Demonstration of T-transferase deficiency in Tn-polyagglutinable blood samples.
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Biomedical subjects
Publications and source records attributed to A Gerbal.
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Using the indirect immunofluorescence technique, P antigen is not detected on skin fibroblasts from Pk1, Pk2 or p donors. P antigen is reexpressed on polykaryon cells obtained by fusion of Pk x p fibroblasts. On the contrary, P antigen is not expressed on polykaryon cells from Pk x Pk or p x p fusion. All these data suggest that in Pk and p fibroblasts, at least two distinct genes are modified, one in the p phenotype, and the other in the Pk phenotype. The two Pk phenotypes studied seem to be homogeneous from a genetic point of view, as do the five p phenotypes.
An investigation of Cad phenotypes in the French population had been carried out in 1973, in four Blood Transfusion Centers (Mulhouse, Nancy, Paris, Versailles), B and O red cells were tested with the Dolichos Biflorus lectin. Out of 78.528 donors, 56 were found to have the Cad antigen on their red cells. The mean frequency was 0,07%. Nevertheless, this frequency varied among the four above mentioned Blood Transfusion Centers: the observed differences were probably due to the preparation procedure of the Dolichos biflorus extract. The family investigation permitted the analysis of four families with at least three Cad individuals. The independence of the Cad system and of Auberger, Gc, Hp, C'3, PGM, Pac and ADA was demonstrated. A quantitative agglutination study on these Cad samples using the Dolichos biflorus lectin,and a selected AB serum showed a high variability of the erythrocyte Cad Strength, even within one family. Most Cad samples were found polyagglutinable when a sensitive technique and selected AB sera were used. All adult sera contained an anti-Cad1 antibody, except Cad1 individuals. Although strong Sda reactivity was always found in Cad red cells, the anti-Cad and anti Sda specificities were not identical: this was demonstrated by the absorption and inhibition tests of anti-Cad and anti-Sda reagents absorption and inhibition tests of anti-Cad and anti-Sda reagents with Sda material. From thf Cad red cells, there was no evidence of the existence of separable anti-A1 and anti-Cad agglutinins in the Dolichos biflorus lectin.
Many cases of acquired B antigens, always observed in group A subjects have been so far reported. Most of them were found in patients with digestive tract disease, essentially colonic cancer. An investigation on 200 patients in a gastroenterology department showed that this B-like antigen was quite frequent (10,6%); it occurred only in A1 individuals and was related to infectious syndrome. Immunological and serological studies of many cases had shown that this B-like antigen differs from that of normal B cells. Groupe A1 cells transfused to patients acquired B activity; on the contrary group A2 and O cells remained unchanged. Likewise, only A1 cell became active when incubated in vitro with C. Tertium A., known to contain a deacetylase. In 1970, we postulated that a deacetylase enzyme could be responsible for this B-like antigen: this enzyme could transform the N-acetylgalactosamine (A specific sugar) into galactosamine, which could cross react with anti-B sera. The relationship between the acquired B antigen and a deacetylase was recently confirmed: A1 acquired B cells, chemically acetylated lost their B reactivity and enhanced their A1 activity. A polyagglutinability, different from that associated with T, Tn, Cad, Hempas has been always found in acquired B cells; nervertheless, because of its weakness, it could sometimes be unnoticed. Besides, it disappeared prior to B reactivity in case of recovery. Like acquired B activity, it decreased in low pH medium of after acetylation of the cells. Nevertheless, this polyagglutinability appears, contrarly to acquired B antigen, in vitro, on all the cells, irrespective of their ABO phenotypes. A deacetylation of N-acetyl-neruaminic acid could explain such a phenomenon.
A new antigen has been recently discoverd in patients with congenital dyserythropoietic anemia type II. The acronyme Hempas was proposed for this disease as a remind of the main morphological feature of erythroblasts (hereditary erythroblastic multinuclearity) and the characteristic serological findings (positive acidified serum test). The patients red cells are agglutinated and lysed by an IgM cold reacting antibody present in the serum of most normal subjects and not previously recognized. This behaviour is thus reminding of cells carrying antigens such as T, Tn, Cad or acquired B. As for T and Tn cells, sialic acid and electrophoretic mobility are reduced, but in contrast, agglutinability of Hempas cells is enhanced by enzyme treatment. Agglutination by anti H and anti Pr specific reagents is reduced. I and mainly i activity are strongly increased. The relationship between the membrane abnormalities of Hempas red cells and the failure of normoblasts to divide their cytoplasm i still largely unknown.
The chemical acetylation of RBC bearing the acquired B antigen led to the disappearance of the agglutinability by anti-B and restored the A1 specificity. The same results are obtained using RBC transformed in vitro by a Clostridium Tertium filtrate, where a deacetylase was reported.
Various anti-Cad and anti-Sda reagents were tested against a panel of Cad red cells in order to determine whether Cad and Sda antigens were identical. According to the results, although strong Sda reactivity was always found in Cad red cells, the two antigen specificities are not identical. The polyagglutinability of Cad red cells seems to be related to an anti-Cad present in all human sera but Cad, and not with anti-Sda antibody.
Fifteen samples of cis AB bloods belonging to six unrelated families were tested by serological and thermodynamic assay techniques. The B and H antigens of cis AB bloods differ significantly from those of trans AB bloods. Differences were found among unrelated samples, but identical results were obtained within a given family : this could mean that there had been as many mutations as there were families.
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Members of six unrelated families from Japan, France, Belgium and Poland were studied in parallel. Major immunological features characteristic of the phenotype produced by the Cis AB complex are the following: 1) The red cell A reactivity is close to normal, is beyond the values of agglutination scores by Helix and by anti-A from B; likewise, with percent agglutination measurements, A reactive appears hiher than that of A2B cells; one sample only is slightly detected by anti-A from Dolichos. 2) The B reactivity, on the contrary, is lower than that of normal AB cells. A single sample is detected by anti-B from A1. All samples are well detected by anti-B from AW, Aend, Ax, Am but none is detected by anti-B from ABx, Cis AB, or by an auto-anti-B. Under standard conditions, percent aggutination is around 80, very close to that of normal AB cells, thus differentiating Cis AB from AB3 (some of which only reach this figure), and from ABx which are very far from this value. 3) An abnormally high reactivity to anti-H antibody is observed, higher than that of normal A2B, similar to that of A2 red cells. 4) Among secretors, A substance is found to be normal or in excess, H substance is in excess, while B substance is only detected by Cis AB red cells inhibition. 5)An anti-B antibody was identified in the samples studied; however, we recently received from Germany a Cis AB samples, the serum of which did not contain anti-B antibody. By these main characteristics, the studied samples seem to be identical; however, agglutination kinetics and thermodynamic methods show that they differ by their reaction with a same anti-B antibody in standard conditions. The reactive structures of the various samples are indeed different from one family to another. The main point is that identical values were observed in all samples within a same family. Thus, the various Cis AB can be considered as different families mutants.
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50 Cad (+) blood samples were tested using various anti-Cad reagents. Results demonstrated a wide Cad antigenic expression, even among the members of one family. In particular experimental procedures, almost all Cad (+) cells were polyagglutinable.
The study of the alpha-N-acetylgalactosaminyltransferase in the sera of 19 individuals belonging to the rare Am blood group makes it possible to confirm the heterogeneity of this phenotype established on genetical and immunological criteria. Two groups of subjects, Am and Ay, can be distinguished. For the individuals of the first group, named Am, 15 samples (7 families) have been studied, the phenotype is inherited as an allele at the ABO locus. 14 of these subjects, have an alpha-N-acetylgalactosaminyltransferase whose kinetic properties were similar to those of A1 subjects. In one family, however, the A transferase detected is of the A1 type. On a quantitative level, the enzyme activities of these sera only reached 30-50 percent of the average value observed for A1 or A2 subjects, respectively. These facts suggest the existence of a genetic inhibitor, possibly linked to the ABO locus, preventing either an A1 or A2 gene from acting at the level of some cellular lines and leading therefore to the recognition of phenotypes named A-m-A1 and A-m-A2. On the contrary, under the experimental conditions used, no alpha-N-acetylgalactosaminyl-transferase activity was detected among the four individuals of the second group, named A-y by Weiner et al. (37), and whose appeareance in siblings results from the action of a recessive modifying y-A gene.
Nine cases of acquired B antigen were studied. By quantitative methods the variation of the A reactivity and of the number of A sites was found to be inversely related to the variation of the B reactivity. By agglutination kinetics using an immune anti-B, the acquired B reactive structure was found to differ from that of a normal B. Agglutination variation, with pH, points out to the part played by one electrically charged chemical group. No transferase galactosyl activity was found in the serum, and no B substance in the plasma. According to these results the B reactive structure must have been formed at the expense of the A reactive structure. This "acquired B" type may have arisen as a result of the action of a bacterial deacetylase, transforming the A reactive N-acetyl galactosamine into galactosamine.