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[Effect of autologous marrow transplantation on bone marrow recovery following large-field fractionated irradiation of dogs].

After large-field fractionated irradiation of dogs at a cumulative dose of 54 Gy, a stable bone marrow depletion occurs persisting for a year following irradiation. The automyelotransplantation after the end of the exposure elicits a transient recovery of the exposed bone marrow, 1.5-2 months after the beginning of irradiation, followed by a secondary depletion of the exposed haemopoietic sites. The control and the automyelotransplanted animals exhibited bone marrow recovery one year and six months after irradiation, respectively, the cellularity being maintained at a high level for 3 years of observation.

Animals↗

Dynamic scintigraphy of bone and bone marrow in multiple myeloma patients with bone-marrow transplants.

PURPOSE: To determine whether dynamic registration at bone and bone-marrow scintigraphy produces additional information compared to subsequent static registrations of bone-marrow transplants in multiple myeloma patients. MATERIAL AND METHODS: In a prospective study, 8 dynamic bone and 6 dynamic bone-marrow scintigraphies were performed in 10 patients. The dynamic scintigraphies were compared with conventional radiography, MR images, and static scintigraphies of bone and bone marrow. RESULTS: No additional information was revealed by the dynamic registration method; on the contrary, 4 of the 8 known lesions were not discerned at dynamic registration. An incidental observation was that the time-activity curves of both radiopharmaceuticals had a specific pattern. CONCLUSION: Dynamic registration at bone and bone-marrow scintigraphy was not useful for detecting disease in multiple myeloma lesions.

Adult↗

[The central registry of bone marrow donors and the program for bone marrow transplantation from unrelated donors--20 months' experience].

The authors describe their 20 months' experience with the establishment of a bone marrow register in the Czech Republic and its practical association with the programme of bone marrow transplantations from non-related subjects. During the 20 months of activity 5455 voluntary bone marrow donors were examined and at present more than 500 new voluntary donors are examined every month. For 21 patients a HLA identical unrelated donor is sought. For four patients a HLA compatible donor was found. The first bone marrow transplantation from a unrelated donor was performed and another patient is prepared for transplantation within the next few days. The authors discuss professional, organizational, moral and ethical problems associated with the elaboration of this medical programme.

Adult↗

Current status of bone marrow transplantation in humans: report from the International Bone Marrow Transplant Registry.

Bone marrow transplantation is being used with increasing frequency as therapy for a number of life-threatening diseases. Data reported to the International Bone Marrow Transplant Registry are presented to show the pattern of growth of bone marrow transplantation worldwide as well as current results in leukemia and aplastic anemia. Risk factors are identified that may predict the development of major complications after bone marrow transplantation.

Anemia, Aplastic↗

Bone marrow separation, purification and cryopreservation: application for autologous bone marrow transplant (ABMT).

Autologous bone marrow transplantation is used in the treatment of patients suffering from malignant diseases. The intention of this study was to develop a technique for cryopreservation of bone marrow stem cells. A technique of purification described elsewhere (2), gave recoveries of 80% of nucleated blood cells and 87% of mononucleated blood cells after apheresis and centrifugation of buffy-coat, but when applied to bone marrow only 59% of NBC and 32% of CFUc have been harvested. We therefore modified the apheresis technique and replaced the centrifugation step by sedimentation with hydroxy-ethyl-starch. This change resulted in a small but significant increase in the yield of bone marrow cells and improved the asepsis of the procedure. From a second group of bone marrow handled with this modified protocol, final yields of 46% of NBC and 62% of MNC were obtained. After cryopreservation 87% of NBC were recovered as assessed by counting.

Adolescent↗

Ancillary techniques in bone marrow pathology: molecular diagnostics on bone marrow trephine biopsies.

Pathologic examination of trephine bone marrow (BM) biopsies plays a central role in the diagnosis and staging of haematological neoplasms and other disorders affecting haematopoiesis. Haematopathology has been profoundly influenced by the advent of molecular genetic techniques suitable for paraffin-embedded tissues, and certain applications, such as the determination of B- and T-cell clonality, belong to its standard diagnostic repertoire. Many of these molecular tests can be performed successfully with nucleic acids extracted from BM trephine biopsies, if some technical aspects specific to this template source such as various fixation and decalcification procedures are taken into consideration. The current indications for molecular BM diagnostics range from the confirmation of lymphoma involvement with gene rearrangement analysis, demonstration of tumor-specific translocations in lymphoid and chronic myeloproliferative disorders along to the detection of microorganisms or marrow involvement by soft tissue sarcomas. The availability of quantitative polymerase chain reaction techniques for the investigation of allelic imbalances and gene expression levels in paraffin-embedded material also open new avenues for research and advanced diagnostics. The molecular detection of minimal residual disease in haematological neoplasms, especially in the context of new treatment strategies, will provide future challenges. This article summarizes the current state of the art in molecular diagnostics applied to paraffin-embedded BM biopsies.

Biopsy↗

Simultaneous injection of bone marrow cells and stromal cells into bone marrow accelerates hematopoiesis in vivo.

We have previously demonstrated that stromal cells can support the proliferation and differentiation of hematopoietic cells in vitro and in vivo and that a major histocompatibility complex restriction exists between hematopoietic stem cells and stromal cells. We have also found that intra-bone marrow (IBM) injection of allogeneic bone marrow cells (BMCs) leads to more rapid reconstitution of hematopoietic cells than intravenous injection. In the present study, we examine the effect of simultaneous injection of stromal cells and BMCs into the same bone marrow on the recovery of donor hematopoietic cells and demonstrate that simultaneous IBM injection of BMCs plus stromal cells is more effective in reconstituting recipients with donor hematopoietic cells than intravenous injection of BMCs plus stromal cells or IBM injection of BMCs alone.

Bone Marrow Cells↗

Janus kinase 3 inhibitor WHI-P131/JANEX-1 prevents graft-versus-host disease but spares the graft-versus-leukemia function of the bone marrow allografts in a murine bone marrow transplantation model.

The purpose of the present study was to evaluate the effects of graft-versus-host disease (GVHD) prophylaxis with the Janus kinase 3 (JAK3) inhibitor WHI-P131/JANEX-1 on the graft-versus-leukemic (GVL) function of marrow allografts in mice undergoing bone marrow transplantation (BMT) after being challenged with an otherwise invariably fatal dose of BCL-1 leukemia cells. GVHD prophylaxis using WHI-P131 markedly improved the survival outcome after BMT. The probability of survival at 30 days after BMT was 11% +/- 6% for vehicle-treated recipients (median survival time, 25 days) versus 63% +/- 12% for recipients treated with WHI-P131 (median survival time, 36 days; P <.0001). Because WHI-P131 is devoid of antileukemic activity against BCL-1 leukemia cells, this marked improvement in survival outcome was due to reduced incidence of GVHD-associated fatalities combined with sustained GVL function of the allografts in the WHI-P131 group. Notably, adoptive transfer experiments demonstrated that the spleens of WHI-P131-treated allograft recipients contained less than 0.001% BCL-1 cells. Notably, GVHD prophylaxis with WHI-P131 plus methotrexate resulted in 100% survival of mice receiving allotransplants challenged with an otherwise invariably fatal dose of BCL-1 leukemia. Taken together, our results provide strong experimental evidence that GVHD prophylaxis using WHI-P131 does not impair the GVL function of the allografts and consequently contributes to an improved post-BMT survival outcome of the recipient mice.

Animals↗

Bone marrow stromal function from patients after bone marrow transplantation.

A two-stage long-term bone marrow culture (LTBMC) has been used to assess haemopoietic supporting ability from patients following allogeneic bone marrow transplantation (BMT). Irradiated confluent LTBMCs derived from recipients following BMT were recharged with flow cytometry sorted CD34+/CD38- cells from normal BM donors. The output of granulocyte/macrophage colony-forming cells (GM-CFC) in the non-adherent fraction of the cultures was monitored weekly until output ceased. Patient stromal cell function was determined by comparing the percentage of the total cumulative GM-CFC output to the output from normal control stroma. Neutrophil recovery was rapid and complete early in the post-transplant period. However, in contrast, GM-CFC recovery was significantly reduced in all patients up to 5 years after BMT. In 58% of patients studied following BMT the BM stroma is considerably compromised in its ability to support early uncommitted haemopoietic cells contained within the CD34+/CD38- cell fraction of normal BM. Bone marrow stroma from BMT recipients exhibited severe deficits in their abilities to support primitive haemopoietic cells and in some patients this effect lasted for several years. While the growth of stroma from patients following BMT was severely delayed and often incomplete, characterisation of cells types using APAAP techniques did not show significant differences in proportion when compared with normal cultures.(ABSTRACT TRUNCATED AT 250 WORDS)

ADP-ribosyl Cyclase↗

Chromosomal analysis of bone marrow stromal fibroblasts in allogeneic HLA compatible sibling bone marrow transplantations.

Karyotyping was performed on bone marrow stromal fibroblasts and marrow haemopoietic cells on six patients who received bone marrow transplants from siblings of the opposite sex. Three patients with severe aplastic anaemia received unmanipulated donor bone marrow cells. Three other patients with leukaemia and conditioned with high dose chemotherapy (+ total body irradiation in two patients) received T-cell depleted marrow mononuclear cells. Marrow chromosomal analysis was performed on samples obtained between 6 weeks and 1 yr post-transplant. All marrow fibroblasts studied were of recipient origin, whereas all haemopoietic cells were of donor origin. These results demonstrate that in allogeneic sibling marrow transplantation, recipient bone marrow stromal cells regenerate and repopulate the bone marrow and this is not influenced by the conditioning regimen used, the type and dose of marrow cells given and the pre-existing disease.

Anemia, Aplastic↗

Bone marrow cell count per cubic millimeter bone marrow: a new parameter for quantitating therapy-induced cytoreduction in acute leukemia.

A new technique is introduced for determining the number of bone marrow cells per cubic millimeter marrow, providing an accurate and objective means for quantitating therapy-induced cytoreduction. The method requires a correction for admixed peripheral blood in bone marrow aspirates to measure the fraction of remaining pure marrow. While cell kinetic differences between blood, aspirates, and biopsies identify the proportion of contaminating blood cells, the ratio of red cell hematocrits in blood and aspirate gives the volume of trapped blood. By combining both procedures, bone marrow cell counts per unit volume pure marrow result (BMC/cu mm BM), which were found highly reproducible. Blast cell counts (BMBC/cu mm BM) were obtained by additional morphological differentiation. BMC and BMBC/cu mm BM were monitored in 16 patients with acute nonlymphoblastic leukemia treated with daunorubicin, cytosine arabinoside, and 6-thioguanine in combination and in 4 patients with end-stage acute leukemias and non-Hodgkin's lymphomas during high-dose thymidine therapy. Total and daily therapy-induced cytoreduction rates were significantly greater (P less than 0.01) in responders than nonresponders to either regimen. Changes in BMC/cu mm BM were also found representative for changes in BMBC/cu mm BM, since the majority of bone marrow cells were blasts. In acute leukemia. BMC/cu mm BM thus provides accurate and objective measurements of treatment efficacy in vivo and after short periods of drug exposure. Differences in cytoreduction rates within the group of responders also suggest possible prognostic implications.

Acute Disease↗

MRI "road-map" of normal age-related bone marrow. I. Cranial bone and spine.

We retrospectively reviewed 733 cranial and 250 spinal T1-weighted MR images of patients younger than 24 years to evaluate the bone marrow changes. The signal intensity of the bone marrow on short-TR/TE images was compared with that of fat and normal muscles in the contiguous region and graded. The signal intensity of all anatomic segments was as low as that of muscle, or inferior, in all patients younger than 3 months because of hematopoietic tissue and probably greater amounts of trabecular bone. The first anatomic segments of cranial bone to become hyperintense were the zygomatic bone and mandibular symphysis, followed by the presphenoid bone, basisphenoid, basiocciput, calvaria, and the petrous apex. After 3 years of age, most patients demonstrated pneumatization of the sphenoid sinus. We describe the most interesting changes in the developing spine, which occur in the first 2 years of life. The morphology of the vertebral bodies was evaluated. The variability of the signal and the morphology of the disks were also evaluated. Regional patterns of bone marrow signal intensity and age-related differences should not be misinterpreted as a pathologic condition.

Adolescent↗

Immunological reconstitution after bone marrow transplant with Campath-1 treated bone marrow.

Immunological reconstitution after allogeneic bone marrow transplant (BMT) was studied in 20 patients who received Campath-1 treated bone marrow. The peripheral blood lymphocyte phenotype was analysed with a panel of monoclonal antibodies at 3, 6 and 12 months. T cell proliferative capacity was evaluated by stimulation with PHA and Con A and in the mixed lymphocyte reaction (MLR). Natural killer (NK) cell activity was analysed against the K562 cell line at specified times after BMT in nine patients. Absolute numbers of T lymphocytes were reduced in all patients at 3 and 6 months. A marked decrease in the number of CD4+ cells persisted beyond 12 months. CD8+ cells regenerated more rapidly and reached normal at 6 months. No correlation was found between changes in lymphocyte subpopulations and the presence of graft-versus-host disease or cytomegalovirus infection. B cells recovered rapidly and maintained normal numbers throughout the study. A moderate increase in HNK1+ (Leu7) cells was observed at 3 and 6 months simultaneously with a low expression of NK15 (Leu11) and OKM1 antigens at 3 and 6 months, suggesting the presence of immature NK cells early after the transplant. A profound decrease of T cell proliferative capacity was observed both after mitogen stimulation and in the mixed lymphocyte reaction. NK cell activity was raised during the first month after transplant in all but one patient but no correlation was found with the presence of GVHD or cell marker analysis.

Adolescent↗

Autologous bone marrow support and bone disease in metastatic breast cancer.

High-dose chemotherapy (HDCT) and autotransplantation of hematopoietic cell is being investigated as a therapy for either metastatic or localized high-risk breast cancer. Breast cancer has a tendency to metastasize to the bones and the bone marrow (BM) and therefore the probability of harvesting malignant cells when collecting stem cells for autotransplantation appears high. Thus, the elimination or decrease of this contamination in the transplanted product appears mandatory. Autologous peripheral blood stem cell transplantation (PBSCT) has shown significant advantages over autologous bone marrow transplantation (ABMT) in improving the feasibility of HDCT, while possibly limiting the BM contamination. The transplantation of only CD34+ products may even be a further advance. The role of ex vivo purging of cancer cells has not been established in ABMT or PBSCT. The question remains if the positive selection of CD34+ products is sufficient for controlling cancer cell contamination or if this product should be purged as well. The review of the literature suggests that contamination of the bone marrow could have an impact in terms or risk of relapse and could thus play a role as a pejorative prognostic factor. These data, although not totally adequate for the autotransplantation setting, are raising concerns over the probability of reinfusing malignant cells at time of autotransplantation following HDCT. There is a tremendous need to address these concerns in the laboratory along with prospective clinical trials. Until further data is available, this risk must be taken into consideration when patients with breast cancer are treated with curative intent.

Antigens, CD34↗