Seldon E. Bernstein

Proof of whole-cell implant in therapy of W-series anemia.

In mice one particular type of severe hereditary macrocrtic anemia (genotype W/WV) can be cured regularly by the injection of hemopoietic cells from adult marrow or fetal liver of normal (+/+) donors (1). When this happens the circulating erythrocytes shift from a low mean number (6-7 X lo6 RBC/mm3) and large mean cell volume (mean, 55-70 p3) to a high mean number (l&11 X lo6 RBC/mm3) and smaller mean cell volume (40-45 p3) characteristic of adult normal (+/+) mice. Recipients retain these normal levels through the remainder of their lifetimes (2). Therapy of this sort, through injection of normal hemopoietic cells, has been used extensively in analysis of W-series gene action (3). In our laboratory it has been effective only when tissues of donor and recipient were highly histocompatible, though not necessarily identical (4), an observation which indicates the importance of at least short-term survival of the injected cells in successful experiments. Seller and Polani (5) succeeded in curing small numbers of WV/WV anemic mice with normal hemopoietic liver cells from fetuses from unrelated inbred strains. From the description of the genetic origin and maintenance of their WV/WV mice, it appeared to us possible that histocompatibility genes

Implantation of normal blood-forming tissue in genetically anemic mice, without x-irradiation of host.

Summary Successful implantation, without x-irradiation or other pretreatment of host, of isologous blood forming tissue from normal (ww) mouse fetal liver in viable severely anemic recipients (20 of 24 adult WW v mice, 9 of 14 juvenile WW v mice) and in lethally anemic recipients (4 of 15 juvenile WW mice) has been demonstrated. Criteria of success were permanent increase in rbc/mm3 to near-normal levels, and decrease of mean cell volume from macrocytic to normocytic levels. These changes appeared in WW v adult anemic hosts longer after cell injection than in similar transplant experiments with WW v hosts subjected to 200-r whole-body irradiation. These findings fit well with the hypothesis of competition between slow-acting indigenous blood forming tissue of genetically anemic hosts and initially small implants of rapidly functioning blood forming tissue from genetically normal donors. If methods for circumventing homografts can be found, cell implantation may be generally useful for therapy of genetic defects in erythropoiesis.

Influence of non-h-2 genetic factors on success in implantation therapy of w series hereditary anemias of the mouse.

The severe macrocytic anemia of nonirradiatcd adult W/Wv mice be permanently cured by implantation of isogoneic normal hematopoietic cells. In one particularly favorable H-2 compatible allogencic combination the anemia of W/Wv adults has also been completely cured by injection of allogeneic normal (+/+) fetal liver cells. The extremely severe macrocytic anemia of WC/Re-W/W surviving adults was also greatly ameliorated by implantation of allogeneic normal WB/Re-+/+ marrow cells. Allogeneic skin grafts between these strains were rejected. Particular characteristics of W series anemias, which favor implantation and proliferation of injected normal blood-forming cells, may account for some of the cross-barrier successs obtained with these two allogeneic combinations. Similar and larger injections of normal fetal liver cells into adult anemic recipients had no beneficial effects in four other H-2 compatible allogencic combinations. Under the conditions of these experiments, successful implantation of normal blood-forming tissue must depend upon matching of H-S and several other sets of histocompatibility alleles. The histocompatibility barrier between WB/Re and WC/Re differs quantitatively or qualitatively from that involved in the four unsuccessful combinations. Between these two strains only, the histocompatibility difference prevents permanent acceptance of skin grafts but allows acceptance of blood-forming tissue. The failure of implant therapy of Sl/Sld mouse anemia, and relation between intrinsic nature of each hereditary anemia and the possibility of its cure by implantation of normal cells, are also discussed.

Pyrrole pigments in normal and congenitally anaemic mice (+/+, W/Wv, ha/ha, nb/nb, mk/mk, f/f and sla/Y)

Abstract 1. Values for faecal urobilinogen (UBG), serum bilirubin, and free erythrocyte protoporphyrin (FEP) are reported for normal and anaemic mice with six different hereditary anaemias. 2. Greatly increased faecal UBG and elevated serum bilirubin were found in mice with severe haemolytic anaemia (nb/nbandha/ha). 3. Mice with microcytosis (mk/mk) and sex-linked anaemia (sla/Y), both showed increased FEP and only moderate increase of faecal UBG. 4. Values for W/Wv mice fell within the normal ranges. 5. FEP levels in normal 15-day foetal red cells were extremely high. Levels for 15-day flexed anaemic foetuses (f/f) were lower, but markedly aboved adult level.

Limitations of whole-body irradiation for inducing acceptance of homografts in cases involving a genetic defect.

Experiments are described and discussed in which normal isologous and homologous hematopoietic tissue implants in genetically anemic mice were exposed to various doses of x rays. The findings suggest considerable limitation to the use of radiation in inducing acceptance of homografts in mice or humans with specific genetic defects. (T.R.H.)

Inborn anemias in mice. Progress report, 1 May 1976 to 31 July 1977

Hereditary anemias of mice have been the chief objects of investigation. At present under study are four macrocytic anemias, four hemolytic anemias, nonhemolytic microcytic anemia, transitory siderocytic anemia, sex-linked iron-transport anemia, and the autoimmune hemolytic anemia of NZB. Each of these blood dyscrasias is caused by the action of a unique mutant gene, which determines the structure of different intracellular molecules, and thus controls a different metabolic process. Thus our wide range of different hereditary anemias has considerable potential for uncovering many different aspects of hemopoietic homeostatic mechanisms in the mouse. Each anemia is studied through: characterization of peripheral blood values, determinations of radiosensitivity under a variety of conditions, measurements of iron metabolism and heme synthesis, histological and biochemical study of blood-forming tissue, functional tests of the stem cell component, examination of responses to erythroid stimuli, and transplantation of tissue between individuals of differently affected genotypes. Considerable effort is devoted to perfection of hematologic, cell culture, and transplant methods to make these techniques useful in dealing with special problems associated with abnormal function.