factor on committed stem cells, long-term culture initiating cells and bone marrow stroma in a one-step long-term bone marrow culture.

Long-term bone marrow culture (LTBMC) supports both differentiation and conservation of hematopoietic progenitor cells. During the culture period, the frequency and proliferative potential of early repopulating stem cells that give rise to progenitors detectable after 5 weeks in LTBMC--so-called long-term culture-initiating cells (LTC-IC)--can be investigated. The adherent stroma cell layer was modified by interleukin-3 (IL-3) + granulocyte-macrophage colony-stimulating factor (GM-CSF)+ stem-cell factor (SCF) with an increased cellularity and higher percentage of differentiated myeloid cells and a reduced percentage of stroma cells. We have studied the effects of stimulative cytokines, such as GM-CSF, SCF, and IL-3, on proliferation of committed colony-forming unit cells (CFU-C), LTC-IC, and stroma cell formation in LTBMC of unseparated bone-marrow cells. IL-3, GM-CSF, and SCF significantly stimulated the cumulative proliferation of nucleated cells and committed CFU-C. Weekly stimulation of LTBMC during the culture period did not exhaust the proliferative capacity of stem cells as seen in maintenance of LTC-IC after 5 weeks in LTBMC. In contrast, no LTC-IC were seen in limiting dilution analyses from LTBMC cultured 5 weeks without cytokine supplementation. Our data indicate that an increased proliferation of committed stem cells can be achieved by the addition of stimulatory growth factors, and maintenance of LTC-IC is possible over a period of 5 weeks. A net expansion of LTC-IC if compared with the starting bone-marrow suspension could not be obtained after a 5-week LTBMC period.

Bone Marrow↗
PubMed1987

Expression of tartrate-resistant acid phosphatase in bone marrow macrophages.

Bone marrow macrophages were found to express tartrate-resistant acid phosphatase (TRAP) under pathological conditions. In chronic granulocytic leukemia and metastatic carcinoma in the bone marrow this phenomenon was striking, all or almost all of the marrow macrophages being reactive. In other conditions, such as hypertransfusion or chemotherapy-induced marrow aplasia, the phenomenon did occur but was clearly a minor one. These observations indicate that tissue macrophages may become TRAP positive under the effect of unknown stimuli operating in certain pathological conditions. The results further suggest that the synthesis of the isoenzyme of acid phosphatase resistant to tartrate inhibition is a marker of macrophage activation rather than of differentiation towards particular subsets of the mononuclear phagocyte system.

Acid Phosphatase↗