Culture conditions for the detection of allergen‐specific T‐cell reactivity in cord blood: Influence of cell number

Raised T‐cell proliferation of cord blood mononuclear cells (CBMC) in response to various ingestant and inhalant allergens has been reported in newborns, suggesting a prenatal allergen contact. In general, for in vitro proliferation assays a concentration of 50 × 10³ or 100 × 10³ cells/well are used. The aim of this study was to analyze whether cell concentration influences T‐cell reactivity in cord blood cells and to study differences of T‐cell reactivity triggered by inhalant and ingestant allergens. CBMC from 51 neonates (34 females: 22 with and 29 without a family history of allergy, i.e. FH+ or FH–) were incubated with interleukin‐2 (IL‐2), β‐lactoglobulin (β‐LG), ovalbumin (OVA), house dust mite allergen Dermatophagoides pteronyssinus (Der p 1), and timothy grass allergen Phleum pratense (Phl p 1) for 7 days. The cell concentration ranged from 62.5 × 10³ to 100 × 10³ cells/well. Proliferation was assessed by incorporation of [³H]‐thymidine and was expressed as counts per minute (c.p.m.). In unstimulated cells, a decreasing cell concentration paralleled a steep drop of background activity. In response to IL‐2, a decreasing cell concentration led to a slow decrease of c.p.m. The corresponding mean stimulation indices (SI) were 9, 32, 77, 47, and 21 for 100 × 10³, 50 × 10³, 25 × 10³, 12.5 × 10³, and 62.5 × 10³ cells/well, respectively. In addition, the highest number of positive proliferative responses to specific allergens were obscured at lower cell concentrations. For β‐LG, the maximal number of positive responses were obtained between 25 × 10³ (n = 44) and 12.5 × 10³ (n = 46) cells/well, for OVA at 25 × 10³ (n = 3) cells/well, for Der p 1 at 50 × 10³ (n = 5) cells/well, and for Phl p 1 between 25 × 10³ and 12.5 × 10³ (n = 5) cells/well. Positive proliferation in at least one of the tested assays was observed in 100% of samples in response to β‐LG, in 22% in response to Phl p 1, and in 14% in response to OVA and Der p 1. T‐cell reactivity did not differ between samples of newborns with or without a family history of atopy. Therefore, sensitivity of T‐cell proliferation measurement is highly influenced by background proliferation of unstimulated cells. Hence, proliferation assays with lower cell numbers unmask T‐cell reactivity in response to ingestant and inhalant allergens. We suggest the use of concentrations of 12.5 × 10³–50 × 10³ cells/well in proliferation experiments.     

Marked increase of peripheral blood myeloblasts following G-CSF therapy in a patient with acute lymphoblastic leukemia

A 9 year old boy with acute lymphoblastic leukemia (ALL) received recombinant human granulocyte colony-stimulating factor (rhG-CSF) and showed a marked increment of myeloblasts in the peripheral blood. He was administered repeated courses of intermediate-dose cytosine arabinoside (Ara-C) therapy (1500 mg/m², days 1–5) for frequent central nervous system (CNS) relapse of ALL. The peripheral white blood cell nadir was less than 1000/μL, so he was treated with rhG-CSF. A marked increment of peripheral blood blasts was noted 3–5 days after rhG-CSF treatment. These cells decreased with the appearance of mature myeloid cells and disappeared about 2 weeks after the start of treatment. These findings suggested that the blasts might have the ability to differentiate into mature myeloid cells. A control patient with repeated CNS relapse of ALL showed no increment of peripheral blood blasts after similar repeated courses of Ara-C without rhG-CSF treatment. Cultured peripheral blood blasts obtained from the present patient showed differentiation into mature myeloid cells by morphological studies and surface marker analysis. These findings indicate that the peripheral blood blasts drawn by G-CSF were not leukemic blasts but normal myeloblasts.     

Significant correlation between peripheral blood CD34+ cell count in children prior to aphaeresis and CD34+ cell yield following aphaeresis: A single‐center experience

Numerous adults’ studies demonstrated that preaphaeresis CD34+ cells significantly correlate with the number of CD34+ cells collected by the aphaeresis procedure. Equivalent studies in children are scarce. We studied retrospectively 92 aphaeresis procedures performed following chemotherapy (44) or in steady state (48) in 60 pediatric patients (40 males, 20 females), median age of 7.5 years. Aphaeresis procedures were performed using a SPECTRA Optica (TERUMOBCT) continuous flow cell separator. CD34+ cell concentrations were assessed using flow cytometry. A highly significant correlation between peripheral CD34 cell count on the day of aphaeresis and CD34 cell yield per kg (R² = .824, P < .0001) was demonstrated. A higher preaphaeresis CD34 cell count was demonstrated in patients with higher preaphaeresis white blood cell count, in patients with brain tumors, and in patients who received chemotherapy as part of their mobilization protocol. A threshold number of 20 peripheral CD34+ cell/μL was found to predict harvesting of 3 × 10⁶ stem cells/kg, and 30 peripheral CD34+ cell/μL for harvesting of 5 × 10⁶ stem cells/kg. This significant correlation between peripheral CD34 cell count and CD34 cell yield, and the threshold number of peripheral CD34 found to predict adequate harvesting can be useful in planning the optimal time for aphaeresis in children.     

Massive splenic infarction in children with sickle cell anemia and the role of splenectomy

Background

Massive splenic infarction (MSI) is a very rare condition. Few reports of splenic infarction of various etiologies including hematological and non-hematological causes have been published. On the other hand, MSI in patients with sickle cell anemia (SCA) is extremely rare. This report describes our experience with 15 children with SCA and MSI outlining aspects of presentation, diagnosis and management.

Patients and methods

The records of all children with MSI were retrospectively reviewed for age at diagnosis, sex, clinical features, precipitating factors, investigations, management and outcome.

Results

15 children (11 M: 4 F) with SCA were treated for MSI. Their mean age was 10.9 years (6–17 years). All presented with severe left upper quadrant abdominal pain. In nine, this was associated with nausea and vomiting. Three were febrile and all had a tender splenomegaly. Their mean hemoglobin was 8.2 g/dl (5.7–11.3 g/dl), mean WBC was 10.97 × 10³ mm^?3 (3.6 × 10³–22.3 × 10³ mm^?3) and mean platelet count was 263.3 × 10³ mm^?3 (40 × 10³–660 × 10³ mm^?3). In seven, there was a precipitating cause including high altitude in two, acute chest syndrome in two, septicemia in two and severe vasooclusive crisis in one. Abdominal ultrasound and CT scan confirmed the diagnosis of MSI which involved more than half of the spleen in 12 and whole spleen in 3. All were treated with IV fluids, analgesia and blood transfusion where appropriate. Eleven had splenectomy because of persistent abdominal pain, three developed splenic abscess and underwent splenectomy and one settled on conservative treatment. Histology confirmed the diagnosis of splenic infarction in 11 and infarction with abscess in the remaining 3.

Conclusion

MSI is extremely rare in children with SCA. It can develop spontaneously or precipitated by other factors namely high altitude, acute chest syndrome and severe stress. Most reported cases of splenic infarction are small in size, focal and can be treated conservatively. MSI, on the other hand, may necessitate splenectomy for persistent symptoms or in case of complications, such as abscess formation.     

Hematological alterations and thymic function in newborns of HIV-infected mothers receiving antiretroviral drugs

Objectives

To investigate the effects of antiretroviral (ARV) drugs on hematological parameters and thymic function in HIVuninfected newborns of HIV-infected mothers.

Study design

Cross sectional study.

Setting

Chiang-Mai University Hospital, Chiang-Mai, Thailand.

Participants/Patients

49 HIV-uninfected and 26 HIV-infected pregnancies.

Methods

Cord blood samples of newborns from HIV-uninfected and HIV-infected mothers were collected. Hematological parameters were measured using automatic blood cell count. T-cell receptor excision circles (TRECs) levels in cord blood mononuclear cells (CBMCs), CD4⁺ and CD8⁺ T-cells were quantified using real-time PCR.

Main Outcome Measures

Hemotological parameters and thymic function.

Results

Newborn of HIV-infected mother tended to have lower mean levels of hemoglobin than those of HIV-uninfected mother (137 ± 22 vs 146 ± 17 g/L, P = 0.05). Furthermore, mean of red blood cell (RBC) counts and hematocrit and median of TRECs in CD4+ T-cells in the newborns of the former were significantly lower than those of the latter [3.6 ± 0.7 vs 4.8 ± 0.6 × 10₁₂ cells/L, P <0.001; 0.40 ± 0.07 vs 0.46 ± 0.05 L/L, P <0.001 and 0.53 (IQR: 0.03–5.76) vs 13.20 (IQR: 2.77–27.51) × 10^?3 pg/μL, P = 0.02, respectively].

Conclusion

ARV drugs altered hematological parameters and thymic function (TRECs CD4⁺ T-cells) in HIV-uninfected newborns of HIV-infected mothers.     

A PILOT TRIAL OF TANDEM AUTOLOGOUS PERIPHERAL BLOOD PROGENITOR CELL TRANSPLANTATION FOLLOWING HIGH-DOSE THIOTEPA AND CARBOPLATIN IN CHILDREN WITH POOR-RISK CENTRAL NERVOUS SYSTEM TUMORS

This is a pilot study performed to determine the maximum tolerated number of courses of high-dose thiotepa and carboplatin with autologous peripheral blood progenitor cell (PBPC) transplantation in poor-risk pediatric central nervous system (CNS) tumor patients. Twelve patients were enrolled and a total of 24 PBPC transplants were performed. The median age was 7.7 years. All patients had CNS tumors: 4 relapsed CNS PNET, 2 high-risk PNET in first remission, 2 relapsed/progressive brainstem tumor, 2 relapsed/progressive anaplastic astrocytoma, 1 relapsed GBM, and 1 recurrent ependymoma. The regimen consisted of thiotepa 250 mg/m²/day × 3 days and carboplatin 400 mg/m²/day × 3 days. No toxic deaths occurred. All patients were hospitalized for a median duration of 17 days. The median number of CD34 cells infused was 5.4 × 10⁶/kg (2.1–29.7 × 10⁶/kg) per course. Median time to ANC >0.5 × 10⁹/L was 9 days, and platelets >20 × 10⁹/L was 13.5 days. Four patients came off protocol after only one course of PBPC (2 had tumor progression, 2 parental choice); 4 patients underwent two, and 4 patients three courses of PBPC. Major nonhematologic complications were mucositis that necessitated infusion of narcotics (11/24 courses), fever of unknown origin (12/24), documented infection (9/24), and hemorrhagic cystitis (3/24). TPN was administered during 22 of 24 courses with a median duration of 15 days. It is feasible to administer 2–3 courses of tandem high-dose thiotepa and carboplatin with PBPC transplant with prompt engraftment and manageable toxicities in pediatric CNS tumor patients.     

Use of Romiplostim for Primary Immune Thrombocytopenia in Children

Very little has been published on the use of romiplostim to treat primary immune thrombocytopenia (ITP), refractory to previous treatments, in children. The objective of this study was to determine its efficacy and safety in pediatric patients in a university general hospital. Retrospective, longitudinal observational study of pediatric patients on treatment with romiplostim. The principal efficacy variable was platelet count. Safety was evaluated by recording possible adverse reactions to the medication, monitoring the appearance of thrombosis, thrombocytopenia during dose reduction, hemorrhage, and myelodysplastic syndromes. Three patients in the authors’ center have been treated with romiplostim (subcutaneous [SC], initial dose: 1 μg/kg/week) for ITP refractory to various treatments: 1 with newly diagnosed ITP and 2 with chronic ITP. Patients were followed up for 27 to 39 weeks after starting treatment. Responses were achieved in 7 to 28 days, and complete responses were maintained for 37% to 91% of the follow-up period, with median platelet counts between 40 × 10³/μL and 215 × 10³/μL. The adverse reactions observed during follow-up were headache and asthenia in one patient and mucocutaneous bleeding after dose suspension in another one. With regard to effectiveness, the response in the 3 patients was varied. The drug was considered to be safe, as there were only mild adverse reactions. Although further studies and long-term follow-up are required, these results show that romiplostim could be considered an alternative to immunosuppressive therapies, such as rituximab, or splenectomy in refractory chronic ITP.     

Chronic myelomonocytic leukaemia in infancy: A case report

The case of an infant with the clinical and haematological features of chronic myelomonocytic leukaemia is reported. The infant presented with a peripheral blood monocyte count of 12,000 cell/mm³ (12.0 × 10⁹ cell/L) but no lymphadenopathy, hepatomegaly, nor splenomegaly. No treatment was given for 14 months during which time the monocytosis persisted. Myeloblasts then appeared abruptly in the peripheral blood reaching a peak of 1,500,000 cell/mm³ (1,500 × 10⁹/L). Cytotoxic Chemotherapy was initially successful but the blast soon became resistant and the child died. Chronic myelomonocytic leukaemia is usually associated with the elderly and the significance of this case is discussed.     

Autologous bone marrow transplantation with 4-hydroperoxycyclophosphamide-purged marrow for acute lymphoblastic leukemia

Ten patients age 13–52 years with acute lymphoblastic leukemia (ALL) in first complete remission (CR) (1), second CR (6), or relapse (3) were treated with cyclophosphamide 50 mg/kg daily × 4 and total body irradiation 3 Gy daily × 4 followed by infusion of autologous marrow purged with 4-hydroperoxycyclophosphamide (4-HC) at 100–120 μg/mL. The patients transplanted in relapse also received 2 mg vincristine and corticosteroids. All marrows were harvested while patients were in CR. The nucleated marrow cell dose was 3.5 ± 0.7 × 10⁸/kg, and the CFU-GM content of the transplant was 0.4 ± 0.5 × 10³/kg after purging with 4-HC. Median time from transplantation is 48 months. The neutrophil count (ANC) exceeded 1.0 × 10⁹/L at a median of 26 days and platelets exceeded 50 × 10⁹/L at 34 days. All patients were in remission after transplantation (ABMT). Five patients relapsed 2–9 months after ABMT (actuarial rate 60%). Three patients are alive and in CR at 17+, 43 +, and 54+ months after ABMT. Purging marrow with 4-HC did not adversely affect engraftment, but it is not clear if the high relapse rate was due to incomplete ex vivo purging or inadequacy of the conditioning regimen.     

Lack of CD4+CD25+FOXP3+ regulatory T cells is associated with resistance to intravenous immunoglobulin therapy in patients with Kawasaki disease

The aim of this study was to investigate changes in CD4⁺CD25⁺FOXP3⁺ regulatory T cells (Tregs) throughout the clinical course of Kawasaki disease (KD) and correlations with response to intravenous immunoglobulin (IVIg) therapy. Participants comprised 18 patients who fulfilled the diagnostic criteria for KD and 20 healthy subjects. Expressions of CD25 and FOXP3 among all CD4⁺ T cells in peripheral blood mononuclear cells were analyzed by flow cytometry before and 7 and 30 days after IVIg therapy. Before treatment, percentages of CD4⁺CD25⁺FOXP3⁺ Tregs among total CD4⁺ Tregs were significantly lower among KD patients (4.19 %; range, 0.16–8.11 %) than among healthy subjects (7.32 %; 4.18–13.42 %; P?=?0.0001). Both percentages and absolute numbers of CD4⁺CD25⁺FOXP3⁺ Tregs on day?7 after IVIg therapy were significantly increased compared with values before treatment (8.02 % (range, 0.51–12.6 %) vs. 4.19 % (range, 0.16–8.11 %), P?=?0.0005; 93.25/?μL (range, 6.67–258.05) vs. 41.85/?μL (range, 0.44–160.62), P?<?0.0001, respectively). Moreover, percentages and absolute numbers of CD4⁺CD25⁺FOXP3⁺ Tregs before treatment were significantly lower in the IVIg-resistant group than in the IVIg-sensitive group (0.18 % (range, 0.16–3.34 %) vs. 4.52 % (range, 2.8–8.11 %), P?=?0.0022; 0.68/μL (range, 0.44–53.81) vs. 51.66/μL (range, 2.88–160.62), P?=?0.0098, respectively). The frequency of CD4⁺CD25⁺FOXP3⁺ Tregs in four of the five IVIg-resistant patients at diagnosis was more than 3 standard deviations below that in healthy subjects. Two of these four patients displayed coronary abnormalities, and one of these two patients developed coronary aneurysm. Conclusion: Lack of CD4⁺CD25⁺FOXP3⁺ Tregs before treatment may predict resistance to IVIg therapy in patients with KD.     

Immunodiagnosis of childhood all: Problems associated with the use of peripheral blood alone

The immunologic classification of acute lymphoblastic leukemia (ALL) based solely on peripheral blood (PB) cell phenotypes may lead to conflicting results. This was demonstrated by the simultaneous assay of five immunologic markers on PB and bone marrow (BM) cells from 13 children with untreated ALL. We assayed erythrocyte (E) rosettes at 4°C and 37°C, presence of membrane Ig(mIg), and binding of antisera raised against thymus (T), and E^? ALL blasts, respectively. At diagnosis, the PB of these children contained > 90% lymphoid cells with 0–48% E rosettes and 1-84% cells with T antigen(s). Of 7 children with WBC < 10,000/cu mm there were 4 who had 20% or more E rosettes and T-antigen-positive cells. Of 6 children with WBC > 10,000/cu mm there were only 2 who had more than 20% E rosettes and T-antigen-positive cells. Based on examination of PB alone, six children may have been classified as having T-like ALL. However, these results were due to the presence of circulating normal T lymphocytes, and assay of BM cells established that only one of the 13 children had T-like ALL and none had B-cell ALL. Bone marrow blasts from 12 patients did not form rosettes at 37°C, did not have mIg, and did not react with anti-T serum. A high proportion of BM blasts from these 12 patients (39-96%) did react with antiserum against E^? ALL blasts. Of these 12 patients 11 had a higher proportion of E^? ALL antiserum-positive blasts in the BM than PB. Thus, immunologic classification of ALL should be based on the study of BM blasts, or both PB and BM cells.     

Infliximab regulates monocytes and regulatory T cells in Kawasaki disease

Background

The effect of infliximab (IFX ) on immune cells has not been fully reported in Kawasaki disease (KD ). To investigate the mechanism of IFX in KD , we examined changes in the abundance of CD 14⁺CD 16⁺ activated monocytes, regulatory T cells (T_reg) cells, and T‐helper type 17 (Th17) cells following treatment with IFX .

Methods

We collected peripheral blood from patients with i.v. immunoglobulin (IVIG )‐resistant KD and analyzed absolute CD 14⁺CD 16⁺ monocyte, T_reg (CD 4⁺CD 25⁺FOXP 3⁺) and Th17 cell (CD 4⁺IL ‐17A⁺) counts on flow cytometry. We also measured changes in serum soluble interleukin (IL )‐2 receptor (IL ‐2R), IL ‐6, and tumor necrosis factor (TNF )‐α on enzyme‐linked immunosorbent assay.

Results

T_reg cells and Th17 cells significantly increased after IFX treatment compared with baseline (126 ± 85 cells/μL vs 62 ± 53 cells/μL, P < 0.01; 100 ± 111 cells/μL vs 28 ± 27 cells/μL, P < 0.05, respectively). In contrast, in a subgroup of patients with CD 14⁺CD 16⁺ monocytes above the normal range before IFX , the CD 14⁺CD 16⁺ monocytes significantly decreased following IFX treatment (72 ± 51 cells/μL vs 242 ± 156 cells/μL, P < 0.05).. Serum TNF ‐α did not change, but soluble IL ‐2R and IL ‐6 decreased after IFX treatment.

Conclusion

IFX could downregulate activated monocytes and upregulate T_reg cells towards the normal range. IFX treatment thus contributes to the process of attenuating inflammation in KD .

     

Imerslund-Grasbeck Syndrome Coexisting with P-Thalassemia Trait

A 9-year-old female patient with Imerslund-Gräsbeck syndrome and heterozygosity for β-thahsemia i presented.At admission the hemoglobin (Hb) was 7.2 g/dL; reticulocytes,0.2%; red blood cell count (RBC),2.3 × 10¹² /L; mean corpuscular volume (MCV),80 fL; hemoglobin A₂ (HbA), 4.3%; fetal hemoglobin intervening sequence (IVS) (HbF),1.9%.In the bone marrow aspiration smear, megaloblastic changes were obseroed; the Schilling test was compatible with mahbsorption.DNA anulysi revealed the presence of heterozygosity for the IVS-I-110 type of β-thahsemia mutation.Five months after treatment with vitamin B₁₂, Hb was found to be 12.8 g/dL; RBC,5 × 10¹²/L; MCV,63 fL.     

COAGULOPATHY AFTER SPIDER BITES TO A 6-YEAR-OLD BOY

A 12-month-old Caucasian female with a history of recurrent ear infections presented to the emergency room with petechiae, severe thrombocytopenia (4000/μL), and abnormally prolonged activated partial thromboplastin time. Further autoantibody investigation detected antinuclear antibodies, anti-double-stranded DNA, and antiphospholipid antibodies. Platelet count, in response to intravenous immunoglobulin infusion, increased to more than 100 × 10³ plt/μL. At 6-month follow-up, no positive autoantibodies were detected.     

Clinical and hematological profile in a newborn cohort with hemoglobin SC

Objectives

Hemoglobin SC is the second most common variant of sickle-cell disease worldwide, after hemoglobin SS. The objectives of the study were to describe the clinical and laboratory characteristics of hemoglobin SC disease in children from a newborn screening program and treated at a blood center.

Treatment of high-risk acute lymphoblastic leukemia in children using the AL851 and ALHR88 protocols: A report from the Kyushu-Yamaguchi children cancer study group in Japan

A total of 125 children, who were diagnosed as having high-risk acute lymphoblastic leukemia (ALL), were treated with two consecutive protocols designated as AL851 (1985–1988) and ALHR88 (1988–1990). All patients received induction therapy consisting of vincristine (VCR), prednisolone (PSL), daunorubicin (DNR), and l-asparaginase (l-Asp). In the ALHR88 protocol, the patients whose blasts in the bone marrow (BM) were ≥25% on day 14 of induction therapy and who were classified into T-cell type received additional cytosine arabinoside (AraC). After consolidation with intermediate-dose methotrexate (MTX), reinduction therapy including VCR, dexamethasone, and adriamycin followed by high-dose AraC was done for all patients. Intrathecal MTX and 24Gy of cranial irradiation were used to prevent central nervous system leukemia. A maintenance therapy consisting of 6-mercaptopurine, cyclophosphamide, MTX, DNR, VCR, and AraC was administered for 3 years after achieving a complete remission (CR). CR was achieved in 51/55 (92.7%) for AL851 and 68/70 (97.1%) for ALHR88. The 5-year event-free survival rates were 49.1 ± 6.7% in AL851 and 62.5 ± 6.1% in ALHR88. The factors related to a poor prognosis were a high initial leukocyte count of greater than 50 × 10⁹/L (P < 0.001), an L2 morphology of leukemic cells by FAB classification (P = 0.009), the chromosomal abnormality (P = 0.004) and high residual leukemic cells in BM (≥25%) on day 14 of induction therapy (P < 0.001). Taking these factors into consideration, more intensive protocols were started in 1990 for the patients with high-risk ALL. © 1996 Wiley-Liss, Inc.     

Childhood T-cell acute lymphoblastic leukaemia expressing “Ia-like” antigen: A case report

A 4-year-old girl presenting with vomiting, abdominal pain, and renal failure was found to have gross hepatosplenomegaly, a renal mass, and bilateral pleural effusions. A diagnosis of acute lymphoblastic leukaemia (ALL) was suggested by a peripheral white cell count (WCC) of 119,000 × 10⁶/mm³, 57% blasts, 22% lymphocytes, and confirmed by bone marrow examination. Lymphocyte surface marker studies at diagnosis enabled classification as a T-ALL, with a significant proportion of the T cells also bearing receptors for the third component of complement (C3). Seventy-two percent of the peripheral blood mononuclear cells reacted with anti-Ia monoclonal antibody (FMC 4), and a smaller proportion (25%) carried receptors for the Fc portion of IgG. The T-classification of this ALL was verified at central nervous system (CNS) relapse and at a subsequent nodal relapse. Double-marker studies on cells from the infiltrated lymph node prepared in suspension confirmed the presence of Ia-positive T cells. The Ia marker is usually a useful discriminant between T and non-T cells in normal and ALL cell populations. The case described here highlights the need for a panel of markers to be used in classification of childhood ALL and supports the suggestion that there is a distinct subtype of Ia-positive T-ALL.     

Autologous cord blood transplantation for metastatic neuroblastoma

Auto‐SCT is a common approach for metastatic neuroblastoma with the intention to rescue hematopoiesis after megadose chemotherapy. PBSC or BM is the usual stem cell source for auto‐SCT. Auto‐CBT for neuroblastoma has very rarely been performed. Currently, case reports are available for two patients only. We performed 13 auto‐SCTs for high‐risk neuroblastoma from 2007 to 2013, including four cases of metastatic neuroblastoma aged 11–64 months treated with auto‐CBT. All four patients had partial or CR to upfront treatments before auto‐CBT. Nucleated cell dose and CD34+ cell dose infused were 2.8–8.7 × 10⁷/kg and 0.36–3.9 × 10⁵/kg, respectively. Post‐thawed viability was 57–76%. Neutrophil engraftment (>0.5 × 10⁹/L) occurred at 15–33 days, while platelet engraftment occurred at 31–43 days (>20 × 10⁹/L) and 33–65 days (>50 × 10⁹/L) post‐transplant, respectively. There was no severe acute or chronic complication. Three patients survived for 1.9–7.7 yr without evidence of recurrence. One patient relapsed at 16 months post‐transplant and died of progressive disease. Cord blood may be a feasible alternative stem cell source for auto‐SCT in patients with stage 4 neuroblastoma, and outcomes may be improved compared to autologous PBSC or BM transplants.     

rhG-CSF在小儿ANLL强烈化疗中的应用

为探讨rhG-CSF对小儿ANLL强烈化疗后粒细胞缺乏的疗效，采用AAE方案（ADM、Ara-C、VP(16)或VM(26)），化疗后当WBC＜1×109/L或ANC＜0．5×109/L时，给予rhG-CSF200μg/m2·d（5～10μg/kg·d），皮下注射，一般连续5～10天。本文15例ANLL，用rhG-CSF30例次。用rhG-CSF前，WBC平均0．78×109/L、ANC0．15×109/L。用rhG-CSF后，平均6．5天WBC升至＞3×109/L、ANC升至＞1×109/L。粒细胞恢复时间与对照组相比明显缩短（P＜0．01）。骨髓复查未见原始细胞增多或复发。rhG-CSF有促进强烈化疗所致骨髓抑制和粒细胞缺乏的恢复，但未见骨髓原始细胞增多和白血病复发。     

Allogeneic peripheral blood stem cell transplant in children

Allogeneic peripheral blood stem cell (PBSC) transplant has recently been introduced for the treatment of hematological malignancies. As the data were limited mainly to adult patients, this study aimed to assess the feasibility and safety of this procedure in pediatric patients and donors. Eleven children aged 2–16 years received allogeneic PBSC transplant for acute lymphoblastic leukemia (n = 4), acute myeloid leukemia (n = 1), myelodysplastic syndrome (n = 1), severe aplastic anemia (n = 3), and thalassemia (n = 2). Nine donors were human leukocyte antigen (HLA)-identical siblings and the other two were one antigen mismatched family members. Eight donors were younger than 18 years old (10 months to 17 years). Donors were primed with granulocyte colony-stimulating factor (G-CSF) at 10–16 μg/kg for 4–5 days. Aphereses were performed on 1 or 2 consecutive days, and the patients received a mean of 14.4 × 10⁸/kg nucleated cells, 6.9 × 10⁶/kg CD34 cells, and 6.9 × 10⁸/kg T cells. All patients achieved neutrophil counts of >0.5 × 10⁹/l at a median of 16 days. Nine patients achieved platelet counts of >20 ×10⁹/l at a median of 13 days. Grade II acute graft vs. host disease (GVHD) occurred in only one patient. Chronic GVHD was not observed in the seven patients with follow-up of more than 3 months. Eight patients remained in continuous complete remission after transplant ranged from 2 to 26 months. Allogeneic PBSC transplant appears safe in pediatric patients and donors, and it seems not to be associated with increase of acute GVHD or chronic GVHD. Med. Pediatr. Oncol. 30:147–151, 1998. © 1998 Wiley-Liss, Inc.