We retrospectively analyzed the correlation between HLA class II DNA typing, MLR, rejection and acute GVHD in 14 patient-donor pairs, who had undergone HLA serologically identical related bone marrow transplantation, in order to evaluate the histocompatibility tests for the clinical outcome after transplantation.
Severe acute GVHD (≥grade II) were seen in 7, rejection in 2 and MLR (rejection side)-positivity were in 1 out of 14 patients. However, the HLA class II genotypes of DRB1, DRB3, DQB1 and DPB1 were completely identical between them.
Among the 7 patients with severe acute GVHD, one patient “F” died day 15 of grade IV GVHD after transplantation. In this case, there were no considerable risk factors such as ABO or sex mismatch between the patient and donor, and the donor had not alloimmunized by previous transfusion.
Two patients, J and K, rejected the donor's bone marrow and only the patient K showed recipient anti-donor reactivity in MLR before the transplantation. In this case, the donor was male and the patient was female who might have been already alloimmunized by pregnancy and transfusion.
These observations strongly indicate the importance of minor histocompatibility antigens in HLA genotypically identical allogeneic bone marrow transplantation.

View full abstract

Forty-seven orthopaedic donor-patients in eight different institutions were evaluated as an early phase II open study to determine the efficacy and safety of once a week subcutaneous (SC) administration of recombinant human erythropoietin (rHuEPO) for weekly blood collection of 400ml. Of the 47 patients, 10 were excluded from efficacy evaluation for not meeting age (7) or hemoglobin (Hb) (2) criteria, or rHuEPO administration schedule (1). The remaining 37 patients, aged between 38 and 75, were divided into three groups by weekly SC dosage; 200IU/kg (12), 400IU/kg (13) and 600IU/kg (12). Weekly blood collection of 400ml began one week after an initial SC administration.
The 37 patients were able to donate autologous blood of 800ml/2 wks or 1200ml/3 wks and required no homologous blood during or after operation. At the time of the first phlebotomy, Hb was significantly increased in all three SC groups. One week after the second phlebotomy, Hb was not significantly decreased in the 600 group (101±6%), while it was decreased significantly in both the 200 group (96±6%) and the 400 group (97±4%) against the preadministration level. The Hb decrease in the 400 group was smaller than that in the 200 group, and it was clinically acceptable. Compared to previous studies at two institutions, equal dosages of rHuEPO at equal frequencies appeared to be more effective with SC administration than with intravenous (IV).
None of the 47 patients showed any adverse effects or abnormal laboratory data attributable to rHuEPO administration.
Our results indicated that a once a week SC administration of rHuEPO at a dose level of 400IU/kg or more could improve post-phlebotomy anemia as effectively as higher frequency IV administration.

View full abstract

A transiently occurring anti-Kell (K1) antibody was observed in a 4-year-old girl who had acute lymphocytic leukemia and upper respiratory tract infection. The patient had nerve received a blood transfusion. Testing with dithiothreitol and anti-immunoglobulin-coated bead (Immunobead_??_) showed the antibody to be IgM class. The “naturally-occuring” antibody was demonstrated for 5 weeks, disappeared from the serum at 9th week after the first detection and never detected for 79 weeks afterward.

View full abstract

In order to moniter the accuracy of the HTLV-I antibody test, assessment of the same specimen was performed using 2 gelatin particle agglutination (PA) methods, 2 Western blot (WB) methods, and an indirect immune fluorescence (IF) method. There were 67 specimens in all, and 55 were positive with the Serodia-ATLA reagent and 12 yielded a ± negative reaction at 16×.
The 16 specimens in which the two PA methods did not agree (23.9%) were 16-64×, low-titer cases and Serodia-ATLA positive and Serodia-HTLV-I negative cases. Although all the specimens were negative in the IF method, 4 (25%) were anti-nuclear antibody positive, and 12 (75%) were anticytoplasmic antibody positive.
For 8 specimens which were Serodia-HTLV-I low-titer positive, judgment was reserved when the WB method was used. When the IF method was used, although 7 specimens (87.5%) were negative, they were either anti-nuclear or anti-cytoplasmic antibody positive. In anti-cytoplasmic antibody positive specimens, these seems to be a substance in the cytoplasm which gives rise to a weakly positive reaction with the Serodia-HTLV-I reagent. Since the IF method serves as a reference for the presence of anti-nuclear antibodies and anti-cytoplasmic antibodies, it was useful as a test to confirm PA low-titer speciments.

View full abstract

We have measured residual leukocytes in leukocyte-depleted blood products using Nageotte and Hausser counting chambers. For testing, one part of platelet concentrate (PC) and red cell concentrate (RCC) were added to 7.5 parts of propidium iodide solution containing RNase, sodium citrate and Triton X-100 or added to 9 parts of Türk's solution containing gentian violet and glacial acetic acid. Stained leukocytes were measured in 2 areas (100μl) with the Nageotte chamber or in 2 areas (6.44μl) with the Hausser chamber. The leukocytes in pre-filtered PC and RCC had first been measured in a Neubauer. hemocytometer and the samples were serially diluted with leukocyte-free PC or RCC prepared by repeated filtration. The number of leukocytes measured with the Nageotte and Hausser chambers as a function of cell number expected from the Neubauer hemocytometer count could be expressed by the regression lines: Y=-3.367+1.199X(r=0.968) for PC and Y=-0.800+0.844X(r=0.906) for RCC. The limit of sensitivity for the counting method using the Nageotte and Hausser chambers was 85cells/ml and 1300cells/ml, respectively. Thus, the hemocytometers, the Nageotte and Hausseri chambers, which have volumes of 50μl and 3.2μl, respectively, and which are greater than that of the Neubauer counting chamber (0.9μl), can provide more sensitive and reliable cell counting than the conventional counting methods such as automated electronic cell counters. The Nageotte and Hausser counting chambers should be useful to count the residual leukocytes in leukocyte-depleted blood products as a standard method for the purpose of quality-controlling them when flow cytometers are not available in blood centers.

View full abstract

We discribe an experimental study of the strept avidin-biotin (SAB)-alkaline phosphatase (ALP) technique with new fuchsin using mouse monoclonal anti-A and B antibodies to A and B antigens of erythrocytes on air-dried smears.
This method is useful for accurate identification of A or B antigens of erythrocytes in peripheral blood and bone marrow, and is very helpful in the diagnosis of chimera or mosaic. It may be also usuful to investigate when A and B antigens on human red blood cells express.

View full abstract

Two types of polyester filters were evaluated in vitro for their effectiveness in removing leukocytes from red cell-MAP stored for 5 and 10 days. Filtration using a new Pall RC400 filter reduced in the number of leukocytes to 4×10⁴cells/bag or less regardless of the number of days the blood was stored, or the filtration rate. On the other hand, the number of leukocytes remaining after intravenous drip filtration (6ml/min) using the Sepacell RC500A II filter was 920.6 (405.4-1929.2)×10⁴cells/bag after 5 days of storage, and 75.8 (15.7-262.1)×10⁴cells/bag after 10 days of storage. However, when the clamp was opened and filtration was performed while allowing a higher flow rate by gravity, the number of residual leukocytes was reduced to 30×10⁴cells/bag regardless of the number of days the blood was stored. The ranges of red cell recovery rate and number of residual platelets were 87-94% and 0.3-0.9×10⁹cells/bag, respectively, for the Pall RC400 filter, and 92-95% and 0.7-1.2×10⁹cells/bag, respectively, for the Sepacell RC500AII filter. Therefore, the new Pall RC400 filter was effective for removing leukocytes from red cell-MAP.

View full abstract

HLA antibodies were detected in 53% of patients subjected to frequent platelet transfusion (100 units or more). The antibodies had polyspecif is characteristics, including A-locus antibodies (23.7%), B-locus antibodies (31.6%), antibodies of A+B loci (28.9%), and multispecific antibodies (15.8%). No significant relation was observed between HLA-antibody production and disorders. In relation to the HLA-antibody production of the patients and HLA antigens of class I, it is suggested that the patients' HLA-A (2, blank) or homozygous A2 antigens raised HLA-antibody production, and that the blank (or homozygous) A antigens raised the production of A-locus-related antibodies. Regarding class II antigens, which greatly contribute to immune response, the patients' with HLA-DR2 antigens frequently produced HLA-antibody and then immune high-response of HLA-DR2 antigens to HLA antigens was suggested.

View full abstract

We reported the experience of peripheral blood stem cell transplantation (PBSCT) performed in patients with lymphoid malignancies.
After the myelosuppressive chemotherapy, the peripheral blood stem cells were collected using the Blood Cell Separator (CS-3000) during bone marrow recovery and subsequently cryopreserved. Nineteen apheresis were performed in 11 patients with lymphoid malignancies. The collected number of granulocyte/macrophage progenitors (CFU-GM) was more than 5×10⁵/kg in 13 aphersis and ranged between 2 and 5×10⁵/kg in 3 apheresis. Seven patients with malignant lymphoma and one patient with acute lymphoblastic leukemia underwent PBSCT following myeloablative chemotherapy. The infused number of CFU-GM ranged between 1.9 and 18.1×10⁵/kg. The median time to reach 500neutrophils/μl or 50, 000platelets/μl was 9 (range; 8-12) and 16 (range; 12-27) days, respectively. Seven patients have been now alive in complete remission for 6-18 months after PBSCT. When the infused number of CFU-GM is more than 2×10⁵/kg, PBSCT following myeloablative chemotherapy seems to be safe and useful treatment in patients with lymphoid malignancies.

View full abstract

Our ability to meet patient needs for blood product replacement falls short of our expectations in some instances. Fortunately, recent scientific developments suggest ways in which these problems can be addressed. The application of this research can assure an adequate supply of safe blood and blood-related products, and can improve the effectiveness of this important medical therapy.
While there are now good methods to sterilize plasma derivatives, much more work is needed to assure comparable freedom from transmission of disease by whole plasma, red cells, and platelet concentrates. Plasma treated by the solvent-detergent method is expected to be a significantly safer product, but the treatment of cellular blood products remains a difficult problem for which there are no immediate solutions. We need to better understand the interaction of viruses and bacteria with red cells and platelets if there is to be progress in addressing this concern.
A second important safety concern is the apparent immunosuppression produced by some blood products. The magnitude of this problem needs to be determined and the role of leukocytes needs to be examined more carefully.
The effectiveness of some transfused blood products is now limited by the development of alloimmune reactions. This can be a serious complication, especially for platelet transfusion. A better understanding of alloimmunization and the identification of ways to prevent this problem will markedly improve the practice of transfusion medicine. At the same time it is important to develop improved blood product storage conditions and to develop new blood products.
Fortunately, progress in biomedical research is providing the necessary tools to address these needs. For example, it is likely that conditions will be identified that support the production of cellular blood products in tissue culture. In addition, recombinant technology may permit production of essential proteins for therapy even though they are difficult to prepare from plasma because of low concentration or instability. Scientists will also seek to develop modified “second generation” products that avoid limitations inherent in some natural proteins. Finally, recombinant growth factors have already had an important effect in transfusion medicine, and additional products are anticipated.

View full abstract