Tomoko Ishida

Platelets expressing P-selectin and platelet-derived microparticles in stored platelet concentrates bind to PSGL-1 on filtrated leukocytes.

The levels of interleukin-6 and platelet-derived microparticles (PMPs) were measured in the blood of 137 pa tients with side effects from platelet concentrate (PC) transfu sion with leukocyte removal filtration, P-selectin-expressing platelet and PMPs in stored PC before and after the filtration, and filtered leukocytes positive for P-selectin glycoprotein li gand-1. The side effects, which were observed in 203 transfu sions for 84 patients with hematologic disease and 53 patients with nonhematologic disease with no significant difference be tween the two groups, included urticaria (75.9%), erythema (18.7%), and fever (17.2%), but no anaphylactic reactions. The levels of interleukin-6 and PMP correlated in both groups, and were significantly higher in the hematologic disease group than in the nonhematologic disease group. The level of PMP, but not interleukin-6, was significantly higher for patients testing posi tive for allergic reaction than for those testing negative. In the stored PC prior to filtration, the level of interleukin-6 was normal. The level of P-selectin-expressing platelets and PMPs was elevated before filtration, but was significantly lower after filtration. Taken together, the results suggest that PMP is in volved in the generation of transfusion reactions, and indicate that both platelets and PMP displaying P-selectin bind to P- selectin glycoprotein ligand-1 of leukocytes retained by the leukocyte filter.

Clinical Evaluation of Transfusion of Prestorage-Leukoreduced Apheresis Platelets

With increased use of platelet concentrate (PC) in recent years, adverse reactions to PC transfusion have received much clinical attention. Most of these reactions stem from white blood cells (WBC) contaminating the transfused PC. Several are thought to be preventable by removing WBC before PC storage. Methods: We routinely filtered apheresis PC either during collection or immediately afterwards and monitored various indicators of platelet quality during storage. After transfusion to patients, transfusion efficacy and the type, severity, and frequency of posttransfusion side effects were compared with those of a control group in which PC was filtered at the bedside. Results: Prestorage-filtered PC contained an average of 3.1±0.7 × 1011 platelets and 0.9±1.2 × 106 contaminating leukocytes. Measurement of platelet function and metabolic indicators revealed normal values throughout the storage period. CD62 measurement revealed no undue platelet activation after filtration or during the storage period. Cytokine, histamine, bradykinin, and complement levels showed no significant increase after filtration or during storage. Transfusion efficacy and overall side effect incidence rates were similar for the prestorage- and bedside-filtered groups, but reactions of the bedside-filtered group included serious reactions such as breathing difficulties and shock. No serious reactions were noted in the prestorage-filtered group. Conclusion: Filtering PC before storage has no adverse effect on PC quality and may reduce the severity of post transfusion side effects.

A Japanese propositus with D-- phenotype characterized by the deletion of both the RHCE gene and D1S80 locus situated in chromosome 1p and the existence of a new CE-D-CE hybrid gene

AbstractIn a family study of a Japanese propositus with the D-- phenotype, the serological data of her D-- phenotype and those of her parents were discrepant. Gene analysis of the propositus showed a gross deletion of the RHCE gene and a new rearrangement of RHCE to yield the CE-D-CE hybrid. It was demonstrated that the hybrid CE-D-CE gene consisted of exon 1 from the RHCE gene, followed by exons 3 to 7 from the RHD gene and exons 8 to 10 from the RHCE gene. However, whether or not exon 2 of the RHD or the RHCE gene was contained in the CE-D-CE gene remained unclear. Moreover, spacer analysis between both RH genes and the family study suggested that the D-- gene complex from the paternal and maternal sides consisted of only the CE-D-CE hybrid gene and a single RHD gene, respectively. For the purpose of confirming the parent-child relationship, a paternity test using DNA fingerprint and polymerase chain reaction (PCR) analysis at the D1S80 locus were performed. DNA fingerprints with two kinds of DNA minisatellite probes (33.15 and 33.6) confirmed that the parent-child relationship in the D-- propositus was compatible. However, in the present case, at the D1S80 locus, the PCR product derived from the mother was lacking, thereby negating a parent-child relationship. It is probable that the RH genes and D1S80 locus exist in close proximity, because they are situated in chromosomes 1p 34.3–36.1 and 1p 36.1–36.3, respectively. These data suggested that at the stage of gametogenesis, both the RHCE gene and the D1S80 locus from the maternal side may have been deleted, thereby producing the D-- gene complex.

HLA and HPA Typing in Idiopathic Thrombocytopenic Purpura Patients Treated with Kami-kihi-to

We performed human leukocyte antigen (HLA) and human platelet antigen (HPA) in patients with Kami-kihi-to-responsive idiopathic thrombocytopenic purpura. The HLA-A2, A61 and Cw1 were significantly increased in responders compared with nonresponders, as were HLA DRB1 *0901, DRB1 *1502, and DPB1 *0501. In contrast, HLA DPB1 *0201 and DPB1 *0901 were significantly decreased in responders. The a/b genotype of HPA-2 and a/a genotype of HPA-3 were markedly increased in nonresponders, and anti-GPIb antibody was also increased. These results suggest that HLA, HPA, and anti-GP antibody studies may predict the response of idiopathic thrombocytopenic purpura to Kami-kihi-to.

Platelet Procoagulant Activity During,Peripheral Blood Stem Cell Harvest

We used flow cytometry to measure platelet-derived microparticle levels in plasma obtained from 16 patients during peripheral blood stem cell harvest (PBSC) and in platelet concentrates prepared by apheresis from 10 normal controls. We also studied the binding of an anti-P-selectin antibody and annexin-V to platelets. When all 60 harvests were assessed, we noted a significant difference in microparticle levels between patients with a platelet count >10 x 104/μl and those with a platelet count <10 X 10 4/μl (12.3 ± 4.4 vs. 75 ± 3.9%). In both the first and total harvests, the percentage of platelets and microparticles positive for anti-P-selectin and annexin-V were significantly higher than the normal control levels. These results suggest that patients undergoing mobilization by granulocyte colony-stimulating factor (G-CSF) who have a platelet count >10 X 10 4/μl are at risk of increased procoagulant activity after retransfusion following PBSC harvest. Key Words: Platelet-derived microparticle— Peripheral blood stem cell harvest—Granulocyte colony-stimulating factor.

HLA Typing of a Family with Systemic Lupus Erythematosus

Although systemic lupus erythematosus (SLE) is a representative collagen disease, the etiology remains unclear. However, both genetic and environmental factors seem to be involved. Based on serological studies, associations between certain human leukocyte antigens and many autoimmune diseases have long been suggested. Regarding the genetic aspects of SLE, the HLA haplotype is regarded as a potentially important factor, although its role remains controversial. Recently, we examined three family members who had SLE with an identical HLA haplotype.

Evaluation of the Bitterness-Masking Effect of Powdered Roasted Soybeans

The masking of bitterness is considered important because many pharmaceutical compounds have a bitter taste. The bitterness-masking effect of powdered roasted soybeans (PRS) was investigated using a bitter taste sensor. PRS was revealed to significantly suppress the bitterness of quinine hydrochloride and denatonium benzoate. Furthermore, the bitterness-masking mechanism of PRS extracts was evaluated using dynamic light scattering. These results showed that the extracted suspension consisted of particles that were several hundreds of nanometers in size. Analysis of the PRS extracts by nuclear magnetic resonance spectroscopy indicated that denatonium benzoate was entrapped in the PRS extracts. Thus, PRS may be useful as a bitterness-masking agent in orally administered pharmaceuticals.

Flow cytometric analysis of Tk-polyagglutination.

We used flow cytometry to investigate the Tk-polyagglutination that was observed in a patient with metastatic lung cancer (59-year-old man). We analyzed the fluorescent patterns of three types erythrocytes obtained from a patient, healthy type-0 subject, and neuraminidase-treatment using four kinds of FITC-conjugated lectins (Griffonia simplicifolia, GS-II; Dolichos biflorus, DBA; Glycine max, SBA; Arachis hypogaea, PNA). Furthermore, the reactivity of normal serum to the patients erythrocytes was analyzed by FITC-conjugated anti-human IgG and IgM antibodies. The patients erythrocytes bound to FITC-GS-II and -PNA, but neuraminidase-treated erythrocytes reacted to FITC-SBA and -PNA. These results were in accordance with those obtained using the aggregation method. However, the reactivity of normal serum to the patients erythrocytes according to flow cytometric analysis was different from that determined by the aggregation method. In particular, anti-human IgM antibody showed the well-reactive tendency. Our results suggest that flow cytometry was useful for the heterogeneous analysis of Tk-polyagglutination in a patient with malignant tumors.

Report on two cases having anti-Wra antibody with special reference to the background of anti-Wra production.

Here reported are two patients having anti-Wra antibody, which have been uncommon in Japan. Further, we have found 38 such cases so far reported in our country. The 40 cases in all have been examined on their backgrounds which might be related to the anti-Wra production.1) Case report: Case No. 1: A 78-year-old female, complaining of dizziness and palpitation, was found to have autoimmune hemolytic anemia.She had never received blood transfusion. Her blood group was B, CcDEe, Wr (a-). Direct antiglobulin test (DAT) using anti-IgG was positive, and the eluate from RBC reacted against all of the panel red cells, showing presence of panreactive warm-type anti-RBC autoantibody. The patients serum, even after its autoantibody was absorbed by her own RBC, still showed presence of anti-Wra.Case No. 2: A 68-year-old male had been suffering from the myelodysplastic syndrome, which thereafter transformed to myelocytic leukemia.His blood group was A, Rh (D)-positive. The screening test for unexpected anti-RBC antibodies was initially negative, which turned to positive along with DAT after frequent blood transfusions. Further laboratory tests detected panreactive warm-type anti-RBC autoantibody in the eluate from the patients RBC as well as alloimmunized anti-E and anti-Wra in his serum.2) Results obtained by the survey on the anti-Wra in Japan: In the majority of the 40 cases with anti-Wra in Japan, including our above-mentioned two, the anti-Wra seemed to be a natural antibody, while, in 3 of them, anti-Wra was coexistent with the warm-type anti-RBC autoantibody. Besides, other 2 cases seemed to be drug-induced. These findings raise interests in the etiology of anti-Wra. However, the anti-Wra itself may be clinically innocent here in Japan, because any individuals having the blood group antigen Wra have not yet been found among Japanese. In addition, if Wra-positive RBC will be hereafter more commonly included in panel red cells for screening unexpected anti-RBC antibody, anti-Wra may, probably, be more frequently detected in Japan.

Anti-glucocorticoid antibodies found in three cases, and their clinical significance.

Anti-glucocorticoid (GCS) antibody or antibody-like substances were detected in our three patients with malignant lymphoma, SLE, and SLE combined with AIHA, respectively, by means of the hemagglutination test (HA). The incidence of positive HAs among our patients with collagen diseases was as high as 4.5% (2/44 cases). It was possible to neutralize all antibodies by anti-IgM serum, and the corresponding antigen determinant was supposed to be the common structural portion of the GCSs themselves. Since the results of the HA did not always correspond to those of drug-induced lymphocyte stimulation test (DLST) or the patch test, or to such allergic manifestations as urticaria, a plurality in the mechanisms of antibody production and of allergic manifestations against the GCSs was supposed.