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PubMed2000

Ex vivo expansion and characterisation of CD34+ cells derived from chronic myeloid leukaemia bone marrow and peripheral blood, and from normal bone marrow and mobilised peripheral blood.

Ex vivo culture of CD34+ has the potential to provide large numbers of cells for clinical use in autologous and allogeneic transplantation and for experimental research involving genetic manipulation. We evaluated the ex vivo expansion of CD34+ cells obtained from bone marrow (BM) and peripheral blood (PB) of untreated patients with chronic myeloid leukaemia (CML) in the chronic phase and compared these results with those obtained from BM from normal volunteers (NBM) and peripheral blood after mobilising chemotherapy from patients with non-haematological disorders (MPB). Selected CD34+ cells were stimulated with interleukin 1(beta), interleukin IL-3, interleukin IL-6 and stem cell factor. The proliferation observed in patients with CML was similar to that seen in normal donors. CD34+ cells derived from patients with CML are more differentiated than their normal counterparts, as shown by the coexpression of CD34 and CD33 antigens on day 0 (85.6% for CML-BM and 76.8% for CML-PB). The culture conditions allowed a significant expansion of granulocyte-macrophage colony-forming units (CFU-GM) from NBM (33-fold increase) and MPB (22-fold increase), in contrast with CML-derived BM and PB CD34+ cells (2.3-fold increase). These results indicate that the optimal time to harvest ex vivo expanded cells is dependent on a critical compromise between cell numbers and successful retention of their repopulating potential.

Antigens, CD34↗
PubMed1983

Bone marrow aspiration biopsy and bone scanning in the staging of prostatic cancer.

Random bone marrow aspiration biopsies of iliac crests and bone scans/skeletal X-rays were evaluated in 163 new cases of carcinoma of the prostate in order to assess the incidence of bone metastases and to compare the sensitivity of these two methods. Bone marrow biopsy revealed cancer cells in 18.4% of all cases. The detection rate of skeletal metastases of bone scan/skeletal X-ray was 32.5%. In this series all patients with positive biopsies had also positive scans. The superior sensitivity of bone scans compared with bone marrow biopsies in detecting metastatic spread of prostatic cancer is shown.

Acid Phosphatase↗
PubMed1993

Interleukin-10 inhibits the osteogenic activity of mouse bone marrow.

Murine bone marrow cells synthesize bone proteins, including alkaline phosphatase (ALP), collagen type I, and osteocalcin, and form a mineralized extracellular matrix when cultured in the presence of beta-glycerophosphate and vitamin C. Interleukin-10 (IL-10) suppressed the synthesis of these bone proteins and mineralization without affecting cell proliferation. In addition, mRNA levels for the latter proteins were reduced in IL-10-treated cultures. This inhibitory effect was most outspoken when IL-10 was added before ALP activity peaked, eg, day 15 of culture. No significant effect was observed when IL-10 was added at later time points. This finding suggests that IL-10 acts at osteogenic differentiation stages that precede ALP expression but is ineffective on cells that progressed beyond this maturation stage. Likewise, IL-10 appeared to be unable to block both ALP activity and collagen synthesis in the preosteosteoblastic cell lines MN7 and MC3T3 that constitutively synthesize these proteins. Whereas IL-10 did not alter the number of fibroblast colony-forming cells of the marrow, it significantly reduced their osteogenic differentiation potential. In contrast to control cultures, IL-10-treated stroma was unable to either synthesize osteocalcin or to mineralize when subcultured over a 25-day period in the absence of IL-10. The inhibitory activity of IL-10 coincided with significant changes in stroma morphology. Whereas control cultures contained mainly flat adherent polygonal cells, significant numbers of rounded semiadherent to nonadherent cells were observed in the presence of IL-10. Scanning and transmission electron microscopy showed that, in contrast to control cultures, IL-10-treated stromas completely lacked a mineralized extracellular matrix. Collectively, these data suggest that IL-10 may have important regulatory effects on bone biology because of its capacity to downregulate early steps of osteogenic differentiation.

Alkaline Phosphatase↗
PubMed2001

Avascular necrosis in the femoral head secondary to bone marrow infarction in a patient with graft-versus-host disease after unrelated bone marrow transplantation.

We previously reported a case of bone marrow infarction attributable to acute graft-versus-host disease (GVHD) in a patient with acute lymphoblastic leukemia after unrelated bone marrow transplantation (BMT). Although the bone marrow infarction-induced arthralgia in this patient improved, severe arthralgia appeared again with exacerbation of chronic GVHD, and the arthralgia was strongly correlated with the clinical course of chronic GVHD, i.e., the course of symptoms such as dermal and hepatic GVHD and ocular dryness. Finally, the patient developed avascular necrosis (AVN) in the right femoral head. Serum interleukin (IL)-6 and IL-10 levels were high at the onset of arthralgia but low during remission, and levels of interferon-gamma were undetectable throughout the period of arthralgia. Based on the clinical course and these data, chronic GVHD was thought to have been the major cause of the AVN. Since IL-10 antagonizes various other cytokines that induce GVHD, the increase in IL-10 might have inhibited the development of GVHD.

Adult↗
PubMed1994

Bone marrow immunoscintigraphy versus conventional bone scintigraphy in the diagnosis of skeletal metastases in urogenital malignancies.

Bone marrow immunoscintigraphy using a 99mTc-labelled anti-NCA-95 monoclonal antibody and conventional bone scintigraphy with 99mTc-methylene-diphosphonate were compared in the diagnosis of skeletal metastases in 58 patients with urogenital tumours. In 13 patients with metastatic disease, bone marrow immunoscintigraphy proved to be superior to bone scintigraphy, detecting 431 metastatic lesions on immunoscans as compared to 261 lesions on bone scans. However, on a patient-by-patient basis bone marrow immunoscintigraphy did not demonstrate a clinical advantage over conventional bone scanning, as in this study no patient was identified with metastatic disease still confined to the marrow space, with bone scintigraphy and bone X-ray being normal.

Adult↗
PubMed1974

The Piromen Test as an assay of bone marrow granulocyte reserves in the calf. I. Studies on bone marrow and peripheral blood leukocytes.

The use of Piromen, a polysaccharide complex of Pseudomonas aeroginosa, has been investigated in 21 calves by a test of marrow granulocyte reserves. The maximal increase in peripheral granulocytes (DeltaG) was determined following multiple intravenous and sub-cutaneous injections of Piromen at various time intervals and was correlated with the mature marrow granulocytes on bone marrow smear differentials. Five ug Piromen per kg body weight, by the subcutaneous route, was found to give a mean DeltaG of 5200/mm(3), very similar to the DeltaG of 5300/mm(3) obtained in man with 0.1 ug/kg intravenous Piromen injections. Clinical effects in calves were minimal with the subcutaneous route as compared to the response following intravenous Piromen injections.

Animals↗
PubMed1989

Growth of xenografted human bone marrow: comparison with hemopoietic reconstitution in patients after allogeneic bone marrow transplant and response to granulocyte macrophage colony stimulating factor.

Normal human bone marrow was grown as xenografts in mice immune-suppressed by thymectomy and total body irradiation. Mononuclear cell fractions isolated from marrow harvests from 17 donors all gave rise to subcutaneous nodules which grew to a variable maximum size and then regressed. Human granulocyte/macrophage progenitors (CFU-GM) were recovered from xenografts up to 20 days postimplantation. Xenograft growth, measured by maximum nodule volume, area under the growth curve, and rate of regression, did not correlate with the speed of neutrophil or platelet recovery in bone marrow transplant patients infused with the same marrow. Assay of numbers of stromal fibroblastoid colony forming cells (CFU-F) in donor marrow was also not predictive of subsequent hemopoietic recovery in recipients. Treatment of host animals with daily intraperitoneal injections of 100 micrograms/kg human recombinant granulocyte/macrophage colony stimulating factor produced a more rapid growth of subcutaneous nodules. This technique may therefore be of use in determining the in vivo efficacy of human hemopoietic regulatory factors.

Animals↗
PubMed1990

[The humoral nature of the colony-stimulating action of bone marrow cells on stromal colony formation in bone marrow cultures].

In 24 hours adherent marrow cell cultures (AMCC) were represented by single stretched fibroblasts. In non-feeder-supplemented AMCC most of the CFU-f remained single fibroblasts or passed through 1-3 cell doublings [correction of dudlings]. The colony stimulating activity of irradiated marrow cells was found to be diffuse across the Millipore filter, which seems to indicate that haemopoietic marrow cells produce a colony stimulating factor which is required for triggering the CFU-f from the Go-period of the cell cycle into cell proliferation.

Animals↗
PubMed2005

Antileukemic effect of interleukin-7-transduced bone marrow stromal cells in mice following allogeneic T-cell-depleted bone marrow transplantation.

Impaired immune reconstitution following allogeneic bone marrow transplantation (BMT) remains a major obstacle to its clinical application. In this study, interleukin (IL)-7-transduced bone marrow stromal cells (MSC-IL7, 1 x 10(6)/mouse) were transfused into lethally irradiated C57BL/6 recipient mice. By day 40 after transplantation, the recipient mice were challenged with the lymphoma cell line EL4. MSC-IL7 co-transplantation protected recipient mice from leukemic mortality (MST >120 days after BMT vs mean survival time (MST) 70 days in the PBS group) It enhance the PFC count and DTH responses of recipients after transplantation. In conclusion, MSC mediated IL-7 gene therapy and may be a more feasible strategy to restore immune function following allo-TCD-BMT.

Animals↗
PubMed1985

Bone lining cells and hematopoiesis: an electron microscopic study of canine bone marrow.

Hematopoietic bone marrow in the dog is enclosed by a nearly complete and rather complex layer of endosteum, consisting of a diverse group of cells collectively called bone lining cells (BLC). Cell types comprising BLC include osteoblasts and osteoclasts, and other cell types, among which are elongated, flat cells with a spindle-shaped nucleus, and small cytoplasmic vesicles. The composition and thickness of the layer of BLC varies along the perimeter of the marrow. The layer may be simple or stratified. Occasionally a zone of tightly packed regularly arranged collagenous fibers lies between the bone lining cells and bone. Hematopoiesis, particularly neutrophilic, often occurs in the bone marrow next to the BLC. Cytoplasmic processes of BLC occasionally extend into the hematopoietic spaces and stromal cells in the hematopoietic compartment may extend processes to the layer of BLC. Occasionally cells of the BLC are similar in appearance to stromal cells within the marrow. Our observations together with the experimental findings of others (that fibroblastic stromal cells contribute to the hematopoietic inductive microenvironment, that hematopoietic stem cells are concentrated subosteally, that cells responsible for regeneration of the marrow stroma are derived from the endosteal layer, and that high concentrations of hematopoietic colony-stimulating factors are produced there) indicate that the hematopoietic capacities of bone marrow may be regulated by BLC.

Animals↗
PubMed2005

Renal involvement in bone marrow transplantation.

Bone marrow transplantation (BMT) is an effective therapeutic strategy for leukaemic malignancies and depressed bone marrow following cancer. However, its side effects on kidneys have been reported. Some drugs and irradiation are also suggested to be nephrotoxic. It is well known that haemolytic uraemic syndrome (HUS) after BMT develops as late-onset BMT nephropathy. Cyclosporine A (CsA) is a possible cause. Radiation nephropathy shows changes that are similar to the histology of HUS. These findings suggest that endothelial damage is closely associated with the pathogenesis of post-BMT nephropathy. Recently, some patients have developed glomerulonephritis accompanied by graft-versus-host disease (GVHD) after BMT. In these patients immune deposits are found mainly in subepithelium and mesangium equal to those of secondary membranous glomerulonephritis. A murine experimental model of GVHD manifests similar symptoms and histological changes to those of actual patients and may suggest the pathogenesis of glomerulonephritis.

Animals↗
PubMed1998

Accelerated recovery of irradiation-induced bone marrow depression by fibroblast-mediated interleukin 6 gene therapy in combination with bone marrow transplantation in mice.

BACKGROUND: Both fibroblast-mediated cytokine gene therapy and bone marrow transplantation (BMT) have proven to be efficient protocols for the recovery of bone marrow depression. In this report, the effects of fibroblast-mediated interleukin (IL)-6 gene therapy, in combination with BMT, on the recovery of irradiation-induced bone marrow depression were investigated. METHODS: NIH3T3 fibroblast cells engineered to secrete IL-6 (NIH3T3-IL-6) or NIH3T3 cells transduced with the neomycin gene (NIH3T3-Neo), in combination with 10(7), 10(6), or 10(5) syngeneic bone marrow cells, were implanted into irradiated mice. RESULTS: The platelets and white blood cells in the peripheral blood of the irradiated mice increased greatly 12 days after implantation of NIH3T3-IL-6 cells and BMT, the white blood cell counts were restored to a normal level 32 days after the combined therapy, and the platelet number was obviously higher than that in mice implanted with NIH3T3-Neo and BMT. Twenty and 25 days after the combined therapy, the mice showed accelerated recovery of colony-forming unit (CFU)-granulocyte/macrophages and CFU-megakaryocytes when compared with the mice implanted with NIH3T3-Neo cells and BMT. Ten days after lethal irradiation with gamma rays, the spleens formed more CFU-spleen in mice implanted with NIH3T3-IL-6 cells and BMT than in mice injected with phosphate-buffered saline or NIH3T3-Neo cells. Combined therapy with NIH3T3-IL-6 cell implantation and BMT delayed the survival period of the hematopoietic-depressed mice significantly when compared with therapy with NIH3T3-Neo cell implantation and BMT. CONCLUSIONS: These data demonstrated that the combined therapy of fibroblast-mediated IL-6 gene therapy and BMT could significantly promote the recovery of irradiation-induced hematopoietic depression.

3T3 Cells↗
PubMed1978

[Regulatory role of bone marrow in immunogenesis. III. Humoral factors stimulating and suppressing antibody formation produced by bone marrow cells in in vitro cultures].

As revealed by the method of cultivation of bone marrow and spleen cells, separated by nucleopore membrane, in two-chamber bottles, the bone marrow cells were capable of producing humoral factor stimulating antibody genesis by the spleen cells. A direct contact of the bone marrow cells with the actively proliferating antigen-stimulated cells of the spleen led to production of a spleen humoral factor suppressing the antibody genesis by the spleen cells. The suppressive action of the bone marrow cells on the antibody genesis in the culture of the spleen cells was mediated through the suppression of the spleen cells proliferation; proliferation of the bone marrow cells is enhanced.

Adjuvants, Immunologic↗
PubMed1988

Serum IgA and the production of IgA by peripheral blood and bone marrow lymphocytes in patients with primary IgA nephropathy: evidence for the bone marrow as the source of mesangial IgA.

The origin of the mesangial IgA1 in primary IgA nephropathy (IgAN) is unknown. The bone marrow, the prime production site of plasma IgA in healthy humans, has not been previously investigated in patients with IgAN. In patients with IgAN, we found an increased percentage of IgA plasma cells containing IgA1 in the bone marrow (89.7% +/- 2.6% v 84.3% +/- 6.6%, P = 0.01), an increased percentage of serum IgA as IgA1 (92.2% +/- 4.9% v 80.2% +/- 6.6%, P less than 0.001), and an increased percentage of IgA1 of the IgA produced by peripheral blood lymphocytes in culture (75% +/- 16% v 54% +/- 19%, P less than 0.01). These findings are compatible with the bone marrow as production site of the mesangial IgA1. The data on IgA1 polymers are more difficult to interpret because their role in the pathogenesis of IgAN is still controversial. We found an increased concentration of both polymeric and monomeric IgA1 in the sera and supernatants of cultures of bone marrow of patients, although the ratio of polymeric to total IgA1 remained normal. In our opinion, the mucosa of the digestive tract is an unlikely source of the mesangial IgA because the absence of IgA2 in the deposits contrasts to the high percentage of IgA2 plasma cells in the intestinal mucosae. Moreover, in normal individuals, the digestive mucosae contribute very little to plasma IgA. Although the respiratory tract contains a higher percentage of IgA1 plasma cells, the 25% fraction of IgA plasma cells containing IgA2 is still very substantial. This percentage argues against the respiratory mucosae as a source of the mesangial IgA.(ABSTRACT TRUNCATED AT 250 WORDS)

Bone Marrow↗
PubMed1981

Histological study of the bone marrow in chronic granulocytic leukaemia in blast transformation. II. Bone marrow fibre content before and after autografting.

A serial study of the extent of bone marrow (BM) fibrosis was carried out in 14 patients with chronic granulocytic leukaemia (CGL) in blast transformation (BT) who received intensive treatment followed by autografts to restore marrow haemopoiesis. Some degree of marrow fibrosis (MF) was identified in 13 of the 14 patients when blast transformation was diagnosed. A transient increase in the amount of marrow fibrosis was observed in most cases immediately following intensive chemoradiotherapy or chemotherapy alone but this had regressed by 4-6 weeks when chronic phase haemopoiesis was established in the marrow. The presence of marrow fibrosis when blast transformation was diagnosed did not appear to impair the effectiveness of the autograft.

Antineoplastic Agents↗
PubMed1986

Suppressor cells detected in murine bone marrow chimeras.

Bone marrow cells from BALB/c mice were transplanted into lethally X-irradiated C3H/He mice, either intravenously (i.v.) or intrasplenically (i.s.). Chimeras created by intrasplenic transplantation (designated i.s. chimeras) showed a higher survival rate than chimeras created by intravenous transplantation (designated i.v. chimeras). During the course of the study on the mechanisms which caused such a difference, suppressor cells were detected in the spleen of i.s. chimeras by means of MLR co-culture assay and CTL assay. Although these cells were also detected in i.v. chimeras, time course study of the suppressor activity against donor anti-host MLR in i.v. and i.s. chimeras showed some difference. Namely, the suppressor activity in the i.s. chimeras was more constant than that in the i.v. chimeras. This difference may induce a beneficial effect of i.s. bone marrow transplantation.

Animals↗