Infectivity of HBV DNA positive donations identified in look-back studies in Hyogo-Prefecture, Japan.

AIMS/OBJECTIVES: To clarify transfusion incidence of hepatitis B virus (HBV) infected blood negative for mini pool-nucleic acid amplification testing (MP-NAT). BACKGROUND: Japanese Red Cross (JRC) blood centres screen donated blood to avoid contamination with HBV. However, a low copy number of HBV may be overlooked. METHODS/MATERIALS: In Hyogo-Prefecture, JRC blood centres screened 787 695 donations for HBV from April 2005 to March 2009. Of these, 685 844 were donations from the repeat donors. To detect the donors with HBV, serological tests, MP-NAT and/or individual donation (ID)-NAT were performed. To detect the recipients with transfusion-transmitted HBV infection (TTHBI), serological analysis and/or ID-NAT were performed. RESULTS: In this study, 265 of the 685 844 repeat donations were serologically and/or MP-NAT positive for HBV. Their repository samples from the previous donation were examined in a look-back study; 13 of the 265 repository samples proved ID-NAT positive. Twelve recipients were transfused with HBV-infected blood components derived from 10 of the 13 HBV-infected donors. Only 1 of the 12 recipients was identified as TTHBI case. Seven of the 12 recipients escaped from our follow-up study and 4 recipients were negative for HBV during the observation period. CONCLUSION: On the basis of the look-back study among the repeat donors in Hyogo-Prefecture, Japan, donations with HBV-infected blood negative for MP-NAT occurred with a frequency of 13 in 685 844 donations (∼1/53 000 donations). However, more than half of the recipients transfused with HBV-infected blood negative for MP-NAT could not be followed up. It is necessary to establish a more cautious follow-up system.

Acute non-hemolytic transfusion reactions and HLA class I antibody: advantages of solid phase assay compared with conventional complement-dependent assay.

To evaluate the specific reactivity of HLA Class I antibodies (HLA-I Abs) in acute non-hemolytic transfusion reactions (ANHTRs) using solid phase assays (SPAs) and conventional complement-dependent lymphocyte cytotoxicity test (LCT). ANHTRs are major issues in transfusion medicine. Anti-leukocyte antibodies have been implicated as one of the causative agents of transfusion-related acute lung injury (TRALI) and febrile reaction. Antibodies to HLA Class I and/or Class II (HLA Abs) have been intensively studied using SPAs for TRALI, but not for febrile reaction. About 107 patients and 186 donors associated with ANHTRs were screened for HLA Abs by SPAs such as enzyme-linked immunosorbent assay (ELISA) and the Luminex method. When HLA-I Ab was detected, its specific reactivity was evaluated by comparing its specificity identified by the Luminex method using recombinant HLA molecules and cognate HLA antigens (Ags), as well as LCT with or without anti-human globulin (AHG). The incidences of HLA Abs were as high as 32.7% of patients' serum samples and 16% of donors' serum samples. The incidence of HLA-I Abs did not differ significantly between cases of febrile and allergic reactions. However, HLA-I Abs associated with febrile reaction showed a significantly higher rate of possessing specific reactivity to cognate HLA Ags than those associated with allergic reactions. In addition, the Luminex method enabled the detection of HLA-I Abs much earlier than AHG-LCT in serum samples from a patient with febrile reaction and platelet transfusion refractoriness (PTR). SPAs seem more useful than AHG-LCT for evaluating reactivity of antibodies in ANHTR cases.

Intensification of chemotherapy using block therapies as consolidation and reinduction therapies for acute lymphoblastic leukemia during childhood.

Between April 1994 and March 1997, 143 children (age range, 1-15 years) with newly diagnosed acute lymphoblastic leukemia (ALL), except for those patients with t(9;22), were treated according to protocol-94 of the Osaka Childhood Leukemia Study Group. In this trial, the intensity of chemotherapy was enforced in the consolidation and reinduction phases by introducing AML-type block therapies consisting of concentrated administration of 4 to 6 drugs during 5 or 6 days. For patients in the higher risk groups, rotational combination chemotherapy was introduced following the early phase. A total of 124 children with B-cell precursor ALL (B-pre ALL) were classified into 3 groups, the ultrahigh-risk group (UHRG) (15 patients), the high-risk group (HRG) (61 patients), or the standard-risk group (SRG) (48 patients), based on age. leukocyte count, immunophenotype, central nervous system leukemia, response to treatment, and selected chromosomal abnormalities. The complete remission rate was 93%, and the 6-year event-free survival (EFS) rate was 79%+/-4%. EFS rates for the UHRG, HRG, and SRG groups were 67%+/-12%, 80%+/-6%, and 81%+/-6%, respectively. Nineteen patients with T-cell ALL were treated with the protocol for the UHRG. Thirteen patients (68%) attained complete remission, and the 6-year EFS rate was 55%+/-12%. Thus, intensification of chemotherapy improved the EFS rate and AML-type block therapies appeared to be effective as the consolidation and reinduction therapies for B-pre ALL.

[CD34+ progenitor cell transplantation from HLA-mismatched donors to two patients with chronic active Epstein-Barr virus infection].

We report two boys with chronic active Epstein-Barr virus infection (CAEBV) refractory to conventional chemotherapy, who received HLA-mismatched allografts of CD34-positive progenitor cells from their fathers. One patient developed veno-occlusive disease (VOD) of the liver on day 18 after transplantation and died on day 26. The other patient received the allograft during partial remission. Although he suffered recurrent infections due to Streptococcus viridans, he is now doing well 23 months after transplantation. CAEBV refractory to chemotherapy is considered to be a fatal EBV-infected T/NK-cell lymphoproliferative disease, and our experiences suggest that CD34-positive progenitor cell transplantation for patients with CAEBV lacking HLA-matched donors may be a feasible and useful treatment. However, the timing of transplantation is considered to be critical, and should be performed when the patient is in good clinical condition.

Mosaic variegated aneuploidy with multiple congenital abnormalities: homozygosity for total premature chromatid separation trait.

Separation of chromatids of all mitotic chromosomes, here called total premature chromatid separation (total PCS), was observed in 67 to 87.5% of repeated cultures of peripheral blood lymphocytes from two unrelated infants. Also noted was a variety of mosaic aneuploidies, especially trisomies, double trisomies, and monosomies, to be called mosaic variegated aneuploidy. The infants both showed severe pre- and postnatal growth retardation, profound developmental retardation, uncontrollable seizures, severe microcephaly, hypoplasia of the brain, Dandy-Walker anomaly, abnormal facial appearance, and bilateral cataract. Patient 1, a girl, in addition had a cleft palate, multiple renal cysts, and Wilms tumor of the left kidney. Whereas patient 2, a boy, had ambiguous external genitalia. They both died within 2 years of age. In the two families of the infants, their parents and three other members showed 2.5 to 47% lymphocytes with total PCS but without mosaic variegated aneuploidy or phenotypic abnormalities. Another 10 relatives studied showed 0 to 1% cells with total PCS and so were judged negative for the total PCS trait. It was deduced that the total PCS trait in the two families was transmitted in an autosomal-dominant fashion, and the two affected infants were homozygous for the trait.

[MR imaging in patients with bone marrow metastases of neuroblastoma].

Sixty patients with neuroblastoma or ganglioneuroblastoma underwent MRI 87 procedures to detect and follow up bone marrow metastases. The detection of bone marrow metastases by MRI was evaluated with aspiration cytology. Relationships were investigated between bone metastases and bone marrow metastases that were morphologically classified as nodular and diffuse by MRI findings. The usefulness of MRI was also studied in evaluating the effect of chemotherapy. MRI showed bone marrow metastases in all of 17 patients, but aspiration cytology proved metastases in only 7 of 17. Bone metastases were proven by bone scintigraphy or bone X-ray in 5 of 15 patients, and only diffuse bone marrow metastases were accompanied with bone metastases. After or during chemotherapy, 10 patients were examined by MRI more than two times. Bone marrow metastases disappeared in 9, decreased in size in 3, did not change in one and recurred at another site in one. It was observed from MRI findings that in the early stage, bone marrow metastases are nodular, but along with progress of the disease, they become diffuse and invade the bone cortex. If chemotherapy was effective, bone marrow metastases were decreased in size or disappeared completely.

Expression of c-ErbB2 in human neuroblastoma tissues, adrenal medulla adjacent to tumor, and developing mouse neural crest cells.

We have examined the expression of c-ErbB2 in primary neuroblastoma tissues, mouse neural crest-derived tissues, and human adrenal gland adjacent to neuroblastoma tissue and of age-matched controls. c-ErbB2 expression was observed in approximately 60% of cases analyzed, and there were two staining patterns; one showed focal and cytoplasmic and the other showed diffuse and membrane staining patterns. The expression of c-ErbB2 in neuroblastoma tissues was confirmed by reverse transcription polymerase chain reaction and Western blot analysis. Diffuse and membrane staining of c-ErbB2 was well correlated with high urinary catecholamine secretion. In mouse tissues, cytoplasmic expression of c-ErbB2 was observed in immature peripheral neurons and adrenomedullary cells. In mature neurons, the immunoreactivity was confined to the plasma membrane. These results suggest that the expression of c-ErbB2 in neuroblastoma reflects the phenotype of developing peripheral neurons. Postnatal human and mouse adrenomedullary cells lacked c-ErbB2 immunoreactivity, although apparently normal adrenomedullary cells adjacent to neuroblastoma tissues showed strong cytoplasmic expression of c-ErbB2. It is not known whether the phenotypic conversion of adjacent adrenal medullary cells had occurred before or after tumor progression at present.

Effect of recombinant human granulocyte-colony stimulation factor (rhG-CSF) on immune system in pediatric patients with aplastic anemia.

To determine the effect of recombinant human granulocyte-colony stimulation factor (rhG-CSF) on the immune system, serum immunoglobulins, lymphocyte subsets, and serum cytokines were analyzed in eight pediatric patients with aplastic anemia (AA) during 8-week rhG-CSF therapy. The rhG-CSF was administered either subcutaneously (200 micrograms/m2 x 4 weeks, followed by 400 micrograms/m2 x 4 weeks) or intravenously (400 micrograms/m2 x 4 weeks, followed by 800 micrograms/m2 x 4 weeks). In response to rhG-CSF therapy, neutrophil counts exceeded the pretreatment counts by twofold during the first week except for one case that did not attain twofold increase until day 41. While serum IgG and IgA were not affected, serum IgM was elevated during treatment in six of the eight cases to more than 1.2-fold basal levels (P < 0.04); however, there was no increase in serum interleukin (IL)-6 and interferon-gamma levels. On the other hand, CD56 positive NK cells significantly dropped from 7.7% to 4.5% (P < 0.02). These results indicate that systemic administration of rhG-CSF affects not only the neutrophil count, but also serum IgM levels and the natural killer cell population in patients with AA.

Heterogeneous origin of established non-T,non-B cell lines from two adolescent patients with acute lymphoblastic leukemia.

Three leukemic cell lines were established from the bone marrow cells of two adolescents with non-T,non-B acute lymphoblastic leukemia (ALL) at relapse. Two cell lines from a 14-year-old girl and one from an 11-year-old boy were designated as KH-3A, KH-3B and KH-4, respectively. Leukemic cells started to grow attached to the bone marrow stromal (BMS) cells. KH-3A was positive for OKIa1 and positive at low percentages for B1, Leu-1 and J5 antigens; KH-3B reacted with OKIa1 and J5. Except for OKIa1, these two cell lines showed no surface marker change after 12-O-tetradecanoylphorbol-13-acetate (TPA) treatment. On TPA treatment, clones (KH-3A-2 and KH-3A-3) isolated from KH-3A in agarose showed the induction of differentiation into T and B cell lineage. KH-4 was positive for OKIa1 and positive at low percentages for B1 and J5, and showed a strong reaction with OKIa1, B1 and J5 after TPA treatment. T cell receptor (TCR) beta-chain gene and immunoglobulin gene (JH and C mu) rearrangements were found in KH-3A, KH-3B, and sublines isolated from KH-3 (KH-3A-2 and -3) simultaneously. These findings indicate that BMS cells are useful for the establishment of leukemic cell lines and that some common ALL (cALL) cell populations may be heterogeneous.

Four cases of t(4;11) acute leukemia and its myelomonocytic nature in infants.

Four cases of acute infantile leukemia with translocation (4;11) (q21;q23) are reported. Although leukemia with this chromosomal abnormality has been classified as L2 acute lymphoblastic leukemia by the FAB classification, two of our cases appeared to be of myelomonocyte origin as demonstrated by cytochemical, immunologic, and electron microscopic studies and differentiation induction by 12-tetradecanoyl-phorbol-13-acetate and methylformamide. This chromosomal change is associated with poor prognosis.

Establishment and cytological characteristics of two in vitro T-cell lines derived from a child with acute lymphatic leukemia and a man with adult T-cell leukemia in Japan.

Two permanent T-cell leukemia lines designated KH-1 and KH-2 were established from the peripheral blood of a 9-year-old boy with acute lymphoblastic leukemia and a 47-year-old man with adult T-cell leukemia (ATL). No T-cell growth factor was used. KH-1 cells grew as single cells and KH-2 cells formed clusters in suspension culture. Erosette formation, the absence of immunoglobulin determinants and Epstein-Barr-virus-associated nuclear antigen, and the presence of T-cell antigens revealed by monoclonal antibodies were characteristics of these cell lines as in other established T-cell leukemia lines. Chromosome analysis at the beginning revealed mosaic presence of cells with 46, XY, t(8q+; 15q-) and 46, XY which was later completely replaced by the latter karyotype in KH-1, and abnormal karyotype, 47, XY, +3, t (8q-; 10p+) was maintained throughout the period of in vitro passage in KH-2. The donor patient of KH-2 formerly lived in the south-western part of Japan where ATL is considered endemic and numerous type-C virus particles were detected electron microscopically, in KH-2 cell pellets.