We investigated whether a commercial kit for the detection of anti-platelet antibody, anti-PLT·OLYBIO·MPHA (OLYBIO·MPHA) can detect anti-HLA antibodies. Fifty-seven sera which were positive by lymphocyte cytotoxicity test (LCT) were used for this study and the results obtained by OLYBIO·MPHA were compared with those by LCT and by flow cytometry (FCM) using platelets as target cells. OLYBIO·MPHA could detect multi-specific or high-titered anti-HLA antibodies, but it failed to detect some monospecific or low-titered anti-HLA antibodies. Six sera which showed discrepant results between LCT and OLYBIO·MPHA were analyzed by FCM. The discrepancies in two sera were due to IgM anti-HLA antibodies and in the other sera were considered to be due to low titer of anti-HLA antibodies.
We conclude that OLYBIO·MPHA is a useful kit for the detection of anti-HLA antibodies since this kit can detect a common anti-HLA antibodies and can be easily performed without special equipments.

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As to the method of detecting anti-HLA antibody by using in the serum of patients who were frequently transfused of platelet, and by allowing it to react with lymphocyte by means of previous LCT (lymphocyte cytotoxicity test) method or AHG-LCT (antihuman immunoglobulin-LCT) method, there had been a problem that this method was difficult to be made popular as a general rutin test, because it was difficult to secure and preserve the panel lymphocytes. The authors developed a method in which HLA antigen was extracted from platelet into a physiological saline solution of 3% sucrose and it was employed as the platelet soluble antigen and fixed as a layer on the U-type Teraksaki plate, and then the screening of anti-HLA antibody was conducted by the use of this plate using the MPHA (mixed passive haemagglutination test). The authors consider that this method is useful because it has such an advantage that the platelet soluble antigen can be constantly prepared for a long period at -80°C, and further that the platelet specific antigen is also detectable, and because this method is capable of use like a commercial screening panel of erythrocyte antibody.

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A 65-year-old female MDS patient Y. M showed refractoriness to transfusions of HLA-matched platelets in 4 out of 72 occasions. Analysis of Y. M serum by a monoclonal antibody specific immobilization of platelet antigens (MAIPA) assay revealed that the serum contained anti-HPA-5b antibody directed against an epitope on platelet glycoprotein (GP) Ia/IIa. The Y. M serum was tested for its reactivity against ineffective as well as effective platelets by MAIPA assay. The serum was reactive to the platelet antigen against an epitope on GP Ia/IIa of ineffective platelets, but not against an epitope on GP IIb/IIIa, GPIb or GPIV. The serum was not reactive to any of GPs of effective platelets. Platelet typing of patient Y. M and four ineffective HLA-matched donors showed that patient Y. M was HPA-5a/a, whereas four ineffective donors were HPA-5a/b. The results indicate that HPA-5 antigen is responsible for the refractoriness to the platelet transfusion.

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Recently, antigen-specific assays for detecting anti-platelet alloantibodies have been available: immunobead assay, modified antigen capture ELISA (MACE) and monoclonal antibody-specific immobilization of platelet antigens (MAIPA). These assays are based on essentially the same principle. In this study, using anti-HPA-1a and anti-HPA-4a antibody, we have compared the sensitivity of MACE using avidin-biotin-complex method (ABC-MACE), MACE, MAIPA, and flowcytometry (FCM) as a conventional method. The sensitivity of MACE for detecting anti-HPA-1a antibody was twofold better than that of MAIPA, while the sensitivity of MAIPA for detecting anti-HPA-4a antibody was twofold better than that of MACE. Using ABC-method, the sensitivity of MACE for detecting anti-HPA-la and anti-HPA-4a increased 8-fold and 64-fold, respectively. In comparison with FCM, ABCMACE showed almost the same sensitivity for detecting anti HPA-4a antibody, and showed better sensitivity for detecting anti HPA-1a antibody. These data demonstrate that ABC-MACE is much more sensitive than MACE or MAIPA for detecting anti-platelet alloantibodies, and that the sensitivity of ABC-MACE is comparable to that of FCM.

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HCV related antibody was screened among 3291 voluntary blood donors by two test kits (Imucheck for screening and Imunojudge for immunobloting) using two recombinant antigens (C7 in NS3 and C11 in core region) cloned from the plasma of Japanese non-A, non-B hepatitis patient. The positive rates of Imucheck and PHA (DAINABOT) were 3.04% and 0.55%, respectively. 88 out of 100 Imucheck positive samples were tested by Imunojudge, which resulted in 53 (60.2%) samples positive to C7 and/or C11 antigen; 12 samples were positive to C7, 23 to C11 and 18 to both. HCV-RNA was detected in 11 samples positive with Imunojudge by PCR using 5′ non-coding primer, all of which showed high C. I. O. above 8.00 by Imucheck and had both C7 and C11 antibodies by Imunojudge. These samples with HCV-RNA showed more than 2¹² in PHA titer and RIBA-II positive. Imucheck detected 5.5 times more positive samples than PHA did, but one third of them was considered non-specific reaction by Imunojudge. The samples which had both C7 and C11 antibodies were mostly PHA positive, and covered the all samples with HCV-RNA. Although Imucheck and Imunojudge would be useful in screening blood donors for HCV related antibodies, it remains to be clarified how to consider only either C7 or C11 positive samples.

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The ability of leukocyte removal filters was investigated with three types of concentrated red blood cells (CRC); i. e., CRC prepared by a standard method (N-CRC), CRC suspended in Map (Map-CRC) and buffy coat depleted CRC (BD-CRC). Pall RC50 filters with the same lots were used, and the same procedures were done in filtration to avoid the variations in filters as much as possible. Two units of each CRC stored for 1, 6 and 10 days was filtered through Pall RC50 filters. The mean residual leukocyte counts (n=12), which were calculated from the sum of each CRC stored for 1, 6 and 10 days, were as follows. For the 1st unit of N-CRC, Map-CRC and BD-CRC were 2.66±1.28(×10⁵), 1.55±0.25(×10⁵) and 1.44±0.25(×10⁵), respectively. For the 2nd units of BD-CRC, the mean count was 1.84±0.91(×10⁵), and 1.42±1.72(×10⁶) for Map-CRC, while it was 6.58±10⁵ to 1.38×10⁸[2.28±4.02(×10⁷)] for N-CRC, showing much variation. The filter was obstructed with much microaggregate in one N-CRC sample stored for 10 days. The residual leukocyte count after filtration with this filter was high (8.16×10⁷). These data suggest that the individual differences in the donor have a strong influence on the ability of leukocyte removal in N-CRC. BD-CRC and Map-CRC are more useful than N-CRC to bring out the ability of a leukocyte removal filter.

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The acquired immunodeficiency syndrome (AIDS) is caused by human immunodeficiency virus (HIV) that is possibly transmitted by blood transfusion, direct exposure to blood (and blood product) or through sexual contact, and in children, transmitted virtically from their infected mothers. It has been more and more important to detect HIV-1 in blood and plasma products. HIV-1 antibody has been measured by gelatin particle agglutination assay (PA), by enzyme immunoassay (EIA), and more confirmatively by western blot assay (WB).
However, there become evident the presence of the second type of HIV, HIV-2, and recently the assay detecting both HN-1 and HIV-2 has become required.
For this purpose, the HIV-1/2 antibody EIA and RPHA (red blood cell particle hemagglutination) tests were introduced.
In this paper, we examined the sensitivity of UBI EIA-1/2, the immunoassay uniquely employing synthetic peptides for the detection of both antibodies. Our results showed that the use of these synthetic peptides offered the advantage of minimize the incidence of non-specific reactions caused by the antibody cross reactivities between virus antigens and the host cell antigens eluting from the cell lysate.

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Hepatitis C virus (HCV) genomes were recently detected in biological materials, and were divided into types I, II, III and IV according with their nucleotide sequences variations.
In this study, typing of the HCV genomes was performed in 84 PHA-positive and HCV-RNA-positive blood donors in Yamanashi Prefecture.
A typing method developed by Dr. Okamoto was used, depending on the amplification of a C gene sequence by polymerase chain reaction (PCR) using a universal primer (sense) and a mixture of four type-specific primers (antisense). Type I was found in 1 sample (1.3%), type II was found in 56 samples (71.8%), type III was found in 12 samples (15.4%) and type IV was found in 9 samples (11.5%) out of 78 blood donors, the rest of 6 samples were not specified.
The prevalence of HCV-subtype was surveyed in Yamanashi Prefecture. The result suggests that there are geographical differences of HCV subtype.
Also, those samples were divided into c-100-positive group and c-100-negative group. And we compared between HCV subtype and c-100 positivity. In c-100-positive group, Type I was found in 1 sample (1.9%), type II was found in 42 samples (80.0%), type III was found in 6 samples (11.5%) and type IV was found in 3 samples (5.8%) out of 52 individuals. In c-100-negative group, Type II was found in 14 samples (53.8%x), type III was found in 6 samples (23.1%) and type IV was found in 6 samples (23.1%) out of 26 individuals.

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Twenty-eight normal bone marrow donors aged 24 (4-53) years received autologous blood transfusion during the marrow harvest for the allogeneic bone marrow transplantation. The median volume of marrow removed from the donors was 1042±234ml. The median volume of autologous blood transfused was 611±200ml. No donors required homologous blood transfusion. Autologous blood transfusion is useful for marrow donors to avoid the risks of homologous blood transfusion.

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We studied antibody response against three different recombinant antigens (5-1-1/C100-3, c33c and c22-3) by RIBAII and HCV RNA detection by RT-PCR in serum samples of anti-HCV positive donors by 2nd generation HCV antibody screening test (PHA). We also compared reactivity of the two agglutination methods (PHA & PAII) with the serum samples.
In HCV antibody (PHA) positive donors, Core (c22-3) antibody was detected in 100% of PCR positive group and in 96.1% of PCR negative group whereas NS3 (c33c) antibody was detected in 98.6% of the former and in only 22.6% of the latter. The seropositivity of NS3, consequently, showed the highest correlation with PCR results among three kinds of antibodies. NS4 (5-1-1/c100-3) antibody was detected in 63.4% of PCR positive group and in 4.6% of PCR negative group. Since the majority of the elevated ALT cases were NS4 antibody positive specimens, NS4 antibody seemed to associate with some abnormality of liver function.
In comparison of PAII reactivity with PHA, PAII didn't react with a part of PHA positive (c22-3 only) PCR negative specimens, but detected all PCR positive specimens due to it's high sensitivity for NS3.

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Blood transfusion, injury with a blood-contaminated needle, and sexual contact with chronic carriers of hepatitis C virus (HCV) seem to be important routes of HCV transmission. We previously reported the house hold contact seems to be a route of transmission. In this study, the comparison of the incidences among the HCV RNA in serum and saliva of the anti-HCV positive patients, by the method of polymerase chain reaction. HCV RNA was detected not only in the serum, but also in the saliva, especially in the saliva of patients with active liver disease, and post-transfusion hepatitis. The viral titres in the saliva seemed to be lower than in the serum. HCV RNA in the serum and saliva disappeared after the treatment with interferon-alpha. It is suggestive that the HCV genome positive saliva might be one of the routes of transmission in sporadic HCV infection.

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A case of partial D patient who formed anti-D by blood transfusion was reported.
The patient was a 54-year-old Japanese man suffering from cerebral infarction. His blood group was B, Rho (D) positive and no irregular alloantibodies were detected in pretransfusion screening and crossmaching tests. He received 3 units of ABO-compatible Rho (D) positive concentrated red cells in treatment of anemia due to gastritis on May. 25-27, 1988.
Anti-D was found in his serum on Nov. 16, 1989 when he was again hospitalized with sequelae of cerebral infarction. His red cells were shown to react neither with 2 samples of monoclonal anti-D (OSK3 and OSK-1, both of which were prepared by Osaka Red Cross Blood Center) nor 2 samples of commercially purchased monoclonal anti-D. But they did react with 4 samples of polyclonal anti-D by both anti-globulin test and indirect immunof luoresence flow cytometry.
His anti-D did not react with D category VI red cells, but reacted with D category IVb and Va cells. The results obtained indicated that he has D category VI red cells and formed anti-D by the transfusion of D-positive red cells.

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A 73-year-old man was admitted to our hospital for femur fracture by traffic accident. Anti Sd^a antibody was detected in his serum by the examination for transfusion. Thirty-three units of RBCs, which were Sd^a positive, were given over three days on operation and post operation period. About two weeks later, the titer of anti Sd^a antibody rose from ×4 to ×128. The activity of his serum lactic dehydrogenase was 531U/1. Total serum bilirubin concentration was 1.5mg/dl. Reticulocyte count was 24‰. Occult blood in urine became positive. Although clinical symptom of hemolytic transfusion reaction was not obviously recognized, the clinical laboratory data might exhibit mild in vivo hemolysis of transfused RBCs.

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