[Advantages and difficulties of an erythrocytapheresis program for sickle-cell patients: a pediatric experience]. / Intérêt et difficultés de l'érythraphérèse chez les patients drépanocytaires : à propos d'une expérience pédiatrique.

Transfusion programs are sometimes necessary to take care of severe sickle-cell patients. Treatment of cerebrovascular disease in sickle-cell disease is the most common indication. Periodic automated red blood cell exchange (erythrocytapheresis) is an alternative treatment. Sixteen patients less than 20 years old have been treated with chronic erythrocytapheresis since 2004 in the pediatric hematology and oncology department of the University Hospital of Rouen, 10 patients for cerebrovascular disease (1 was on secondary prevention and 9 were on primary prevention), 5 patients for pain crisis recurrence, and the last one for mild psychocognitive deficit disorder. This treatment was unsuccessful for 4 patients, 3 on primary prevention and 1 treated for pain crisis recurrence. These failures were caused by alloimmunization for 2 patients and venous access problems for 2 patients. For the other 12 patients, 5 of the 6 patients on primary prevention showed clear improvement (normalization of transcranial Doppler ultrasound or improvement on magnetic resonance angiography), the patient on secondary prevention had stability on cerebral MRI after 2 years of treatment, the 5 patients with pain crisis recurrence had good improvement, and psychocognitive abilities improved for the last patient. One hundred and ninety-nine erythroexchange sessions were performed for the 9 patients treated over a period of 10 to 30 months. Erythrocytapheresis sessions ran on average less than 1.5h. Three patients showed high ferritin levels at the beginning of erythroexchange, which normalized 2 to 10 months later. All patients reported better quality of life. Periodic erythroexchanges are an effective treatment for complicated sickle-cell anemia and iron overload. It requires human, material, and financial support, but not as much as simple transfusion or manual erythroexchange. Practical experience shows problems of venous access because of coagulation when sampling.

Mycoplasma hominis wound infection after a vascular allograft.

Cases of Mycoplasma hominis infections after allograft are rare. We report a case of M. hominis wound infection after a vascular allograft. The allograft was positive before having any contact with the recipient, and our investigation suggests that M. hominis may have been transmitted from the donor to the recipient. It is not clear, however, whether specific diagnosis of M. hominis should be performed on tissue before grafting in order to prevent such donor-to-host transmission.

[Transfusions of rhesus-incompatible platelet concentrates in Rouen University Hospital: procedures and consequences]. / Transfusions de concentrés plaquettaires Rhésus incompatible au CHU de Rouen: pratiques et conséquences.

INTRODUCTION: Guidelines for distribution and use of blood products have been established for both blood transfusion institution and hospitals, in particular for the use of Rh (D)-incompatible platelet concentrates. The aim of this study was to evaluate: 1) the rate of attribution for the Rh (D)-incompatible platelets concentrates, 2) the immunisation prophylaxis practices, 3) the immunological consequences using short and medium term follow-up of transfused patients. METHODS: Patients with Rh (D)-incompatible platelets concentrate administered during the year 2003 at Rouen University Hospital were retrospectively selected. Patients on transfusion were described. The relationship of various factors with the injection as well as the appearance of allo-immunization was statistically tested. RESULTS: During a year, 280 Rh (D)-incompatible platelets concentrates were administered to 67 patients. Immunisation prophylaxis by injection of Ig anti-D was not systematically performed. Four immunizations in the Rhesus group system were identified: 2 against D antigen (Ag), 1 against E Ag and 1 against C Ag. Immunisations against D Ag occurred for two younger women considered as immunodeficient. Immunization prophylaxis was more frequent in poly-transfused patients. However no difference was observed for the other factors. CONCLUSION: Compatibility concerning Rhesus (D) is not always possible. The immunization against red cells persists, in particular against the antigens of the Rhesus group system and moreover for the immunodeficient patients. Recommendations for immunization prophylaxis by injection of specific anti-D immune-globulin (Ig) could be reconsidered.

Transient anti rhesus alloantibody produced by graft after non-myeloablative allogeneic stem cell transplant.

BACKGROUND: We report the case of a patient who received an allogeneic transplant with peripheral blood compatible ABO, Rhesus mismatched progenitor cells and who developed an asymptomatic transient anti Rhesus alloimmunisation. CASE REPORT: A 56-year-old man with renal cell carcinoma received a non-myeloablative allogeneic PBPC ABO compatible graft from his HLA-identical brother. Graft-versus-host disease prophylaxis consisted of cyclosporine alone. On day + 59, prior to any transfusion, a positive direct antiglobulin test (IgG++, C3d-) was detected. The indirect antiglobulin test (IAT) was considered doubtful, and IAT identification revealed the presence of an active anti Rhesus antibody (anti D specificity) in the patient's serum. This immunisation had no clinical consequence, with no acute hemolytic episode. Further monitoring showed negative antibody screening tests on day + 78. CONCLUSION: To our knowledge this is the first reported case of transient anti Rh (D) allo-immunisation after non-myeloablative allogeneic peripheral blood progenitor cell (PBPC) transplant. The period of occurrence and the specificity of this antibody strongly suggest a donor cell origin.

Bacterial resistance of refrigerated and cryopreserved aortic allografts in an experimental virulent infection model.

PURPOSE: The bacterial resistance of refrigerated and cryopreserved aortic allografts in a highly virulent infection in a dog model was studied. METHODS: The infrarenal aorta of 12 dogs was replaced with either a cryopreserved aortic allograft (group I, n = 6) or a refrigerated aortic allograft (group II, n = 6) in infected sites. Allografts were harvested from dogs and stored for 1 week, either by cryopreservation (-140 degrees C) or refrigerated method (4 degrees C), in a preservation medium. At the time of implantation, induction of infection was achieved with an infected piece of knitted Dacron placed just beneath the allograft. The Dacron was contaminated in vitro by soaking it in a solution with Staphylococcus aureus PR209. All 12 dogs received no adjunct antibiotic or antithrombotic therapy. Four weeks after implantation, the animals were killed to recover the grafts for bacteriological and histological analyses. Bacterial results were expressed as colony-forming units (CFU)/cm2 of graft material. RESULTS: In group I, only one allograft grew bacteria at 2. 16 x 10(6 )CFU/cm2, with a blood culture positive for S aureus. In group II, one dog died at 3 weeks from a false septic aneurysm rupture, all the allografts were infected (P <.05) with a mean bacterial count of 9.41 +/- 6.8 x 10(4) CFU/cm2, and three blood cultures were positive for S aureus. The patency of the grafts was analyzed at the time of recovery. Three laminar thrombi without occlusion were present in group I; none were present in group II. A better preserved endothelium in group I was revealed by means of histologic analysis staining with factor VIII antibody before implantation. After 4 weeks of implantation in the infected site, infected allografts presented polynuclear infiltrates in the media with a high degree of inflammatory reaction, and endothelial recovery was more significant in group I, with numerous young plump cells. CONCLUSION: This study demonstrates that cryopreserved allografts implanted in infected sites in a dog model can produce greater bacterial resistance.

[A rare cause of intra-alveolar hemorrhage: a transfusion-related incident with leukoagglutination due to antigranulocyte antibodies (Trali syndrome)]. / Une cause rare d'hémorragie intra-alvéolaire: l'accident transfusionnel avec leucoagglutination par anticorps antigranuleux (syndrome Trali).

INTRODUCTION: Transfusion-related acute lung injury (TRALI) is an infrequent but life-threatening complication of hemotherapy, usually secondary to passive transfer of antibody from the donor's plasma to the recipient. TRALI is a diagnosis of exclusion often masked by underlying factors. EXEGESIS: We report a new case of TRALI in a patient with severe multinevritis associated with Sjögren's syndrome and cryoglobulinemia, who had received intravenous immunoglobulins. CONCLUSION: This case report underlines the difficulty to establish a diagnosis in both acute respiratory failure and intra-alveolar hemorrhage in patients with auto-immune disorders. This case report also emphasizes the necessity of taking precautions in these immunocompromised patients in whom hemoglobin transfusion is required.

Peripheral blood stem cell transplantation for relapsed or refractory aggressive lymphoma in patients over 60 years of age.

Intensive chemotherapy with autologous bone marrow transplantation is now considered the treatment of choice for young patients with sensitive relapse of non-Hodgkin's lymphoma (NHL) but results of this procedure in older patients remain unknown. We evaluated the feasibility of two cycles of salvage therapy followed by an autologous peripheral blood stem cell (PBSC) transplantation in 13 patients aged more than 60 years (median age: 62; range 61-72) suffering from relapsed (n = 10) or refractory (n = 3) aggressive NHL. All patients had previously received first-line treatment containing doxorubicin. An association of ifosfamide, VP16, cytosine-arabinoside with or without high-dose methotrexate was used as salvage and priming therapy prior to collection of PBSC. All patients received G-CSF following salvage therapy. PBSC collection could be performed in 10 patients and yielded a median number of CFU-GM: 98.4 x 10(4)/kg (range 68-369). Nine patients underwent a transplantation using BEAM conditioning regimen. The median time to granulocyte and platelet recovery was 13 days (range respectively: 9-25 and 9-16). One patient died from sepsis after transplantation. The main adverse experience occurring after transplantation was a prolonged decline of performance status. Seven patients achieved a complete remission and one failed to respond. Three patients are still alive in CR. We conclude that PBSC collection was possible in selected patients over 60 years of age with refractory or relapsed aggressive NHL and myeloablative therapy could be used with tolerable toxicity. Hematologic recovery and organ toxicity appears to be similar to those observed in younger patients. Deterioration of performance status after transplantation is the most important factor that could limit this procedure. Further investigations are necessary to determine which patients will be able to benefit by this procedure in terms of survival and quality of life.

[Hemolysis can conceal another hemolysis]. / Une hémolyse peut en cacher une autre.

We report a delayed haemolytic reaction during the course of a severe HELLP syndrome, which required red blood cell transfusions. Haemolysis was mainly ascribed to the erythrocyte alloantibody anti-Fy a. The fact that this antibody was undetectable before transfusions (despite cross-matching and anti-erythrocyte antibody screening test) emphasizes the limits of these tests for low antibody concentrations. Moreover, this patient, who had a remarkable obstetrical history, received red blood cell transfusions matched for Rhesus and Kell antigens. The opportunity to perform a more extended phenotype screening for patients at high-risk to develop multiple alloantibodies is discussed.

Ifosfamide, etoposide, cytarabine, and methotrexate as salvage chemotherapy in relapsed or refractory aggressive non-Hodgkin's lymphoma.

BACKGROUND: Patients with relapsed or resistant non-Hodgkin's lymphoma (NHL) have a poor prognosis and are rarely cured with usual salvage chemotherapy. Intensive treatment with the support of peripheral blood stem cells (PBSC) may be an effective therapy for these patients. We used a combination of ifosfamide, etoposide, cytarabine, and methotrexate (IVAM) with the intention both to reduce tumor burden and collect PBSC prior to transplantation. METHODS: Thirty-one patients (17 with relapsed NHL and 14 with refractory NHL) were treated with 2 courses of chemotherapy: IVAM regimen (ifosfamide, 1500 mg/m2 daily for 5 days plus mesna; etoposide, 150 mg/m2 daily for 3 days; cytarabine, 100 mg/m2 daily for 3 days; and methotrexate, 3 g/m2 on Day 5, with leucovorin rescue). Twenty-three patients had an intermediate grade and 8 patients had a high grade lymphoma. RESULTS: After IVAM therapy, 19 patients (61%) achieved complete response, 8 patients (26%) achieved partial response and 4 patients (13%) failed to respond. The major toxicity of IVAM was myelosuppression, but there were no toxic deaths. PBSC harvest could be performed in 29 patients (94%) with a median granulocyte-macrophage colony-forming unit count of 55 x 10(4)/kg (range, 2-391 x 10(4)/kg). Three patients could not undergo transplantation because of disease progression. One patient received a syngeneic transplant, 25 patients received PBSC transplantation, and 2 patients received a bone marrow transplant. In an intent-to-treat analysis, the overall survival rate at 4 years was 37% for the whole group (95% confidence interval: 22-55). CONCLUSIONS: We conclude that IVAM is an effective salvage chemotherapy for refractory or relapsed NHL and permits PBSC collection in most of these patients.

Treatment of Crohn's disease by lymphocyte apheresis: a randomized controlled trial. Groupe d'Etudes Thérapeutiques des Affections Inflammatoires Digestives.

BACKGROUND/AIMS: Several uncontrolled trials suggest that lymphapheresis improves the clinical course of patients with Crohn's disease; this study was designed to assess the efficacy of lymphapheresis in preventing early relapses of Crohn's disease in patients in clinical remission after steroid treatment for an acute attack. METHODS: Twenty-eight patients in clinical remission at the end of 3-7 weeks of steroid therapy were included in this randomized multicenter prospective trial. Before starting steroid tapering, patients were randomly assigned either to the lymphapheresis group (9 procedures within 4-5 weeks) or to the control group. The primary judgement criterion was the cumulated recurrence rate after steroid discontinuation. RESULTS: All the patients treated by lymphapheresis (12 of 12) were successfully withdrawn from prednisolone and only 10 of 15 in the control group (NS). At the end of the 18-month follow-up period, the cumulated relapse rate was 83% in the lymphapheresis group and 62% in the control group. CONCLUSIONS: Although there was a trend towards a diminished incidence of corticosteroid dependence, lymphapheresis did not prevent the occurrence of early relapses.

[Refractory thrombocytopenia by HLA alloimmunization in a multitransfused patient]. / Thrombopénie réfractaire par immunisation anti-HLA chez une patiente polytransfusée.

A case is reported of 60-year-old woman who developed transfusion refractoriness after having been transfused several times. This patient who had been transfused with 4 standard packed red cell packs (PRC) for a surgical repair of a hiatal hernia, required three further operations within two months for postoperative complications. After the first operation, she had developed anti-JK1 and anti-CW alloantibodies. Seven phenotype compatible PRC and five fresh frozen plasma (FFP) were transfused during the surgery carried out on day 32. Massive haemorrhage occurred during the fourth operation on day 48, and the patient was transfused with 31 phenotype compatible PRC, 21 fresh frozen plasma, 36 standard platelet concentrates (SPC), fibrinogen, factor VIII and anti-thrombin III. Postoperative disseminated intravascular coagulation occurred, with thrombocytopaenia (45 G.l-1). Major thrombocytopaenia persisted for 6 days (12 G.l-1 on day 52), after the other signs of intravascular coagulation had been corrected, and despite the transfusion of 40 SPC. Platelet counts progressively returned to normal (195 G.l-1 on day 56). An HLA alloimmunization was discovered, which may have been induced by leukocytes contaminating the transfused red blood cell and platelet concentrates. A fifth operation carrying a high risk of haemorrhage was therefore prepared by harvesting autologous platelet rich plasma two days before and on the morning of the operation. These were transfused intraoperatively, together with phenotyped and leukocyte-free PCR, thus avoiding massive and expensive homologous platelet transfusions. In patients with a high risk of HLA immunization (previous pregnancies, multiple transfusions), autotransfusion or leukocyte-poor blood products should be used.

Immunological evaluation of harvested stem cells obtained by leukapheresis after chemotherapy.

Some patients suffering from malignancies may benefit of myeloablative chemotherapy followed by hematological reconstitution with autologous peripheral blood reinfusion. A quick evaluation of the number of hematopoietic progenitors present in leukapheresis blood samples is necessary to ensure the collection of a sufficient number of these cells. A study was performed on a series of 25 leukapheresis following initial chemotherapy. The number of granulomonocytic colony-forming unit (CFU-GM) and the number of CD34+ cells were evaluated simultaneously, in each sample. Results have shown a relatively strong linear correlation between both methods of evaluation of hematopoietic progenitors, suggesting that immunophenotyping could be a useful method to estimate the number of progenitors.

Cyclosporin for treatment of life-threatening alloimmunization.

We describe a 16-year-old girl with aplastic anemia who, 1 year after initial diagnosis developed a refractory state to platelet transfusions due to alloimmunization and resulting in severe bleeding. Treatment with cyclosporin, initially prescribed as treatment of the bone marrow failure, resulted in prompt decrease in lymphocytotoxic antibodies, which paralleled a marked improvement in platelet recovery. To our knowledge, such a dramatic effect of cyclosporin on alloimmunization has not been previously reported and merits further attention.

[Preventive treatment of infection caused by gram-negative bacteria using anti-lipopolysaccharide antibodies. Evaluation in patients with aplasia]. / Traitement préventif des infections à bacille à Gram négatif par des anticorps anti-lipopolysaccharides. Evaluation chez l'aplasique.

The prophylaxis of severe Gram-negative infections with human antiserum to lipopolysaccharide (LPS) was evaluated in a randomised study of 60 patients with therapeutic aplasia for leukaemia. The antiserum was found to be ineffective in preventing Gram-negative infections. The levels of anti-LPS antibodies showed that passive immunization was obtained in only one half of the patients. These disappointing results warrant further investigations to evaluate the effectiveness of this prophylactic treatment.

Peripheral blood stem cells collected before and after leukapheresis in the very early remission phase of hematopoietic malignancies.

Human peripheral blood obtained after chemotherapy-induced remission in hemopoietic malignancies has been suggested to be a potential substitute for autologous bone marrow as regards autologous hematopoietic reconstitution. The schedule and consequences of early leukapheresis are, however, still imprecise. We report a study performed in two series of, respectively, 10 and 14 patients where sequential leukapheresis (total number = 84) was evaluated with regard to colony-forming unit (CFU) potency. Our data demonstrate that adequate numbers of progenitor cells can be collected by leukapheresis and that, even when this is performed at an early stage after remission, subsequent hematopoietic reconstitution is not impaired.

Daily evaluation of circulating granulocyte-monocyte progenitors during bone marrow recovery from induction therapy in acute leukemia.

The notable increase of the circulating granulocyte-monocyte progenitors (PB CFU-GM) during bone marrow recovery following chemotherapy is a well known phenomenon. It has led to consider harvesting a large number of autologous stem cells by cytapheresis. Daily assessments have been conducted on the PB CFU-GM level in 9 patients with acute leukemia at the time of bone marrow regeneration after the first induction course in order to identify the circumstances of this rise. The PB CFU-GM maximum peak, of an average of 2142/ml, occurs between day 17 and day 23 after the chemotherapy has ended. A ten-fold increase of the PB CFU-GM level above normal values is maintained between 2 and 10 days. The PB CFU-GM peak coincides with that of the circulating immature myeloid cells and monocytes. The platelet rise above 100 X 10(9)/1 always occurs 2-7 days before the PB CFU-GM peak.

[Anti-Gram-negative bacterial antibodies in alcoholic cirrhosis. Study of 58 patients]. / Les anticorps anti-bacille à Gram négatif au cours de la cirrhose alcoolique. Etude de 58 malades.

Gram-negative bacterial infections are frequent and severe in cirrhotic patients. Existence of endotoxemia in cirrhosis is controversial. The demonstration of Gram-negative bacterial antibodies could be an alternative approach to the pathogenic role of these bacteria. In 58 patients with alcoholic cirrhosis, the immunoglobulin G specifically directed against the Gram-negative bacteria lipopolysaccharide expressed by the J5 mutant of Escherichia coli 0111:B4 was measured. Antibody titres were compared to those of a control group of blood donors. The distributions of antibody titres were similar in cirrhotic patients and in control subjects. No correlation was found between antibody titres and biological parameters of liver function. These results seem to confirm previous reports on the absence of latent endotoxemia in cirrhotic patients, and they suggest that antibody production against Gram-negative bacteria lipopolysaccharides is not enhanced in these patients.