“…The random two-dimensional distribution of the Rho(D) antigen as observed in our experiments may therefore represent an instantaneously fixed arrangement of a component which normally is continuously diffusible in the plane of the erythrocyte membrane. Such a structurally passive role for the Rho(D) antigen in the membrane must be reconciled, however, with the observations that those rare individuals who lack all known Rh-Hr antigenic determinants on their erythrocytes (29,30) often appear to have an unexplained hemolytic anemia (31).…”
A method is described for determining the two-dimensional distribution of specific antigens on cell surfaces, and is applied to the D antigen of the Rh antigenic system. Rh-positive human erythrocytes are allowed to react with purified 1251-labeled human anti-Rh0(D) -yglobulin antibodies, and the sensitized cells are then lysed at an air-water interface. The residual cell membranes are spread flat by surface forces, and are picked up on a carbon-strengthened collodion-coated electron microscope grid. The membranes are then stained with ferritinconjugated goat -antibodies directed against human -yglobulins. Only Rh-positive cells sensitized with antiRho(D) antibodies bind the ferritin-conjugated antihuman -y-globulins. The
“…The random two-dimensional distribution of the Rho(D) antigen as observed in our experiments may therefore represent an instantaneously fixed arrangement of a component which normally is continuously diffusible in the plane of the erythrocyte membrane. Such a structurally passive role for the Rho(D) antigen in the membrane must be reconciled, however, with the observations that those rare individuals who lack all known Rh-Hr antigenic determinants on their erythrocytes (29,30) often appear to have an unexplained hemolytic anemia (31).…”
A method is described for determining the two-dimensional distribution of specific antigens on cell surfaces, and is applied to the D antigen of the Rh antigenic system. Rh-positive human erythrocytes are allowed to react with purified 1251-labeled human anti-Rh0(D) -yglobulin antibodies, and the sensitized cells are then lysed at an air-water interface. The residual cell membranes are spread flat by surface forces, and are picked up on a carbon-strengthened collodion-coated electron microscope grid. The membranes are then stained with ferritinconjugated goat -antibodies directed against human -yglobulins. Only Rh-positive cells sensitized with antiRho(D) antibodies bind the ferritin-conjugated antihuman -y-globulins. The
“…Erythrocytes which lack all the antigens of the human Rhesus blood group system have been described by Vos et al (1961), and are denoted Rhnuil erythrocytes. These cells have an abnormal stomatocytic -shape, and show increased osmotic fragility (Sturgeon, 1970).…”
1. Rhnull human erythrocytes lack the antigens ofthe Rhesus blood group system, have an abnormal shape and an increased osmotic fragility, and are associated with mild chronic haemolytic anaemia. 2. Studies with phospholipase A2 and sphingomyelinase C show that the asymmetric distribution of phosphatidylethanolamine (PtdEtn) in the membrane of these cells differs from that found in control cells. The amount of PtdEtn which can be hydrolysed by phospholipase A2 in the presence of sphingomyelinase C in intact Rhnul cells is twice as high as that in normal erythrocytes. 3. In intact Rh,,,l cells all of the phosphatidylcholine (PtdCho) present in the membrane can be readily exchanged with a PtdCho-specific exchange protein, whereas in control cells 75% is readily exchanged and 25% at a much lower rate. This indicates that PtdCho experiences a relatively fast transbilayer movement in the'
“…In our experiments ME-induced agglutination occurred only with anti-Rh sera of high titer. The possibility remains that treatment of red cells with ME may expose previously blocked antigenic sites, which would explain such diverse phenomena as the weakly-reactive Rh cells (24), (25,26), and the ---(deletion) cells (27). Investigation of this and other possibilities is currently in progress in our laboratory.…”
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