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

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*

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

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*

Good's syndrome (thymoma with immunodeficiency) is a rare cause of combined B-cell and T-cell immunodeficiency in adults. We present here a case of Good's syndrome involving a 52 year-old man with an ABO blood group abnormality. He had undergone surgery for thymoma with myasthenia gravis 27 years ago. He also had a history of pulmonary tuberculosis, herpes zoster and pure red cell aplasia. On admission, he was suspected of having pneumonia, and S. pneumoniae was isolated from blood culture. The immunoglobulin levels were markedly decreased. Lymphocyte subset analysis revealed the absence of CD19+ B cells. The result of ABO typing showed a normal strong reaction on the cell typing, but a relatively weak reaction on the serum typing. Therefore, we performed ABO genotyping to confirm his ABO type, which was revealed to be B/O1 . This case suggests that ABO typing should be performed when the diagnosis of Good's syndrome is made. Moreover, Good's syndrome (thymoma with hypogammaglobulinemia) should be considered and evaluated for in patients with a weak ABO reverse type.
Adult ; B-Lymphocytes ; Herpes Zoster ; Humans ; Immunoglobulins ; Lymphocyte Subsets ; Myasthenia Gravis ; Pneumonia ; Red-Cell Aplasia, Pure ; T-Lymphocytes ; Thymoma ; Tuberculosis, Pulmonary

BACKGROUND: Anti-K is one of the most significant unexpected antibodies that cause hemolytic transfusion reactions. Individuals with anti-K have to be transfused with K antigen-negative red cells. Although Koreans rarely have the K antigen, we have detected three cases of anti-K and we analyzed the Kell blood group genotypes for the KEL*1/KEL*2 alleles at the same time. METHODS: We analyzed the KEL*1/KEL*2 allele genotypes from 261 blood donors at Seoul National University Bundang Hospital. Kell genotyping were carried out using polymerase chain reaction (PCR) and restriction enzyme length polymorphism (RFLP). Identification of anti-K was performed using three kinds of methods; 37degrees C albumin, an anti-human globulin phase tube, a bead cassette and a gel card. Three cases of anti-K also underwent PCR with a sequence specific primer (SSP) for Kell genotyping. For comparison, the KEL*1 allele (698C>T) was synthesized by site-directed-mutagenesis. RESULTS: All 261 donors were KEL*2/KEL*2 homozygotes and a digested KEL*1 allele was not found. The three patients with anti-K were also KEL*2/KEL*2 homozygotes and the reactivities of the anti-K identification test were the same. CONCLUSION: The KEL gene frequency for the KEL*1/KEL*2 allele corresponded with that of the Kell phenotype, as was previously reported. We experienced three cases of anti-K and two out of the three were assumed that they had been transfused with the K antigen-positive blood of foreigners. This study revealed that the possibility of anti-K alloimmunization and hemolytic transfusion reactions cannot be excluded in Koreans.
Alleles ; Antibodies ; Antigens, Bacterial ; Antigens, Surface ; Blood Donors ; Blood Group Incompatibility ; Emigrants and Immigrants ; Gene Frequency ; Genotype ; Homozygote ; Humans ; Phenotype ; Polymerase Chain Reaction ; Tissue Donors