scholarly journals Immunochemical characterization of rhesus proteins with antibodies raised against synthetic peptides

Blood ◽  
1993 ◽  
Vol 82 (2) ◽  
pp. 669-676 ◽  
Author(s):  
P Hermand ◽  
I Mouro ◽  
M Huet ◽  
C Bloy ◽  
K Suyama ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Western Blot ◽  
Red Blood Cells ◽  
Blood Group ◽  
Blood Cells ◽  
Synthetic Peptides ◽  
Rare Variants ◽  
Polyclonal Antibodies ◽  
Rh Proteins ◽  
Rh Blood Group

Rabbit polyclonal antibodies were raised against synthetic peptides corresponding to hydrophilic regions of the human Rhesus (Rh) IX cDNA- encoded polypeptide predicted to be extracellularly or intracellularly exposed in the topologic model of the Rh blood group protein. Four antibodies encompassing residues 33–45 (MPC1), 224–233 (MPC4), 390–404 (MPC6), and 408–416 (MPC8) were characterized and compared with a polyclonal anti-Rh protein obtained by immunization with purified Rh proteins. All antibodies had specificity for authentic Rh polypeptides and reacted on Western blot with Rh proteins immunoprecipitated with human monoclonal anti-RhD, -c, and -E. MPC1, but not the other antibodies, agglutinated all human erythrocytes except Rhnull and Rhmod cells, which either lack totally or are severely deficient in Rh proteins, respectively. Immunoblotting analysis with membrane proteins from common and rare variants showed that MPC1 and MPC8 reacted in Western blot with 32-Kd Rh polypeptides from all common red blood cells except those from Rhnull and Rhmod, indicating that peptide regions 33– 45 and 408–416 may be common to several if not all Rh proteins, whatever the Rh blood group specificity. MPC4 reacted only with membrane preparations from cells carrying the E antigen, whereas MPC6 recognized preferentially the Rh proteins from E and Ee preparations, suggesting that the protein encoded by the RhIXb cDNA carries the E and/or e antigen(s). Immunoadsorption experiments using inside-out or right-side-out sealed vesicules from DccEE red blood cells as competing antigen showed that the MPC6 and MPC8 antibodies bound only to the cytoplasmic side of the erythrocyte membrane, thus providing evidence for the intracellular orientation of the C-terminal 27 residues of the Rh polypeptides. Attempts to transiently or stably express the Rh polypeptides. Attempts to transiently or stably express the Rh cDNA in eukaryotic cells were largely unsuccessful, suggesting that Rh antigen expression at the cell surface requires correct transport and/or folding of the Rh proteins, possibly as a complex with one-membrane proteins of the Rh cluster that are lacking in Rhnull cells.

scholarly journals Immunochemical characterization of rhesus proteins with antibodies raised against synthetic peptides

Blood ◽  
1993 ◽  
Vol 82 (2) ◽  
pp. 669-676 ◽  
Author(s):  
P Hermand ◽  
I Mouro ◽  
M Huet ◽  
C Bloy ◽  
K Suyama ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Western Blot ◽  
Red Blood Cells ◽  
Blood Group ◽  
Blood Cells ◽  
Synthetic Peptides ◽  
Rare Variants ◽  
Polyclonal Antibodies ◽  
Rh Proteins ◽  
Rh Blood Group

Abstract Rabbit polyclonal antibodies were raised against synthetic peptides corresponding to hydrophilic regions of the human Rhesus (Rh) IX cDNA- encoded polypeptide predicted to be extracellularly or intracellularly exposed in the topologic model of the Rh blood group protein. Four antibodies encompassing residues 33–45 (MPC1), 224–233 (MPC4), 390–404 (MPC6), and 408–416 (MPC8) were characterized and compared with a polyclonal anti-Rh protein obtained by immunization with purified Rh proteins. All antibodies had specificity for authentic Rh polypeptides and reacted on Western blot with Rh proteins immunoprecipitated with human monoclonal anti-RhD, -c, and -E. MPC1, but not the other antibodies, agglutinated all human erythrocytes except Rhnull and Rhmod cells, which either lack totally or are severely deficient in Rh proteins, respectively. Immunoblotting analysis with membrane proteins from common and rare variants showed that MPC1 and MPC8 reacted in Western blot with 32-Kd Rh polypeptides from all common red blood cells except those from Rhnull and Rhmod, indicating that peptide regions 33– 45 and 408–416 may be common to several if not all Rh proteins, whatever the Rh blood group specificity. MPC4 reacted only with membrane preparations from cells carrying the E antigen, whereas MPC6 recognized preferentially the Rh proteins from E and Ee preparations, suggesting that the protein encoded by the RhIXb cDNA carries the E and/or e antigen(s). Immunoadsorption experiments using inside-out or right-side-out sealed vesicules from DccEE red blood cells as competing antigen showed that the MPC6 and MPC8 antibodies bound only to the cytoplasmic side of the erythrocyte membrane, thus providing evidence for the intracellular orientation of the C-terminal 27 residues of the Rh polypeptides. Attempts to transiently or stably express the Rh polypeptides. Attempts to transiently or stably express the Rh cDNA in eukaryotic cells were largely unsuccessful, suggesting that Rh antigen expression at the cell surface requires correct transport and/or folding of the Rh proteins, possibly as a complex with one-membrane proteins of the Rh cluster that are lacking in Rhnull cells.

scholarly journals OCCURRENCE OF 19S AND 7S ANTI-IGGS DURING HYPERIMMUNIZATION OF RABBITS WITH STREPTOCOCCI

1972 ◽  
Vol 136 (4) ◽  
pp. 799-815 ◽  
Author(s):  
Viktor A. Bokisch ◽  
David Bernstein ◽  
Richard M. Krause
Keyword(s):  
Red Blood Cells ◽  
Blood Group ◽  
Pneumococcal Vaccine ◽  
Blood Cells ◽  
Genetic Influence ◽  
Group A

All 110 rabbits immunized with Group A, A-variant, and C streptococcal vaccines produced 19S anti-IgG in addition to antibodies to the streptococcal carbohydrates. 19S anti-IgG was detected by hemagglutination of rabbit red blood cells coated with rabbit anti-blood group F antibody. Antisera of 88 of these animals were also tested for 7S anti-IgG with a coprecipitation assay. This assay is based on the coprecipitation of 7S anti-IgG with complexes of streptococcal carbohydrate and anti-carbohydrate antibody. 50 of the 88 anti-Group C streptococcal antisera contained 7S anti-IgGs. In eight antisera the concentration was greater than 5 mg/ml. The data suggest a genetic influence on the occurrence of 7S anti-IgG. The eight rabbits which produced more than 5 mg/ml of 7S anti-IgG belonged to three related families. Moreover, there were families in which almost every member produced 7S anti-IgG and other families in which only 30% of the members manufactured 7S anti-IgG. The streptococcal vaccine was an especially efficient stimulus for the production of 19S anti-IgG, whereas the pneumococcal vaccine was much less effective in this respect. Furthermore, 7S anti-IgGs were not detected in antipneumococcal antisera, although the concentration of anti-capsular antibodies was similar to that of anti-carbohydrate antibodies in antistreptococcal antisera.

HEMOLYTIC DISEASE OF THE NEWBORN DUE TO ANTI-A AND ANTI-B

PEDIATRICS ◽  
1955 ◽  
Vol 15 (1) ◽  
pp. 54-62
Author(s):  
Clare N. Shumway ◽  
Gerald Miller ◽  
Lawrence E. Young
Keyword(s):  
B Cells ◽  
Red Blood Cells ◽  
Blood Group ◽  
Newborn Infant ◽  
Blood Cells ◽  
Osmotic Fragility ◽  
Red Cells ◽  
Hemolytic Disease ◽  
Abo Incompatibility

Ten infants with hemolytic disease of the newborn due to ABO incompatibility were studied. In every case the investigations were undertaken because of jaundice occurring in the first 24 hours of life. The clinical, hematologic and serologic observations in the infants and the serologic findings in the maternal sera are described. Evidence is presented to show that the diagnosis of the disorder rests largely upon the demonstration of spherocytosis, increased osmotic fragility of the red cells, reticulocytosis, and hyperbilirubinemia in a newborn infant whose red blood cells are incompatible with the maternal major blood group isoantibody and against whose cells no other maternal isoantibody is demonstrable. The anti-A or anti-B in each of the maternal sera tested in this series hemolyzed A or B cells in the presence of complement. Other serologic findings in the maternal sera were less consistently demonstrated.

scholarly journals AB0-incompatibility of mother and fetus: the role of anti-glycan alloantibodies in the hemolytic disease of newborns

2021 ◽  
Vol 23 (1) ◽  
pp. 17-34
Author(s):  
P. S. Obukhova ◽  
A. V. Kachanov ◽  
N. A. Pozdnyakova ◽  
M. M. Ziganshina
Keyword(s):  
Red Blood Cells ◽  
Blood Group ◽  
Blood Cells ◽  
Hemolytic Disease ◽  
Disease Condition ◽  
Mother And Child ◽  
Group A ◽  
Group B ◽  
Maternal Igg

The mother and fetus incompatibility due to Rh-factor, blood group or other blood factors can lead to hemolytic disease of the fetus and newborn (HDN). HDN is a clinical disease condition of the fetus and newborn as a result of hemolysis, when maternal IgG alloantibodies cross the placenta and destroy the red blood cells of the fetus and newborn. The child disease begins in utero and can dramatically increase immediately after birth. As a result, hyperbilirubinemia and anemia develop, that can lead to abortions, serious complications, or death of the neonates in the absence of proper therapy. The range of HDN has changed significantly now compared to previous decades. Half a century ago, HDN was considered an almost complete synonym of RhD-alloimmunization, and this was a frequent problem for newborns. By now due to the high effective of Rh-conflict prevention, immunological AB0-conflicts have become the most common cause of HDN. The review aimes to one of the main causes of jaundice and anemia in neonates at present, i.e. HDN due to immunological AB0-conflict of mother and newborn (AB0-HDN). The main participants of the AВ0- incompatibility mother and child are considered, namely A- and B-glycans, as well as the corresponding anti-glycan alloantibodies. Close attention is paid to the structure features of glycan alloantigens on the red blood cells of the fetus and adult. The possible correlation of the frequency and severity of HDN with the blood group of mother and child, as well as with the titer of maternal alloantibodies, has been considered. The influence of immunoglobulin G subclasses on the AB0-HDN development has been evaluated. In most cases, AB0-HDN appear when the mother has the blood group 0, and the fetus has the group A (subgroup A1) or the group B. Other rare incidences of AB0-incompatibility with severe course are occurred. As a whole the etiology of AB0-HDN is complex and the HDN severity is influenced by many factors. The authors have analyzed statistical data, as well as the prevalence of AB0-incompatibility and AB0-HDN in various regions of the world. Current approaches to the diagnosis of AB0-HDN are discussed in addition. By now the problems of AB0- HDN occurrence and developing of ways to overcome this disease remain relevant.

Capture of red blood cells onto optical sensor for rapid ABO blood group typing and erythrocyte counting

2018 ◽  
Vol 262 ◽  
pp. 411-417 ◽  
Author(s):  
Xuemeng Li ◽  
Hanbo Feng ◽  
Yangyang Wang ◽  
Cuiping Zhou ◽  
Wei Jiang ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Blood Group ◽  
Optical Sensor ◽  
Blood Cells ◽  
Abo Blood Group ◽  
Blood Group Typing

scholarly journals Rearrangements of the blood group RhD gene associated with the DVI category phenotype

Blood ◽  
1994 ◽  
Vol 83 (4) ◽  
pp. 1129-1135 ◽  
Author(s):  
I Mouro ◽  
C Le Van Kim ◽  
C Rouillac ◽  
DJ van Rhenen ◽  
PY Le Pennec ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Blood Group ◽  
Blood Cells ◽  
Genomic Rearrangements ◽  
Blood Group Antigens ◽  
Reading Frame ◽  
Mrna Sequencing ◽  
Major Antigen ◽  
Rhesus Blood Group ◽  
Equivalent Region

Abstract The Rh (Rhesus) blood group antigens, D, Cc, and Ee, are carried by three unglycosylated membrane proteins of the human erythrocytes encoded by two highly related genes, D and CcEe. The major antigen, D, is a mosaic composed of at least nine determinants (epD1 through epD9). The lack of expression of some of these D epitopes at the surface of variant red blood cells defines the so-called D category phenotypes. In this report, we have determined the molecular basis of the DVI category phenotype characterized by the lack of epitopes D1, D2, D5, D6/7, and D8. Southern blot analysis and mRNA sequencing showed that the DVI phenotype is associated with two types of rearrangement of the D gene. Of 10 DVI genomes investigated, 8 exhibited a segmental DNA replacement (gene conversion) between the D fragment encompassing exons 4, 5, and 6 and the equivalent region of the CcEe gene. In the two other variants, these three exons are deleted. In both cases, the genomic rearrangement did not alter the reading frame of the variant RhD transcripts that are translated in 417 and 266 amino acid polypeptides, respectively. A heterogeneity of category DVI samples based on variable reactivity of the red blood cells with anti-D antibodies was previously found to be associated with the CDVIe or cDVIE haplotypes. Interestingly, our present results indicated that this serologic subdivision of the DVI category is correlated to two types of genomic rearrangements of the D gene.

Forssman expression on human erythrocytes: biochemical and genetic evidence of a new histo-blood group system

Blood ◽  
2013 ◽  
Vol 121 (8) ◽  
pp. 1459-1468 ◽  
Author(s):  
Lola Svensson ◽  
Annika K. Hult ◽  
Robert Stamps ◽  
Jonas Ångström ◽  
Susann Teneberg ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Blood Group ◽  
Blood Cells ◽  
Human Erythrocytes ◽  
Blood Group System ◽  
Group System ◽  
Glycolipid Antigen ◽  
Human Red Blood Cells ◽  
Key Points ◽  
Forssman Antigen

Key Points A new histo-blood group system was discovered, based on the identification of Forssman glycolipid antigen on human red blood cells. A newly described polymorphism in the GBGT1 gene activates the encoded enzyme to synthesize Forssman antigen.

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