The absolute number of circulating CD34+ cells predicts the number of hematopoietic stem cells that can be collected by apheresis.

We report our single institution's effort to standardize the method of collecting peripheral blood progenitor cells (PBPC) used for autologous transplantation. PBPC were mobilized by different types of chemotherapy followed by G-CSF (24 patients) or G-CSF alone (six patients) in 30 patients with various underlying neoplastic diseases. A median of three aphereses (range: 2-7), using the CS3000 cell separator was performed and a blood volume of 101 was processed. We studied the relationship between the absolute numbers of circulating leukocytes, mononuclear cells and CD34+ cells and the amount of PBPC collected during a single apheresis procedure. CD34+ cells were quantified by leukocyte subset analysis based on flow cytometry. Both CFU-GM and CD34+ cell contents of the apheresis products (69 procedures analyzed) correlated strongly: rho = 0.936, P = 0.0001). Regression analysis showed that the total number of CD34+ cells or CFU-GM content of the apheresis products could be predicted (r = 0.915, P = 0.0001) from the absolute number of CD34+ cells in the blood. A number of 10 circulating CD34+ cells per mm3 blood ensured a minimum of 0.5 x 10(6) CD34+ cells per kg, collected on the same day. Of the 30 patients, 17 received an autologous graft that contained only PBPC in 13. Long-term and complete hematological reconstitution was observed in all patients who had a minimum of 2 x 10(6) CD34+ cells per kg reinfused.

Intracellular inclusion bodies in 14 patients with B cell lymphoproliferative disorders.

Two types of intracytoplasmic inclusion were detected by immunofluorescence microscopy in 12 patients with chronic lymphocytic leukaemia and two patients with a leukaemic phase of well differentiated lymphocytic lymphoma. Further analysis with light- and electron microscopy, showed that most inclusion bodies were rod-like crystalline structures. However, in three patients they consisted of amorphous vesicular precipitates. Immunological studies revealed the presence of immunoglobulins of the same class and type at the cell surface as well as in the inclusion bodies. The monoclonal immunoglobulins were all of lambda type except in two cases. The origin of immunoglobulin inclusion bodies in B cell malignancies is discussed in relation to published data and our own observation in one patient followed during treatment.

Neonatal infections with Pseudomonas aeruginosa associated with a water-bath used to thaw fresh frozen plasma.

In our 15-bed neonatal intensive care unit (NICU), four new-borns were found to be colonized or infected with Pseudomonas aeruginosa within a period of one week. To identify the outbreak source, three independent studies were performed: epidemiological investigation, environmental surveillance and genotypic typing of isolates. Although epidemiological investigation by a case-control study revealed no conclusive results, the transfusion of fresh frozen plasma (FFP) and human albumin (HA) appeared to be the factor with highest risk. Environmental surveillance and random amplification of polymorphic DNA (RAPD) of isolates identified a water-bath used to warm FFP and HA as the likely reservoir for the outbreak. Further spread of the organism did not occur after elimination of this water-bath from the NICU. RAPD identified in addition an isolate from an infant hospitalized in the NICU five months before the outbreak with a pattern matching the one of the outbreak cluster.

Therapeutic leukapheresis in a patient with prolymphocytic transformation of B-cell chronic lymphocytic leukemia.

A 64-year old woman with chronic lymphocytic leukemia (CLL) developed a prolymphocytic transformation with severe anemia, thrombocytopenia and deterioration of the general clinical condition. Neither treatment with chlorambucil and prednisolone, nor a combination chemotherapy (cyclophosphamide, vincristine, prednisolone) were successful. Splenectomy was not considered as the bone marrow presented signs of dyserythropoiesis. Encouraged by the results of therapeutic leukapheresis in different forms of leukemia, it was decided to start such therapy in our patient. During a seven-month period, thirty weekly aphereses were performed. The white blood cell count did not decrease, but the clinical condition of the patient improved dramatically.

Preliminary study on plateletpheresis with the CS-3000 blood cell separator using a single phlebotomy technique.

A coaxial dual flow catheter has been tested for thrombapheresis with a continuous flow blood cell separator in order to render the procedure as comfortable as possible for our donors. Therefore, two groups of five donors underwent a standard plateletpheresis: 3500 ml of blood were withdrawn at a speed of 35-45 ml per minute and were anticoagulated with ACD-A in a 1/9 ratio to whole blood. In the first group one vein puncture was effectuated with the coaxial catheter after local anesthesia. Two 14G catheters were used in the second group. In both groups the platelet yield was studied as well as the half life of 111-Indium-oxinate labeled platelets after autologous transfusion. Although the comfort of the donors was greatly improved, the one needle procedure resulted in a mean platelet yield of 1,28.10 while the two needle procedure yielded 3,46.10 platelets. In contrast, the survival of the platelets after autologous transfusion did not differ significantly in the two groups: t 1/2 in group I was 3,94 d, in group II 3,66 d. It is advocated that the poor efficacy of the single needle procedure might be due to recirculation.

Blood usage review in a Belgian university hospital.

The appropriate use of whole blood, packed red cells (PRC), fresh-frozen plasma (FFP) and platelets was evaluated. The indications, effectiveness and complications of transfusion were determined on the basis of existing criteria described in the literature and prevailing practices in the hospital. A retrospective review of medical records demonstrated that the administration of PRC was unnecessary in 15% of the cases and that the use of FFP was not indicated in 67% of the patients. The result of PRC transfusion was inadequate in 4% of the cases. Haemolysis and circulatory overload occurred in 2% and 4% of the patients, respectively. Major hazards such as Acquired Immunodeficiency Syndrome (AIDS) and hepatitis were not observed. In order to improve the appropriate use of blood components, transfusion guidelines were proposed to all medical departments.

Macromolecular creatine kinase type 1: a serum marker associated with disease.

The prevalence of circulating macromolecular creatine kinase type 1 (macro CK type 1 or CK-immunoglobulin complexes) is significantly higher in a patient population selected for CK isoenzyme assay than in age- and sex-matched blood donors (n = 1304). In greater than 8000 patients studied, 49 individuals with macro CK type 1 were identified, yielding an overall prevalence of 0.61%. Macro CK type 1 complexes occurred more frequently in women and in patients older than 70 years, and were often associated with complications of cardiovascular disease, life-threatening conditions, and poor outcome. These latter clinical associations could arise, at least partly, from the selection of patients for whom CK isoenzyme analysis was ordered.