Beatriz Pérez-Romano

Deficiency of red cell bound CD55 and CD59 in patients with systemic lupus erythematosus

CD55 and CD59 are glycosylphosphatidylinositol-anchored proteins with complement inhibitory properties. Autoimmune hemolytic anemia (AIHA) has been associated with antiphospholipid antibodies (APLA). The aim of this study was to evaluate the presence of APLA and its possible correlation with diminished CD55 and CD59 in red blood cells from patients with primary AIHA or secondary to systemic lupus erythematosus (SLE). Flow cytometric analyses were performed on CD55 and CD59 stained erythrocytes from 24 patients (primary AIHA, n=8; AIHA plus SLE, n=11; and SLE without AIHA, n=5) and 20 healthy controls. Antibodies to several phospholipids were detected in the sera by ELISA. Most patients with AIHA plus SLE and few with primary AIHA showed deficiency of either or both CD55 and CD59 expression and was not associated to the presence of APLA, while SLE patients exhibited a normal expression of these molecules. Although our findings showed CD55 and CD59 deficiency in primary or secondary AIHA, it appears that this defect plays a facilitator rather than a triggering role for the hemolytic process. Additionally, a role of anti-phospholipid antibodies as causative of this acquired defect is questionable.

Non-cryopreserved peripheral blood stem cells autotransplants for hematological malignancies can be performed entirely on an outpatient basis

We have prospectively performed peripheral blood stem cell autotransplants in six patients with hematological malignancies on an entirely outpatient basis. Patients were conditioned with high‐dose melphalan and received a median of 4.2 × 108/kg non‐cryopreserved, non‐purged mononuclear cells, containing a median of 3.9 × 106/kg CD34 + cells. The median time to achieve > 500 granulocytes/μl was 21 days, with a range of 16 to 40, whereas the median time to achieve > 20,000 platelets/μl was 38 days, with a range of 21 to 48. Only three patients were transfused with platelets whereas packed red blood cells were transfused in two. All patients survived 60 days after the autograft and three are alive at 450, 690, and 1,950 days after the autotransplant. One patient was given an allogeneic bone marrow transplant when relapsing after the autotransplant. One patient had to be admitted to the hospital on day +10 because of fever. A median of 6,500.00 USD per patient was calculated as the total cost of each outpatient autotransplant. Since outpatient autologous transplants with non‐frozen PBSC are feasible, restrictions to PBSC autotransplant programs may be overcome and costs may be diminished. Am. J. Hematol. 58:161–164, 1998.

Results of an Autologous Noncryopreserved, Unmanipulated Peripheral Blood Hematopoietic Stem Cell Transplant Program: A Single-Institution, 10-Year Experience

Background: Methods to simplify the stem cell transplantation procedures are needed mainly in developing countries. We have previously shown that unprocessed leukapheresis material is useful to restore hematopoiesis after high-dose chemotherapy. Methods: Over a 10-year period in a private practice setting, we prospectively performed autotransplants using noncryopreserved and unmanipulated peripheral blood stem cells mobilized from the bone marrow to the peripheral blood by means of filgrastim and using a single-day conditioning regimen with high dose (200 mg/m2) melphalan. Results: Forty-six individuals were given 50 autografts. The median age of the patients was 33 years (range 8–69). Twenty-two patients with acute leukemia (13 with myeloblastic and 9 with lymphoblastic leukemia), 4 with chronic myelogenous leukemia, 6 with multiple myeloma, 7 with Hodgkin’s disease, 3 with non-Hodgkin’s lymphoma and 4 with metastatic breast carcinoma were included. The median time to achieve >0.5 × 109/l granulocytes was 14 days (range 0–86), whereas the median time to achieve >20 × 109/l platelets was 25 days (range 0–102). The 3,300-day posttransplant survival was 63%, the median posttransplant overall survival was over 3,300 days, the 3,300-day disease-free survival was 50% and the transplant-related mortality was 2%. The procedure was performed entirely on an outpatient basis in the case of 48 autografts (96%). The approximate cost of each graft was 7,500 USD. Conclusion: This simplified method to autograft patients, avoiding in-hospital stays, purging procedures and cryopreservation of the cells, is feasible and results in a substantial decrease in the cost of the autologous hematopoietic stem cell transplantation methods.

Non-Cryopreserved Unmanipulated Hematopoietic Peripheral Blood Stem Cell Autotransplant Program: Long-Term Results

BACKGROUND Methods to simplify bone marrow transplantation procedures are needed mainly in developing countries. METHODS Between May 1993 and February 1999 in a private-practice setting, we performed 29 autotransplants in 28 patients using non-cryopreserved and unmanipulated peripheral blood stem cells mobilized from the bone marrow to the peripheral blood by means of hematopoietic growth factors. The autografting procedure was performed entirely on an outpatient basis in 19 cases (65%). The median age of the patients was 30 years, with a range of 9-67. There were 15 patients with acute leukemia (9 with acute myelogenous leukemia), 3 with chronic myelogenous leukemia, 2 with multiple myeloma, 3 with Hodgkins disease, 2 with non-Hodgkins lymphoma, and 4 with metastatic breast carcinoma. RESULTS The median time to achieve > 0.5 x 10(9)/L granulocytes was 14 days (range 7-42), whereas the median time to achieve > 20 x 10(9)/L platelets was 20 days (range 5-49). The 64-month post-transplant survival was 38%, whereas the median post-transplant survival was 18 months. The transplant-related mortality was 3.4%. The approximate cost of this simplified procedure was 10.8% for in-hospital procedures and for outpatient autografts, substantially lower than figures reported from the U.S. for autotransplants. CONCLUSIONS This simplified method for autografting patients, avoiding in-hospital stays, purging procedures and cryopreservation of the cells is feasible and results in a substantial decrease of the cost of autologous hematopoietic stem cell transplantation methods.

Protein S deficiency associated to anti-protein S antibodies in a patient with mixed connective-tissue disease and its reversal by danazol

We describe here a female patient with mixed connective tissue disease, secondary antiphospholipid syndrome, pulmonary hypertension and severe acquired, probably autoimmune, antibody-mediated type I coagulation protein S deficiency (total, free and C4bp-bound). No previous instance of anti-PS antibody-associated PS deficiency had been reported. The patient was treated initially with prednisone, but the protein S levels did not rise until danazol was added, and dropped again after its withdrawal.

Donor cell leukemia after non-myeloablative allogeneic stem cell transplantation: A single institution experience

Leukemia relapse occurring in donor cells, so-called donor cell leukemia (DCL), after allogeneic hematopoietic stem cell transplantation has been reported in rare cases, with less than 30 cases bei...

Decreased dopaminergic tone and increased basal bioactive prolactin in men with human immunodeficiency virus infection.

OBJECTIVE The aims of the study were: (1) to assess dopaminergic tone in a group of HIV infected men and the bioactivity and the molecular species of their circulating PRL in comparison with healthy men and (2) to search for a correlation between serum PRL and CD4+ T lymphocytes and viral load.

Treatment of vitiligo with a chimeric monoclonal antibody to CD20: a pilot study

Five patients with active disseminated vitiligo were given 1 g of a chimeric (murine/human) monoclonal antibody to CD20 in a single intravenous infusion and followed‐up for 6 months. Three of the patients showed an overt clinical and histological improvement of the disease, one presented slight improvement and the remaining patient showed no changes. Improvement was neither associated with changes in laboratory parameters nor to a specific human leucocyte antigen D‐related (HLA‐DR) phenotype. We believe that these preliminary results are encouraging, and further clinical trials should be undertaken. An important aim should be the finding of a marker with a good response to this therapeutic approach.

Minimal residual disease testing in acute leukemia by flow cytometry immunophenotyping: prognostic significance.

Two main techniques are being used for the detection of minimal residual disease (MRD) in acute leukemia (AL): immunophenotypic analysis and polymerase chain reaction (PCR). In this paper, we analyze the results of assessing MRD by means of flow cytometry (FC) in a group of 93 patients with AL who were prospectively studied and treated in a single institution over a 9-year period. The presence or absence of MRD was established at a cut-off level of 2%, as judged by FC; a single result above this level was considered to define the positivity. The patients were grouped in 4 subsets: (1) acute lymphoblastic leukemia (ALL) patients with MRD (n = 36); (2) acute myeloblastic leukemia (AML) patients with MRD (n = 13); (3) ALL patients without MRD (n = 31); and (4) AML patients without MRD (n = 13). The relapse rates in these groups were 17%, 8%, 0%, and 0%, respectively, whereas the overall 7-year survival was 65%, 69%, 83%, and 98%, respectively. Our results support the usefulness of serially assessing MRD in patients with AL by means of FC; because this method is applicable to all cases of AL, despite being less sensitive than a molecular biology study; it is a good option to follow-up patients and to decide therapeutic and timely interventions.

Immunophenotypic Analysis of Peripheral Blood Lymphocytes

This unit on basic phenotyping describes two basic and two alternate protocols for the immunophenotypic identification and classification of human peripheral blood lymphocytes. The presented protocols comply with consensus recommendations from professional organizations that regulate the clinical use of such assays. The authors discuss whole blood assay systems as well as enriched systems from several perspectives, including absolute number determination.