We tried to determine an appropriate size for a registration system of unrelated donor candidates for HLA matched (identical, semi-identical as well as semi-semi-identical) bone marrow transplantation (BMT). From the distribution of frequencies of HLA-A, -B, -DR haplotypes in Japan, European Caucasoid (CAU), and North American Caucasoid (NAC) obtained at the 8th International Histocompatibility Workshop, we calculated the distribution of the frequencies of HLA phenotypes of both potential BMT recipients and their HLA matched donors in all three populations.
From these distribution of frequencies of HLA phenotypes, we estimated percent of potential BMT recipients to find at least one HLA matched donor in different donor pool sizes in Japan, CAU and NAC.
To find at least one HLA-identical donor for more than 80% of potential BMT recipients, it is necessary to HLA-type and register about 50, 000 donor candidates in Japan, while a million and four hundred thousand volunteers would be needed in CAU and NAC, respectively.
Considering the Japanese donor pool size being relatively small, a registration system for HLA-matched unrelated BMT will be expected to be feasible with the cooperation of public spirited Japanese in the near future.

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To study a possibility of using recombinant human erythropoietin (r-HuEPO) for the prevention and treatment of anemia associated with surgical operations, rats were phlebotomized and effects of administration of r-HuEPO on subsequent anemia were observed. Injection of r-hEPO prior or subsequent to blood-letting amounting 1% of body weight prevented or accelerated the recovery from anemia. Blood-letting amounting 2% of body weight induced a severe anemia in control rats. Prior injection of large amounts of r-HuEPO (1600U/kg) lessened significantly the degree of anemia. It is suggested that the r-HuEPO can be very useful for the prevention of anemia associated with surgical operations. It also can be used to accelerate the recovery from anemia. Prior injection of r-HuEPO can also be very useful for autologous blood transfusion in surgical operations.

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Removal of macromolecule blood component containing pathogenetic factors as complete as possible is desirable for rapid recovery in some diseases such as acute intoxication of drugs or acute hepatic failure. A method of complete blood cleaning utilizing PFC-containing artificial blood (Fluosol-DA^® 20%, Green Cross, Inc. Osaka, Japan) has been introduced and investigated in animal experiments. Blood was replaced with Fluosol-DA 20% in 6 acute drug intoxication dogs (digoxin: 0.05-0.06mg/kg) as completely as possible until hematocrit lowered down to less than 5%. Subsequently fresh blood was infused while PFC with the specific gravity of 1.8 was retrieved by centrifugation with Haemonetics V-50^®. By this series of the procedures, approximately 96.3% of the reduction in serum digoxin concentration was achieved. Animals well tolerated during and after the blood replacement experiments, and 2 of 6 dogs survived more than 4 weeks. This new procedures is expected to be an effective modality of complete blood cleaning as compensating for the disadvantages of cumbersome procedures in total body wash-out in which patients blood was replaced with electrolytes solution under hypothermic condition using a heart-lung machine.

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A standard method for antibodies to adult T-cell leukemia-associated antigens (anti-ATLA), indirect immunofluorescence method (IF), is not suitable for mass screening of anti-ATLA. Recently, new methods of testing anti-ATLA suitable for mass screening of anti-ATLA, enzyme-linked immunosorbent assay (ELISA) and gelatin particle agglutination method (PA), have been developed and carried out in many institutes in Japan. However, few comparative study on these methods of anti-ATLA was reported. Therefore, we carried out the comparative study on anti-ATLA among ELISA, PA and IF method, furthermore adding Western blotting method (WB), which has been used for the confirmatory test of anti-ATLA, in the sera from healthy residents in the Kagoshima district, one of the highly endemic areas of human T-cell leukemia virus type I (HTLV-I) in Japan.
The results obtained were summarized as follows:
1) Of 517 tested sera from healthy residents in the Kagoshima district, positive rate of anti-ATLA was 28.8% (149 sera/517 sera) by PA, 21.1% (109 sera/517 sera) by ELISA and 18.2% (94 sera/517 sera) by IF.
2) The coincident rate between IF and PA and that between IF and ELISA was very high (98.6% and 99.5%, respectively) in the positive sera for anti-ATLA, decided by each method.
On the other hand, the concordant rate between IF and PA and that between IF and ELISA was not so high (59.7% and 84.4%, respectively) in the negative sera for anti-ATLA.
Therefore, PA and ELISA were revealed to be suitable for mass screening in the negative sera for anti-ATLA.
3) Of 69 sera, tested by WB, the coincident rate between WB and ELISA was high (90.7%). However, especially in the sera having low titer antibodies (1:16-1:64) by PA, there was disagreement between PA and WB. So it might be careful to decide the positivity of anti-ATLA in the sera having relatively lower titer antibodies by PA.
4) Of 69 sera, tested for anti-ATLA by above 4 methods, it was important that there were some sera positive by WB but negative by PA or ELISA. Further studies to clarify the discrepancy between these methods and the developement of a new accurate testing method for anti-ATLA are thought to be necessary.

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Erythrocytes from dogs depleted of buffy coat and plasma were stored in a medium containing citrate, phosphate, maltose, adenine and phosphoenolpyruvate (CPMAP) for 21 or 42 days at 4°C. The 24-hour autologous posttransfusion survival of ⁵¹Cr-labeled erythrocytes exceeded the accepted criterion of 70 percent; 89.0 and 95.7% after storage for 21 days, and 87.4% and 94.0% after storage for 42 days. The half life (T1/2) of erythrocytes stored for 42 days was approximately 20 days. There was a significant correlation between 24-hour viability and half life, and also between ATP levels and viability.
After storage for 42 days, the shape of erythrocytes stored in CPMAP was fairly preserved, and echinocytic transformation was less prominent comparing to those stored in CPD-adenine.
From these results, it is suggested that the addition of PEP to the preservative solution is beneficial to the prolongation of the storage period of erythrocyte concentrates.

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A relatively simple ETA method was applied to detect human class II antigens of cell surface. The antibody-B cell reaction was carried out on Millititer plate (Millipore), followed by washing cells through its bottom filter by suction.
A monoclonal anti-DR framework antibody was reactive to the cultured B cells, whereas it did not react to the cultured T cells. This monomorphism of the anti-DR antibody was confirmed by the inhibition test, where all the B cells inhibited the reaction but T cells did not. Polymorphism detected by DQ specific monoclonal antibodies was also confirmed both by direct and inhibitory EIA. Similarly, with peripheral blood lymphoid cells, polymorphism as well as monomorphism defined by the anticlass II antibodies was detectable by the ETA method.
The ETA method using Millititer plate is sensitive and applicable for screening and monitoring the reactivities of monoclonal antibodies especially when they are not complement fixing.

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The frequency for the platelet antigen, phenotype Yuk (a-b+) and Yuk (a+b+) in the population of Kitakyushu district was estimated to be 97.9% (1151/1176) and 2.1% (25/1176) respectively, whereas the phenotype Yuk (a+b-) was not detected, so the gene frequency for the Yuk^a and Yuk^b was 0.010 and 0.990, respectively. Furthermore, in one family in which Yuk (a+b+) was detected in the 7 out of 10 members, we could detect the Yuk^a antibody in the serum of a woman with the history of pregnancy.
In addition, another anti-Yuk^a antibody was found in the 1689 sera from post partum hemorrhage. The children born from the two mothers with anti-Yuk^a antibodies in their sera did not have the history of neonatal alloimmune thrombocytopenia (NAITP).

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In a screening test for natural antibodies against human T-cell leukemia virus type I (HTLV-I) was performed using a new gelatin particle agglutination assay (PA) and an indirect immunofluoresecence assay (IF) sera with weak positive PA but negative IF were found in healthy donors. To confirm the presence of HTLV-I in these healthy donors, we performed Western blot analysis and Southern blot analysis. The proviral genome of the HTLV-I virus was detected in lymphocytes from 2 healthy donors. These results suggest that some healthy donors with weak positive PA but negative IF may be infected with the HTLV-I virus.

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In order to avoid transfusion of unnecessary components of blood, usage of a a mere component of plasma fraction has become popular and subsequently easier, safer and less expensive ways of plasma collection have been sought. Safety confirmation study of membrane plasmapheresis, which is assumed to be a candidate for a such purpose, was performed paying a special concern on complement activation in healthy voluneteers utilizing three different membrane plasma separators specially designed for donation plasmapheresis. Specification of three separators and plasma collection maneuver was as follows; ‹A›; hollow fiber membrane of cellulose triacetate, 0.25m², intermittent blood flow via a single needle. ‹B›; hollow fiber membrane of polypropylene, 0.20m², continuous blood flow via two needles. ‹C›; flat sheet membrane, 0.22m², continuous blood flow. About 400 ml of plasma was collected within 30min. using ACD as an anticoagulant. No serious symptoms were observed during and after the membrane donation plasmapheresis (MDP) in 25 volunteer donor. Only slight decrease in serum concentration of TP, albumin, IgG, IgA, IgM, and Ca, but normalized in one week. C3a and C5a were contained more in the separated plasma and in the patient' blood after contacting with the separator membrane than in the preoperational serum in all procedures. However, there were no marked changes in coagulation factors during and after the procedures. In conclusion, MDP can be safely done in volunteer donors even though complements are more activated compared to the centrifugal procedure.

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Effects of prevention of plasticizer leakage from poly (vinylchloride) bags on platelet functions were examined. Three kinds of bags, DEHP-bags, glow discharge-treated bags (GD-bags) and bags plasticized by tri-(2-ethylhexyl)-trimellitate (TOTM-bags) were used. Platelets stored in bags from which plasticizer does not leak (GD-bags and TOTM-bags), maintained well aggregability, ability to change the shape, adhesion ability and %HSR. GD-bags, discharged in the presence of H₂ gas, were more effective for maintaining platelet functions than TOTM bags.

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In recent blood banking, a totally improved red cell product is required, such as, leukocyte depletion for decreasing of transfusion reaction and maintenance of both ATP, for posttransfusional viability, and 2, 3-diphosphoglycerate (2, 3-DPG), for oxygen-delivering, are necessary.
Phosphoenolpyruvate (PEP) is known to increase both ATP and 2, 3-DPG levels in red cells. We applied it to the additive solution as a protein-free-medium for the preservation of plasma and leukocyte poor red cell concentrates. Plasma and leukocyte poor red cell concentrates were prepared by re-centrifugation (4, 000G, 6 minutes). Over 80% of leukocytes were removed and 60ml of plasma was collected from 1 unit of concentrated red cells (400ml collection). During 6 weeks storage, ATP and 2, 3-DPG levels of stored red cells in the new additive solution containing maltose, citrate, phosphate, adenine and PEP, were kept very well. After 6 weeks storage, ATP and 2, 3-DPG levels were 80% and 50% of fresh ones, respectively, by incubation at 37°C for 30 minutes. Furthermore, the stored red cell shapes were retained discoid and hemolysis decreased.
This additive solution enabled red cell preparations to be stored at least for up to 42 days and to provide an excellent red cell product, which was advantageous for patients to receive red cells with a normal oxygen affinity, a long shelf life in vivo and decreasing of transfusion reaction.

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Cryofiltration was applied to two severe RA patients with secondary amyloidosis, who had been treated with conventional steroids or analgesics administration in vain, in order to remove pathogenic substances such as immune complex and other abnormal proteins. Continuous cryofiltration with double filtration was performed by using the apparatus of Asahi Plasauto 1000 (Asahi medical, Tokyo, Japan) in extra-corporeal circulation with AP-05H (Asahi medical, Tokyo, Japan) as the first plasma separator and AC-1730 (Asahi medical, Tokyo, Japan) as the second macromolecule filter. One processed plasma volume by cryofiltration was about 4000ml at the averaged plasma flow rate of 30ml per minute and separated plasma after the first separator was led to cooling chamber at 4°C and then filtered at the second filter. Cryogel was eluted by saline and analysed immunochemically. After 4 processes of cryofiltration performed at 5 day interval, clinical symptoms of arthralgia, restriction of joint movement, joint swelling and morning stiffness were fairly relieved. Cryogel precipitated at 4°C in the second filter was analysed by gel and ion exchange chromatography. Cryogel was fractioned into very low molecule protein (MW 20-30K), albumin, IgG and high molecule protein designated as X-RF (MW ca. 800K). Molecular weight of X-RF is similar to that of IgM but X-RF was turned out to possess a different property from IgM by ion exchange chromatography. X-RF is regarded as main pathogenic substance and could be reduced from serum by repeated process of cryofiltration. And moreover, low molecule protein is conceived to be compatible to amyloid protein which was shown in the sera of amyloidosis patients by same chromatography method. This low molecule protein could be also reduced like X-RF by cryofiltration.
We could conclude that cryofiltration was fairly effective method to remove the pathogenic substance from RA patients and could relieve clinical symptoms.

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We have detected 2 donors (0.04%) having anti-Yuk^a antibody out of 4, 506 female blood donors who have a history of delivery.
The frequency of the Yuk^a antigen was studied with those 2 lots of anti-Yuk^a antibodies and we could find 29 examples (2.8%) of Yuk^a positive in 1, 037 donors.

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An autopsy case of 62 years old man who had a fatal complication of graft versus host disease (GVHD)-like syndrome (so-called post-operative erythrodermie) after mitral valve replacement (MVR) was reported.
In 18, July 1986, MVR was performed with Björk Shiley valve prosthesis, during operation and post-operation 2400ml of fresh blood and FFP were used.
The post-operative course was uneventful, but following fever of 37.5°C on the 9th day after operation, various clinical symptoms apperated, ie liverdysfunction, erythrodermie, azothemie, leukopenie, thrombocytopenie and bleeding of GI-tract. He died in the 24th day after operation. The cause of death was suspected septic shock. The postmortom examination revealed candidiasis, and destruction of normal structures of bone marrow, lymphnode and spleen were characteristics. Histology of the skin lesion showed similar findings of GVHD. The type of HLA in the patient showed A11, 33, B35, 44, W4/6, Cw3, but his real HLA type was suspected A26, 33, B35, 44, W4/6, Cw3 from the examination of his family.
This case give us the suspicion that the pathogenesis of so-called post-operative erythrodermie may take a part of GVHD.

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We report a case of rare blood group phenotype P^k₁. The patient was a healthy 29-years-old woman and had a history of 3 first-trimester miscarriages and no successful pregnancies. Her parents were second cousins each other. She had an anti-P in her serum, which was mainly of IgG type and partly of IgM type. The anti-P titers with polyvalent antiglobulin, anti-IgG and anti-IgM sera were 512-1024, 512 and 64, respectively. Her serum contained IgG 1, 2, 3 and 4 anti-P. No restriction of response to IgG3 was revealed. There were no etiologic factors that have been associated with early pregnancy loss except for having anti-P in her serum. Therefore, we considered that IgG anti-P caused her habitual abortions.

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一昨年3月, クームス教授は日本学術振興会 (JSPS) の招聘により来日されましたが, その折に「クームス教授を歓迎する会」(発起人代表 森亘先生, 世話人代表 福岡良男先生) 主催により札幌, 東京, 京都, 福岡の4ヵ所で「Coombs Reaction」と題した来日記念講演会が開催されました. クームス反応は輸血領域において最も重要な検査の一つであり, それは医学, 医療の場に多くの益をもたらしました. 先生の御好意によりその来日記念講演の内容を掲載致しました. なおクームス教授は現在, 英国ケンブリッジ大学の病理学教室の教授で免疫学的部門を担当されております.

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