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Biomedical subjects

H E Heier

Publications and source records attributed to H E Heier.

At least 37 records · Page 2Linked to original sources

[Yersinia enterocolitica septicemia after blood transfusion].

Transfusion-associated transmission of Yersinia enterocolitica was first described in 1982. Since then more than 40 cases have been reported world-wide. The blood units are contaminated from apparently healthy donors who may, however, recently have had an episode of diarrhoea. Y enterocolitica is able to grow in packed red cells at refrigerator temperature. The mortality rate among recipients of contaminated blood is more than 50%. We describe a non-fatal case of a 80 year-old male who received one unit of packed red blood cells contaminated with Y enterocolitica (serogroup O:3, biotype 4). The blood had been collected 14 days before from a Norwegian donor with transient and slight abdominal discomfort. The microbe was isolated both from the patient's blood and from the donor blood bag. The patient was treated with ofloxacin and recovered without sequelae.

Aged↗

Backscatter electron imaging of double immunogold labeled erythrocytes using two primary monoclonal IgM antibodies.

The majority of mouse monoclonal antibodies reacting with blood group epitopes on erythrocytes are of the IgM class, have equal light chain type, and are available as culture supernatants only. To study the interrelationship of the blood group antigens, a method is presented which allows double labeling applying two unconjugated monoclonal antibodies of the same class and species. The method comprises two indirect, sequential labelings using mouse IgM anti-A and anti-H as primary antibodies and two goat anti-mouse IgM conjugated to 30 and 20 nm colloidal gold particles as secondary antibodies. After labeling for the first antigen, free binding sites on the primary antibody are blocked by incubation with an unconjugated goat anti-mouse antibody. The free anti-species on the secondary antibody, conjugated to 30 nm gold particles, are inactivated by silver enhancement. The silver enhancement also enlarges the gold particle for optimal discrimination between the two particle sizes, which are chosen accordingly. Semiquantitations of double labeled cells from subgroup A2 and A3 were found to be in good agreement with the counts of the corresponding single labelings as well as between experiments, irrespective of which of the two antibodies was applied in the first labeling sequence. The results were in accordance with a reciprocal but nonlinear relationship between the A and H antigens and suggest different affinities of the two antibodies for the epitopes in the subgroups investigated, indicating different biochemistry of the antigen determinants.

ABO Blood-Group System↗

Expression of A antigens on erythrocytes of weak blood group A subgroups.

Scanning immune electron microscopy using a monoclonal anti-A antibody which reacts with all type A oligosaccharide chains revealed A antigens on less than 5% of Am and Ael cells, some of which showed extremely strong labelling. This explains why Am and Ael cells can absorb significant amounts of anti-A without being agglutinated. A3 may be a heterogeneous subgroup, since A antigens were found on 82 and 58%, respectively, of the cells of 2 A3 individuals. A antigens were found on 75% or more of Ax cells. In many weak A individuals A-positive cells are apparently best detected if an anti-A is used which reacts strongly with other A oligosaccharide chains than type 2. From hyperimmune pregnancy sera Ax, Am and Ael erythrocytes absorbed antibodies which seemed to have other fine specificities than those absorbed by A2 cells. We conclude that weak subgroups of A may deviate from A2 both by number of erythrocytes expressing A antigens and the biochemical nature of the antigens.

ABO Blood-Group System↗

[External national quality control of blood group typing 1983-92. Significance for quality assurance in transfusion medicine].

A programme for external quality control in blood group serology has been run in Norway since 1983 by the National Institute of Public Health. Uncomplicated AB0 and Rh(D) grouping, including Du typing, has been performed satisfactorily. Rh(D) grouping of presensitized, Rh (D) negative cells is still a problem, but one which has been reduced significantly. Both indirect and direct antiglobulin reactions have given about 15% false negative results, mainly with weak antibodies and weakly sensitized cells. Poor training of technicians in reading weak agglutination reactions may be an important source of error. The importance of post-graduate training of blood bank staff is emphasized.

ABO Blood-Group System↗

[Production and consumption of blood and blood products in Norway. Factors of quality assurance in transfusion medicine].

In Norway self-sufficiency has been achieved with regard to all major blood products, despite a relatively low number of blood donations. There is good balance in the consumption of the various products. The total consumption of erythrocytes has gone down only slightly since pre-HIV times, while consumption of thrombocytes has increased. There is probably an over consumption of erythrocytes, albumin and plasma, and more attention should be given to the consistent application of principles of modern haemotherapy. If each department is given responsibility for its own economy, improvement may be achieved in this respect.

Blood Component Removal↗

Antibody binding to blood group antigens in relation to temperature: scanning electron microscopy of immunogold-labelled erythrocytes.

Binding patterns of mouse monoclonal antibodies (mAb) to P1, Pk, N, I, H, Y or A antigens were visualized in the backscatter electron imaging mode of a scanning electron microscope by indirect immunogold labelling. Experiments were performed at room temperature (RT) and at 4 degrees C. In experiments with anti-P1 and anti-Pk, clusters of immunolabelling particles dominated the immunolabelling pattern much more at RT than at 4 degrees C. By contrast, no clustering was seen with anti-N, even at RT. Clustering was also observed at RT with anti-I, anti-H and anti-Y, and on some Ax and A3 cells with anti-A, but was much reduced at 4 degrees C. Immunolabelling was stronger at 4 degrees C than at RT with all mAb except anti-N and anti-A. The results indicate that glycolipid blood group antigens are more mobile in the membrane of intact erythrocytes at RT than at 4 degrees C, and that the cells bind more antibodies to such antigens at 4 degrees C than at RT. We suggest that antigen immobilization in the cold will reduce cross-linking of antigens and hence increase the number of antibody molecules needed for epitope saturation, leading to increased binding of antibody in the cold. This may be the main reason for cold-enhanced agglutination with human blood group antibodies.

Animals↗

Expression of B and H antigens on red cells from a group B(weak) individual studied by serologic and scanning electron microscopic techniques.

The proposita was classified as B(el), B(y), or B(m), Le(b+) by routine blood grouping and by adsorption/elution studies using anti-A and -B hyperimmune pregnancy sera. Red cells from the proposita adsorbed as much anti-B from the hyperimmune sera as did red cells from normal B individuals, but adsorbed less anti-A,B (group O serum). Saliva contained H, but not B, soluble substance. Red cells from the proposita and a normal B donor were sensitized with monoclonal A and B blood group antibodies immunolabeled with colloidal gold particles, and examined in a scanning electron microscope. B antigens were found on more than 95 percent of normal B cells, but on only 2-3 percent of red cells from the proposita. However, when the same cells were sensitized with anti-A,B that reacted strongly with B oligosaccharides other than type 2 chains, half of the labeled red cells from the proposita were labeled more strongly than any normal B cells. Our results explain why red cells from the proposita adsorb significant amounts of anti-B and anti-A,B without being agglutinated by these antibodies. The results of both adsorption/elution and immunolabeling suggest that the B antigen on her cells differs biochemically from that on normal B cells.

Journal Article↗

Effect of the metal in the reaction between metallothionein and antimetallothionein antibody.

1. An antimetallothionein antibody, raised against Cd-carrying metallothionein, was applied in Western blotting of metallothionein. 2. Treatment of the electroblotted nitrocellulose sheets with metals belonging to the periodic system transition groups Ib and IIb, or with Pb, Ni or Cr, considerably enhanced binding of anti-metallothionein. A similar effect was found when the electroblotted sheets were treated with the strong alkylator N-ethylmaleimide. 3. It seems that the binding of metal to metallothionein modifies the configuration of the antibody binding sites by the formation of metal thiolate complexes. 4. Metal treatment of the nitrocellulose sheets after electroblotting, but before application of the primary antibody, offers a convenient method for use in Western blotting to significantly potentiate the reaction between metallothionein and the antimetallothionein antibody.

Alkylation↗

Mitogenicity and metallothionein induction: two separate effects of zinc ions on human mononuclear blood cells.

The mitogenic effect of elevated concentrations of zinc ions on human lymphocytes was found to be inhibited by the glucocorticoid hormone dexamethasone. The effect decreased progressively from complete block to partial inhibition when the culture period before the addition of dexamethasone was prolonged. In contrast, dexamethasone did not inhibit an induced formation of metallothionein in mononuclear cells. Furthermore, dexamethasone itself was found to induce small amounts of metallothionein. Apparently, the mitogenic effect and the induction of metallothionein by zinc ions in mononuclear cells occur by mutually independent mechanisms.

Cells, Cultured↗

Induction of metallothionein production by zinc in human mononuclear cells.

1. Low mol. wt proteins with high Cd-binding capacity were found to be induced by Zn in cultured monocytes and lymphocytes. 2. In T cell cultures one protein was found to be induced by 125 microM Zn for 6 days, while in monocytes and B enriched cells under these conditions two proteins were found, of which the one with higher mol. wt had similar electrophoretic mobility to the T cell protein on polyacrylamide gels. 3. Mol. wt criteria and crossreactivity towards anti-metallothionein (Mt) antibody identified these proteins to be Mts of about 23 and 27 kDa mol. wt. 4. Metal binding studies indicated that monocyte and lymphocyte Mts had a higher affinity to Zn compared to Cd than rat liver Mt and Mts from Cd-resistant substrains of a human epithelial and a murine fibroblast cell line. 5. The presence of the T cell mitogen phytohemagglutinin (PHA) was found not to be necessary for this Mt induction by Zn.

B-Lymphocytes↗

IgG subclass antibody responses to pneumococcal polysaccharide vaccine in splenectomized, otherwise normal, individuals.

Subclasses of IgG antibodies to pneumococcal polysaccharide serotype antigens 4, 6A, and 23F were measured before and 4 weeks after vaccination with pneumococcal vaccine in young individuals splenectomized because of trauma and in a control group. An ELISA technique was applied. IgG2 anti-pneumococcal antibodies predominated before vaccination, especially against serotypes 4 and 6A. The youngest individuals in the splenectomy group tended to have lower IgG2 anti-pneumococcal antibody levels than the older ones. Vaccination induced antibodies of the IgG1 and IgG2 subclasses, and in some individuals also of the IgG4 subclass. Splenectomy does not seem to influence the IgG subclass pattern of antipneumococcal antibodies.

Adolescent↗

Silver enhancement of gold probes (5-40 nm): single and double labeling of antigenic sites on cell surfaces imaged with backscattered electrons.

Silver enhancement of immunogold-labeled cells was carried out to increase the applicability of colloidal gold probes for visualization in the backscatter electron imaging (BEI) mode of a scanning electron microscope. Optimum conditions were established for single particle discrimination and differential counting of labeling density at low magnifications. Red blood cells double-labeled with 15 + 40 nm and 5 + 20 nm gold probes were silver-enhanced for 6 min and 20 min, respectively, at which times both pairs of labels increased to about 25 + 50 nm. The gold probes still appeared spherical after enhancement and were easily discriminated. Cells were also single-labeled with the above probes and enhanced accordingly. The present method enables visualization of individual particles of any probe size, labeling one, or simultaneously two, antigenic sites on cell surfaces. The silver enhancement procedure thereby allows cells to be labeled with small probes with increased labeling efficiency.

Antigens, Surface↗

Binding patterns of monoclonal anti-B, anti-H and anti-(Leb + Y) on erythrocytes, imaged in the scanning electron microscope.

Binding patterns of monoclonal IgM anti-B, anti-H and anti-(Leb + Y) antibodies on erythrocytes were visualized in the backscatter electron imaging (BEI) mode of a scanning electron microscope by indirect immunogold labelling. The results were independent of secretor status, supporting the view that type 2 oligosaccharides predominate on erythrocytes. There were considerable cell-to-cell variations of amounts of the antigens, presumably because of differences of antigen development. The amount of H antigens may have been overestimated in previous studies. B and H antigens seemed linked mostly to mobile structures, probably lipid, on the convex parts of the cells and to immobile structures, probably protein, on the concave parts. Y antigens appeared bound to mobile structures also on the concave part of the cell membrane. The results therefore confirm that B oligosaccharides on erythrocytes develop from both lipid- and protein-bound H, but suggest that Y develops mainly from lipid-bound H.

ABO Blood-Group System↗

A study of immunogold-labelled blood group A erythrocytes in the scanning electron microscope.

This report presents binding patterns on A1, A2, A3, Ax and Ae1 erythrocytes of a monoclonal anti-A (A 003) antibody which reacts predominantly with difucosylated A oligosaccharides, visualized with colloidal gold particles in the backscattered electron imaging mode of a scanning electron microscope. A relatively weak labelling was found on most A1 cells, while the labelling in subgroups A2, A3 and especially Ax appeared relatively stronger. Very few Ae1 cells were labelled. The results emphasize the qualitative uniqueness of A1 and suggest that many Ax cells have high proportions of difucosylated A oligosaccharides. Labelling variations were found between different A3 and Ax traits, and in all subgroups between cells and even between different parts of the cells. No immunolabelling differences were found in relation to secretor status.

Antibodies, Monoclonal↗

Reactivity with legume lectins of three monoclonal antibodies made against the Lathyrus odoratus lectin.

Three murine IgG1 monoclonal antibodies (MoAb) were produced against the glucose/mannose-specific two-chain mitogen from seeds of Lathyrus odoratus (sweet pea) belonging to the Vicieae tribe of the Leguminosae family. Their antigenic specificities were tested against subunits of two-chain and one-chain legume lectins separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and electrotransferred to nitrocellulose filters. Different binding to native and detergent-treated lectins in dot-blots indicated that two of the antibodies bound to conformational epitopes which by immunoblotting were shown to be expressed on the heavy subunits from all the five tested two-chain lectins and on the six one-chain lectins, including PHA (Phaseolus vulgaris) and Con A (Canavalia ensiformis). However, ELISA cross-inhibition studies showed that these two antibodies bound to different epitopes on the lectin molecules. The third MoAb reacted with a continuous epitope not revealed by classical taxonomy since only three of five heavy subunits of the two-chain lectins stained with the antibody. Comparison of the known amino-acid sequences of the chains indicates eight positions as possible binding sites for this antibody.

Antibodies, Monoclonal↗

Antibody dependent cell mediated cytotoxicity against anti-D sensitized human erythrocytes. Influence of various effector cells and enzyme treatment of the target cells.

Antibody dependent cell mediated cytotoxicity (ADCC) was measured against anti-D sensitized erythrocytes using various mononuclear cell populations from peripheral blood in a chromium release assay. Adherent effector cells gave stronger cytotoxicity than unseparated and non-adherent cells, and were less dependent on pretreatment of the target cells with papain. However, close correlations were found between the cytotoxicity obtained with the various effector cell populations, indicating that either of them might be used for the purpose of establishing in vitro methods to determine the clinical significance of erythrocyte antibodies. The optimal choice of effector cells is discussed.

Adult↗