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Biomedical subjects

A Lubenko

Publications and source records attributed to A Lubenko.

At least 55 records · Page 3Linked to original sources

Western blotting is a sensitive technique for the detection of anti-PlA1.

Western (immuno-) blotting was evaluated as a technique for anti-PlA1 detection, using untreated and chloroquine or low pH-treated platelets. Chloroquine-treated platelet membranes generated the cleanest blots, giving end-point titres of 4000-16,000 for three different anti-PlA1 sera. These titres were between three and eight doubling dilutions higher than those obtained in solid phase, ELISA or indirect immunofluorescence tests. Nitrocellulose strips used for blotting could be stored for at least six months at room temperature without significant loss of PlA1 activity. However, immunoblotting still failed to detect anti-PlA1 in the sera of four of fivePl (A1-) women whose infants were born with neonatal thrombocytopenia of probable alloimmune origin.

Antibodies↗

A new qualitative variant of the RhE antigen revealed by heterogeneity among anti-E sera.

We have recently detected a new qualitative variant of the RhE antigen characterised by negative reactions with a minority of anti-E sera, and an inability of the relevant cells to completely absorb most, if not all, anti-E samples that we tested. In tube tests, using a two-stage enzyme technique, cells from the proposita (V.R.) were completely agglutinated by 8 out of 12 undiluted polyclonal sera, weakly agglutinated (2+ and 3+ reactions) by 2 sera, but not agglutinated by the 2 remaining sera. Cross-absorption tests using 1 non-reacting, 1 weakly reacting and 4 strongly reacting polyclonal anti-E sera showed that V.R.'s cells only slightly reduced their anti-E titres versus r"r cells, whilst completely removing their reactivity against her own cells. Absorptions with R2R2 cells abolished activity against both cell types. The cells of V.R. failed to absorb or elute anti-Ew, and had normal expression of c. Three monoclonal human anti-E reagents were tested by fluorescence flow cytometry; one reagent failed to react with the cells of V.R., a second bound as strongly with V.R.'s cells as with R1R2 controls while the third bound much more weakly with V.R. than with any other E+ sample. The variant of RhE in the V.R. family is therefore distinct from Ew, and lacks an epitope recognised by a major constituent of most anti-E sera. This epitope is unlikely to be ET, since all of 91 samples from E-positive Australian aborigines (many of which are expected to lack ET) were agglutinated by a serum that failed to react with V.R.'s cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Agglutination Tests↗

Should anti-Rh immunoglobulin be given D variant women?

D variant women occasionally form anti-D during or following pregnancy with a D-positive fetus. It is not known whether Rh immunization could be suppressed by using anti-Rh immunoglobulin (Rh Ig), or whether the injected antibodies would be absorbed by the woman's D variant cells. In order to predict the likely outcome, three anti-Rh Ig preparations were absorbed independently with five examples of D variant red cells: R1VIr (n = 3), R1Br (n = 2), and with cells of common Rh-positive and Rh-negative phenotypes: R0, r'r, r"r or rr cells (n = 1 each). The titres of the Ig preparations against all these cells were compared after three, six, nine and 12 absorptions. The titres of the unabsorbed anti-Rh Igs against R0 cells ranged from 2(15) to 2(18). Anti-D activities were hardly affected by absorption with r'r, r"r or rr cells, but were completely exhausted after three to six absorptions with R0 cells. Most of the D variant cells were as ineffective as rr cells in reducing anti-D activity against R0 cells, but one R1VIr variant produced a fall in titre against R0 cells, equivalent to four or five doubling dilutions after three to six absorptions, which could not be reduced further by subsequent absorptions. Therefore, a substantial proportion of the anti-D in the immunoglobulin preparation does not bind to D variant red cells. The component of the anti-D that does not bind to D variant cells would be expected to be present in the circulation of D variant individuals injected with anti-Rh immunoglobulin, and should be effective in suppressing Rh immunization. An increased dose of the anti-Rh Ig might have to be administered in order to allow for the fraction that can bind to D variant red cells and which would not be available for immunosuppression.

Absorption↗

Analysis of the heterogeneity of the binding site specificities of hyperimmune human anti-A and -A, B sera: the application of competition assays using murine monoclonal antibodies.

Monoclonal antibodies PL41 and AL62 have previously been shown to recognize two distinct blood group A epitopes on the red cell surface. Competitive inhibition of the binding of 125I-PL41 and 125I-AL62 to group A1 red cells, by hyperimmune polyclonal human antibodies, has been employed to investigate the binding site specificities of 15 anti-A and 8 anti-A,B sera. Differences in the degree of inhibition of the binding of the two MABs by individual anti-A or -A,B samples indicate that polyclonal reagents are composed of varying proportions of up to 3 (or more) different antibody specificities, each recognizing a distinct epitope: PL41-like, AL62-like and a third (as yet undefined) category of antibody. In general, those anti-A sera with PL41-like specificities are superior agglutinators of A2B cells with weakly expressed A antigens; similarly, the specificity of potent anti-A,B, sera capable of strongly agglutinating Ax cells was likewise directed towards PL41-binding blood group A trisaccharide haptens.

ABO Blood-Group System↗

Kg, a new low-frequency red cell antigen responsible for hemolytic disease of the newborn.

Hemolytic disease of the newborn in a Japanese infant led to studies which indicate that an antibody detected in the maternal serum is recognizing a hitherto unknown red cell antigen. The antigen, which we have named Kg, was found in two generations of the family and it is inherited as a Mendelian dominant character. The maternal serum failed to react with the red cells of more than 600 random Japanese blood donors, 64 red cell samples known to possess low-frequency antigens and all but one of 75 red cell samples known to lack high-frequency antigens without recognized low-frequency antigens. The father's red cells were tested extensively for established low-frequency antigens; only Dia was demonstrated and the maternal antibody was shown not to contain anti-Dia.

ABO Blood-Group System↗

Two examples of a new low-frequency red cell antigen, JFV.

A new low frequency red cell antigen JFV has been found in two families, one of German and one of Dutch origin. JFV is inherited as an autosomal dominant character. Family studies showed that JFV is not controlled by the Rh, MNS, Lu, Fy, or Jk loci and that it is not X- or Y-borne.

Female↗

Expression of blood group H antigen by normal, benign, and carcinoma cells of the oral epithelium: immunohistochemical study using monoclonal antibody RS13.

The H antigen was investigated by an indirect immunoperoxidase method in sections of 65 surgical specimens from the oral mucosa. These comprised 29 squamous cell carcinomas, 28 benign lesions, and 8 specimens of clinically healthy mucosa. A monoclonal antibody (RS13) was used to identify the H antigen. The peroxidase stain was positive at high dilutions of the antibody in the epithelium of normal mucosa, and the titers were significantly higher than those within benign lesions (p less than 0.001) and carcinomas (p less than 0.001). However, the titers in the benign lesions varied considerably, with two specimens recorded as negative. In contrast, the reaction for the H antigen was negative in 19 specimens (66%) of the malignant lesions and the endpoint titers of the H positive carcinomas indicated marked loss of the antigen. This loss was significant when compared to the benign lesions (p less than 0.001). The results show that the loss of the H antigen on malignant epithelial cells may be a valuable marker for primary squamous cell carcinoma.

ABO Blood-Group System↗

Epitope specificity of blood-group-A-reactive murine monoclonal antibodies.

Monoclonal antibodies (MABs) towards blood groups A manifested three broad patterns of binding distinguished on the basis of affinities for A1 or A2B phenotype red blood cells and blood group substances. Competitive inhibition of binding of radiolabelled MABs by other unlabelled antibodies and absorption studies with synthetic haptens suggested that GpA specificity was expressed as two topographically related structures, one being the terminal GpA trisaccharide and the second probably involving part of the oligosaccharide backbone. The enhancement of binding of an extremely low-affinity antibody by higher affinity antibodies recognising either epitope indicated a topographic, possibly conformational relationship between these epitopes. It is suggested that only those antibodies that recognise terminal GpA trisaccharides can agglutinate weak A2B cells.

ABO Blood-Group System↗

Serology and genetics of an MNSs-associated antigen Dantu.

Dantu, a previously undescribed low-incidence red cell antigen, is inherited as a Mendelian dominant character. The Dantu antigen is associated with very weak s antigen, protease resistant N antigen and either very weak or no U antigen. Two of the propositi had previously been shown to have an unusual hybrid MNSs sialoglycoprotein, and it is probably this which carries these unusual N, s and U antigens as well as the Dantu antigen. A study of the family of one propositus suggests, by conventional genetics, that Dantu is not controlled by the MNSs locus; a possible explanation is given. Several examples of anti-Dantu are known, one was found to cause a positive direct antiglobulin reaction on neonatal red cells.

ABO Blood-Group System↗