William J. Kuhns

HYPERTRANSFUSION REGIMEN IN PATIENTS WITH Cooley's ANEMIA*

The clinical manifestations of Cooley’s anemia (severe homozygous 8thalassemia) can be explained in light of our present knowledge of the pathogenesis of this disease, which has been summarized previously in this annal. In Cooley’s anemia, the grossly unbalanced Hb synthesis results in intense intramedullary hemolysis. In these patients erythropoiesis is thus abortive, and the hyperactivity of the bone marrow only leads to the output of few and short-lived red cells. Anemia further stimulates the overgrowth of hemopoietic tissue. This accentuates the hepatosplenomegaly and the severe bone alterations that ultimately develop in the characteristic Cooley’s facies. As a consequence of the severe chronic anemia, cardiomegaly develops and, finally, the patient succumbs to congestive heart failure. It is obvious that all these effects could be prevented if the anemia could be corrected. At the present time the only treatment available to correct the anemia is the use of blood transfusions. These have been traditionally restricted in these patients to the amount required to maintain minimal physical activity, for fear of inducing iron overload. The most common cause of death for these patients is, in fact, intractable heart failure associated with Fe deposition in the myocardium.’ In 1964, at the first Conference on The Problems of Cooley’s Anemia, Wolman reported that patients maintained at a higher baseline level of Hb were, in general, in better health and had smaller organs.? His observations have been confirmed by several other It was suggested that the intractable heart failure of Cooley’s anemia patients could be due primarily to prolonged anoxia and cardiomegaly and that iron deposition in the myocardium may be only a secondary and/or a contributory effect. In 1968, at the Second Conference on the Problems of Cooley’s anemia, we presented preliminary observations on four very young patients maintained at a baseline hematocrit as nearly normal as practicable. ( “Hypertransfusion regimen”).5 The objects of the present report are to provide a follow-up on these patients and to summarize our current experience with this treatment.

Cell adhesion-related proteins as specific markers of sponge cell types involved in allogeneic recognition

Sponge immunocyte identification is of interest to comparative immunologists since characterizing these cells will allow investigations into the mechanisms of non-self recognition in the oldest animal phylum. Here, we report that polyclonal antibodies raised against the core protein of a proteoglycan involved in cell adhesion in the marine sponge Microciona prolifera are specific markers for archaeocytes, the totipotent sponge cells. Archaeocytes are mobilized upon allogeneic contact and they accumulate in the contact zone. A second type of cell, the gray cells, are specifically recognized by monoclonal antibodies raised against CD44, a hyaluronan receptor. Gray cells do also accumulate in the contact area. Specific staining of a third sponge cell type, the rhabdiferous cells, shows that these do not accumulate upon allografting. These specific cell markers allow tracking of archaeocytes and gray cells, and show that they play an active role in sponge allogeneic reactions.

SERUM ANTIBODY RESPONSE TO TISSUE TRANSPLANTATION ANTIGENS IN MAN

There is general agreement that the rejection of homologous tissue transplants is achieved through a n immunological response undertaken by the h0st.l Some of the mechanisms invoked in homograft reactions appear to bear a similarity to responses observed in the delayed type of hypersensitivity.2 However, the possibility also exists that some form of serum antibody response is associated with homograft rejection in certain animal species3 In man, however, the one fully documented report of a serum antibody response to skin homograftsZ was based on studies of a severely burned child, in whom many factors other than the applied grafts may have been operative, and, on the results of Van Rood, in subjects also sensitized by injections of blood ~ l a t e l e t s . ~ Colombani et al. have been unable to demonstrate a leukoagglutinating serum antibody response in normal skin homograft recipients,6 and the attempt to transfer skin homograft sensitivity with serum obtained from hypersensitized human white graft reactors has also been un~uccessful .~ Thus, within the limitations of experimental models used, specific serum antibody response to skin homograft rejection in man has been a rather rare finding. The systematic study of human subjects sensitized to skin homografts in our laboratories by the injection of various preparations of human blood leukocytes afforded a further opportunity to study this problem at various stages following sensitization and homograft rejection. For this purpose, each individual studied in the course of attempts to localize human transplantation antigens present in blood leukocytes was also investigated from this standpoint.

Hyaluronic acid-receptor binding demonstrated by synthetic adhesive proteoglycan peptide constructs and by cell receptors on the marine sponge Microciona prolifera.

The aggregation factor (MAF) of Microciona sponge is depicted as a species specific, adhesive proteoglycan molecule configured as a sunburst-type structure with a central ring, or core, from which numerous elongated arms project (1). Although the carbohydrate content of MAF is very high, the core is considered to be a protein (2). Fernandez-Busquets et al. have demonstrated, in a cloned peptide MAF fragment, a putative hyaluronic acid (HA) binding sequence bearing a chemical similarity to some other HA binding proteins on vertebrate cell membranes (3, 4) that are known to be involved in growth, and cell cycle and motility events (5, 6). The sequence of predominantly basic amino acids is generally designated by the formula B (X7) B, where B is arginine or lysine, and X is any non-acidic amino acid; there is also at least one additional basic amino acid within, or adjacent to, the basic unit (7, 8). By these criteria, the deduced MAF peptide sequence is shown to be B (X6) B, and is thus analogous to its vertebrate counterparts. In an earlier study, we used a biotin-labeled HA (BHA) probe to demonstrate HA receptors on Microciona cells; we also used antibodies against RHAMM (receptor for HA mediated motility, having the chemical formula shown above) to demonstrate that RHAMM is present on some of these cells (9). These results led us to hypothesize that HA, known to be present on MAF, might serve as a ligand for these chemically similar binding peptides, one on the MAF core peptide, the other on cell membranes, and that it might, thereby, be capable of signal transduction, as is known to occur in higher organisms. The aim of this study was to provide evidence in support of these notions.

Studies of Immediate Wheal Reactions and of Reaginic Antibodies in Pregnancy and in the Newborn Infant.

Summary Eight Schick negative pregnant women were given repeated injections of fluid diphtheria toxoid during pregnancy to stimulate formation of reaginic antibodies as judged by IWR. Although this occurred in most cases, it was not mirrored in the cord bloods of the corresponding infants which were examined for reagins at term. In addition, the skin of all infants was unreactive in the presence of intradermal toxoid. On the other hand, IWR were elicited in these infants and their mothers when they served as recipients for passive transfer (P.K.) tests using reaginic antibodies derived from an alternate source.

Hemagglutinins in the Plasma of Catfish (Ictalurus nebulosus) Injected with Saliva from Human Secretors of Various A‐B‐O Blood Groups

Serums from non‐immunized catfish contain hightitered thermolabile hemolytic activity for all human red cells, and for red cells of other species such as rabbits. However, after heat inactivation, these sera only weakly agglutinate human red cells.

Historical Milestones Blood Transfusion in the Civil War

Two cases of transfusion of whole blood during the Civil War are described.

Detection and Characterization of Blood Group Antigens on Untransformed Human Amnion Cells

Amnion cells, primary and in culture, along with the infants own erythrocytes, were tested by mixed agglutination for blood groups A, B, H, M, N, Tja, Rho(D), Yta, I and i. Studies were carried out on cell cultures ranging in age from one day to 39 days. Only A, B, H, Tja, I and i could be demonstrated on amnion cells. Decrease in the number of cells positive for Tja, and possibly for A and B, was observed in the course of cell passages. In cultures positive for H, the percentage of positive cells, although small, generally continued as long as the culture was studied.

Experimental studies on the transfusion of blood obtained postmortem.

A decrease in the posttransfusion survival of Cr51‐labeled rabbit erythrocytes obtained postmortem became more pronounced as the time after death increased. This effect became proportionately greater when rabbit cadavers were maintained at 37 C, as compared with 4 C or 20 C, prior to the procurement of blood for survival studies. Decreased survival of normal control cells incubated with postmortem plasma or subjected to anoxia for eight hours was not observed. The following postmortem changes in blood were observed: increase in red cell osmotic fragility, alteration of morphology with appearance of spherocytes and crenated cells, increased plasma hemoglobin, decrease in pH and intracellular ATP. These changes became more marked as the time interval between death and blood collection increased. Normal and postmortem specimens possessed equivalent antigenicity as judged by immunization experiments in rats.

Disappearance of human diphtheria antitoxin from human passive transfer skin sites.

Summary This paper has described further experiences with a passive transfer technic which depends upon the ability of intradermal antitoxin to neutralize Schick toxin introduced simultaneously or later at the same sites. Partial, total or no inhibition of toxic reactions indicates the amount of antitoxin remaining at the skin sites. The amount of antitoxin which has disappeared may be calculated by difference, assuming that 1 Lf toxin = 1 Unit of antitoxin. On the basis of this test 50% of antitoxin introduced at skin sites was generally found to disappear in 10 to 14 hours. No difference in disappearance from the skin was observed between precipitating antitoxin, non-precipitating antitoxin and skin sensitizing antitoxin.