W. L. Marsh

The I Antigen and Antibody

The red cells of an English blood donor, Mr. M., are of the phenotype i: the anti‐I which is present in his serum is presumably a “naturally occurring” antibody for he has never been a recipient. The red cells of Mr. M. are extraordinarily weak in their reaction with anti‐H and anti‐O sera but not unusual in any other respect.

Chronic Granulomatous Disease and the Kell Blood Groups

Summary. Fifteen antigenic determinants are known to be related to the Kell blood group. Some boys with X‐linked chronic granulomatous disease have the very rare McLeod or K0 phenotype on their red cells. Serological studies of the McLeod type suggest that the weak Kell antigens that are present differ qualitatively and quantitatively from those on red cells of common Kell type. A new antigen, Kx, has been characterized and shown to be present on red cells and neutrophil leucocytes. Lack of red‐cell Kx is associated with the McLeod phenotype, lack of leucocyte Kx is associated with chronic granulomatous disease.

Mapping human autosomes: evidence supporting assignment of rhesus to the short arm of chromosome No. 1.

Rh-negative erythrocytes were found in the blood of an Rh-positive man suffering from myelofibrosis. Nucleated hemopoietic precursors were also circulating in his blood, and these cells had an abnormal chromosome complement from which identifiable chromosome segments had been deleted. Correlation of the serological and cytogenetic findings, combined with previous data, indicates that the Rhesus blood group locus is on the distal portion of the short arm of chromosome No. 1.

Inhibition of anti-I sera by human milk.

Summary. Human milk has been found to contain a high concentration of water‐soluble I blood‐group substance. Tests with 24 different anti‐I sera showed that to a variable extent all of them could be inhibited by milk and some could be inhibited by strong I secretor saliva. The susceptibility of I antibodies to inhibition was not related to titer, and the results suggest that qualitative differences in the antibody‐antigen reaction are responsible.

The definition of two I antigen components.

Abstract. The I blood group antigen has two main components named IFetal (IF) and IDeveloped (Id). The IF component is present on all human red cells including those of icord and iadult persons, and also on Rhesus monkey red cells. The ID component develops slowly on the red cells before birth and to a greater extent in the 18 months after birth. Inhibition studies with human milk show that strongly‐inhibitable anti‐I sera are of the anti‐ID variety, but only a minority of such sera are inhibitable. Comparative studies on the red cells of 12 newborn infants suggest that the same sequence of I antigen development occurs in them all, but different infants vary in the amount of I development that they have achieved at the time of birth. I antigen variants affect mainly the ID component, and it is this part of the antigen that may be weakened on the red cells of people with leukemia.

The Recognition of Water-Soluble I Blood Group Substance

Abstract. A powerful cold auto anti‐I has been found that is strongly inhibited by hydatid cyst fluid and inhibited to a varying extent by all of the 181 human saliva samples tested. The investigation showed that the inhibition effect was specific for I and that water soluble I blood group substance must be responsible. No correlation could be found between the concentration of I substance in the saliva and the presence of ABH, Lewis or Sda substances. The amount of salivary I substance secreted appears to be a constant feature of the individual. Infants at birth and i adults have high concentrations of I substance in their saliva and the amount of I substance present does not reflect the I status of the red cells.

Biochemical studies on McLeod phenotype red cells and isolation of Kx antigen

Summary Red cells of the McLeod blood group phenotype have weak Kell antigens, lack Kx antigen and have acanthocytic morphology. We have immunoprecipitated Kell antigens from McLeod red cells and show that they are markers on the same 93 kDa membrane protein that carries Kell antigens on normal red cells. However, as determined by Western immunoblotting, McLeod red cells have a marked deficiency of this protein. We have also studied the near‐neighbour relationship of McLeod and common Kell red‐cell membrane proteins by cross‐linking intrinsic sulphydryl groups by oxidation, catalysed with orthophenanthroline and copper, or by cross‐linking amino groups with dimethyl 3,3‐dithiobispropionimidate. Results were analysed by diagonal mapping in two‐dimensional gels. No abnormalities of membrane protein inter‐relationship were detected in McLeod red cells.

Acquired loss of red cell Kell antigens

Summary. A 19‐year‐old patient with a long history of idiopathic thrombocytopenic purpura developed a potent antibody against a high‐incidence antigen in the Kell blood group system. The direct antiglobulin test on his red cells was negative. His cells exhibited profound depression of Kell blood group antigens, but antigens of other blood groups were normal. Transfusion of incompatible blood was well tolerated and differential agglutination tests, using selected Rh antisera, showed in vivo survival of the transfused red cells for more than 8 weeks. However, the transfused red cells also showed acquired loss of Kell antigens. Five months after the initial findings, Kell‐related antibody disappeared and Kell antigens reappeared on his red cells. The patients serum stored from the initial investigation now reacted with his freshly collected red cells. These data suggest that an environmental agent in the patients plasma was responsible for the temporary loss of Kell antigens from red cells in his circulation.

Kx antigen, the McLeod phenotype, and chronic granulomatous disease: further studies.

Abstract. Leukocytes of nine unrelated boys with X‐linked chronic granulomatous disease lack Kx antigen. In three of these cases, the red cells also lack Kx and have the McLeod phenotype and abnormal morphology. X‐linked chronic granulomatous disease CGD can thus be separated into two types. Type I cases have an antigenic deficiency that is restricted to the phagocytic leukocytes while in type II, the deficiency involves both leukocytes and red cells. Red cells of type II CGD patients have enhanced i antigen activity suggesting that they are under hemopoietic stress. Normal Kx synthesis is directed by an X‐linked gene named X1k. Three rare variants, X2k, X3k, and X4k order the different permutations of leukocyte and red cell Kx antigen production that have been recognized.

Biochemical studies on McLeod phenotype erythrocytes.

Abstract. Red cells of the McLeod phenotype in the Kell blood group system have an acanthocytic morphology. The membrane protein composition analyzed on sodium dodecylwlfate‐polyacrylamide gel electrophoresis, the ATP level and the activities of a large number of intracellular enzymes appear to be normal. Membranes prepared from McLeod red cells incubated with γAT[32P] and MgCl2 incorporated twice as much radioactivity into spectrin and also showed a slight elevation of phosphorylation in band 3 protein when compared to membranes from normal cells. Intact normal red cells incubated with carrier‐free [32P] incorporated radioactivity into several proteins. with most incorporation in spectrin and band 3 protein. In comparison, McLeod cells incorporated three times more radioactivity into spectrin and band 3 protein but increased phosphorylation also occurred in other, but not all, membrane proteins. Intact McLeod red cells also showed increased phosphorylation of membrane phospholipids, but they incorporated [32P] into intracellular nucleotide phosphates in a normal manner.