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Biomedical subjects

A Lubenko

Publications and source records attributed to A Lubenko.

At least 37 records · Page 2Linked to original sources
PubMed1995

Prediction of the severity of haemolytic disease of the newborn. Quantitative IgG anti-D subclass determinations explain the correlation with functional assay results.

Sera containing anti-D, taken from 44 RhD-negative women with RhD-positive infants, were tested in antibody-dependent cellular cytotoxicity (ADCC) and monocyte monolayer assays (MMA) which used similar target and effector cell populations. In addition, the anti-D concentration was measured in the Auto Analyzer and the number of IgG1 and IgG3 anti-D molecules bound to the target red cells was measured by flow cytometry. The results of the functional assays and Auto Analyzer quantitation were examined for correlation with IgG subclass quantitation and all results were compared for their ability to predict the severity of haemolytic disease of the newborn (HDN). ADCC correctly predicted HDN in 39/44 (88.6%) cases, Auto Analyzer quantitation in 35/44 (79.5%) and the MMA in 32/44 (72.7%). For all three assays, the number of correct predictions was highest when the maternal serum contained both IgG1 and IgG3 anti-D. ADCC activity and HDN were correlated with the number of cell-bound IgG1 molecules (r > or = 0.58), but MMA activity was most closely correlated with the number of cell-bound IgG3 molecules (r = 0.68). Hence the superior predictive value of ADCC is due to its ability to reflect the IgG1 component of maternal anti-D, which has a better correlation than IgG3 anti-D with the severity of HDN.

Autoanalysis↗
PubMed1994

A survey of the incidence of Miltenberger antibodies among Hong Kong Chinese blood donors.

BACKGROUND: The ready availability of red cells of the Miltenberger (Mi) class III phenotype (6.28%) prompted the study of Mi antibodies among Chinese blood donors in Hong Kong, 98 percent of whom are descended from inhabitants of Guangdong Province in southern China. STUDY DESIGN AND METHODS: Red cells of the Mi class III phenotype were used to conduct a survey of the frequency of Miltenberger antibodies in 56,161 random Chinese blood donors, over a period of 12 months, using a microplate technique. RESULTS: Sera from 32 donors (0.057%) were found to contain Mi antibodies: sera from 22 contained anti-Mur + Hut; sera from 4 contained anti-Vw + Mur + Hut; sera from 4 had monospecific anti-Mur; and sera from 2 had monospecific anti-Hil. The immunoglobulin isotypes of 24 sera were mixtures of IgM and IgG, 4 were pure IgM, and 4 were pure IgG. CONCLUSION: The majority of Mi antibodies detected were naturally occurring. This survey proved useful for mass screening of random donors for the procurement of valuable Mi antisera.

Blood Donors↗
PubMed1994

In vitro culture of colony forming unit-megakaryocyte (CFU-MK) in fetal alloimmune thrombocytopenia.

Perinatal alloimmune thrombocytopenia (PAITP) causes intracranial haemorrhage in the fetus and neonate. However, the severity of the thrombocytopenia correlates poorly with maternal anti-platelet antibody titres. To test the hypothesis that reduced platelet production contributes to fetal thrombocytopenia in PAITP, maternal sera from three HPA-1a-negative mothers whose pregnancies were complicated by anti-HPA-1a (two severe cases, one mild case) were added to colony forming unit-megakaryocyte (CFU-MK) cultures from HPA-1a positive and negative individuals. Sera from the two severely affected pregnancies containing anti-HPA-1a caused 66-100% inhibition of HPA-1a-positive fetal and neonatal CFU-MK, whereas CFU-MK from two HPA-1a-negative mothers were not inhibited by the anti-HPA-1a-containing sera. Maternal serum from the case of mild PAITP caused only mild inhibition of HPA-1a-positive cord and adult CFU-MK and did not inhibit HPA-1a-positive fetal CFU-MK. Taken together, these findings suggest that reduced megakaryocyte production contributes to fetal thrombocytopenia due to maternal anti-HPA-1a antibodies and also that the degree of CFU-MK inhibition correlates with severity of fetal thrombocytopenia.

Adult↗
PubMed1994

Transplacental IgG subclass concentrations in pregnancies at risk of haemolytic disease of the newborn.

The relationship of haemolytic disease of the newborn (HDN) to the transplacental passage of the four IgG subclasses was assessed at various gestational ages by comparing the maternal and fetal IgG subclass concentrations in 34 pregnancies at risk of HDN with those in 30 pregnancies not at risk. Higher maternal and fetal IgG1 levels were attained in pregnancies at risk of HDN than in pregnancies not at risk. In contrast, a slight decrease in maternal IgG2 and IgG4 levels occurred in pregnancies at risk of HDN, as compared with a slight rise in maternal IgG2 and IgG4 levels in pregnancies not at risk of HDN. Changes in fetal IgG2 and 4 concentrations in either type of pregnancy were very similar, showing only slight increases between the 19th and 34th week of gestation. A slight decrease in maternal IgG3 occurred in both types of pregnancy. In contrast, higher and fairly steady levels of fetal IgG3 were observed in fetuses not at risk of HDN throughout gestation, when compared with those in 'at risk' pregnancies. However, the statistical reliability of these results is not clear since only small numbers of samples were tested and because wide variations in IgG concentrations were observed. The IgG subclass concentrations in 50 paired maternal and cord blood samples were also measured and revealed that IgG1 levels were substantially higher in cord rather than maternal blood; cord and maternal IgG2, 3 and 4 levels, on the other hand, were fairly similar.

Erythroblastosis, Fetal↗
PubMed1994

Prenatal determination of human platelet antigen type using DNA amplification following amniocentesis.

OBJECTIVES: To demonstrate that fetal human platelet antigen (HPA1) type can be determined, without the need for fetal blood sampling, by amplification of fetal DNA from amniotic fluid cells using polymerase chain reaction and allele specific oligonucleotide hybridisation. DESIGN: Oligonucleotide DNA primers were designed to amplify a portion of the platelet glycoprotein GpIIIa gene which spans the site of the single base change which differentiates HPA1a from HPA1b. Specific oligonucleotides were designed to hybridise either to the amplified HPA1a allele or to the HPA1b allele. Amniotic cells were used as the DNA template both directly and following formal isolation of DNA. Fetal HPA1 type, determined by this method in fifteen pregnancies not at risk of perinatal alloimmune thrombocytopaenia, was compared to typing of fetal blood obtained following cordocentesis. The methodology was then used to HPA type the fetus in two pregnancies at risk of the disease. SETTING: Department of Molecular Biology and Centre for Fetal Care, Queen Charlotte's Hospital. SUBJECTS: Fifteen women undergoing amniocentesis and fetal blood sampling for other indications and two women at risk of perinatal allo-immune thrombocytopaenia whose partners were heterozygotes. RESULTS: In the 15 control cases and the two clinical cases, determination of fetal HPA1 type from amniotic fluid cells agreed with typing of fetal blood. There was no difference in the efficiency of amplification from amniotic fluid cells directly or from isolated DNA. CONCLUSIONS: Fetal HPA type may be reliably determined by amplification of DNA from amniotic fluid cells, eliminating the need for fetal blood sampling or immunoglobulin administration when the fetus is HPA1a negative.

Amniocentesis↗
PubMed1994

Monocyte isolation by flow cytometer-monitored centrifugal elutriation: a preparative tool for antibody-dependent cell-mediated cytotoxicity (ADCC) assays.

The value of isolating monocytes by flow cytometer-monitored centrifugal elutriation (CE) after initial density gradient centrifugation (DGC) was investigated. A pooled cell suspension prepared by DGC+CE had a monocyte purity of 90% while a pool prepared by DGC alone had a monocyte purity of only 30%. Monocyte suspensions prepared from 15 separate blood donations demonstrated that, despite wide variation in initial monocyte purity following DGC (i.e. 13-32%), CE consistently increased the purity to 85-92%. Antibody-dependent cell-mediated cytotoxicity (ADCC) studies showed that the most sensitive and reproducible assays were those performed with the DGC+CE pool. At a monocyte concentration of 2 x 10(6)/ml the DGC+CE pool was almost twice as active as the DGC pool (60.2% lysis vs 31.2% lysis, respectively). Considerable variation in ADCC activity was seen when using monocytes from individual donors. We conclude that centrifugal elutriation is a valuable tool for preparing effector cells for use in monocyte driven ADCC assays.

Antibody-Dependent Cell Cytotoxicity↗
PubMed1993

Low-incidence red cell antigen ELO (700.51): evidence for exclusion from thirteen blood group systems.

Genetic studies of the families of two unrelated propositi prove that the low-incidence antigen ELO (ISBT No. 700.51) is not part of the MNS, Rh, Duffy, Kidd, Xg, Kx and Gerbich blood group systems and indicate that it is probably not part of the Lutheran, Scianna, Dombrock or Colton systems. Serological studies indicate exclusion from the Kell and Chido/Rodgers systems. Although the ELO antigen is rare, anti-ELO is not uncommon in sera containing at least one specificity for a low-incidence antigen.

Blood Group Antigens↗
PubMed1992

Molecular biology for platelet alloantigen typing.

Hitherto, full investigation of patients with alloimmunization to platelet-specific antigens has been difficult due to the limited availability of both typing reagents and panels of typed platelets. Following recent advances in the understanding of the molecular and genetic basis of platelet alloantigens, it is now possible to genotype individuals for the alleles coding for the epitopes of four platelet antigen systems (HPA-1-4). This is based on the finding that the two alleles differ by only a single base pair substitution, resulting in one amino acid difference in the relevant platelet glycoprotein. The technique involves amplification of the relevant segments of genomic DNA from any nucleated cell by the polymerase chain reaction, followed by restriction fragment length polymorphism analysis. The technique allows investigation of thrombocytopenic individuals and fetuses/neonates, and can be readily applied to large-scale typing of platelet donors.

Alleles↗
PubMed1992

Haemolytic disease of the newborn probably due to anti-ELO, an antibody to a low frequency red cell antigen.

Investigation of mild bilirubinaemia and a positive direct antiglobulin test in a 2 day old baby revealed that the mother's serum contained an antibody against the low frequency antigen ELO, which was present on the father's red cells. Family studies showed that the ELO antigen segregates from Rh, Gc, ADA and PGM1. The ELO antigen is enzyme resistant and therefore not likely to be part of the MNS or Duffy systems. No abnormalities were detected in immunoblotting studies. Although insufficient samples were available to attempt elution of the antibody from the cord cells, it is probable that this case represents the first reported haemolytic disease of the newborn due to anti-ELO.

Adult↗
PubMed1992

Severe haemolytic disease in an infant born to an Rh(null) proposita.

A 35-year-old Brazilian woman (gravida 4, para 2) was delivered of a severely anaemic child whose cord red blood cells had a strongly positive direct antiglobulin test and who required two exchange transfusions within 24 h of birth. Because of the emergency of the situation and the lack of a local immunohaematology reference laboratory, the phenotype of the mother and the specificity of the relevant antibody could not be determined. Hence, compatible blood was not immediately available and the infant had to be given repeated exchange transfusions with incompatible group 0 Rh-negative blood. The infant is now healthy and thriving. The mother's red cells were subsequently found to lack all the antigens of the Rh system, and her serum reacted with all red cell samples except those of two unrelated Rh(null) individuals. Her serum gave high titres (i.e. 1,024-4,096) by the indirect antiglobulin test against red cells of normal Rh phenotype, as well as against cells with partially deleted Rh phenotypes (titres = 128-512 with -D-/-D- and .D./.D. samples, respectively), and was extremely active in antibody-dependent cell-mediated cytotoxicity and monocyte monolayer assays against red cells of normal Rh phenotypes.

Adult↗
PubMed1992

TSEN: a novel MNS-related blood group antigen.

We report an antibody (anti-TSEN) that recognizes an antigen (TSEN) at the unique amino acid sequence that results from the junction of GPA58 to GPB27 if the GPB carries the S antigen. Red cells from several unrelated donors that possess this specific GP(A-B) hybrid molecule were agglutinated by anti-TSEN. Since a synthetic peptide with the amino acid sequence at this junction (Pro-Glu-Glu-Glu-Thr-Gly-Glu-Met-Gly-Gln-Leu-Val-His-Arg) specifically inhibited anti-TSEN, it must detect an antigen within this novel amino acid sequence. The TSEN antigen has been provisionally assigned the MNS blood group system number 002.033 on behalf of the ISBT Working Party on Terminology for Red Cell Surface Antigens.

Adult↗