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Biomedical subjects

D J Anstee

Publications and source records attributed to D J Anstee.

At least 145 records · Page 8Linked to original sources
PubMed1975

Separation of ABH, I, Ss antigenic activity from the MN-active sialoglycoprotein of the human erythrocyte membrane.

The erythrocyte sialoglycoprotein was purified from cells with complementary ABO(H), MN and Ss phenotypes. Serological examination of the sialoglycoprotein preparations demonstrated that this molecule does not carry the ABO(H), I or Ss antigens. The results also suggest that Ss activity may be associated with a minor erythorcyte glycoprotein. The characteristics of the ABO(H) and I antigens are consistent with earlier suggestions that these antigens are carried on complex glycolipids.

ABO Blood-Group System↗
PubMed1974

The distribution of blood-group antigens on butanol extraction of human erythrocyte 'ghosts'.

The distribution of protein and blood-group-antigen activity obtained after butanol extraction of erythrocyte ;ghosts' under various conditions is described. Butanol extraction under low-ionic strength conditions results in the recovery of membrane protein in high yield in the aqueous phase. Blood-group-A activity is found in both the aqueous and butanol phases, whereas blood-group-P activity is confined to the butanol phase and blood-group-I and blood-group-MN activity are restricted to the aqueous phase. Much lower yields of protein are obtained in the aqueous phase when high-ionic-strength conditions are used. An appreciable amount of material is precipitated at the interface. Under these conditions blood-group-P activity is found only in the butanol phase, blood group-A activity in the butanol phase and interface material and only blood-group-MN activity in the aqueous phase. In contrast with previous reports no correlation could be demonstrated between the secretor status of the donors and the presence of blood-group-A activity in the aqueous phase after butanol extraction under any of the extraction conditions used. By using butanol extraction under high-ionic-strength conditions it is possible to isolate the blood-group-MN-active sialoglycoprotein in high yield from erythrocyte ;ghosts' by a simple procedure.

ABO Blood-Group System↗
PubMedDate not supplied

Studies on the blood of an MiV/Mk proposita and her family.

An individual (J-1) was shown to be heterozygous for the MiV and Mk genes. Her red cells typed as M+(weak), N-, S-, s+(strong), U+, Hil+, Wr(a-b-), En(a+weak). Polyacrylamide gel electrophoresis analysis of her red cell membranes revealed absence of PAS-staining bands corresponding to normal MN and Ss sialoglycoprotein (SGP), and presence of a hybrid MNSs SGP [(alpha-delta)MiV] similar but not identical to that reported for an MiV homozygote. However, J-1 cannot be homozygous for MiV since the red cells of two of her children are Hil- and s-, carry only a single dose of M antigen, and have a sialic acid content that is consistent with the presumption that they are Mk heterozygotes. J-1's hybrid MNSs SGP is considered to be gene-fusion product resulting from unequal crossover between a normal alpha M and delta gene, and her red cells lack that portion of the Ena antigen that is resistant to ficin. Her hybrid MNSs SGP differs, therefore, from that reported for the MiV homozygote, which probably arose from unequal crossover between alpha N and delta genes. Further, the red cells of the MiV homozygote carry the ficin-resistant Ena determinant.

Aged↗
PubMedDate not supplied

NBTS/BRIC 8. A monoclonal anti-C3d antibody.

A stable mouse hybridoma line secreting monoclonal anti-C3d antibody (NBTS/BRIC 8) was produced. Ascitic fluid containing BRIC 8 can be used at a final dilution of 1 to 5000 to provide an excellent antiglobulin reagent when mixed with a conventional polyclonal rabbit anti-human IgG serum.

Animals↗
PubMedDate not supplied

Evidence that the Lub blood group antigen is located on red cell membrane glycoproteins of 85 and 78 kd.

A murine monoclonal antibody of specificity anti-Lub was produced. Immunoblotting of the electrophoretically separated components of membranes from Lu(b+) red cells with the monoclonal antibody identified two glycoproteins of relative molecular mass 85 and 78 kd, respectively. The expression of Lub antigenic activity on these glycoprotein components was shown to be dependent on the presence of one or more N-glycosidically linked oligosaccharides and on the presence of disulphide bonding.

Antibodies, Monoclonal↗
PubMedDate not supplied

A severe transfusion reaction associated with anti-EnaTS in a patient with an abnormal alpha-like red cell sialoglycoprotein.

A 71-year-old woman (Ped) received 3 units of red cells (RBCs), compatible by the indirect antiglobulin test but strongly (4+) incompatible by direct agglutination at 37 degrees C. The next day, her plasma hemoglobin was 1252 mg percent and the direct antiglobulin test (DAT) was weakly positive (IgG and C3). Less than 5 percent of the transfused cells could be detected 48 hours posttransfusion. Her clinical condition deteriorated and renal failure developed. The patient died of pulmonary embolism. Her serum contained a strong (4+) IgM agglutinin and a weakly reactive (microscopically positive) IgG antibody, with anti-EnaTS specificity. EnaFS and EnaTS antigens were severely depressed or absent from the patient's RBCs; the ficin-resistant Ena antigen (EnaFR) appeared to be present. Pretransfusion RBC sialic acid level was 53 +/- 2 percent of normal. The patient's RBC membranes were shown to contain sialoglycoproteins beta and delta by sodium dodecyl sulphate polyacrylamide gel electrophoresis with periodic acid Schiff's base staining, with weak staining of components in the regions corresponding to alpha, alpha 2 and alpha delta. The nature of these components was not identified, but their presence suggested that the patient's RBCs expressed a previously undescribed sialoglycoprotein alpha variant.

Aged↗
PubMedDate not supplied

Murine monoclonal antibody MB-2D10 recognizes Rh-related glycoproteins in the human red cell membrane.

The human red cell membrane components reacting with monoclonal antibody MB-2D10 were examined by immunoblotting. The antibody bound to a diffusely staining band extending from Mr 30,000 up to the high-molecular-weight region of the gel in normal membranes and in Rhnull U + membranes, but not in Rhnull U - membranes. Treatment of normal red cells with an endoglycosidase F-containing preparation destroyed the epitope recognized by MB-2D10. The reactivity of the antibody with purified preparations of Rh-related glycoproteins D30 polypeptide, D50 polypeptide, R6A32 polypeptide, and R6A45 polypeptide was also examined. Only the purified R6A45 and D50 components reacted with MB-2D10. These results show that MB-2D10 recognizes a carbohydrate-dependent epitope on the R6A45 and D50 group of Rh-related polypeptides. The results also suggest the possibility that the U antigen arises from interaction between glycophorin B and the Rh-related components D50 and R6A45.

Animals↗
PubMedDate not supplied

The Wrb antigen in Sta-positive and Dantu-positive human erythrocytes.

Immunoprecipitation using a monoclonal antibody showed that the Wrb antigen is present on the abnormal (delta-alpha) hybrid sialoglycoprotein of Sta-positive human erythrocytes but not on the abnormal (delta-alpha) hybrid sialoglycoprotein of Dantu-positive erythrocytes. These results provide further information regarding the nature and location of the Wrb antigen on the normal erythrocyte sialoglycoprotein alpha.

Amino Acid Sequence↗
PubMedDate not supplied

The nature and abundance of human red cell surface glycoproteins.

Available evidence suggests that there are at least 30 different proteins at the red cell surface. These proteins vary in abundance from a few hundred copies/cell to more than a million copies/cell. Recently, a new Glycophorin gene (Glycophorin E) has been identified and sequenced. In this paper evidence is presented that a monoclonal anti-M detects the product of the Glycophorin E gene in normal red cells.

Blotting, Western↗
PubMedDate not supplied

Functional factors in the red cell membrane: interactions between the membrane and its underlying skeleton.

Recent studies involving two abnormal red cell phenotypes (South-east Asian ovalocytosis and Leach phenotype) provide novel information concerning the nature and significance of interactions of both the anion transport protein AE-1 (syn. band 3) and Glycophorins C and D with the underlying skeleton. The location of Wra and Dia blood group antigens to mutations on AE-1 at residues 658 and 854 respectively, together with the availability of monoclonal antibodies recognising epitopes dependent upon the integrity of the third extracellular loop of AE-1, have allowed us to study the organisation of the membrane domain of the mutant AE-1 found in South-east Asian ovalocytes (AE-1 SAO). The results suggest that the organisation of the whole membrane domain of AE-1 SAO is abnormal and that the organisation of other integral membrane proteins like those involved in expression of Rh blood group antigens may also be affected. Increased homo- and hetero-associations involving AE-1 SAO and other integral proteins may in turn result in reduced membrane flexibility. Purified protein 4.1 binds with 50-fold higher affinity to protein 4.1 depleted normal red cell membranes than to protein 4.1 depleted red cell membranes of Leach phenotype which lack Glycophorin C (GPC) and Glycophorin D (GPD). Experiments using purified protein 4.1 and p55 together with synthetic peptides corresponding to different regions of the cytoplasmic domain of Glycophorins C and D (GPC/D) demonstrate that protein 4.1 interacts directly with GPC through residues 82-98. They also show that p55 binds to GPC through residues 112-128. Since p55 also binds directly to protein 4.1 it is clear that protein 4.1 can bind to GPC through two different sites either directly through residues 82-98 or indirectly through p55. These results show that GPC and GPD provide major attachment sites for the red cell skeleton via protein 4.1 and that p55 is part of this complex.

Anion Exchange Protein 1, Erythrocyte↗
PubMedDate not supplied

Studies on the blood of an MiV homozygote.

An individual, whose parents are third cousins, has been shown to be homozygous for the rare Mi.V. condition. The proposita's red blood cells type as M-, N+(weak), S-, s+(strong), U+, Mi(a-), Vw-, Hil+; Wr(a-b-). The cells react, albeit less strongly than most other samples, with anti-Ena. However, from studies on the red blood cells of the proposita and on those of another person of the En(a+), Wr(a-b-) phenotype, it is apparent that the term "anti-Ena" actually describes a number of antibodies of differing specificities. Inhibition studies with sialoglycoprotein (SGP) isolates, and tests on protease-modified red blood cells illustrate some of the differences in specificity. Biochemical analyses of the SGPs of the red blood cells of the MiV homozygote and those of her parents confirm that the Mi.V condition is associated with the absence of normal MN SGP (alpha) and normal Ss SGP (delta), the appearance of a hybrid SGP molecule comprised of a portion of the MN SGP at its NH2 terminal end, and a portion of the Ss SGP at its C terminal end.

Animals↗