Vesna Najfeld

A pilot study of allogeneic bone marrow transplantation using related donors stimulated with G-CSF.

Growth factor administration to donors prior to bone marrow (BM) harvesting results in an enrichment of the graft for myeloid precursors. In animals, growth factor-primed BM has a higher repopulating ability than untreated BM. Ten patients received an HLA-identical sibling, allogeneic transplant using granulocyte colony-stimulating factor (G-CSF)-stimulated BM. Stimulation consisted of G-CSF at 10u2009μg/kg/day for 2 days prior to harvest. Patients were transplanted for various benign and malignant hematological conditions. The GVHD prophylaxis consisted of cyclosporine, methotrexate and/or prednisone. Compared to untreated historical control BM, stimulated BM infusions contained similar number of nucleated cells (meanu2009±u2009s.d.: 3.5u2009±u20091.5 vs 4.0u2009±u20090.9u2009×u2009108/kg), CD34+ cells (meanu2009±u2009s.d.: 7.5u2009±u20093.0 vs 9.4u2009±u20096.7u2009×u2009106/kg), and CD3+ cells (meanu2009±u2009s.d.: 129u2009±u200930 vs 190u2009±u200959u2009×u2009106/kg) but higher numbers of granulocyte–macrophage colony-forming units (meanu2009±u2009s.d.: 20u2009±u200912 vs 96u2009±u200934u2009×u2009104/kg). Patients receiving stimulated BM had prompt and stable engraftment of white cells and platelets. On average they attained an ANC of ⩾1u2009×109/l 9 days earlier and a platelet count of ⩾20u2009×u2009109/l 6 days earlier than historical controls receiving unstimulated HLA-identical sibling BM. Hospitalization was shortened by a mean of 10 days and transfusion requirements were modest. None of the patients developed severe GVHD or disease relapse. Two patients died of severe VOD post-BMT and thus were unevaluable for platelet engraftment. A third patient died of TTP on day 76 post-BMT. Seven patients are alive and well 49–585 days post-BMT. Stimulated BM may provide a valuable alternative to allogeneic BM and PBSC transplants. Ideal stimulation regimens need to be investigated.

Interphase FISH analysis of sex-mismatched BMT utilizing dual color XY probes

Interphase FISH analysis, utilizing dual color XY probes, was performed on 27 patients following allogeneic sex-mismatched bone marrow transplantation and on 31 controls. Of the 123u2009167 examined interphase nuclei, 63u2009318 were from 19 of the 21 patients (54 specimens) who engrafted, 31u2009827 from five of the six patients (29 specimens) who relapsed (four) or failed to engraft (one) and 24u2009703 from the 31 control specimens. In patients who engrafted, the mean percentage of host cells was 0.26% between day 29 and 5 years following BMT. Microchimerism of 0.7% or less than 1–5 years following BMT was not predictive of relapse. Interphase FISH analysis predicted relapse or failure of engraftment in five of the six evaluable patients. In three of five patients both conventional cytogenetics and interphase FISH of bone marrow cells provided important information regarding engraftment status and degree of chimerism.

Identification of subpopulations of human macrophages through the generation of human macrophage hybridomas

We have generated a series of human macrophage hybridomas by fusing an HGPRT-deficient promonocytic cell line, U937, with macrophages obtained by allowing monocytes to mature into macrophages in teflon bags. The fusions were documented as true hybrids by the acquisition of donor class I molecules, as well as donor derived macrophage surface antigens. The hybridomas represent clonal expansion of individual macrophages, retaining the surface antigen expression and functional capacity of the normal donor cells including cytokine production and stimulation in a mixed lymphocyte reaction. These cell lines differentially express Vmax antigens found on normal macrophages, potentially identifying subpopulations of macrophages. These lines may be useful not only to study normal macrophage function but may be relevant to a variety of disease states where expansion of subpopulations of macrophages identified by Vmax antigens may be important in disease pathogenesis.

Transformation of polycythemia vera to acute nonlymphocytic leukemia accompanied by t(1;3)(p36;q21) karyotype

The case of a patient with a history of polycythemia vera and 46,XX karyotype who transformed into acute nonlymphocytic leukemia with 46,XX,t(1;3)(p36;q21) rearrangement is reported. Significant percentages of the circulating blasts in the peripheral blood were positive on immunocytochemical stains for megakaryocyte markers, anti-Factor VIII, and antiglycoprotein IIb/IIIa. On the basis of our studies and published reports, trilineage hematopoietic abnormalities and a poor response to therapy may represent typical features of patients with t(1;3).

Acquisition of the Ph chromosome and BCR-ABL fusion product in AML-M2 and t(8;21) leukemia: cytogenetic and FISH evidence for a late event

A patient with the M2 subtype of AML who had a 45,X,−X,t(8;21) karyotype at diagnosis was found to have the Ph chromosome in one out of 37 evaluated cells 18 months after the initial diagnosis. Interphase FISH studies utilizing a BCR-ABL dual-color probe did not detect a fusion product 4 months prior to the appearance of one Ph-positive cell. Nineteen months post diagnosis and 5 months after clinical relapse all evaluated cells had the Ph chromosome in a clone characterized by t(8;21). These observations suggest that late appearing Ph is a secondary event which may be either therapy-related or consistent with one of the later events in a multistep pathogenesis of AML.

An immunological syndrome featuring transverse myelitis, Evans syndrome and pulmonary infiltrates after unrelated bone marrow transplant in a patient with severe aplastic anemia.

A patient with severe aplastic anemia underwent a matched unrelated bone marrow transplant, following which he developed a complex autoimmune syndrome. This featured transverse myelitis, immune mediated Coombs positive hemolytic anemia and immune thrombocytopenia (Evans syndrome), pulmonary infiltrates, eosinophilia, muscle pains and cramps and lichenoid dermatitis all of which may represent manifestations of graft-versus-host disease as they showed response to immunosuppression. Thus, although immune-mediated cytopenias after an allogeneic bone marrow transplant are rare, they should be considered as a possible cause of cytopenia in post-transplant patients. Bone Marrow Transplantation (2000) 26, 1225–1228.

Syngeneic stem cell transplant for spent-phase polycythaemia vera: eradication of myelofibrosis and restoration of normal haematopoiesis

Summary. We report a patient with spent‐phase polycythaemia vera (S‐PV) and massive splenomegaly who failed to engraft after a syngeneic granulocyte colony‐stimulating factor‐primed peripheral blood stem cell transplant (SCT), but later engrafted after splenectomy. Bone marrow (BM) showed resolution of myelofibrosis (MF) and absent endogenous erythroid colonies. This case demonstrated that (1) normal haematopoiesis can be restored after syngeneic SCT despite extensive MF, and (2) fibrosis can regress following a total body irradiation‐containing regimen and syngeneic SCT. As a graft‐versus‐BM stroma effect is non‐existent in syngeneic transplants, there may be a role for autologous SCT to obliterate MF in S‐PV.

A new variant translocation 11;17 in a patient with acute promyelocytic leukemia together with t(7;12)

Bone marrow cells from the majority of patients with acute promyelocytic leukemia (APL) are characterized by t(15;17)(q22;q11-12). At least 12 variant translocations have both also reported, and in each case, either abnormal chromosome 15 or del(17q) or both were involved in complex rearrangements. We report a patient with APL showing two translocations without apparent involvement of chromosome 15 and without del(17q). The karyotype was 46,XY,t(7;12)(p15;p13),t(11;17)(q13;q12). Rearrangement involving t(11;17) is probably associated with APL, while t(7;12) appears to be therapy related.

del(2)(p23): A new recurrent abnormality in acute myeloid leukemia

In two patients with acute myeloid leukemia, we found a new chromosome abnormality: del(2)(p23) which was detected in 13% and 50% of studied bone marrow cells, respectively. In patient one, del(2p) was an additional abnormality to t(8;21), while patient two had del(2p) as the sole abnormality. Based on our observation and a review of the literature we propose that del(2p) is a new recurrent chromosome abnormality for acute myeloid leukemia.

Fluorescence In Situ Hybridization (FISH) and Conventional Cytogenetics for Hematology and Oncology Diagnosis

Over the past 35 years, cytogenetic analysis of malignant hematological disorders has been one of the most rapidly growing areas in cancer. More than 45,000 cytogenetically abnormal neoplastic disorders have been reported and the evidence accumulated clearly demonstrates that karyotype information privide both biological and significant clinical value. While improved cell culture methods and the application of chromosome banding techniques had advanced our understanding of disease-specific abnormalities, molecular cytogenetics has now made it possible to identify genes involved at translocation breakpoints in specific chromosomal rearrangements. This knowledge is at the forefront of our understanding of the molecular pathogenesis of cancer. The goal of this chapter is to illustrate specific cytogenetic events and to delineate molecular phenotypes, which are key to the diagnosis and prognosis of leukemia, lymphoma, breast cancer, bladder cancer, and sarcoma. Thus, the hypothesis put forward by Boveri at the turn of the century, namely, that an abnormal chromosome pattern is intimately associated with the malignant phenotype of the tumor cell has proved correct for many malignant disorders.