HLA class II antibodies in transfusion-related acute lung injury (TRALI). A case report.

TRALI, a serious complication of blood transfusion, is underdiagnosed. Anti-granulocyte and anti-HLA class I molecules in donors or recipients and very recently, lipids in stored blood as well as anti-HLA class II have been associated with the syndrome. We present a TRALI case which occurred in a 56 year old woman after plasma transfusion. HLA class II antibodies were identified in the donor and were correlated with the recipients' HLA antigens. The presence of HLA class II antibodies without anti-HLA class I has been reported in very few cases and may facilitate the understanding of the pathogenesis of the syndrome.

Merocyanine 540 mediated photolysis of normal bone marrow, committed hemopoietic progenitors and neoplastic cells. implications for bone marrow purging.

The effect of merocyanine 540 (Mc 540) mediated photoirradiation on both neoplastic and normal hemopoietic progenitor cells was studied. Bone marrow (BM) cells from children with acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML) at initial diagnosis, ALL in remission, neuroblastoma and normal children as well as cells of Reh-6 and HL-60 cell lines were incubated with Mc 540 in the presence of human albumin (HA) and exposed to different argon laser 514 nm doses. Cell survival was estimated using Trypan Blue supravital stain following a 24-h incubation and leukemic cell lines were studied in continuous cell cultures of 4 weeks duration. Our results showed that HA protects normal BM cells from Mc 540 mediated phototoxicity. A 99.9999% inhibition of Reh-6 and HL-60 was noted at irradiation doses where the corresponding mean survival of normal BM cells was 77.4 +/- 12 and 70.3 +/- 10%, respectively. BM leukemic cells from children with ALL and AML were also very sensitive to Mc 540 photoirradiation in contrast to neuroblastoma cells where only a three-fold reduction was observed. Finally, the survival of normal BM progenitors was 38% for colony forming unit erythroid CFU-E, 37% for burst forming unit erythroid BFU-E, 55% for CFU-GM and 29% for CFU-GEMM. In conclusion it seems that Mc 540 mediated photoirradiation in neoplastic cells exerts selective cytotoxicity and can be used in ex vivo purging of malignant cells in BM.

Bone marrow purging by photodynamic treatment in children with acute leukemia: cytoprotective action of amifostine.

In order to evaluate the combined effect of Amifostine and Merocyanine 540 during photoirradiation in neoplastic cells, bone marrow cells from children with acute leukemia (AL), age-matched controls as well as HL-60 cell line were studied. Cell suspensions were incubated with Amifostine, then with MC 540 and they were subsequently exposed to different irradiation doses by Argon Laser 514 nm. Cell survival was estimated by trypan blue supravital stain following a 24-h incubation. The leukemic cell line was studied in continuous liquid cell cultures for 4 weeks. The survival of normal bone marrow progenitors has been estimated by colony formation assay in methylcellulose cultures. Our results showed that Amifostine enhances the photokilling effect of MC 540 on leukemic cells and significantly protects bone marrow nucleated and committed progenitors (BFU-E and CFU-GM) from children with AL under chemotherapy. In conclusion, Amifostine seems to be a promising cytoprotective agent in the clinical use of purging with MC 540 mediated phototherapy.

Phenotypic characteristics of cord blood hemopoietic cells.

Bone marrow transplantation is limited due to the lack of HLA-matched donors and to the frequent occurrence of GvHD. Hemopoietic transplants using cord blood cells are being increasingly used in pediatric patients. In this study, the immunophenotypic characteristics of cord blood cells have been investigated by one or two-color flow cytometric analysis. The CB cells were characterized by a low proportion of CD3+ T-cells, increased CD4/CD8 and CD45RA/CD45RO ratios, minimal expression of HLA-DR, increased proportion of CD5CD19 double positive B-cells, while CD3- CD8+ and CD3- CD7+ subsets, not usually found in adult PB, were detected. These data reflect the immaturity of CB cells as assessed by immunophenotypic analysis suggesting that it could be a valuable alternative source of transplantable hematopoietic progenitor cells and might alleviate some of the problems associated with bone marrow or peripheral blood transplantation.

Merocyanine 540 mediated photoirradiation of leukemic cells. In vitro inference on cell survival.

In order to evaluate the selective killing of merocyanine 540 (MC 540) mediated photoirradiation in neoplastic cells, bone narrow cells from children with leukaemia or neuroblastoma and normal children as well as peripheral blood cells and Reh-6 and HL-60 cell lines were studied. Cell suspensions were incubated with MC 540 and exposed to various argon laser 514 nm doses. Cell survival was estimated with trypan blue supravital stain following a 24 h incubation and has been followed in continuous cell cultures of 4 weeks duration. Our results showed that the inhibition of survival of neoplastic haemopoietic cells by laser in the presence of MC 540 is proportional to the MC 540 and photoirradiation doses. A 99.9999% inhibition of Reh-6 and HL-60 was noted at irradiation doses where the corresponding mean survival of normal bone narrow cells was (33.6 +/- 15.5)% and (50.6 +/- 10.7)% respectively. Peripheral blood mononuclear cells were not sensitive to MC 540 mediated photoirradiation. The inhibition of survival of bone marrow metastatic neuroblastoma cells was (69.9 +/- 4.1)%. In conclusion, it seems that MC 540 mediated photoirradiation in neoplastic cells exerts selective cytotoxicity and can be used in ex vivo purging of malignant cells in the bone marrow.

Treatment with recombinant human erythropoietin in children with malignancies.

The effect of recombinant human erythropoietin (rHuEPO) on the anemia of cancer was examined in 15 children with hematologic malignancies (group I) and solid tumors (group II), whose hemoglobin (Hb) was under the third percentile for sex and age. The response to rHuEPO was defined as an increase of Hb to above the 10th percentile following 8 weeks of therapy. The rHuEPO caused an increase in the Hb and hematocrit (Hct) in 46% of children of both groups at a dose of 150 IU/L, in 28.5% of children at a dose of 250 IU/L and in 25.5% of children at a dose of 400 IU/L. Leukocyte and platelet counts were not influenced by the rHuEPO treatment. The red cell transfusion requirement decreased to 66% in both groups after rHuEPO treatment. Erythropoietin (EPO) levels were measured prior to the treatment and then every 4 weeks during rHuEPO treatment. Children who responded to EPO had an initial EPO level of < 100 IU/L, while those who did not respond had an initial EPO level of > 100 IU/L. Erythropoietin was well tolerated in all children, with no side effects.

Isolated intracranial metastasis of neuroblastoma 2 years after completion of therapy.

Metastatic neuroblastoma in the brain without evidence of intracranial or extracranial disease elsewhere, is a very rare event. The pattern of spread is believed to be via the hematogenous route or cerebrospinal fluid. A child with a cystic neuroblastoma mass in the right temporal lobe 23 months following completion of therapy for the initial disease is described here. Chemotherapy with the protocol 'eight drugs in 1 day' was initially effective for this patient.

Efficacy of recombinant human granulocyte colony-stimulating factor and recombinant human granulocyte-macrophage colony-stimulating factor in neutropenic children with malignancies.

The difference between the effects of administration of recombinant human granulocyte colony-stimulating factor (rhG-CSF) and recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) was studied in 39 children with neutropenia secondary to chemotherapy (absolute neutrophil count (ANC) less than 1,500/microliters. The children were divided into two groups. The first group (G-CSF) included 25 children (12 with acute lymphoblastic leukemia [ALL]-non-Hodgkin's lymphoma [NHL] and 13 with solid tumors) and the second group (GM-CSF) included 14 children (5 with ALL-NHL and 9 with solid tumors). All 39 children received of either G-CSF or GM-CSF (5 micrograms/kg/day) subcutaneously at the end of each chemotherapy course for a maximum duration of 14 days. The effect of G-CSF and GM-CSF on the ANC, the antibiotic therapy administration, and the length of hospital stay were studied for both groups at two cycles of chemotherapy. During both cycles a faster rise of ANC was observed in the children of the first group (G-CSF) compared with those of the second group (GM-CSF), but there was no difference in either the incidence of antibiotic therapy administration between the two groups (26% vs 25%) or the length of hospitalization. Both growth factors were well tolerated by all children studied with minimal side effects observed (including bone pain with G-CSF in 2 of 25 children and pruritus with GM-CSF in 1 of 14). We conclude that G-CSF reduces the duration of neutropenia more than does GM-CSF, but the incidence of severe infection and the duration of hospitalization do not differ between children receiving either G-CSF or GM-CSF.

Altering interface detector positioning in combination with prestorage filtration to achieve a better quality of single donor platelet concentrates using the CS 3000 Plus blood separator.

The interface detector (ID) is an optical density sensor that affects the quality of single donor platelet collection using the CS 3000 Plus blood separator. The purpose of this study was to evaluate the effect of altering ID position on platelet yields and the contamination of leukocytes (WBC) in platelet concentrates (PCs). Dual-needle apheresis procedures (n = 93) were performed using an A35 collection chamber. Plateletpheresis products were separated according to interface detector offset (IDO) positioning into four groups: A: IDO = 6 (n = 33), B: IDO = 10 (n = 28), C: IDO = 12 (n = 18), D: IDO = 18 (n = 14). For 32% of the collections, the closed system apheresis kit with integral Sepacell filter (Baxter) was used and 33% of them were leukodepleted using the LRP-6 (PALL) filter. Our results showed that: (1) Although the mean blood volume and the time of apheresis were significantly higher, the mean platelet (PLT) yields were significantly lower in PCs of group A as compared to all other groups (P < 0.0001). (2) The mean WBC content was significantly higher in PCs of group D as compared to all other groups (P < 0.0001). (3) With the LRP-6 filter, a significantly higher WBC reduction as well as PLT loss in PCs was observed as compared to Sepacell leukapheresis filter. A higher PLT loss was observed with both filters when leukoreduction was performed within the first 6 hours as compared to 24 hours after the procedure. Conclusively, an IDO setting of 10 or 12 results in better platelet yields in PCs without increasing the WBC contamination. An IDO positioning of 18 or higher must be avoided or should be always combined with PCs leukodepletion. Finally, the best timing for leukoreduction is 24 hours after the plateletpheresis.

Congenital systemic Langerhans cell histiocytosis (report of two cases).

Two cases of congenital systemic Langerhans cell histiocytosis (LCH), diagnosed and treated in our department from June 1995 until May 1996, are described. The cases concern two neonates (one female and one male) born with necrotic lesions and skin nodules. The diagnosis was confirmed by skin biopsy which showed diffuse infiltration by CD1 antigen and S-100 protein positive histiocytes. The babies didn't present with anemia, hepatosplenomegaly or lymphadenopathy. Hepatic and renal function were normal. In both infants skeletal survey showed no lytic lesions but chest X-rays and high resolution computerized tomography (HRCT) scan revealed diffuse mottling of both lung fields. Bone marrow aspiration showed the presence of histiocytes in percentages of 6% and 10%, respectively. Both babies were treated with prednisolone 1 mg/kg body weight for three months. The first child who is 20 months old, is now well with resolution of skin and pulmonary lesions occurring within one month of the initiation of steroids, while the second, who presented spectacular resolution of skin lesions within the first three weeks of therapy, is also in excellent condition five months after completion of treatment. We conclude that congenital LCH has to be suspected in neonates with persisting skin lesions. If the disease is systemic but without organ dysfunction, treatment with steroids may be beneficial.

Effect of granulocyte colony-stimulating factor on chemotherapy-induced neutropenia in children with cancer.

We investigated the effects of recombinant granulocyte colony-stimulating factor (G-CSF) administration on duration of neutropenia, antibiotic therapy, and hospitalization days in 25 children with malignancies (Group A: 12 leukemia and lymphoma; Group B: 13 tumors) who were undergoing chemotherapy. We compared the effect of G-CSF with a control group of 21 children with equivalent diseases and chemotherapy that did not receive G-CSF treatment. All 25 children received 5 micrograms/kg/day of G-CSF at the end of chemotherapy courses when absolute neutrophil counts were < or = 1000/mm3. The effect of G-CSF on median neutrophil profiles, antibiotic therapy, and hospitalization days was studied for both groups at the 1st and 4th cycle of chemotherapy. During both cycles, children who received G-CSF showed a faster rise of absolute neutrophil count (P < 0.001) and fewer hospitalization days (P < 0.05), and not as many received systemic antibiotic therapy (P < 0.0001). We conclude that G-CSF accelerates neutrophil recovery in chemotherapy-induced neutropenia in childhood malignancies.

Ondansetron and tropisetron in the control of nausea and vomiting in children receiving combined cancer chemotherapy.

Ondansetron (Zofron, Glaxo) and tropisetron (Navoban, Sandoz) are selective serotonin (5HT3) antagonists that have proven very effective in the prevention of vomiting and nausea in adults and children receiving cancer chemotherapy. This study compared the efficacy of the two agents in the prevention of vomiting and nausea in children receiving chemotherapy for solid tumors and blood malignancies. A total of 23 children were studied in 205 chemotherapeutic cycles (116 one-day regimens and 89 multiple-day regimens). In 102 chemotherapeutic cycles the children received ondansetron as an antiemetic agent in a dose of 5 mg/m2 30 min before chemotherapy was given and then 4 mg/m2 every 8 h i.v. (group A) and in 103 cycles they received tropisetron in one dose of 0.2 mg/kg 24 h-1 i.v. (max dose 5 mg) 30 min before cytotoxic drugs administration every day they received chemotherapy (group B). The response was defined as complete in the absence of nausea and vomiting per 24 h of chemotherapy, as partial given the presence of 1-4 events of vomiting and/or nausea less than 5 h per 24 h, and as failure if there were more than 4 events of vomiting and/or nausea for more than 5 h per 24 h of chemotherapy. The response of the two groups was studied independently and depending on the degree of emetogenicity of the chemotherapeutic agents, which were divided into mildly, moderately, and highly emetogenic. The comparison of the two groups not taking into consideration the emetogenicity of the chemotherapeutic agents showed that ondansetron was more effective in 1-day regimens (P = .023), whereas the two agents were equally effective in multiple-day regimens (P = .2). The statistical analysis depending on the emetogenicity of the chemotherapeutic agents showed increased efficacy of ondansetron in mild (P = .017) and moderately emetogenic chemotherapeutic agents, whereas there was no difference in the highly emetogenic drug group. Ondansetron is found to be more effective than tropisetron in controlling acute nausea and vomiting in children receiving mild and moderately emetogenic chemotherapeutic drugs, although there is no difference in the efficacy of both antiemetic agents when highly emetogenic drugs are administered.

Expression of the Epstein-Barr virus encoded latent membrane protein in tumor cells of Hodgkin's disease occurring in childhood.

Paraffin sections from 21 cases of Hodgkin's disease (HD), 28 cases of non-Hodgkin's lymphomas (NHL) and 34 cases of non-specific reactive lymphadenitides occurring in childhood were examined for the presence of the Epstein-Barr Virus (EBV)-encoded Latent Membrane Protein (LMP) using a double layer immunohistochemical method. LMP was detected in 12/21 (57%) cases of HD but not in NHL or reactive lymph nodes. LMP reactivity was restricted to Reed-Sternberg and Hodgkin's (HRS) cells in 4 of 9 (45%) cases of nodular sclerosis (NS), 6 of 9 (66%) cases of mixed cellularity (MC) and 2 of 2 (100%) cases of lymphocyte depletion (LD) while it was undetectable in the single case of lymphocyte predominance (LD) subtype. These results provide further evidence for an association between EBV and Hodgkin's disease, and they show that LMP expression occurs more frequently in the clinically more aggressive subtypes of HD. Furthermore, in view of the in vitro transforming potential of the LMP protein, the exclusive immunolocalization of LMP in HRS cells, suggests that EBV may be involved in the pathogenesis of a proportion of cases of HD.

Immediate free flap mandibular reconstruction in osteosarcoma of the mandible in childhood.

Mandibular osteogenic sarcoma (OS) is a very rare entity in childhood. Adequate surgical rejection with a wide margin of normal tissue is the mainstay of treatment of this site, while the role of adjuvant chemotherapy remains uncertain. A case is presented of a 15 1/2-year-old male with a huge OS of the mandible. The boy underwent surgical resection of the mandible with immediate fibula free flap reconstruction and is alive and free of disease 6 1/2 years following unitial diagnosis. This case suggests that immediate bone reconstitution with vascularized grafts have good functional and morphological results for osteosarcoma of the lower jaw.