Manipulation, and cryopreservation of autologous peripheral blood stem cell products in Italy: A survey by GITMO, SIDEM and GIIMA societies.

There is considerable heterogeneity in manipulation and cryopreservation of hematopoietic stem cells (HSC) for autologous HSC transplantation across Europe and Italy. To better address this point, three Italian Scientific Societies (GITMO- Gruppo Italiano per il Trapianto di Midollo Osseo; SIDEM- Società Italiana Emaferesi e Manipolazione Cellulare; and GIIMA- Gruppo Italiano Interdisciplinare Manipolazione e Aferesi per Terapie Cellulari), in collaboration with the Competent Authority "National Transplant Center" (CNT) sent to 85 Italian transplant centers (TC) a survey, which included 12 questions related to the most critical elements in graft processing. Fifty-nine centers (70 %) responded to the questionnaire. Overall, this survey demonstrates that the majority (>90 %) of responding TC used standardized procedures for HSC processing; however, an intercenter heterogeneity was clearly documented in several standard operating procedures adopted by different TC. These results seem to suggest that further standardization and efforts are needed to provide recommendations and guidelines on HSC manipulation, cryopreservation and functional assessment of cryopreserved material for autologous HSCT.

Rapid reconstitution of functionally active 6-sulfoLacNAc(+) dendritic cells (slanDCs) of donor origin following allogeneic haematopoietic stem cell transplant.

The role of dendritic cells (DCs) and macrophages in allogeneic haematopoietic stem cell transplant (HSCT) is critical in determining the extent of graft-versus-host response. The goal of this study was to analyse slanDCs, a subset of human proinflammatory DCs, in haematopoietic stem cell (HSC) sources, as well as to evaluate their 1-year kinetics of reconstitution, origin and functional capacities in peripheral blood (PB) and bone marrow (BM) of patients who have undergone HSCT, and their presence in graft-versus-host disease (GVHD) tissue specimens. slanDCs were also compared to myeloid (m)DCs, plasmacytoid (p)DCs and monocytes in HSC sources and in patients' PB and BM throughout reconstitution. slanDCs accounted for all HSC sources. In patients' PB and BM, slanDCs were identified from day +21, showing median frequencies comparable to healthy donors, donor origin and kinetics of recovery similar to mDCs, pDCs, and monocytes. Under cyclosporin treatment, slanDCs displayed a normal pattern of maturation, and maintained an efficient chemotactic activity and capacity of releasing tumour necrosis factor (TNF)-α upon lipopolysaccharide (LPS) stimulation. None the less, they were almost undetectable in GVHD tissue specimens, being present only in intestinal acute GVHD samples. slanDCs reconstitute early, being donor-derived and functionally competent. The absence of slanDCs from most of the GVHD-targeted tissue specimens seems to rule out the direct participation of these cells in the majority of the local reactions characterizing GVHD.

Immunohematological aspects of bone marrow transplantation.

Allogeneic bone marrow transplantation (BMT) is an effective treatment for some severe hematologic or nonhematologic diseases. The blood group antigen mismatch between donor and recipient may cause immunohematological complications during or after BMT. In this review, we analyze the ABO, Rh and other red cell antigen mismatches between donor and recipient, the main immunohematological complications and the techniques to prevent them. The data reported are derived from the experience of the authors and from the medical literature. The clinical implications of the immunohematological aspects of BMT emphasize the importance of close immunohematological monitoring in patients undergoing allogeneic BMT with ABO, Rh or other red cell antigen mismatches between donor and recipient.

Unrelated allogeneic bone marrow donation: short- and long-term follow-up of 103 consecutive volunteer donors.

Between 1992 and 1999, 105 unrelated allogeneic bone marrow collections from 103 volunteer donors (65 males and 38 females; median age 33 years) were carried out in three northern Italian centers (Verona, Bolzano and Padova) affiliated with the Italian Bone Marrow Donor Registry (IBMDR). The average volume of BM collected was equivalent in both genders (1143.1 ml for males and 1054.2 ml for females; P = 0.1), although the average volume collected for unit of body weight and the average post-collection blood volume depletion was higher in females (respectively 17.1 ml/kg and 14.2% in females, 14.8 ml/kg and 12% in males; P= 0.01 and 0.03). There was no statistically significant difference between males and females in the total number of nucleated cells collected. We did not record any acute life-threatening event during or after the bone marrow collections. The most frequent complaint was pain at the collection site (77%) followed by the onset of fatigue (38%) and nausea and vomiting (25%); all of these were short-term problems. Hospitalization was short (average 20.2 h) and donors started their normal daily activities after an average of 5.4 days. We also monitored Hb, serum ferritin levels, WBC and platelet counts in the post-collection period (average follow-up 40.1 months). All donors signed a written informed consent for a further bone marrow collection, if needed. Our findings confirm the short- and long-term safety of allogeneic bone marrow collection in volunteer donors.

Safety and efficacy of subcutaneous bolus injection of deferoxamine in adult patients with iron overload.

We compared 48-hour urinary iron excretion after a twice-daily subcutaneous bolus injection of deferoxamine and after 12 hours of subcutaneous continuous infusion of the drug in 27 patients with iron overload (mean age, 55.7 years). In most patients, the iron overload was due to multiple transfusions administered during chemotherapy or as part of supportive care for a hematologic or oncologic disorder. One patient had sickle cell anemia and 1 had hereditary hemochromatosis and spherocytosis. Similar urinary iron excretion was observed with the 2 methods of administration; mean +/- SD values were 6935.3 +/- 3832.3 microg/48 hours with subcutaneous bolus injection and 6630.4 +/- 3606.9 microg/48 hours with subcutaneous continuous infusion (P =.3). Twenty-six patients (96.3%) chose to continue therapy with bolus injection. The long-term efficacy of bolus injection was evaluated by measuring the serum ferritin concentration at regular intervals for a follow-up time of 20.1 +/- 4.5 months. Ferritin concentration decreased to below 1000 microg/L in 73% of the patients and to below 500 microg/L in 42% and became normal in 26%. Best results were obtained in patients who were no longer receiving blood transfusions when chelation therapy was initiated. Three of 26 patients (11.5%) had mild, transient side effects after bolus injection. Larger prospective, randomized studies must be conducted before deferoxamine bolus injection can be routinely recommended for patients with iron overload. (Blood. 2000;95:2776-2779)

Detection of an anti-RhD antibody 2 years after sensitization in a patient who had undergone an allogeneic BMT.

We describe an HLA matched bone marrow transplantation with minor ABO incompatibility and RhD mismatch (donor RhD negative and recipient RhD positive). GVHD appeared on day +96 and therapy with steroid and cyclosporin was started. When GVHD disappeared and immunosuppressive therapy was stopped (2 years after BMT), an anti-RhD antibody was detected in the patient's serum. The delayed appearance of this antibody may have been associated with the prolonged immunosuppression that was required for treatment of the patient's GVHD.

Subcutaneous bolus injection of deferoxamine in adult patients affected by onco-hematologic diseases and iron overload.

BACKGROUND AND OBJECTIVE: Chelation therapy is often necessary for patients who undergo chronic transfusion therapy for myelodysplastic syndromes. In these patients, deferoxamine, the most widely used chelating agent, has been reported to be effective in reducing the iron burden and the transfusion requirement. Unfortunately, compliance with the drug, that is usually administered by slow subcutaneous infusion via a battery operated pump, is often poor, especially in elderly patients. DESIGN AND METHODS: To verify efficacy and tolerability of deferoxamine by subcutaneous bolus injection as compared to the conventional pump-driven slow infusion, eleven patients affected by oncohematologic diseases were given 2 g of deferoxamine diluted in 10 mL of distilled water over twelve hours by continuous infusion, or by bolus injection in two divided doses. RESULTS: Mean urinary excretion was comparable with the two methods, being 9,183 +/- 4,349 micrograms/48 h after two daily subcutaneous bolus injections and 8,291 +/- 3,970 micrograms/48 h with the slow infusion. The bolus injection was preferred by all eleven patients, who chose to continue chelation therapy by this method. INTERPRETATION AND CONCLUSIONS: The iron excretion induced by bolus injection is not statistically different from that induced by subcutaneous infusion. The side effects are acceptable. Subcutaneous bolus injection of deferoxamine is an acceptable alternative to slow, pump-driven infusion.

Transmission of an anti-RhD alloantibody from donor to recipient after ABO-incompatible BMT.

We report a bone marrow transplant which was HLA matched, with major and minor ABO and minor RhD incompatibility (anti-RhD antibody) between the donor and recipient. When engraftment occurred, the recipient developed an anti-RhD antibody of donor origin detected by direct and indirect antiglobulin tests (DAT, IAT) and showed signs of mild hemolytic anemia. With the disappearance of the recipient RBCs, the DAT became negative and the hemolysis disappeared, while the anti-RhD alloantibody persisted in the patient's serum. This case emphasizes the importance of close immuno-hematological monitoring in patients undergoing allogeneic BMT with ABO-RhD incompatibility between recipient and donor.

Acute leukemia and pregnancy. Case report.

In this case report we describe a case of acute myeloid leukemia (AML; FAB M4) diagnosed in a 27-year-old female at the 20th week of gestation. After informed consent, the patient chose to undergo anti-leukemic treatment without therapeutic abortion. Complete remission was obtained following standard chemotherapy for AML (doxorubicin, cytosin-arabinoside, 6-thioguanine). The patient successively underwent an autologous bone marrow transplant (ABMT). No fetal malformation was observed. Urgent cesarean section was necessary at the 29th gestational week because of the onset of foetal sufferance. Fourteen months after diagnosis and seven months after ABMT the patient died due to relapse of AML. The child is presently 3.5 year old and well. In our opinion, the care of a pregnant woman with acute leukemia is feasible and it needs a multi-specialist effort that is easier to be achieved in a tertiary care institution.

Circulating levels of soluble CD23 reflect clinical and biological features of leukemic B-cell chronic lymphoproliferative disorders.

One hundred and twenty-four sera from patients with various leukemic B-cell chronic lymphoproliferative diseases were investigated at diagnosis by ELISA for their soluble CD23 content. Immunophenotyping was carried out in all patients, and in a selected subset the mean number of membrane-bound CD23 molecules per cell was also investigated. Seventy-three patients had typical B chronic lymphocytic leukemia, 41 leukemic B-cell disorders with atypical morphological and/or immunophenotypic features, 5 had low-grade follicular cell lymphoma in the leukemic phase, and 5 had splenic lymphoma with villous lymphocytes. Soluble CD23 levels were significantly higher than in normal sera (mean +/- SD: typical B chronic lymphocytic leukemia 3,650 +/- 4,654 U/ml, atypical B chronic lymphocytic leukemia 3,440 +/- 4,671 U/ml, follicular cell lymphoma 3,200 +/- 1,511 U/ml, splenic lymphoma with villous lymphocytes 8,236 +/- 7,294 U/ml, controls 137 +/- 128 U/ml; P < 0.001). More advanced Rai's stages were related to higher soluble CD23 levels (P < 0.01), both in typical and atypical B chronic lymphocytic leukemias, the highest levels and the best correlation with the absolute number of circulating CD19+ cells (r = 0.50) being observed in the typical form. The number of membrane-bound CD23 molecules per cell was significantly higher in typical than in atypical B chronic lymphocytic leukemias (mean number 156,727 +/- 94,668 vs. 12,010 +/- 10,643, P < 0.001). Our data suggest that soluble CD23 levels correlate with the clinical and biological features of leukemic B-cell lymphoproliferative disorders.

Serum levels of soluble CD30 are elevated in the majority of untreated patients with Hodgkin's disease and correlate with clinical features and prognosis.

PURPOSE: To evaluate the serum levels of the soluble form of the CD30 molecule (sCD30) in patients with Hodgkin's disease (HD) to establish whether there is a correlation with clinical features at presentation and prognosis. PATIENTS AND METHODS: The sCD30 serum levels of 117 patients were measured at diagnosis with a commercial sandwich enzyme-linked immunoadsorbent assay (ELISA) test kit, and in 78 of these patients the sCD30 levels were also recorded during the follow-up period. RESULTS: sCD30 levels at diagnosis were increased (> 20 U/mL) in a high proportion of patients (87.2%; mean +/- SD, 108 +/- 134 v 5.3 +/- 5.7 U/mL in controls, P < .0001) and correlated with stage (stages I + II, 73 +/- 97 U/mL; III + IV, 162 +/- 165 U/mL; P < .0001), with presence of B symptoms (stage A, 69 +/- 82 U/mL; stage B, 162 +/- 171 U/mL; P < .0001), and, to some extent, with tumor burden (bulky presentation, 141 +/- 129 U/mL; nonbulky, 91 +/- 133 U/mL; P = .058). Patients with sCD30 levels greater than 100 U/mL at diagnosis had a significantly higher rate of poor outcome in terms of failure to achieve a complete remission (CR) or disease relapse after CR achievement. In fact, the event-free survival (EFS) duration of patients with sCD30 levels greater than 100 U/mL was significantly worse (P = .0016). Using multivariate analysis, an sCD30 level greater than 100 U/mL retained its significance after adjustment for other prognostic parameters. CONCLUSION: sCD30 in HD at presentation strictly correlates with clinical features. Serum levels greater than 100 U/mL at diagnosis entail a significantly higher risk of treatment failure, a factor that is independent of other prognostic parameters.