No abstract available.

The T cell antigen receptor (TcR) in combination with costimulatory signals triggered by accessory molecules present on the surface of the antigen-presenting cells (APC) regulates the activation and growth of T lymphocytes. Calyculin A and Okadaic acid is known to be an inhibitor of serine/threonine phosphatase and RK-682 specifically blocks functions of tyrosine phosphatase. To investigate roles of these inhibitors in TcR-mediated signaling cascade, chimeric molecule CD8-5 which contains the extracellular and transmembrane domains of the human CD8a molecule and the cytoplasmic tail of TcR 5 chain were stably expressed in Jurkat cell line. CD8-5 chimeric protein induced tyrosine phosphorylation of various cytoplasmic substrates and IL-2 gene expression in a NFAT dependent manner by stimulation with anti-CD8 mAb OKT8 as seen in TcR stimulation. When CD8-5 transfectants were preincubated with Okadaic acid, Calyculin or RK682, they differentially affected tyrosine phosphorylation of signaling mediators including CD8-5 molecule. When Jurkat Tag cell line was used where SV40 T antigen is stably expressed and the expression of p-galactosidase is driven by the multiple NFAT binding sites plus minimal IL-2 promoter, these phosphatase inhibitors -RK682, Calyculin A, Okadaic acid- effectively inhibited IL-2 gene expression at the concentration of 1.2832 x 10 ' M, 3.9924 x 10 M, 7.2707 x 10 M respectively. These results suggested that Okadaic acid, Calyculin or RK682 modulate TcR-proximal as well as TcR-distal signaling events during T cell activation.
Antigen-Presenting Cells ; Antigens, Viral, Tumor ; Binding Sites ; Cell Line ; Cytoplasm ; Gene Expression ; Humans ; Interleukin-2 ; Jurkat Cells ; Okadaic Acid* ; Phosphorylation ; Receptors, Antigen, T-Cell ; T-Lymphocytes ; Tyrosine

We report a case of hemolytic disease of the newborn caused by anti-Mia (Miltenberger) antibody. Full term male infant was admitted due to hyperbilirubinemia on second day of life. Total serum bilirubin level was 8.6 mg/dL at 12 hours of age and 12.3 mg/dL at 24 hours of age. The blood group of patient and his mother were both RhD positive B type. Direct antiglobulin test was strongly positive in the patient, and testing of maternal serum and patient's serum against a red cell panel including cells known to carry the antigenic determinants of some Miltenberger phenotypes revealed the presence of anti-Mia . Testing of paternal red cells and patient's red cell against anti-Mia serum revealed positive reaction. This report documents the first case of hemolytic disease of the newborn due to anti-Mi a in Korea.
Bilirubin ; Coombs Test ; Epitopes ; Humans ; Hyperbilirubinemia ; Infant ; Infant, Newborn* ; Korea ; Male ; Mothers ; Phenotype

OBJECTIVE: The purpose of this study was to investigate the incidence of apoptosis and expression of bcl-2 in the placenta of normal pregnancy, Pregnancy Induced Hypertension, and Intrauterine Growth Restriction. METHODS: Placenta samples were collected from 15 cases of normal full-term pregnancies, 15 cases of second trimester pregnancies, 17 cases of Pregnancy Induced Hypertension, and 13 cases of Intrauterine Growth Restriction. Hematoxylin and eosin staining and TUNEL (terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate marker nick end-labeling) staining were used to quantify the incidence of apoptosis and the electron microscopy was used to confirm it. Expression of bcl-2 was confirmed by using immunohistochemical stain in relation to apoptosis. RESULTS: 1. In TUNEL staining, quantification of apoptosis was 1.05 in 2nd trimester (n=15), 3.65 in pregnancy induced hypertensive pregnancy (n=17), 2.92 in intrauterine growth restrictive pregnancy (n=13) and 1.93 in normal full-term pregnancy (n=15). The incidence of apoptosis was significantly higher in placental tissues from full-term pregnancies than second trimester pregnancies (p<0.05, t test), and higher in pregnancy induced hypertensive pregnancy and in intrauterine growth restrictive pregnancy than in normal full-term pregnancy (p<0.05, t test). 2. Bcl-2 expression was significantly higher in placental tissues from second trimester pregnancies than full-term pregnancies (p<0.05 t test). But, there was no statistically significant difference between pregnancy induced hypertension and normal full-term pregnancy (p>0.05, t test), and between intrauterine growth restriction and normal full-term pregnancy (p>0.05, Mann-Whitney U test). CONCLUSION: These data suggest that apoptosis increases with gestational age, and in pathophysiologic states such as pregnancy induced hypertension and intrauterine growth restriction, and that bcl-2 expression is lower with gestational age.
Apoptosis* ; Deoxyuridine ; Eosine Yellowish-(YS) ; Female ; Gestational Age ; Hematoxylin ; Humans ; Hypertension, Pregnancy-Induced* ; In Situ Nick-End Labeling ; Incidence ; Microscopy, Electron ; Placenta ; Pregnancy Trimester, Second ; Pregnancy*

OBJECTIVE: The purpose of this study was to investigate the incidence of apoptosis and expression of bcl-2 in the placenta of normal pregnancy, Pregnancy Induced Hypertension, and Intrauterine Growth Restriction. METHODS: Placenta samples were collected from 15 cases of normal full-term pregnancies, 15 cases of second trimester pregnancies, 17 cases of Pregnancy Induced Hypertension, and 13 cases of Intrauterine Growth Restriction. Hematoxylin and eosin staining and TUNEL (terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate marker nick end-labeling) staining were used to quantify the incidence of apoptosis and the electron microscopy was used to confirm it. Expression of bcl-2 was confirmed by using immunohistochemical stain in relation to apoptosis. RESULTS: 1. In TUNEL staining, quantification of apoptosis was 1.05 in 2nd trimester (n=15), 3.65 in pregnancy induced hypertensive pregnancy (n=17), 2.92 in intrauterine growth restrictive pregnancy (n=13) and 1.93 in normal full-term pregnancy (n=15). The incidence of apoptosis was significantly higher in placental tissues from full-term pregnancies than second trimester pregnancies (p<0.05, t test), and higher in pregnancy induced hypertensive pregnancy and in intrauterine growth restrictive pregnancy than in normal full-term pregnancy (p<0.05, t test). 2. Bcl-2 expression was significantly higher in placental tissues from second trimester pregnancies than full-term pregnancies (p<0.05 t test). But, there was no statistically significant difference between pregnancy induced hypertension and normal full-term pregnancy (p>0.05, t test), and between intrauterine growth restriction and normal full-term pregnancy (p>0.05, Mann-Whitney U test). CONCLUSION: These data suggest that apoptosis increases with gestational age, and in pathophysiologic states such as pregnancy induced hypertension and intrauterine growth restriction, and that bcl-2 expression is lower with gestational age.
Apoptosis* ; Deoxyuridine ; Eosine Yellowish-(YS) ; Female ; Gestational Age ; Hematoxylin ; Humans ; Hypertension, Pregnancy-Induced* ; In Situ Nick-End Labeling ; Incidence ; Microscopy, Electron ; Placenta ; Pregnancy Trimester, Second ; Pregnancy*

The systematic study of products from bacteria and fungi has led to the development of two immunosuppressive drugs, cyclosporin A and FK 506 (tacrolimus) which are useful to suppress adaptive immune responses to the grafted tissue. However, they affect all immune responses indiscriminately and are both toxic to kidneys and other organs. To facilitate the development of immunosuppressor to block the T cell receptor (TcR)-mediated signaling cascade specifically, a novel Jurkat T cell transfectants, JK NFAT-SEAP were generated in which the expression of the secreted alkaline phosphatase (SEAP) is driven by the multiple NFAT binding sites plus minimal IL-2 promoter. Upon stimulation with ionomycin or anti-TcR mAb OKT3 in the presence of PMA, these transfectants secreted high level of SEAP into the medium, which was conveniently analyzed by SEAP analysis. The secretion of SEAP was effectively inhibited by cyclosporin A or FK 506 at the concentration of [10 ' ug/ml], [10 ug/ml] respectively. JK NFAT-SEAP transfectants will provide two major advantages for the development of a novel immunosuppressor. First, analysis of SEAP secreted into the culture medium by SEAP analysis enables us to test a large number of samples within a short period of time. Second, Usage of IL-2 promoter for the expression of SEAP makes us identify bioproducts to target specifically on TcR-mediated signaling pathway.
Alkaline Phosphatase ; Bacteria ; Binding Sites ; Cell Line* ; Cyclosporine ; Fungi ; Interleukin-2 ; Ionomycin ; Kidney ; Mass Screening* ; Muromonab-CD3 ; Receptors, Antigen, T-Cell ; T-Lymphocytes* ; Tacrolimus ; Transplants

Posttransfusion purpura is a rare but potentially fatal disorder, characterized by the development of severe thrombocytopenia following transfusion. We report the first case of posttransfusion purpura in Korea in a 39-year-old multiparous woman. Nine days after transfusion of two units of red blood cells during cesarian section operation, she developed sudden onset of purpura, gum bleeding, and severe thrombocytopenia. Serological analysis revealed that the patient had antibodies against HPA-3a (Baka) and HLA. She was treated with plasmapheresis and the platelet count dramatically normalized. Though posttransfusion purpura is a self-limited disorder, early diagnosis and proper treatment can shorten the duration of the clinical course.
Adult ; Antibodies ; Early Diagnosis ; Erythrocytes ; Female ; Gingiva ; Hemorrhage ; Humans ; Korea* ; Plasma Exchange ; Plasmapheresis ; Platelet Count ; Purpura* ; Thrombocytopenia

BACKGROUND: The number of reagent red cells has an effect on the results of an unexpected antibody screening test. We evaluated the effect of the number of reagent red cells on antibody screening and identification using test panels of a DG Gel (Diagnostic Grifols, Barcelona, Spain). METHODS: A total of 310 samples were tested in parallel using SeraScan Diana 2 and SeraScan Diana 4 (Diagnostic Grifols, Barcelona, Spain) and ID-DiaCell (DiaMed, Cressier, Morat, Switzerland). Positive samples as determined by the use of SeraScan and ID-DiaCell were identified on an ID-Dia panel (DiaMed), Identisera Diana and Identisera Extend (Diagnostic Grifolsn). RESULTS: Among the 310 samples, 54, 59 and 59 samples were determined as antibody positive by the use of SeraScan Diana 2, SeraScan Diana 4 and ID-DiaCell, respectively. Unexpected antibodies were identified in 10/59 samples (17%) by the use of SeraScan Diana 4, and were identified in 34/59 samples (57.6%) by the combined use of SeraScan Diana 2 and SeraScan Diana 4. Identification of unexpected antibodies by the use of Identisera Diana or Identisera Extend was not different. CONCLUSION: When the results of SeraScan Diana 2 and SeraScan Diana 4 are integrated, unexpected antibodies could be identified in 57.6% of the screening-positive samples. Therefore, if the number of reagent red cells is increased, some antibodies can be identified by antibody screening tests, and the results can be used to validate those of antibody identification tests.
Antibodies ; Mass Screening*

BACKGROUND: ABO genotyping is being widely used in the case of ABO discrepancies and in forensic medicine. We have designed a method using a multiplex single-base primer extension reaction that has allowed us to detect six single nucleotide polymorphism (SNP) sites in the ABO gene and to determine ABO genotypes. METHODS: Genomic DNA was isolated from the peripheral blood of 75 unrelated Korean subjects. Exon 6 containing nucleotides 261 and 297 and exon 7 containing nucleotides 703, 802, 803 and 1059 were amplified using two pairs of primers. Using the products as templates, a multiplex single-base primer extension reaction was performed with six typing primers of different lengths for the six SNP sites. These reactions were performed on a PTC-200 thermal cycler (MJ Research, Waltham, MA, USA) using the SNaPshot multiplex kit (Applied Biosystems, Foster City, CA, USA), and the products were analyzed using an ABI 3130xl Genetic Analyzer (Applied Biosystems). RESULTS: The ABO genotypes determined by this method (75/75) all matched the genotypes that were determined by the use of the polymerase chain reaction using sequence-specific priming (PCR-SSP). We analyzed the peak pattern detected at each of the six SNP sites for each sample. For the smaller-sized primers, peaks were shifted to the right-side compared with the expected site and for the larger-sized primers peaks was close to the expected site. In addition, the coefficients of variation (CVs) of the smaller-sized primers were higher than the CVs of the larger-sized primers. CONCLUSIONS: We are able to detect six SNP sites in the ABO gene and to determine ABO genotypes using a multiplex single-base primer extension reaction.
DNA ; Exons ; Forensic Medicine ; Genotype ; Nucleotides ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide

BACKGROUND: Blood loss due to laboratory phlebotomy among neonates is correlated with anemia as well as transfusion. In this study, microcolumn agglutination cards for performing ABO & RhD blood typing and direct antiglobulin tests in neonates were evaluated and compared with other established systems. Also, the blood group antibody production rates according to the age were calculated to determine the upper age limit for the new method. METHODS: Eighty subjects were tested by using the DianaGel Neonatal cards (Diagnostic Grifols, Barcelona, Spain), and the results were compared with those of the slide methods for ABO and RhD blood typing, and the DiaMed-ID DC-Screening I test (DiaMed, Morat, Switzerland) for direct antiglobulin tests. A total of 546 subjects who were under 12 months old were tested for the ABO back-typing, and 58 subjects with the AB blood type were excluded. RESULTS: The results of the DianaGel Neonatal card were in agreement with those of the conventional methods for all the subjects. Only one subject showed a discrepant result for the DAT between the DianaGel and DiaMed methods. Blood group antibodies were detected in 29 out of 169 (17.2%) one-day-old neonates, in eight out of 34 (23.5%) infants between one and three months of age and in 81 out of 96 (84.4%) infants between six and twelve months of age. CONCLUSION: The DianaGel Neonatal card showed at least equivalent performance as compared to that of the conventional methods, and it showed advantages for a low blood volume requirement and stronger agglutination grades. The DianaGel card is a suitable alternative for blood typing and DAT in infants under the age of 3 months and who do not necessarily need back-typing of the blood groups due to the low production rate of antibodies.
Agglutination ; Anemia ; Antibodies ; Antibody Formation ; Blood Group Antigens ; Blood Grouping and Crossmatching ; Blood Volume ; Coombs Test ; Humans ; Infant ; Infant, Newborn ; Phlebotomy