Phosphate Partition in the Erythrocytes of Normal Newborn Infants and Infants with Erythroblastosis Fetalis. III. P32 Uptake and Incorporation

The uptake and incorporation of P³²O₄ by the cord blood erythrocytes ofnormal and erythroblastotic infants and the erythrocytes of adults was studiedby incubating specimens in ACD at 37 C. in air. The uptake of p³² from theplasma was slower in the infants than in the adults. It was possible to demonstrate an initial "fast" phase followed by a "slow" phase in all the plasma p³²uptake experiments. The half-value time and the percentage turnover perminute indicated greater activity in the adults compared to the infants. Thebalance of movement of phosphorus was from red cells to plasma in all theexperiments. No striking differences in phosphate metabolism were demonstrated between the normal and erythroblastotic newborns, except for thelarge turnover in µM of P per minute by the red cells of the latter during thefirst 20 minutes of incubation.

Labeling of intraerythrocytic inorganic phosphorus (P_i), adenosine triphosphate (ATP) and 2,3-diphosphoglycerate (2,3-DPG) proceeded more slowlyin the infants than in the adults. The labeling of the erythrocyte 2,3-DPG of allthe infants lagged far behind P_i and ATP whereas in the adults the r.s.a. of2,3-DPG had equalled or exceeded the r.s.a. of P_i and ATP in four hours. Itis suggested that there is decreased synthesis of phosphoglycerates by theerythrocytes of the newborn which may be due to a lower level of glyceraldehyde-3-phosphate dehydrogenase activity.

Accepted on January 11, 1962     

The Lutheran blood groups: a second example of anti-Lu b and three further examples of anti-Lu a

1. An example of the blood group antibody, anti-Lu^b, was found in a patientwho had a mild hemolytic transfusion reaction. It was shown to possess thecharacteristics of an immune antibody and to be able to distinguish between asingle dose and a double dose of the Lu^b gene.

2. Three new examples of the antibody, anti-Lu^a, are presented. All of themwere found in normal blood donors and have properties which indicate that theyare naturally occurring antibodies.

Dr. R. R. Race and Dr. R. Sanger confirmed the presence of anti-Lu^b in Mrs. S.’sserum, and studied other members of her family and the three anti-Lu^a sera. We aregrateful to them for many favors and their kind encouragement.

We are obligated to Miss Marie Cutbush for making available the Lu^aLu^a cells fromMrs. R. and her sister, and for a supply of anti-Lu^b serum.

Thanks are due to Dr. A. E. Mourant who furnished our original supply of anti-Lu^aserum and to Dr. Philip Levine for the anti-Tj^a and anti-Vel sera.

We are indebted to Dr. J. M. Fine of Milwaukee for permission to study Mrs. S. andto the patient and her family for their cooperation.

The sera from 18,613 blood donors were studied by Betty McCarthy, Rosemary Polka,Pearl Lemke, Agnes Molnar, Jeannette Flagstadt and Betty Hutter.

Submitted on March 20, 1957 Accepted on July 1, 1957     

Autoradiography of Diffusible Compounds in Human Nonnucleated Erythrocytes: Studies with Tritiated Glucose and Adenine

A simple method for autoradiography of diffusible substances in erythrocytes is described. It is based on retarding diffusion from the red corpusclesduring fixation and processing by using the bifunctional aldehyde, glutaraldehyde; rapid drying; and applying the nuclear emulsion as a dried, preformedfilm.

The technique was used for studying the changes during storage in theability of red blood cells to utilize glucose. The incorporation of glucose-6-³Hby samples of blood stored for different periods of time was followed by thenumber of silver grains present in the red corpuscles. It was observed thatthe percentage of erythrocytes with fewer than four grains increased progressively during the storage of ACD blood. The incorporation of glucose asobserved by the silver grain pattern was reversible toward that of freshly collected red corpuscles following incubation of the stored samples withadenosine.

Similar studies were performed using adenine-8-³H as the label in bloodstored for six weeks in ACD adenine solution. Incubation of the blood samplescontaining 0.5 mM. adenine for one hour at 37°C resulted in autoradiographicpatterns equivalent in grain counts to those given by red blood cells storedwith the radiolabeled adenine for five weeks. The incubation of fresh sampleswith inosine was found to inhibit the incorporation of adenine, whereas inosineenhanced the number of grains in red blood cells which had "run down"during storage.

The autoradiographic technique presented should be of value for the in vitroevaluation of methods of blood preservation and for studying the metabolicheterogeneity of normal red corpuscles and other tissue cells. It should be useful for investigating hereditary and acquired abnormalities of metabolism.