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D J Anstee

Publications and source records attributed to D J Anstee.

At least 91 records · Page 5Linked to original sources
PubMed1988

Evidence for the occurrence of human erythrocyte membrane sialoglycoproteins in human kidney endothelial cells.

Several mouse monoclonal antibodies recognising different epitopes on human erythrocyte sialoglycoproteins alpha and beta (syn: glycophorins A and C, respectively) react with human renal endothelium. Those monoclonal anti-blood group M and anti-blood group N antibodies which recognised the M/N epitopes on sialidase-treated human erythrocytes also reacted with human renal endothelium whilst those which recognise the M/N epitopes on native, but not on sialidase-treated human erythrocytes were not reactive with human renal endothelium. These results provide evidence for the occurrence of alpha and beta molecules in renal endothelium and suggest that alpha in renal endothelium is incompletely sialylated.

Antibodies, Monoclonal↗
PubMed1988

The Ina and Inb blood group antigens are located on a glycoprotein of 80,000 MW (the CDw44 glycoprotein) whose expression is influenced by the In(Lu) gene.

The Ina and Inb blood group antigens were found to be located on an erythrocyte membrane glycoprotein of 80,000 MW by immunoblotting with human anti-Ina and anti-Inb antibodies under non-reducing conditions. This glycoprotein is shown here to be identical to that defined by monoclonal antibodies to CDw44, and a new murine monoclonal antibody (BRIC 35) is added to this cluster. Experiments with endo-beta-galactosidase and Endo F preparations suggest that the glycoprotein contains one or more N-glycans but that these oligosaccharides do not contain extensive poly-N-acetyllactosaminyl sequences. Experiments using membranes prepared from sialidase-treated normal erythrocytes, from Tn erythrocytes and from Cad erythrocytes suggest that the glycoprotein does not contain a substantial content of O-glycans. The Inb antigen and the epitope defined by a murine monoclonal antibody (BRIC 35) show reduced expression on Lu(a-b-) erythrocytes which result from the effect of the dominant inhibitor gene In(Lu). Evidence is presented here that the Inb antigen is expressed on normal granulocytes and lymphocytes and on the haemopoietic cell lines HEL, K562 and HL-60, a lymphoblastoid cell line and lymphocytes from two patients with B-CLL.

Antibodies, Monoclonal↗
PubMed1987

Structural relationships between human erythrocyte sialoglycoproteins beta and gamma and abnormal sialoglycoproteins found in certain rare human erythrocyte variants lacking the Gerbich blood-group antigen(s).

The human erythrocyte membrane sialoglycoproteins beta and gamma are important for the maintenance of the discoid shape of the normal erythrocyte. In this paper we show that the human erythrocyte sialoglycoproteins beta and gamma (hereafter called beta and gamma) are structurally related. Rabbit antisera produced against purified beta and beta 1 and rendered specific to the cytoplasmic portion of these proteins also react with the cytoplasmic portion of gamma. Some human anti-Gerbich (Ge) sera react with the extracellular portion of both beta and gamma. This reactivity is shown to be directed towards a common epitope on beta and gamma. However, most anti-Ge sera do not react with beta, but react with an extracellular epitope only present on gamma. All individuals who lack the Ge antigens lack beta and gamma. In some cases abnormal sialoglycoproteins are present in the erythrocytes, and these are shown to be structurally related to beta and gamma. Rabbit antisera raised against the purified abnormal sialoglycoprotein from a Ge-negative erythrocyte type reacted with the cytoplasmic portion of both beta and gamma. Unlike normal beta and gamma, the abnormal sialoglycoproteins found in Ge-negative erythrocytes migrate as a diffuse band on SDS/polyacrylamide-gel electrophoresis. Studies using endoglycosidases suggest that the diffuse nature of these bands results from carbohydrate heterogeneity and that the abnormal sialoglycoproteins contain N-glycosidically linked oligosaccharides with repeating lactosamine units. Such polylactosamine chains are not present on normal beta or gamma.

Blood Group Antigens↗
PubMed1987

Normal membrane function of abnormal beta-related erythrocyte sialoglycoproteins.

Red cells totally deficient in beta and gamma sialoglycoproteins (the Leach type of Gerbich-negative) are elliptocytic and have altered membrane physical properties as evidenced by marked decreases in both membrane mechanical stability and membrane deformability. Red cells from individuals who are of the Gerbich and Yus phenotypes of Gerbich-negative are also deficient in beta and gamma sialoglycoproteins, but possess abnormal beta-related sialoglycoproteins. In order to determine if these beta-related sialoglycoproteins can functionally substitute for normal sialoglycoproteins, we measured membrane deformability and stability of red cells of the Gerbich and Yus phenotypes. In contrast to the red cells of the Leach phenotype, cells of Gerbich and Yus phenotypes were found to have normal membrane deformability and stability. Moreover, flow cytometric analysis using a monoclonal anti-beta antibody revealed that the Gerbich and Yus phenotype red cells expressed the beta-related sialoglycoprotein to the same extent as its normal counterpart on normal cells. Based on these data, we suggest that the abnormal beta-related sialoglycoproteins can functionally substitute for normal beta and gamma sialoglycoproteins.

Erythrocyte Deformability↗
PubMed1987

Estimation of the number of binding sites for a murine monoclonal anti-Lub on human erythrocytes.

The number of Lub antigen sites on the human erythrocyte membrane, as recognised by the murine monoclonal antibody BRIC-108, has been determined. The number of antibody molecules bound per cell on one example of cells of the phenotype Lu(a-b-) with recessive inheritance was an average of around 200 in replicate determinations, which probably represents non-specific antibody binding. A similar number of antibody molecules to this bound to trypsin- or pronase-treated normal cells or cells of the Lu(a-b-) phenotype associated with the inheritance of the X-borne gene, XS2. The number of binding sites on three examples of cells of the phenotype Lu(a-b-) with dominant inheritance was from 440 to 690. A variation in the number of binding sites per cell from 1,640 to 4,070 was found in five individuals with the phenotype Lu(a-b+) and from 850 to 1,820 in four individuals with the phenotype Lu(a+b+). Four individuals with the Lu(a+b-) phenotype had an average of 480 binding sites per cell. The Lub antigen therefore appears to a have low-site density and a variable level of expression on the erythrocyte surface.

Animals↗
PubMed1987

Inheritance of human-erythrocyte Gerbich blood group antigens.

The blood group-antigenic determinant Gerbich was first described greater than 25 years ago, but its mode of inheritance has not been established. We performed protein immunoblotting by means of anti-beta sialoglycoprotein (SGP) and anti-gamma SGP reagents. The anti-beta SGP was a monoclonal antibody that reacts with normal beta SGP and with the abnormal beta-related SGPs associated with Gerbich and Yus types of Ge-negative red cells. In the families studied, we have shown that the products of the Ge alleles are inherited in an autosomal codominant manner.

Alleles↗
PubMed1987

A human cell-surface glycoprotein that carries Cromer-related blood group antigens on erythrocytes and is also expressed on leucocytes and platelets.

A new human erythrocyte glycoprotein has been identified by immunoblotting with murine monoclonal antibodies under non-reducing conditions. The glycoprotein has a MW of 70,000 and carries Cromer-related blood group antigens. The monoclonal antibodies also react with normal peripheral blood leucocytes and platelets and several haemopoietic cell lines. The glycoprotein has a reduced MW after sialidase treatment. The MW is markedly reduced in Tn erythrocyte membranes and slightly increased in Cad erythrocyte membranes. These results suggest that the glycoprotein has a substantial content of O-glycans. The glycoprotein appears to be absent from, or grossly altered in, the erythrocytes of two individuals with the rare Inab phenotype.

Antibodies, Monoclonal↗
PubMed1986

Identification and partial characterization of the human erythrocyte membrane component(s) that express the antigens of the LW blood-group system.

Rhnull human erythrocytes lack the antigens of the Rhesus blood-group system, have an abnormal shape, have an increased osmotic fragility, and are associated with mild chronic haemolytic anaemia. Rhnull erythrocytes also lack all antigens of the LW blood-group system, but the functional significance of this deficiency is unknown. We have identified, by immunoblotting with two mouse monoclonal antibodies (BS46 and BS56), the LW-active component(s) in normal human erythrocytes as a broad band of Mr 37 000-47 000 on SDS/polyacrylamide-gel electrophoresis. Treatment of intact human erythrocytes with endoglycosidase F preparation destroyed the epitopes recognized by antibodies BS46 and BS56, suggesting that one or more N-glycosidically linked oligosaccharides are required for the formation of the LW antigens. Estimation of the number of LW antigen sites per erythrocyte by using radioiodinated purified antibody BS46 gave average values of 4400 molecules/cell for Rh(D)-positive adult erythrocytes and 2835 molecules/cell for Rh(D)-negative adult erythrocytes. Like the Rh(D) polypeptide, the LW polypeptide(s) is (are) associated with the cytoskeleton of normal erythrocytes. These results suggest the possibility that the absence of the LW polypeptide may also contribute to the functional and/or morphological abnormalities of Rhnull erythrocytes.

Acetylglucosaminidase↗
PubMed1986

The use of monoclonal antibodies to quantify the levels of sialoglycoproteins alpha and delta and variant sialoglycoproteins in human erythrocyte membranes.

By using radioiodinated monoclonal antibodies we have estimated that there are about 600 000 copies of sialoglycoprotein alpha (synonym glycophorin A) and 80 000 copies of sialoglycoprotein delta (synonym glycophorin B) per normal human erythrocyte. Erythrocytes expressing the product of only one alpha gene contain about 300 000 copies of alpha/cell. Two erythrocyte types containing alpha-delta hybrid molecules were studied. Those with heterozygous expression of the (alpha-delta)Mi.V gene contain about 100 000 alpha-delta copies per cell, whereas those with heterozygous expression of the En(UK) gene contain about 80 000 alpha-delta copies/cell. Erythrocyte types containing delta-alpha hybrid molecules were also studied. About 200 000 copies of (delta-alpha)Dantu were measured in cells with heterozygous expression of the (delta-alpha)Dantu gene (donor M.P.), whereas about 315 000 copies of the putative (delta-alpha)Dantu hybrid were found on the erythrocytes of donor J.O. [which also have heterozygous expression of the putative (delta-alpha)Dantu gene]. The erythrocytes of donor M.P. have normal levels of alpha, whereas those of donor J.O. have only about half-normal levels. It is proposed that the hybrid sialoglycoprotein of donor J.O. is of alpha-delta-alpha composition [(alpha-delta-alpha)Dantu] rather than delta-alpha and results from a double cross-over analogous to that which gives rise to haemoglobin Parchman.

Antibodies, Monoclonal↗
PubMed1986

Inhibition of malarial invasion by intracellular antibodies against intrinsic membrane proteins in the red cell.

It has previously been shown that antibodies against the transmembrane proteins, band 3 and glycophorin A, inhibit entry of the merozoite into the red cell and, in the case of band 3, it was established that attachment of the parasite to the cell is not prevented. We have found that antibodies against the cytoplasmic domains of band 3 and of glycophorin A, when present in the interior of resealed ghosts of human red cells, also inhibit invasion by P. falciparum. It is inferred that attachment of the merozoite to the red cell causes structural effects that are transduced to the membrane cytoskeleton and the antibodies against transmembrane proteins interfere with the invasion sequence at this level.

Animals↗
PubMed1986

A family demonstrating inheritance of the Leach phenotype: a Gerbich-negative phenotype associated with elliptocytosis.

The proposita of a Canadian family and 2 of her sibs have the rare Leach phenotype and provide the first evidence that the Leach phenotype is an inherited condition. Their red cells are Ge: -2, -3 and fail to react, by the antiglobulin test, with the monoclonal antibodies GERO, BRIC 4 and BRIC 10. The minor sialoglycoproteins beta, beta 1 and gamma are apparently absent from their red cell membranes. A proportion of their red cells are elliptocytes indicating that beta and/or beta 1 and/or gamma have a function in maintaining normal red cell shape.

Antibodies, Monoclonal↗
PubMed1985

Abnormal minor human erythrocyte membrane sialoglycoprotein (beta) in association with the rare blood-group antigen Webb (Wb).

Individuals whose erythrocytes are positive for the rare blood-group antigen Webb (Wb) have an altered form of the minor sialoglycoprotein beta (synonyms glycophorin C and glycoconnectin). This altered sialoglycoprotein beta (beta Wb) has an Mr about 2700 lower than that of normal sialoglycoprotein beta. Treatment of normal sialoglycoprotein beta with endo-beta-N-acetylglucosaminidase F decreased its Mr by about 3600, but similar treatment of sialoglycoprotein beta Wb had no effect. These results suggest the possibility that sialoglycoprotein beta Wb lacks the N-glycosidically linked oligosaccharide found on normal sialoglycoprotein beta.

Blood Group Antigens↗
PubMed1985

The phospholipid organisation in the membranes of McLeod and Leach phenotype erythrocytes.

The phospholipid composition, the distribution of phospholipids over the two membrane layers as well as the phosphatidylcholine-specific transfer protein-mediated exchangeability of phosphatidylcholine from the membrane, has been investigated in two types of abnormal erythrocytes--the McLeod phenotype and the Leach phenotype. The acanthocytic McLeod cells appeared to have a normal phospholipid composition and distribution, but the exchangeability of phosphatidylcholine was found to be markedly enhanced. Unlike control erythrocytes, in which 75% of all of the phosphatidylcholine can be exchanged during an 8 h incubation, the McLeod cell showed a complete exchange of this phospholipid within the same time period. This obviously indicates an enhanced transbilayer mobility of phosphatidylcholine in the membrane of McLeod cells. Erythrocytes of the Leach phenotype showed an elliptocytic shape and increased osmotic fragility, but no abnormalities were observed as to the composition and organisation of the phospholipid complement of their membranes.

Erythrocyte Membrane↗
PubMed1984

The use of functional and quantitative assays to study glycoprotein Ib in platelets stored under various in vitro conditions.

There is much evidence to suggest that platelet membrane glycoprotein Ib is involved in the haemostatic function of platelets and it has been suggested that loss of this glycoprotein may occur during in vitro storage of platelet concentrates. In this study two quantitative radioimmunoassays were developed to measure the content of glycoprotein Ib in platelets stored under a range of conditions used in blood banks. One assay involved the use of iodinated Maclura pomifera lectin and the other the binding of a monoclonal antibody (AN51) specific for glycoprotein Ib. The results showed that there was no significant reduction in the glycoprotein Ib content of platelets under the storage conditions used. These results suggest that any loss of haemostatic effectiveness which occurs on in vitro storage of platelet concentrates is not attributable to a selective loss of glycoprotein Ib from the platelet surface.

Antibodies, Monoclonal↗
PubMed1984

The Rhesus (D) polypeptide is linked to the human erythrocyte cytoskeleton.

Cytoskeleton preparations derived from lactoperoxidase-radioiodinated human erythrocytes were found to be enriched in a labelled component with the same apparent molecular mass as the Rhesus (D) (Rh(D] antigen polypeptide. Immune precipitation from the cytoskeleton preparations confirmed that this component is the Rh(D) polypeptide. The results suggest that the Rh(D) polypeptide may be linked to the erythrocyte skeletal matrix. The possibility that the Rh(D) antigen is involved in maintaining the shape and viability of the erythrocyte is discussed.

Erythrocyte Membrane↗
PubMed1984

Rhnull human erythrocytes have an abnormal membrane phospholipid organization.

Rhnull human erythrocytes lack the antigens of the Rhesus blood group system, have an abnormal shape and an increased osmotic fragility, and are associated with mild chronic haemolytic anaemia. Studies with phospholipase A2 and sphingomyelinase C show that the asymmetric distribution of phosphatidylethanolamine (PtdEtn) in the membrane of these cells differs from that found in control cells. The amount of PtdEtn which can be hydrolysed by phospholipase A2 in the presence of sphingomyelinase C in intact Rhnull cells is twice as high as that in normal erythrocytes. In intact Rhnull cells all of the phosphatidylcholine (PtdCho) present in the membrane can be readily exchanged with a PtdCho-specific exchange protein, whereas in control cells 75% is readily exchanged and 25% at a much lower rate. This indicates that PtdCho experiences a relatively fast transbilayer movement in the Rhnull cells. The observation that the loss of two membrane polypeptides in the Rhnull cells leads to abnormal shape, increased osmotic fragility, abnormal PtdEtn distribution and enhanced transbilayer mobility of PtdCho strongly suggests that one or both polypeptides are essential for the maintenance of a proper membrane-membrane skeleton interaction.

Erythrocyte Membrane↗