Prothrombin fragment F1 + 2: correlations with cardiovascular risk factors.

Plasma prothrombin fragment F 1 + 2 (F1 + 2), thrombin-antithrombin complexes (TAT), fibrinogen and factor VII were related to variables associated with increased cardiovascular risk in 86 plasma donors (49 male and 37 female). F1 + 2 had a log-normal distribution and increased significantly with age and body mass index (BMI). Significantly, higher F1 + 2 levels were found in smoking compared with non-smoking males and in indolent males compared with males taking regular exercise. Higher levels were found in subjects with a parental history of ischaemic heart disease than in those lacking such a history. F1 + 2 correlated strongly with increasing cholesterol in males. Fibrinogen was significantly higher in male smokers than male non-smokers but did not vary with age or BMI. Factor VII correlated strongly with cholesterol and to a lesser extent with fibrinogen, F1 + 2 and BMI, but not with smoking. F1 + 2 correlated more closely with risk factors for cardiovascular disease than fibrinogen and factor VII, and consistently reflected the difference in cardiovascular risk when correlated with risk factors which have markedly different effects between the sexes. It promises to be a useful predictive marker of ischaemic heart disease.

Autologous bone marrow transplantation for myeloma patients using PNA- and CD19-purged marrow rescue.

A new method of in vitro bone marrow purging using a lectin and monoclonal antibody in combination has been used for the first time in vivo. Two patients with advanced myeloma were treated with high-dose melphalan and total body irradiation and then rescued with autologous bone marrow which had been purged in vitro to remove malignant cells by using a combination of a plasma cell-binding lectin (peanut agglutinin, PNA) and the anti-B lymphocyte monoclonal antibody anti-CD19, bound to magnetised microspheres. Both patients showed rapid engraftment of the purged bone marrow and remain well 36 and 46 months later with normal bone marrow morphology, although one patient still has a low level of circulating paraprotein. This is a promising form of therapy for what has been an invariably fatal condition.

A method for clinical purging of myeloma bone marrow using peanut agglutinin as an anti-plasma cell agent, in combination with CD19 monoclonal antibody.

Previous studies have shown that the lectin peanut agglutinin (PNA) binds bone marrow plasma cells in the majority of patients with myeloma and does not bind to normal haemopoietic progenitors. This lectin has been used in combination with anti-CD19 monoclonal antibody (moAb) in a system for purging myeloma bone marrow. This has now been scaled up for application to ex vivo treatment of large volumes of bone marrow suitable for autologous bone marrow transplantation. Four bone marrow harvests from patients with myeloma containing 9.5 +/- 4.9% plasma cells were depleted of erythrocytes and mature granulocytes by Ficoll separation using the Haemonetics V50 cell separator. The mononuclear fraction was then purged with magnetic beads coated with PNA and anti-CD19 moAb. The system proved highly efficient with removal of all detectable plasma cells and CD19+ cells. Average mononuclear cell recovery following purging was 71% of the concentrated marrow with 78% yield of CFU-GM. Normal progenitor recovery related to patients' weight is predicted to be adequate for haemopoietic reconstitution following ablative chemoradiotherapy. This system is therefore feasible for large-scale clinical purging.

Effective concentration of bone marrow mononuclear cells using density gradient separation within an automated cell separator.

An effective method for concentrating bone marrow is described. Concentration was achieved using an intermittent flow cell separator. Elimination of mature haemopoietic cells was enhanced by the addition of a density separation medium (Ficoll-metrizoate) which was then removed by washing. All procedures were undertaken using the cell separator, this allowed for standardization of procedure and less manipulation which is associated with enhanced mononuclear cell recovery and less risk of microbial contamination. Mature granulocytes were depleted by 86.8% and red cells by 97.7% whilst mononuclear cells showed a 49% recovery. Marrows processed in this way and subsequently purged and/or cyropreserved were shown to successfully engraft when reinfused.

Continuous flow cell separator use for bone marrow processing.

Many techniques have been described for processing bone marrow prior to transplantation, purging or cryopreservation. Effective techniques incorporate centrifugation and, or, density separation to produce an ideal marrow concentrate. We report on the use of a continuous flow cell separator (COBE Spectra) for marrow processing. Preliminary results indicate that the improved technology incorporated in this machine together with the new algorithm control of its collection functions allows for rapid collection of an ideal marrow concentrate. The addition of an inert sedimenting agent prior to processing enhances differential mononuclear cell collection and elimination of red blood cells and granulocytes. By this technique a volume depletion of 87% was achieved with recovery of 76.4% mononuclear cells and 86.5% CFU-GM progenitor cells. Marrow processed in this manner has been successfully transplanted; patients receiving such marrow show no delay in engraftment and their grafts have been sustained.

Red cell antigens and renal transplantation.

Donor-specific transfusion was performed with and without cyclosporine between haplomismatched relatives prior to living-donor renal transplantation. Red cell antigen mismatching was not taken as a contraindication to DST. Of 80 patients included in the trial; eleven were ABO-mismatched, 15 were Rh(D)-mismatched, and a further 11 were transfused in the presence of atypical red cell antibodies (anti-D, -C, -Fya, -Kell -N, -H/I -I, -P1, -Wra). Patients were randomized to receive cyclosporine (10 mg/kg) daily during DST or not (control group). The presence of atypical red cell antibodies, with the exception of Rh anti-D, did not appear to influence DST or renal transplantation. DST did not act as a primary stimulus to Rh anti-D production but stimulated preexisting anti D levels. ABO mismatching did not appear to influence DST or subsequent renal transplantation except in one group A [corrected] patient who received group O [corrected] blood and cyclosporine. This patient developed a severe, but self-limiting, autoimmune hemolytic anemia due to auto-anti A antibodies. A similar group A patient in the control group developed an auto-antibody with no clinical sequelae. The influence of cyclosporine on the development of this auto-antibody is uncertain. We conclude that, with the exception of preexisting anti-D antibodies, minor red cell antigen disparities should not preclude pretransplant conditioning with donor-specific transfusions.

Autologous Bone Marrow Transplantation in Hodgkin's Disease-A Single Centre U.K. Experience.

Twenty-five patients with relapsed or primary resistant Hodgkin's Disease were treated with high dose combination chemotherapy comprising cyclophosphamide, carmustine (BCNU) and etoposide followed by autologous bone marrow transplantation (ABMT). Sixteen (67%) of the twenty-four patients who survived the treatment schedule attained complete response (CR) and eleven of these remain disease free at a median time post ABMT of 16 months (range 6 to 27 months). Two of the patients who relapsed died at 7 and 17 months and the remaining three patients are alive with persistent disease at 12, 14 and 18 months. Four patients showed partial response. Three of these developed disease progression soon after the procedure and all died within ten months. The remaining patient achieved CR following further chemotherapy. There was a tendency for patients with bulky mediastinal disease, extra-nodal disease and heavy pre-treatment to fail to achieve CR.

Skin infiltration associated with chronic myelomonocytic leukaemia.

Chronic myelomonocytic leukaemia (CMML) is typically associated with a prolonged clinical course and is not usually very responsive to chemotherapeutic intervention. Skin infiltration has not been recognized previously as a feature of this illness. We have seen four patients recently with CMML, who during the the course of their illness developed marked skin infiltration. Whilst sensitivity to chemotherapy could be demonstrated in the peripheral blood cell population, skin infiltration was quite resistant to treatment. Skin infiltration heralded a more aggressive phase of the disease although no discernible change in morphology, cytochemistry or membrane marker analysis of the leukaemic cell population could be demonstrated in three of the patients studied; one patient, however, transformed to an acute leukaemia shortly thereafter.