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PubMed1990

The use of recombinant cytokines for enhancing immunohematopoietic reconstitution following bone marrow transplantation. II. The influence of lymphokines on CFU-GM colonies from human untreated, ASTA-Z or Campath-1M treated bone marrow.

Patients undergoing bone marrow transplantation (BMT) are subjected to the risk of pancytopenia in the immediate post-BMT period. Recipients of bone marrow (BM) autografts purged in vitro by chemical agents such as mafosfamide (ASTA-Z) are even more likely to develop a delayed engraftment. In a previous study in mice, we showed earlier immunohematopoietic reconstitution after syngeneic marrow grafting with BM cells precultured with single or combined cytokines. In this work we determined optimal culture conditions for the use of single and various cytokine combinations in order to activate progenitor cells following in vitro cultures of untreated, ASTA-Z purged or T cell-depleted human BM preparations prior to BMT. The best single cytokine for enhancing CFU-GM was found to be GM-CSF (0.1 mg/ml) which produced an increase of up to 8-fold over controls after 3 days' incubation. Addition of recombinant human interleukin 3 (rhIL3) (0.1 mg/ml) to rhGM-CSF had an additive effect. The same enhancing effect of in vitro CFU-GM by both cytokines was observed following ASTA-Z purging of BM cells obtained from patients undergoing autologous BMT. Similarly, depletion of lymphocytes from BM using the monoclonal rat anti-human lymphocyte antibody Campath-1M and human complement did not diminish the beneficial influence of rhIL3 and rhGM-CSF. When combined with rhGM-CSF and rhIL3, rhIL1 and rhIL2 had no appreciable enhancing effect on CFU-GM and occasionally even reduced it. We suggest that enhancement of differentiation of hematopoietic progenitor cells may be accomplished by in vitro incubation of BM cells without continuous administration of cytokines in vivo following BMT.

Antineoplastic Agents↗
PubMedDate not supplied

[Bone marrow transplantation in aplastic anemia. Experience at a Mexican institution. Bone marrow transplantation group of the Salvador Zubirán National Institute of Nutrition].

During the period of May 1986 through February 1991, nine allogeneic bone marrow transplants (BMT) on eight severe aplastic anemia (SAA) patients were performed at the Instituto Nacional de la Nutrición Salvador Zubirán in Mexico City. Mean age at BMT was 18 years (age interval 12-30); seven were men; all patients had a clinical history of multiple blood transfusions; six individuals were infected at the time of the transplant. The conditioning regimens were: cyclophosphamide (Cy) in three patients; Cy+ total nodal radiation in five; and total nodal radiation only in the second transplant of one patient. Graft versus host disease (GVHD) prophylaxis was attempted with methotrexate plus cyclosporin A (CsA) in six patients, methylprednisolone plus CsA in two, and prednisone + CsA in the patient retransplanted. All procedures were carried out under single reverse isolation without gut decontamination. Seven of the nine procedures grafted (two cases died on days +8 and +25 due to infection). In the surviving, the median time for reaching > 1.0 white blood cells x 10(9)/L was 22 days (time interval 11-31); > 0.5 neutrophils x 10(9)/L in 27 days (time interval 15-42) and the same lapse to reach > 50 platelets x 10(9)/L. Length of hospital stay was 42 days (time interval 15-61). Acute GVHD was seen in one of the seven patients surviving the period of bone marrow aplasia (14%). Of six long term survivors (including one patient with a second transplant) chronic GVHD was present in four (67%): chronic GVHD was fatal in one individual but was well controlled in three.(ABSTRACT TRUNCATED AT 250 WORDS)

Actuarial Analysis↗
PubMed2003

Increase of bone marrow lymphocytes in systemic mastocytosis: reactive lymphocytosis or malignant lymphoma? Immunohistochemical and molecular findings on routinely processed bone marrow biopsy specimens.

AIMS: To clarify the nature (reactive or neoplastic) of lesional, perifocally aggregated lymphocytes in bone marrow infiltrates of systemic mastocytosis (SM), the histopathology of which can resemble malignant lymphoma with focal bone marrow involvement, particularly low grade malignant B cell lymphoma of lymphoplasmacytic immunocytoma subtype, which frequently exhibits increased mast cell (MC) numbers. METHODS: Thirteen cases of SM and three of lymphoplasmacytic immunocytoma with predominant focal bone marrow infiltration were investigated. Immunostaining of formalin fixed, paraffin wax embedded bone marrow specimens was performed using antibodies against CD2, CD5, CD20, CD23, and CD25; kappa and lambda immunoglobulin light chains; and MC markers chymase, tryptase, and CD117 (KIT). Monoclonal rearrangements of IgH and TCRgamma were studied using seminested polymerase chain reaction (PCR). c-kit point mutation Asp816-Val was detected by PNA mediated PCR clamping and hybridisation probes. RESULTS: The lymphocytic clusters in SM contained nearly equal numbers of mature T and B cells, the latter with no coexpression of aberrant antigens, such as CD5 or CD23. Most MCs in SM cases constantly coexpressed tryptase, CD25, and CD117. No monoclonal rearrangements were seen for IgH or TCRgamma. In contrast, B cells from immunocytomas showed light chain restriction and monoclonal rearrangement for IgH, confirming their neoplastic nature. c-kit point mutation Asp816-Val was found in ten of 13 SM cases, but in none of the three immunocytomas. CONCLUSIONS: Focal accumulations of lymphocytes in the bone marrow of SM are reactive in nature and could be termed lymphocytosis. A diagnosis of SM-AHNMD/immunocytoma should not be made.

B-Lymphocytes↗
PubMed1994

Effects of interleukin-4 on granulocyte-macrophage-colony formation from murine bone marrow cells and hematopoietic reconstitution following murine allogeneic bone marrow transplantation.

We investigated the effects of mouse recombinant IL-4 on hematopoiesis in vitro and in vivo. IL-4 alone was found to be incapable of stimulating colony formation, but it inhibited both IL-3 and GM-CSF-induced colony formation by murine hematopoietic progenitor cells. In contrast, colony formation induced by G-CSF was enhanced in the presence of IL-4. We also studied the influence of IL-4 on hematopoietic reconstitution after allogeneic bone marrow transplantation in a murine model, and found that IL-4 had significant inhibitory effects on neutrophil recovery and that neutrophil recovery accelerated by IL-3 and G-CSF was significantly suppressed by IL-4. The combination of IL-4 and GM-CSF caused a significant decrease in the absolute number of neutrophils.

Animals↗
PubMed1994

Understanding the mechanism of cure of acute myeloid leukemia by allogeneic bone marrow transplantation: toward the application of interleukin-2 in autologous bone marrow transplantation.

Analysis of the results of different methods of post remission therapy in acute myeloid leukemia (AML) has been used to study their impact on outcome, with a particular emphasis on antileukemic effects by contrasting the risk of relapse. We have compared consolidation chemotherapy (CT), autologous bone marrow transplantation (ABMT) with and without subsequent interleukin-2 (IL-2) and allogeneic bone marrow transplantation (BMT). Additionally we have studied the immunological consequences of each of these maneuvers with particular regard to the cellular components having proven or suspected antileukemic properties and the secondary cytokines released by these cells in vitro and in vivo.

Acute Disease↗
PubMed1991

Quantitative determination of non-haem iron and ferritin iron in bone marrow using flameless atomic absorption spectrophotometry. A comparative study on the cytological and chemical determination of the bone marrow iron content.

A method for non-haem iron analysis in bone marrow aspirates using graphite furnace atomic absorption spectrophotometry has been developed. Bone marrow aspirates were obtained from patients with various disorders. A good correlation is observed between chemical and cytological assessment of total non-haem iron in bone marrow. An intra-assay coefficient of variation of 9.0% was observed. The ferritin-iron concentration was also determined and a CVduplo of 11% was found. The ferritin iron concentration increased with an increasing total iron content until saturation of ferritin appeared to be reached at about 3 g ferritin per kg protein. It was concluded that the quantitative determination of bone marrow iron can be of value in the diagnosis and investigation of both hypo- and hyper-ferraemic disorders.

Bone Marrow↗
PubMed1986

Enrichment for GM-CFU from human bone marrow using Sambucus nigra agglutinin: potential application to bone marrow transplantation.

A new lectin, purified from black elder-berries, Sambucus nigra L. (SNA I, hereafter called SNA), was used for fractionation of normal human marrow cells. The stem cell enrichment capability of SNA was investigated by comparing colony formation (GM-CFU) in unagglutinated cell fractions (SNA-) following agglutination with SNA with that with soybean agglutinin (SBA-). GM-CFU recovery in SNA- was equal or superior to that in the SBA- fraction. A modified procedure was developed to combine stem cell enrichment and depletion of E-rosette-forming T-lymphocytes. Bone marrow cells were exposed in one step to SNA and to untreated sheep red blood cells (SRBC). Nonagglutinated, nonrosetting cells were collected after Ficoll-Hypaque-gradient separation. The procedure, a modification of Reisner's multistep procedure involving the agglutination of SBA+ cells; separation of the unagglutinated SBA- cells from the top of a 5% bovine-serum-albumin gradient; the formation of E-rosettes with SRBC; and the separation of rosettes over a Ficoll-Hypaque gradient, is faster, simpler, and as effective for T-cell depletion and stem cell enrichment.

Bone Marrow Transplantation↗
PubMed2004

Cytokines transduced bone marrow stromal cell lines promote immunohematopoietic reconstitution in mice after allogeneic bone marrow transplantation.

Impaired immune reconstitution following allogeneic T-cell depleted bone marrow transplantation (allo-TCD-BMT) is a major obstacle to its clinical application. Stromal cell line QXMSC1, established from bone marrow cells of BALB/c(H-2d), was transfected with murine IL-3 and/ or IL-2 gene, and injected into lethally irradiated C57BL/6(H2b) mice. We evaluated its effects on immunologic and hematopoietic reconstitution after allo-TCD-BMT. The results showed that QXMSC1-IL-3 + IL-2 could significantly increase the numbers of hematopoietic primitive progenitors (CFU-S), committed progenitors (CFU-GM, and BFU-E), and lymphocytes (CD8+ cells, CD4+ cells, and B cells). Similarly, immune functions of recipient mice were significantly enhanced in the QXMSC1-IL-3 + IL-2 group. In addition, QXMSC1-IL-3 or QXMSC1-IL-2 also exerted apparent effects on accelerating immune reconstitution, but these effects were far less than that of QXMSC1-IL-3 + IL-2. Our results demonstrated that stromal cell-mediated IL-3 and IL-2 gene therapy may be a potent approach in promoting immunologic and hematopoietic reconstitution after allo-TCD-BMT.

Animals↗
PubMed1988

Allogeneic bone marrow transplantation in chronic lymphocytic leukemia: report from the European Cooperative Group for bone marrow transplantation (8 cases).

Allogenic bone marrow transplantation (BMT) was performed in 8 patients with B chronic lymphocytic leukemia (CLL): 6 males and 2 females with a median age of 41 years (37 to 46). Seven patients were resistant to previous therapy and were grafted in the evolutive phase. Only 1 patient was grafted without any previous treatment. At the time of BMT 5 patients were classified according to the Rai classification in stage IV, 1 in stage III and 2 in stage 1. The delay between diagnosis and BMT was 47 months (5 to 68). Graft versus host disease (GVHD) prophylaxis was methotrexate (1 patient), cyclosporine (2 patients), methotrexate + cyclosporine (3 patients), cyclosporine + physical removal of T cells (2 patients). The conditioning regimen was standard for 5 patients, reinforced for 3 patients with Chlorambucil (1 patient) and etoposide (2 patients). A successful engraftment was obtained in all cases. The lymphocytosis decreased slowly and disappeared from day 0 to day 28. All patients developed an acute GVHD 5 of which resolved. Three patients died: 2 of acute GVHD and 1 of intracerebral hemorrhage. Five patients are alive in persistent complete remission without clinical symptoms and with normal peripheral blood and normal bone marrow. The median follow-up is 27 months (13-53). This small series suggests that allogeneic BMT should be investigated in selected patients as possible curative treatment of CLL.

Acute Disease↗
PubMed1977

Haemopoietic recovery in spleen and marrow after transplantation of bone marrow from either normal or hydroxyurea treated mice.

Haemopoietic regeneration was studied following x-irradiation and transplantation of bone marrow from either normal or hydroxyurea-treated donor mice, to ascertain the contribution of proliferating progenitor cells to regeneration. With transplantation of equivalent numbers of CFU-S, total DNA and 3HTdR uptake into DNA in spleen and femoral bone marrow and the erythroid, granulocytic and mononuclear cell populations were not significantly different between normal (NBM) and hydroxyurea-treated (HUBM) marrow. The response of hypertransfused x-irradiated mice to erythropoietin (EPO) administration was also not significantly different in spleens of mice receiving normal or hydroxyurea-treated marrow.

Animals↗
PubMed1992

Myelodysplastic syndrome associated with bone marrow fibrosis.

Bone marrow biopsy (BMB) in myelodysplastic syndrome (MDS) frequently reveals a slight alteration in the reticulin stroma which does not have any clinical significance. However, in a minority of cases, full-blown bone marrow fibrosis (BMF) can be found. Primary MDS patients with BMF show distinct clinico-pathological features and an unfavourable prognosis mainly attributable to complications deriving from pancytopenia and continuous transfusions, while leukemic transformation occurs only rarely. Since BMF may characterize other hematological disorders, primary MDS with BMF should be included in the differential diagnosis particularly with malignant myelofibrosis (MM) and idiopathic myelofibrosis (IMF). Secondary MDS with BMF represent a variety of preleukemic conditions in subjects treated for previous neoplasias. Unlike the primary forms, they do not form a clearcut clinico-pathological entity.

Humans↗
PubMed1991

[Acute unclassified leukemia with bone marrow necrosis].

Massive bone marrow necrosis was seen in a 42-year-old male with acute leukemia. In December, 1988, on admission, laboratory data revealed pancytopenia and a high level of serum LDH and ALKP. Bone marrow aspiration resulted in dry-tap and showed bone marrow necrosis in the bone marrow biopsy specimen. A bone marrow scintigraphy with 111In faintly visualized the bone marrow but visualized area was expanded in the extremities compared with normal subjects. The second bone marrow biopsy showed proliferation of blasts. In the middle of March, blasts began to appear in peripheral blood. The blasts were cytochemically negative for POX, Es, PAS, AcP, TdT and had surface markers CD3-, CD19-, CD33-, CD13-, LCA-, HLA-DR-. Even by investigation on rearrangement of the immunoglobulin heavy chain region, an origin of the blasts could not be determined. In April, the number of blasts in peripheral blood increased and hepatosplenomegaly developed rapidly. Therefore, he was put on the chemotherapy with vincristine and prednisolone, but he died of cerebral hemorrhage. The autopsy revealed widespread bone marrow necrosis. It has rarely been reported that massive bone marrow necrosis is found prior to the occurrence of acute unclassified leukemia.

Acute Disease↗
PubMed1993

Autologous bone marrow transplantation with alkyl-lysophospholipid-purged marrow.

Autologous bone marrow transplantation (ABMT) appears to offer clinical benefit to leukemia patients, but the major reason for failure is relapse. This may be related to the presence of residual leukemic cells in the harvested marrow. To circumvent this problem, various procedures have been developed to purge ex vivo residual leukemic cells from the marrow. The alkyl-lysophospholipids are a new group of anticancer drugs that target membranes as their major site of action. They are unique in that they are relatively selectively toxic to neoplastic cells and spare normal marrow stem cells and progenitor cells. The most active compound is edelfosine. Twenty-nine patients with acute leukemia in second or subsequent remission or early relapse or in first remission, either with a history of treated extramedullary relapse, or requiring more than one induction program to achieve remission, underwent ablative therapy followed by infusion of autologous marrow which had been purged by a 4-hour exposure to edelfosine prior to cryopreservation. Thirty-one percent of the patients are alive and free of leukemia for a median of 630 days (range 185-1,613). These results in this high-risk group of patients warrant further investigation.

Acute Disease↗
PubMed2003

Contamination of bone marrow products with an actinomycete resembling Microbacterium species and reinfusion into autologous stem cell and bone marrow transplant recipients.

Bacterial contamination of bone marrow or peripheral blood stem cell transplant products typically occurs with skin flora or, rarely, gram-negative organisms. We describe a clonal outbreak of contamination in transplant products caused by contamination with an aerobic actinomycete that occurred at our institution during the summer of 2001. From 1 July through 12 September 2001, 73 peripheral blood or bone marrow stem cell products were obtained from 39 patients, and 34 products were found to be contaminated with the outbreak strain. Fourteen patients were reinfused with contaminated cells, and the outbreak strain was isolated from the blood cultures for one patient. Investigation revealed multiple potential sources for contamination during the product cryopreservation process. The outbreak of contamination was aborted upon modification of the cryopreservation process.

Actinobacteria↗
PubMedDate not supplied

Autologous bone marrow transplantation in non-Hodgkin's lymphomas: prediction of mononuclear cell yield in bone marrow harvests.

We retrospectively analyzed the factors influencing the mononuclear cell (MNC) yield on bone marrow (BM) harvests in a cohort of 15 non-Hodgkin's lymphoma patients undergoing autologous bone marrow transplantation. All the patients were treated with the F-MACHOP regimen and four of them also received radiotherapy on bulky disease. Before harvesting, the patients underwent complete peripheral blood (PB) count, BM biopsy and aspirate. WBC and MNC/microL were determined on the pre-harvest PB and BM aspirate samples using an automated counter. The following variables were examined in univariate and multivariate regression analysis for possible influence on the MNC yield in the harvests: age, sex, number of cycles of CT, previous radiotherapy, state of the disease at the time of harvest, interval between the end of therapy and BM harvest, cellularity of the BM biopsy, absolute WBC and MNC counts in the PB before harvesting, absolute WBC and MNC counts in the BM aspirate performed before harvesting. The amount of marrow harvested was constant for all patients: 21.2 +/- 0.24 microL/kg B.W. Among the factors analyzed, two correlated strongly with the MNC yield/kg B.W. in the harvests: the MNC count in pre-harvest PB (r = 0.823; p = 0.0001) and the MNC count in pre-harvest BM aspirate (r = 0.802; p = 0.0003). A regression equation was generated that allows calculation of the MNC/kg yield prior to harvesting.

Antineoplastic Combined Chemotherapy Protocols↗