Yasusuke Onozawa

Adenovirus Is a Key Pathogen in Hemorrhagic Cystitis Associated with Bone Marrow Transplantation

Late-onset hemorrhagic cystitis (HC) is a well-known complication of bone marrow transplantation (BMT) that is mainly attributed to infection with BK virus (BKV) and adenovirus (AdV). From 1986 through 1998, 282 patients underwent BMT, and 45 of them developed HC. Urine samples tested positive for AdV in 26 patients, of which 22 showed virus type 11. Among patients who underwent allogeneic BMT, logistic regression analysis revealed acute graft-versus-host disease (grade, > or = 2) to be the most significant predictive factor for HC (P < .0001). In addition, a total of 193 urine samples regularly obtained from 26 consecutive patients who underwent allogeneic BMT were examined for BKV, JC virus (JCV), and AdV by means of polymerase chain reaction. Of patients without HC, approximately 30% of the specimens tested positive for BKV (58 samples) and JCV (55 samples), whereas 5 (3%) tested positive for AdV. Of the 3 samples obtained from patients with HC, the numbers of positive results for BKV, JCV, and AdV were 3, 1, and 1, respectively; the numbers of positive results increased to 14 of 17, 9 of 17, and 10 of 17, respectively, when we added another 14 samples obtained from 14 patients with HC (P < .0001, P = .026, and P < .0001, respectively). In conclusion, there was significant correlation between AdV and HC in the patients we studied.

Invasive infection due to Trichosporon cutaneum in patients with hematologic malignancies

Invasive infection due to Trichosporon is a rare but often fatal disease in immunocompromised patients. In this study, data on patients with hematologic malignancies who had an invasive infection due to Trichosporon diagnosed at the Tokyo Metropolitan Komagome Hospital in Tokyo, Japan, were analyzed.

Effects of interleukin‐12 on natural killer cell cytotoxicity and the production of interferon‐7 and tumour necrosis factor‐a in patients with myelodysplastic syndromes

The effects of Interleukin 12 (IL‐12) on natural killer (NK) cell cytotoxicity and on the production of interferon‐7 (IFN‐7) and tumour necrosis factor‐a (TNF‐a) were examined in 15 patients with myelodysplastic syndromes (MDS), which are well known to have immunologic defects, and in 11 normal subjects. The NK cell cytotoxicity of all of the normal subjects was augmented by incubation with IL‐12 alone, and co‐incubation with interleukin 2 (IL‐2) further augmented it (type A response). The MDS patients showed varied responses to IL‐12/IL‐2. Seven patients showed the type A response, resulting in augmented NK cell cytotoxicity which was similar to that in the normal subjects. In five other patients the cytotoxicity was not increased by IL‐12 alone, but the combination of IL‐12 and IL‐2 did augment the cytotoxicity (type B response). The augmented cytotoxicity in these type B patients was lower than that in the normal subjects. In the final three MDS patients the cytotoxicity was low and not affected by IL‐ 12 and/or IL‐2 (type C response). AH patients with refractory anaemia with excess blasts (RAEB) and patients with RAEB in transformation showed a type B or C response. Conversely, six of eight refractory anaemia patients showed a type A response. In MDS patients there was a positive correlation between the percentage of CD3CD56+ cells in pre‐incubated cells and the cytotoxicity of cells incubated with IL‐12/IL‐2. The combination of IL‐12 and IL‐2 augmented IFN‐7 and TNF‐Q production by nonadherent mononuclear cells in a synergistic or cumulative manner, respectively, in most patients. These results suggest that IL‐12, alone or with IL‐2, may modulate these important immunologic functions in most MDS patients.

Normalized natural killer (NK) cell activity in long-term remission of acute leukaemia

NK cell activity was studied in 28 cases of acute leukaemia in remission. It was found that patients in long‐term remission for 5 years or more had normal NK cell activity but those in remission for less than 5 years had significantly lowered activity. This suggests that patients with acute leukaemia had not been constitutionally deficient in NK cell activity. Low activity was augmented but not fully restored by interferon pretreatment in vitro. Therefore, the low NK cell activity observed during remission is not solely attributable to inadequate secretion of interferon in the host. It is suggested that the production of NK cells in bone marrow is impaired, or the function of NK cells is suppressed, by the leukaemia itself or by chemotherapeutic agents. Increased consumption of NK cells is also a possibility.

Plasma soluble interleukin‐2 receptor level in patients with primary myelodysplastic syndromes: a relationship with disease subtype and clinical outcome

To assess the hypothesis that the plasma soluble interleukin‐2 receptor (sIL‐2R) level may have predictive value for morbidity/mortality in patients with myelodysplastic syndromes (MDS), we determined the plasma sIL‐2R level of 80 MDS patients and examined their subsequent clinical course. Compared with low‐risk MDS (refractory anaemia (RA) and RA with ringed sideroblasts) patients and normal subjects, the plasma sIL‐2R level was significantly elevated in high‐risk MDS (three other MDS subtypes and acute leukaemia following MDS) patients (high‐risk MDS versus low‐risk MDS, P < 0.01; high‐risk MDS versus normal subjects, P < 0.01). 14/40 low‐risk MDS patients developed at least one of the following during the follow‐up period: erythrocyte transfusion dependence, infections requiring hospitalization, disease progression or MDS‐related death. The plasma sIL‐2R level was higher in these eventful subjects than in event‐free low‐risk subjects (P < 0.0001), and all of 10 low‐risk subjects with a plasma sIL‐2R level > 540 U/ml experienced at least one event. By logistic regression analysis of various parameters in these 40 low‐risk subjects, the plasma sIL‐2R level was identified as the strongest independent parameter for predicting eventful subjects (P < 0.0047). The plasma sIL‐2R level did not show a predictive value in high‐risk MDS. This study revealed that the plasma sIL‐2R level is significantly elevated in high‐risk MDS and suggested that the plasma sIL‐2R level is a valuable predictive factor for the clinical outcome in low‐risk MDS.

Elevated plasma soluble interleukin 2 receptor level correlates with defective natural killer and CD8+ T-cells in myelodysplastic syndromes

The plasma soluble interleukin 2 receptor (sIL-2R) level and its relationships with haematologic and immunologic data were examined in 40 patients with myelodysplastic syndromes (MDS). The plasma sIL-2R level was significantly higher in the high-risk MDS group (refractory anaemia with excess blasts (RAEB), RAEB in transformation and chronic myelomonocytic leukaemia) than in the low-risk MDS group (refractory anaemia (RA) and RA with ringed sideroblasts) or in normal subjects, although there was considerable variation in the plasma sIL-2R level within each MDS group. The plasma sIL-2R level correlated positively with the bone marrow cellularity and bone marrow blast mass, but not with the absolute number of CD25+ lymphocytes. This may support the idea that plasma sIL-2R is derived from malignant MDS cells in the bone marrow. The plasma sIL-2R level correlated negatively with the absolute numbers of the CD8+, CD3-CD16+, and CD3-CD56+ cell populations in freshly isolated lymphocytes, the percentage of CD3-CD56+ cells in lymphokine (interleukin 2)-activated killer (LAK) cells, and the cytotoxicity of LAK cells. We conclude that MDS patients having a high plasma sIL-2R level often have a defect in natural killer and CD8+ T-cells.

Assessment of therapeutic potential of interleukin 2 for myelodysplastic syndromes

Summary. The therapeutic potential of interleukin 2 (IL‐2) for myelodsplastic syndromes (MDS) was evaluated in vitro, IL‐2‐induced lymphokine‐activated killer (LAK) cells were prepared from 38 MDS patients and 20 normal subjects. The cytotoxicity of LAK cells against K562 and Raji cell lines and MDS blasts was significantly reduced in high‐risk MDS (refractory anaemia with excess blasts (RAEB). RAEB in transformation, and leukaemic transformation of MDS), but was relatively well‐preserved in low‐risk MDS (refractory anaemia (RA) and RA with ringed sideroblasts). Examination of the immunophenotypes of freshly‐isolated lymphocytes showed that the percentage of CD4+ cells in low‐risk MDS and the percentage of CD3+, CD4+ and CD8+ cell populations in high‐risk MDS was significantly reduced compared with these populations in normal subjects. After cultivation with IL‐2, these three cell populations were still reduced in the corresponding MDS groups and the percentage of CD3‐CD56+ cells were significantly reduced in high‐risk MDS. There was a positive correlation between the percentage of K562 cells lysed by MDS LAK cells and the percentage of CD3‐CD56+ lymphocytes in MDS LAK cells. These aberrant lymphocyte subpopulations appeared to explain, at least in part, the reduced LAK cell cytotoxicity in MDS. These results present a possibility that IL‐2 and LAK therapies are ineffective for most high‐risk MDS patients, whereas they have potential value for low‐risk MDS patients whose lymphocyte cytotoxicity is usually preserved.

Defective natural killer (NK) cell-mediated cytotoxicity does not imply clonal involvement of NK cells in myelodysplastic syndromes

SUMMARY. The clonality of purified cells was examined in 10 myelodysplastic syndromes (MDS) patients by analysing the restriction fragment length polymorphism and methylation pattern of the phosphoglycerate‐kinase gene. Natural killer (NK) cell‐mediated cytotoxicity was also examined. The granulocytes and monocytes were monoclonal or oligoclonal in all cases, except for the monocytes in one case. Conversely, the NK and T cells had a polyclonal pattern in most cases, including all cases who had defective NK cellmediated cytotoxicty. The hypothesis that reduced NK cellmediated cytotoxicity in MDS is caused by a clonal involvement of NK cells was not supported by the present study.

Deficient natural killer (NK) cell activity in paroxysmal nocturnal haemoglobinuria (PNH)

Summary. Natural killer (NK) cell activity was studied in nine patients with paroxysmal nocturnal haemoglobinuria (PNH) and 16 healthy subjects. NK cell activity was deficient in PNH and pretreatment of mononuclear cells with human leucocyte interferon in vitro failed to normalize this abnormality. The deficient NK cell activity may result from an abnormality of a lymphocyte subpopulation not described previously.

Etoposide with/without G‐CSF with busulfan and cyclophosphamide as conditioning for bone marrow transplantation

To increase the efficacy of bone marrow transplantation (BMT), we have tried to add etoposide (VP‐16) to busulfan/cyclophosphamide (BU/CY). Twelve patients received 16 mg/kg of BU and 120 mg/kg of CY with 15–30 mg/kg of VP‐16. Another two patients received 5 μg/kg of G‐CSF with 30 mg/kg of VP‐16. Patients tolerated escalating doses of VP‐16 without any significant hepatotoxicity. Their maximal level of bilirubin was 37.6 μmol/L (2.2 mg/dl), and there was no significant skin toxicity or mucositis. By contrast, two patients who received G‐CSF with 30 mg/kg of VP‐16 developed hyperbilirubinemia and veno‐occlusive disease, which terminated this phase I study. VP‐16 can be safely combined with BU/CY ≤30 mg/kg in three divided doses, and its effect on survival should be evaluated. G‐CSF added to this regimen, however, should be used with great caution.