Ryuji Nakagawa

Analysis of maternal and neonatal factors that influence the nucleated and CD34+cell yield for cord blood banking

BACKGROUND: It would be beneficial to be able to predict the cord blood (CB) cell yield from volunteer donors before cell processing.

Pulsed wave Doppler tissue echocardiography assessment of the long axis function of the right and left ventricles during the early neonatal period

Objective: To assess the long axis function of both ventricles during the early neonatal period by using pulsed wave Doppler tissue (PWDT) echocardiography. Design: PWDT echocardiography was recorded from the lateral sites of the mitral and tricuspid annuluses and the tip of interventricular septum in 130 neonates within 24 hours after birth (day 0 group), in 135 neonates 1–7 days after birth (day 1–7 group), and in 131 healthy children (children group). Results: Peak systolic motion velocity (Sw) of the three ventricular walls positively correlated with the number of days after birth (p < 0.005). Compared with the children group, in neonates Sw in the right ventricle and peak early diastolic motion velocity (Ew) and peak atrial systolic motion velocity in the interventricular septum were lower than in the remaining two walls (p < 0.0005, p < 0.0001, and p< 0.0001, respectively). Although peak mitral and tricuspid flow velocities during early diastole (E) correlated with the number of hours after birth in the day 0 group, there was no significant change in the Ew of either ventricle. The E:Ew ratio of both ventricles was significantly higher in both neonate groups than in the children group (p < 0.001). The E:Ew ratio of the left ventricle was higher in the day 0 group than in the day 1–7 group (p < 0.005). Conclusions: The two ventricles differ in their normal PWDT echocardiographic values and in the parameter change after birth during the early neonatal period, which may reflect differences in ventricular adaptation after birth.

Factors associated with granulocyte colony-stimulating factor-induced peripheral blood stem cell yield in healthy donors

Background and Objectives  Poor collection results are a clinical problem in granulocyte‐colony stimulating factor (G‐CSF)‐induced peripheral blood stem cell (PBSC) collection in healthy donors. It would be beneficial to be able to predict the PBSC yield from allogeneic donors before mobilization or harvesting.

Tendency toward atopy in Kawasaki disease

Abstract To evaluate the possible trend towards developing allergic disease in children who had suffered from Kawasaki disease (KD), we evaluated data related to allergy that were collected by parental questionnaire on 1,165 children who had suffered from KD. Comparisons were made with 5,825 sex- and age-matched control children. The incidence of household pets and of cigarette smoking in the family were significantly lower in the children with a history of KD than in those of control children. A family history of allergy was significantly more common in the children with a history of KD (71%) versus the controls (56%) (P < 0.001). The incidence of atopic dermatitis and of allergic rhinitis was significantly higher (by approximately 1.7 times) in the KD children versus the control group (P < 0.01), even in subgroups with no family history of allergy. Conclusion We suggest that a genetic predisposition to atopy may be associated with a susceptibility to KD. Patients with KD tended to develop atopic dermatitis and allergic rhinitis.

Low numbers of megakaryocyte progenitors in grafts of cord blood cells may result in delayed platelet recovery after cord blood cell transplant.

Delayed platelet recovery is an inherent problem with cord blood cell transplantation (CBCT). To investigate this problem, the number of human megakaryocyte (MK) progenitor cells in cord blood (CB; n = 24) was measured and compared with that in G‐CSF‐mobilized peripheral blood stem cells (PBSC; n = 25). The median numbers of colony‐forming units for MK (CFU‐MK) that were detected by a serum‐free assay system in CB and peripheral blood (PB) were 26 (range, 6‐102)/105 nucleated cells (NC) and 37 (2‐540)/105 mononuclear cells (MNC), respectively. The numbers of colony‐forming units for granulocyte/macrophage (CFU‐GM) were 88 (33‐241)/105 NC in CB and 138 (6.3‐1,250)/105 MNC in PB. The frequencies of CD34+ cells in CB and PB were, respectively, 0.44% (0.10‐1.07) and 0.98% (0.05‐20.8). The numbers of CFU‐MK in CB and PBSC were correlated with those of CD34+ cells. The estimated number of infused CFU‐MK in CBCT was 1/15 that of PBSC transplantation (PBSCT), based upon the above data and the widely used standard doses for both types of transplants. Further, the numbers of infused CFU‐MK in patients who received allogeneic PBSCT at our institute were inversely correlated with the speed of platelet recovery. These data indicate that delayed platelet recovery after CBCT is simply due to the low number of CFU‐MK contained in grafts.

Cryopreservation of mobilized blood stem cells at a higher cell concentration without the use of a programmed freezer

Cryopreservation of peripheral blood stem cells (PBSC) mobilized by chemotherapy combined with or without granulocyte colony-stimulating factor (G-CSF) is an essential part of procedure for anti-cancer strategies. We evaluated whether a higher cell concentration (2×108/ml) without the use of a programmed freezer was acceptable for the storage of mobilized PBSC in an autologous setting. Mobilized PBSC were enriched to mononuclear cells (MNC) by Percoll separation and then frozen at cell concentrations of 2–5×107/ml (group I, n=20) or 2×108/ml (group II, n=44) without the use of a programmed freezer using 5% DMSO, 6% hydroxy ethyl starch, and 4% autologous serum or human albumin. CD34+ cells purified by ISOLEX300 were frozen at 2×107/ml (group III, n=22) using the same method. The median recovery rates of CD34+ cells and CFU-GM were, respectively, n.d. (not determined) and 88% in group I, 103 and 64% in group II, and 98 and 53% in group III. There was a statistical significance between the recovery rate of CFU-GM in group III and that in group I (p=0.02). The median percentage of cell viability after thawing in each group was 89, 87, and 75%, respectively. The median numbers of days after PBSCT to achieve a WBC of >1.0×109/l, an absolute neutrophil count of >0.5×109/l, and a platelet count of >50×109/l were, respectively, 11, 11 and 15 in group I; 12, 12 and 16 in group II; and 12, 12 and 27 in group III. These results suggest that enriched MNC from mobilized PBSC could be frozen at a higher cell concentration (2×108/ml) without the use of a programmed freezer, leading to reduction of the toxicities associated with infusion of thawed cells and of costly space required for cell storage.

Successful unrelated cord blood transplantation in an infant with Wiskott-Aldrich syndrome following recurrent cytomegalovirus disease.

We describe successful unrelated cord blood transplantation in a 14-month-old boy with Wiskott-Aldrich syndrome. He had been suffering from recurrent cytomegalovirus (CMV) pneumonia. Ganciclovir was given pretransplantation and posttransplantation, and CMV antigenemia was monitored as a marker of reactivation. The conditioning regimen was cyclophos-phamide, busulfan, and antithymocyte globulin. The patient received an HLA 1-locus-mismatched cord blood unit, and the total number of infused nucleated cells was 9.0 X 107/kg. Neutrophil engraftment was achieved on day +20, and a platelet count greater than 50 X 109/L was achieved on day +51. A normal lymphoproliferative response to phytohemagglutinin mitogen was detectable 7 months posttransplantation. Long-term use of ganciclovir prevented CMV reactivation and did not compromise engraftment.

Intra-apheresis recruitment of blood progenitor cells in children

BACKGROUND: Determination of the optimal duration of apheresis requires a careful examination of blood progenitor cell (BPC) kinetics during apheresis. Intra‐apheresis recruitment of BPCs should be evaluated.

Effect of granulocyte colony-stimulating factor on bone metabolism during peripheral blood stem cell mobilization.

Granulocyte colony—stimulating factor (G-CSF) has been shown to affect the biochemical markers of bone metabolism, including serum bone alkaline phosphatase (BALP), serum osteocalcin, and urine deoxypyridinoline. To determine the association between bone resorption and formation and the G-CSF—induced mobilization of peripheral blood stem cells (PBSC), we examined these markers during mobilization in 19 healthy donors. The average (± SEM) serum BALP level before treatment was 81.6 ± 17.0 IU/dL, and the level increased significantly to 117.7 ± 15.8 IU/dL on day 5 of G-CSF administration (P < .0001). The urine deoxypyridinoline level before treatment was 12.3 ± 2.4 nmol/mmol creatinine, and this level also increased significantly to 19.4 ± 3.0 nmol/mmol creatinine on day 5 of G-CSF administration (P < .0001). In contrast, the average level of serum osteocalcin significantly decreased from 8.07 ± 2.88 ng/mL to 1.53 ± 0.18 ng/mL on day 5 (P = .0353). During G-CSF administration, we also studied the serum levels of various cytokines (IL-lβ, osteoclastogenesis inhibitory factor [OCIF], IL-6, tumor necrosis factor a, transforming growth factor β, interferon-γ, macrophage colony—stimulating factor) related to bone metabolism. Only the kinetics of OCIF were significantly affected. The serum level of OCIF increased immediately after the start of G-CSF administration and remained high during G-CSF administration. These results demonstrate that high-dose G-CSF affects bone metabolism and that OCIF may play a role in bone metabolism. Consistent with the notion that G-CSF affects bone metabolism, a significant correlation was observed between CD34+ cell yield and the increase in urine deoxypyridinoline but not for the changes in serum BALP and osteocalcin levels. This result suggests that bone resorption is either directly or indirectly related to the mobilization of PBSC by G-CSF.

Autoimmune hepatitis following allogeneic PBSCT from an HLA-matched sibling

Summary:A 7-year-old boy with acute lymphoblastic leukemia (ALL) in second remission received an allogeneic PBSCT from his HLA-matched sister. Acute grade II graft-versus-host disease (GVHD) resolved with corticosteroids. Chronic GVHD in the skin and oral mucosa at around day 60 responded to corticosteroids and cyclosporin A. At 6 months after the transplant, he developed hepatic dysfunction with elevated serum transaminases and gamma-globulin. Liver biopsy revealed chronic inflammation with lymphocytes and plasma cells in portal areas without destruction of bile ducts, suggesting autoimmune hepatitis. While rare, autoimmune hepatitis should be considered a potential long-term complication in patients with hepatic dysfunction in the late post-transplant phase.