Cytochemical characteristics of leukopoietic differentiation in murine erythroleukemic (Friend) cells.

The 3BM-78 murine Friend erythroleukemia cell line was obtained by in vitro transformation of bone marrow cells of DBA/2J mice by the polycythemic Friend virus complex. Thirty-five subclones have been isolated and tested for their ability to express various markers of blood and bone marrow cells. Upon dimethyl sulfoxide treatment, the cells differentiated along the erythroid pathway as shown by morphological evidence and by their increased synthesis of hemoglobin and spectrin. In addition, a high proportion of dimethyl sulfoxide-induced cells stained positive for specific esterase, a marker characteristic of granulocytic cells. Of these cells, about 20% stained positive for both hemoglobin-peroxidase and specific esterase. Analysis of the subclones showed that the expression of these markers for erythroid and leukopoietic differentiation was uncoordinated. Further dissection of expression was obtained by the use of two potent tumor promoters, 12-O-tetradecanoylphorbol-13-acetate and phorbol-12,13-benzoate, which in general inhibited specific esterase more than hemoglobin peroxidase expression. Inhibition was related to the structure of the phorbol diester and was unrelated to toxicity. No evidence was found for other markers characteristic of different pathways of differentiation, such as Fc and C3 receptors, cell surface immunoglobulins, theta antigen, or the capacity to phagocytose inert particles. All cells stained positive for nonspecific esterase activity in both the presence and the absence of dimethyl sulfoxide. This staining was only partially fluoride sensitive. In unstimulated cultures, a few cells also reacted for myeloperoxidase, Sudan black staining and, very rarely, alkaline phosphatase staining. These findings support the view that 3BM-78 cells are leukemic cells which, despite a prevalent commitment to erythroid differentiation, retain the genetic determinants for some traits of leukopoietic differentiation. These traits may be expressed under suitable culture conditions.

Meningitis due to penicillin-resistant Streptococcus pneumoniae in patients with chronic graft-versus-host disease.

Two episodes of meningitis due to penicillin-resistant Streptococcus pneumoniae occurring in two patients with chronic graft-versus-host disease (GVHD) are reported. Both patients were treated with ceftazidime. The first patient died, unresponsive to therapy. The second patient showed clinical improvement, reverting to her baseline mental status. This report draws attention to the fact that in chronic GVHD patients: (1) bacterial prophylaxis does not ensure protection against encapsulated bacteria; (2) rapid microbiological investigation is recommended with any upper respiratory tract infections.

Amebiasis after bone marrow transplantation.

A patient undergoing BMT for acute non-lymphocytic leukemia (ANLL) developed bloody diarrhea due to amebiasis. The infection was successfully treated with intensive and prolonged antiparasitic therapy.

Growth and endocrine function following bone marrow transplantation for thalassemia.

Twenty two patients with thalassemia major who received successful bone marrow transplantation (BMT) were followed to verify the impact of the transplant procedure on subsequent growth and development. The transplant preparative regimen consisted of busulphan and cyclophosphamide. Growth and endocrinological function were assessed during the first 4 years following BMT. At the time of transplant most patients showed growth retardation. The median difference between chronological age and bone age was -9.5 months for the boys and -8.5 months for the girls. Patients > 7 years old at the time of BMT showed a significant worsening of their growth delay at 48 months following BMT compared with 12 months before transplantation. Patients < 7 years at the time of BMT had their growth retardation constant over time span after transplantation. Moreover six of 11 younger patients showed an improvement of their growth delay compared with one of 11 older patients. The outcome of height standard deviation score at 24 and 48 months following BMT was strictly correlated with the level of serum transaminases and ferritin. Sixteen patients had impaired growth hormone secretion after a provocative test evaluated at 24 months after transplant. At 48 months there was no significant increase in the mean peak GH levels. This study confirms that the growth retardation of patients with thalassemia major is multifactorial.

Reconstitution of normal neutrophil function in chronic granulomatous disease by bone marrow transplantation.

Allogeneic bone marrow transplantation was carried out on an 11-year-old boy with chronic granulomatous disease and severe chronic pulmonary insufficiency of restrictive type. After preparative regimen with busulfan (13 mg/kg) and cyclophosphamide (200 mg/kg), the patient received marrow cells from his HLA-identical and MLC-nonreactive sister. Durable sustained engraftment of donor hematopoietic and lymphoid populations occurred, as documented by analysis of genetic markers and complete reversal of the neutrophil function defect. No episode of infection occurred in the post-transplant course and, currently, 40 months after transplantation the patient is in excellent health and growing normally and showing an increasing improvement of his respiratory capacity. The successful outcome in this patient demonstrates that marrow transplantation is at present the only curative approach for this congenital disorder of neutrophil function.

Cytogenetic survey of 80 patients with acute nonlymphocytic leukemia.

The authors report a cytogenetic survey of 80 patients with acute nonlymphocytic leukemia. The prognostic value of chromosome aberrations has been evaluated with three methods. The first one showed that patients with NN or AN bone marrow cellularity have a significantly better prognosis than those with AA cellularity; the second method confirmed the relatively good prognosis for patients with t(8;21) and abnormal 16 and a poor one for those with rearrangements of chromosomes 5 and/or 7. The authors also noted, surprisingly, that patients with hyperdiploidy had a significantly poorer prognosis than those with hypodiploidy and especially pseudodiploidy. The third method showed that patients with very complex karyotypes and a worse outcome than those with simple changes. Finally, they discuss the prognostic value of unusual and/or undeciphered chromosome changes detected in 18 patients, with a mean survival of 9.6 months, showing that these changes have a negative prognostic significance.

Leukemic evolution in three patients with myelodysplastic syndrome and unusual chromosome changes.

The authors describe three patients with myelodysplastic syndrome without a history of exposure to chemical agents and who showed many chromosome rearrangements not previously reported in this hematologic disorder, and a rapid outcome of the disease. The authors discuss the significance of the chromosome changes, suggesting, in agreement with others, that patients with complex rearrangements have a poor prognosis.

Cytogenetics and bone marrow transplantation.

The authors report hematologic and cytogenetic data on 19 patients treated with allogeneic bone marrow transplantation (BMT) for severe hematologic disorders: 8 patients with chronic myelogenous leukemia, 6 with acute leukemia, 3 with severe aplastic anemia, 1 with refractory anemia, and 1 with beta-thalassemia major. Cytogenetic assays were performed on marrow cells before conditioning, 30 days after BMT, and at subsequent times. The authors discuss the role of cytogenetic studies in the evaluation of bone marrow engraftment, leukemic transformation of the graft, and disease relapse.

Detection of minimal residual disease by polymerase chain reaction in patients with different hematologic diseases treated by bone marrow transplantation.

Thirteen male patients affected by different hematologic diseases who underwent bone marrow transplantation (BMT) with female donors were investigated by cytogenetic analysis and polymerase chain reaction (PCR) amplification of a DNA sequence specific for the Y chromosome. In six of these patients, PCR showed the presence of the Y chromosome-related sequence; in only three of these did cytogenetic analysis confirm the presence of mixed chimerism. In the remaining three patients, the results of the PCR were confirmed by in situ hybridization on cell nuclei with a probe for the alpha-satellite of the Y chromosome. We compare results obtained with the two methods and discuss the meaning of the minimal residual disease detected by PCR in patients submitted to BMT.

Early hematopoietic reconstitution after autologous transplantation with blood-derived stem cells in a patient with advanced lymphoma.

A 30-year-old man with advanced non-Hodgkin lymphoma underwent repeated leukaphereses for harvesting blood-derived hemopoietic stem cells. Collection was started 8-10 days after the end of L-VAMP therapy (3 cycles). Nine procedures were performed and a total of 65.4 x 10(9) mononuclear cells (0.87 x 10(9)/Kg) were collected, processed, cryopreserved and stored in liquid nitrogen. The yields of CFU-GM, BFU-E and CFU-GEMM were respectively 964 x 10(4) (12.4 x 10(4)/Kg), 249 x 10(4) (3.2 x 10(4)/Kg) and 798 x 10(4) (10.4 x 10(4]. The patient received a myeloblative regimen consisting of fractionated total body irradiation (1200 cGy) and cyclophosphamide (120 mg/kg) followed by infusion of his own thawed cells. Early trilineage hematopoietic recovery was first observed on day +8; 1 x 10(9)/l WBC were reached on day +11, 0.5 x 10(9)/l PMN on day +13 and 50 x 10(9)/l platelets on day +11. Course was uneventful and the patient was discharged from hospital on day +21. Eight months after transplant the patient is in continuous unmaintained complete remission with normal blood cell counts. This reports suggests that complete and sustained engraftment can be achieved with peripheral stem cells recruited after "soft" chemotherapy.

[Efficacy of estrogen-progestin replacement therapy after bone marrow transplantation]. / Efficacia del trattamento sostitutivo con estro-progestinici in pazienti sottoposte a trapianto di midollo osseo.

Ovarian failure is a common consequence of chemotherapy and radiotherapy in women undergoing bone marrow transplantation. The longer survival in these women has raised, during the past years, the need for a better quality of life. The objective of the present study has been to evaluate perspectively the potential benefit of hormonal replacement therapy in 24 women who underwent bone marrow transplantation. The data obtained indicated that hormonal replacement therapy results effective in preventing and/or relieving the multiple manifestations of gonadal failure, including amenorrhea, hot flashes, atrophy of genital apparatus, osteoporosis and cardiovascular disease.

Hereditary haemolytic ovalocytosis with defective erythropoiesis.

Patients belonging to four families with 'atypical elliptocytosis' have been investigated. Clinical, haematological, erythrokinetic and enzymatic characteristics as well as the effect of splenectomy are discussed. These studies appear to define the fundamental features of a particular disorder or a variety of hereditary elliptocytosis; characterized by a genetic autosomal dominant character, moderate degree of RBC eccentricity, erythroid dysplasia with relative marrow failure and incomplete response to splenectomy.