Acute graft-versus-host disease (aGVHD) is a serious complication of allogeneic peripheral blood stem cell transplantation (PBSCT). Patients with severe aGVHD not responding to treatment with steroids have a poor prognosis. We treated three patients with severe aGVHD refractory to steroids with infliximab. Patients (MDS 1, NHL 1, ALL 1) developed grade II-IV GVHD at a median of 13 days (range 9-17) after non-myeloablative PBSCT (HLA mismatched). All patients had received treatment with high-dose steroids for a median of 7 days (range 7-10) in addition to mycophenolate mofetil (MMF) (one). Infliximab was given in 3 weekly doses of 5 mg/kg. In one of three patients a partial resolution of diarrhea and minor improvement of skin were observed. One patient died with refractory GVHD. Infliximab is apparently an effective drug for the treatment of aGVHD, but can be more effective at doses of 5 mg/kg or higher and/or by administering it repeatedly every week.
The bone marrow aspiration test conventionally has been performed by visual methods, using a light microscope, because automatic blood cell analyzers cannot adequately capture erythroblasts and immature granulocytes (IGs) (Tatsumi et al.: Osaka City Med J 1988;34:135-146 by the microscopic method and with the XE2100, and compared the results. A good correlation was found in the nucleated red blood cell (NRBC), white blood cell (WBC), and total nucleated cell (TNC) counts; the myeloid/erythroid ratio (M/E ratio); neutrophils, lymphocytes, eosinophils, and IGs in the immature myeloid information (IMI) channel; and the total cell count. These items can all be analyzed in about 54 sec with the XE2100, which is faster than with the microscopic method. Therefore, analysis of bone marrow aspiration specimens with this apparatus appears to be very useful for clinical screening as well as laboratory testing.
In clinical hematology, the demand for bone marrow aspiration testing is increasing. However, conventional automatic blood cell analyzers cannot completely analyze erythroblasts, and evaluation has mainly been performed by visual examination (the microscopic method). Using the CELL-DYN 4000 automatic blood cell analyzer (CD4000) (Abbott Laboratories, North Chicago, IL), specific recognition and classification of erythroblasts by DNA staining is possible. In the present study, using bone marrow blood collected from normal subjects and patients with hematological malignancy, we classified cells by the microscopic method and with the CD4000, and compared the results. Good correlations were found for total nucleated cell count (TNCC), neutrophils, lymphocytes, erythroblasts, and the myeloid series to erythroid series (M/E) ratio. It is possible to detect blasts that emerge in patients with hematological malignancy using the blast flag system installed on the CD4000. Since all of the items can be analyzed in about 80 sec with the CD4000, cells in bone marrow aspirates can be classified faster with this apparatus than by the microscopic method. Therefore, analysis of bone marrow aspirates with this apparatus appears to be very useful not only for laboratory testing but also for clinical screening.
The R-3000 reticulocyte analyzer uses flow cytometry with an argon laser as its light source. This analyzer stains residual RNA with auramine O to provide a reticulocyte maturation differential. Using the R-3000, we analyzed 119 samples of bone marrow (BM) and peripheral blood (PB) from 111 patients with hematologic disorders. Parameters were reticulocytes, immature reticulocyte fraction (IRF) percentage in BM and PB, BM/PB reticulocyte ratio, and BM/PB IRF ratio. Reticulocytes and IRF percentage in BM were significantly higher than in PB (p < 0.01). There was also a good correlation between reticulocyte percentages in BM and in PB (r = 0.81). Patients were classified into a normal group (without anemia) and an anemia group. Furthermore, the anemia group was classified into three groups: group 1: cases with hematopoietic dysfunction; group 2: cases in bone marrow recovery phase after chemotherapy and hematopoietic stem cell transplantation, and hematologic disorders with bone marrow accelerative phase, and group 3: cases with ineffective hematopoiesis (myelodysplastic syndrome). The mean reticulocyte percentage of the normal group was 2.3 ± 1.1%, which was close to the normal value in BM. The BM/PB reticulocyte ratio of group 3 was statistically higher than that of groups 1 and 2. This indicates that group 3 had ineffective erythropoiesis and that the BM/PB ratio is a useful indicator for the diagnosis of myelodysplastic syndrome.
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