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Investigation of the Rh blood group system in humans and non-human primates
Abstract
The human Rh blood group system plays a central role in transfusion medicine, being responsible for hemolytic disease of the newborn, transfusion incompatibility, and as a target antigen in autoimmune disease. Rh D is the major antigen; the term "Rh positive" indicates inheritance of the D antigen, and "Rh negative" indicates its absence. In addition, other Rh alleles at this locus include C/c and E/e. Rh proteins appear to be important for membrane integrity, as rare individuals who lack all Rh antigens suffer hemolytic anemia. This thesis represents an investigation of the Rh system at the molecular level in humans and non-human primates using serologic and molecular techniques. Two Rh cDNA clones were isolated from human reticulocytes by polymerase chain reaction. An Rh message isolated from the K562 human erythroleukemia cell line revealed a new RhD splicing isoform. These cDNA's were used as probes in the investigation of the Rh gene in genomic DNA from humans, K562 cells, and non-human primates by Southern blot. These studies demonstrated that in humans, the Rh negative condition represents a gene deletion at the Rh locus, which allowed ready detection of the Rh negative phenotype at the DNA level. A RhD polymorphism was detected with the restriction enzyme Dra I in Rh-positive individuals. Southern blot analysis demonstrated that the K562 cell line originated from a Rh-positive individual. Southern blots also showed the presence of Rh-related sequences in anthropoid apes, all New and Old World monkeys, and in most non-primate animals tested. More polymorphism was detected in chimpanzees and gorillas than in humans. A gorilla genomic clone corresponding to the 3$\prime$ end of the human RhD gene was sequenced and characterized. PCR-generated genomic fragments demonstrated that the RhD gene is conserved in gorillas and chimpanzees, and that the increased polymorphism may be due to an additional Rh gene duplication in the gorilla.
Subject Area
Molecular biology
Recommended Citation
Westhoff, Connie Marie, "Investigation of the Rh blood group system in humans and non-human primates" (1993). ETD collection for University of Nebraska-Lincoln. AAI9416003.
https://digitalcommons.unl.edu/dissertations/AAI9416003