Joseph W. Ferrebee

SUPRALETHAL WHOLE BODY IRRADIATION AND ISOLOGOUS MARROW TRANSPLANTATION IN MAN

Leukemia has been studied in two sets of identical two. One leukemic twin was irradiated with 850 r and the other with 1,140 r from Co/sup 60/ sources. Each was then given bone marrow from the respective normal twin. Successful transplantation of this isologous marrow was determined by the return of morrow function, evident after less than two weeks, and by a benign clinical course following radiation. Leukemia recurred after remissions of seven weeks in one case and 12 weeks in the other. From these two patients it was concluded that transplants of isopogous marrow are readily achieved in man; one thousand r of whole-body radiation does not produce troublesome acute radiation sickness in man when given at a rate of 20 to 40 r per hour; whole-body irradiation at the 1,000 r level produces a remission but not a cure of leukemia when followed by isologous marrow. (auth)

Prolonged Storage of Marrow and Its Use in the Treatment of Radiation Injury

Marrow was removed from the femurs of dogs and frozen in 15 per cent glycerol at 1 C. per minute. Marrow from one dog was stored at −80 C. and from the other at −180 C. for a period of 14 months. Each dog was then given 1,200 r of whole body irradiation followed by infusion of his own thawed marrow. Recovery was rapid, and the dogs were living and well two months later.

RADIATION INJURY AND MARROW REPLACEMENT: FACTORS AFFECTING SURVIVAL OF THE HOST AND THE HOMOGRAFT

Excerpt INTRODUCTION Exposure of the entire body to ionizing radiation†in the range of 500 to 1,000 Roentgens (r) is followed by death in from seven to 14 days.1A failure of marrow function is the ...

FUNCTIONING HOMOGRAFTS OF THE LUNG IN DOGS

A lung autograft in the dog can survive indefinitely and perform almost normal pulmonary function.lS2 A lung homograft is destroyed in six days. The first evidence of rejection, the appearance of a few perivascular round cells, occurs two days after transplantation. Moderate pulmonary edema and increasing concentrations of lymphocytes are seen at four days. Hemorrhagic necrosis of the transplanted lung is found after the sixth post-transplant day unless measures are taken to alter the rejection response. This report describes three methods of altering the immune response to homografts of the lung which allow prolonged graft survival.

Passage of the Blue Dye T-1824 from the Blood Stream into the Lymph.

Summary 1. Blue dye T-1824, used in the determination of blood plasma volume, passes from the blood stream into the thoracic and cervical duct lymph of normal dogs within the first hour following its intravenous injection. 2. Under normal physiological conditions correction for the escape of dye from the vascular bed can probably be approximated by extrapolating the curve of dye disappearance. Under abnormal and varying conditions application of this correction may be difficult.

Induction of unresponsiveness to major transplantable organs in adult mammals: a recapitulation of ontogeny by irradiation and bone marrow replacement.

Transplantation of renal allografts obtained from prospectively selected genotypically DLA-identical donors into supralethally irradiated dogs reconstituted with their own stored bone marrow has produced a state of unresponsiveness to these kidneys in the recipients. Eleven of 18 kidneys transplanted at 12 hours after marrow replacement currently survive with normal function and maintain life in the recipients for 757, 800, 825, 978, 1062, 1092, 1136, 1282, 1373, 1380, and 1381 days, respectively. Similar results occurred in eight of 13 allografts transplanted at 28 hours after marrow replacement, which currently survive for 349, 363, 377, 407, 436, 470, 485, and 513 days, respectively, and in eight of 13 kidneys grafted at 36 hours after marrow replacement, which are surviving for 197, 247, 298, 324, 337, 396, 443, and 472 days, respectively. Achievement of optimal results is dependent on the specific timing and sequence of each procedure. Only four of 16 recipients of kidneys transplanted at the time of marrow replacement were unresponsive to their allografts. Similarly, only five of 19 recipients of kidneys placed in irradiated dogs at 40 hours before marrow replacement accepted such allografts. When kidney transplants were placed into the recipients 20 hours before removal of marrow, irradiation, and reconstitution with stored marrow, only three of 21 dogs became unresponsive to such allografts. In five of 12 instances, the recipients were also unresponsive to skin allografts obtained from their respective kidney donors. Such skin grafts currently survive for 606, 673, 687, 701, and 708 days, respectively. The remaining seven skin grafts were rejected at 28, 39, 42, 84, 90, 92, and 115 days, respectively. Second- and third-set skin grafts from the same kidney donor were rejected by six of these dogs at 19, 20, 21, 29, 29, and 30 days, and at 21, 22, 23, 24, 27, and 27 days, respectively. Rejection of these skin grafts had no detectable effect on the function and survival of kidney allografts from the same source. Seven of eight skin grafts obtained from other DLA-identical donors were rejected at 13, 14, 16, 25, 28, 38, and 84 days, respectively; one allograft continues to survive for 708 days. Eleven DLA-incompatible skin allografts placed on the recipients at the same time were rejected within 11–20 days. Supralethal total body irradiation and bone marrow replacement can establish in the adult canine host a privileged phase of immunological reactivity during which exposure to alloantigens produces specific long-term unresponsiveness rather than sensitization. The use of stored autoiogous rather than allogeneic bone marrow for reconstitution of the irradiated recipient eliminates the hazards of GVH complication usually associated with this procedure. This consideration and the apparent capacity of the tolerant host to maintain a long-term state of unresponsiveness without any further immunosuppressive therapy point to the potential relevance of the results to human transplantation.

MARROW TRANSPLANTS IN LETHALLY IRRADIATED DOGS: THE EFFECT OF METHOTREXATE ON SURVIVAL OF THE HOST AND THE HOMOGRAFT.

Twenty dogs were given 1500 r whole-body irradiation and an infusion of homologous marrow. Ten dogs treated with methotrexate were compared with 10 dogs that received no drug therapy. The dogs given methotrexate showed an increased incidence of graft take, a decreased incidence of graft rejection, and an increased number of long-term survivors. (auth)

Induction of Allogeneic Unresponsiveness in Adult Dogs: ROLE OF NON-DLA HISTOCOMPATIBILITY VARIABLES IN CONDITIONING THE OUTCOME OF BONE MARROW, KIDNEY, AND SKIN TRANSPLANTATION IN RADIATION CHIMERAS

Exposure to supralethal total body irradiation and transplantation of bone marrow from a DLA- and pedigree-identical donor have regularly produced successful engraftment and the establishment of stable long-term chimerism in beagles of the Cooperstown colony. Bone marrow allografts performed in pairs of dogs bearing identical DLA haplotypes derived from different pedigree origins (i.e., different classes of the same haplotype) yielded two different results. Depending upon the particular haplotype pedigree combination used, such transplants either led to long-term chimerism or to failures of engraftment, secondary disease, and death of the recipients (i.e., pedigree-incompatible combinations). Radiation chimeras given bone marrow from a DLA-and pedigree-identical donor were challenged within 8-12 h after marrow transplantation with a renal allograft obtained from another DLA- and pedigree-identical donor. The recipients have remained unresponsive to such renal allografts and have survived indefinitely with normal renal function. In contrast, renal allografts obtained from donors bearing the same DLA haplotypes derived from pedigree-incompatible sources were rejected within 25-50 days after transplantation. The long-term surviving recipients have also been unresponsive to skin allografts obtained from their donor of marrow and the kidney donor. Skin grafts obtained from other DLA- and pedigree-identical dogs were rejected within 13-41 days, and grafts from DLA-incompatible donors survived for 10-25 days. These results highlight the potential importance of genetically controlled histocompatibility determinants other than DLA in conditioning allograft reactivity. The determinants uncovered in the present study appear to be linked to the DLA complex, as demonstrated by the ability of the pedigree origins of DLA haplotypes present in individual dogs to serve as an effective marker system for such non-DLA antigen(s). The results also point to the potential usefulness of the early postirradiation period for the induction of allogeneic unresponsiveness in large adult mammals.

LETHALLY IRRADIATED DOGS GIVEN INFUSIONS OF FETAL AND ADULT HEMATOPOIETIC TISSUE.

Lethally irradiated dogs, 1500 r, were given infusions of fetal hematopoietic tissue (16 dogs), adult marrow and fetal hematopoietic tissue (17 dogs), and adult marrow and adult spleen (8 dogs). The incidence of successful engraftment, graft rejection, and secondary syndromes was found to be similar to that observed after adult marrow alone. It is concluded that in this species as in the mouse successful marrow engraftment and survival of secondary syndromes depend primarily on a choice of donor and recipient of reasonable histocompatibility.

Specific allogeneic unresponsiveness in irradiated dogs reconstituted with autologous bone marrow.

Hemopoietic reconstitution of supralethally irradiated adult dogs of the Cooperstown colony with their own stored bone marrow can produce long-term unresponsiveness to DLA-identical kidney allografts with no need for any additional immunosuppression. Eleven of 18 kidneys transplanted 12 hr after replacement of autologous marrow into irradiated recipients currently survive with normal function for as long as 1,417 days; 8 of 13 organs transplanted 28 hr after marrow replacement, and 8 of 13 organs transplanted 36 hr after marrow injection, currently survive up to 502 days, with no further treatment. Alterations in the timing and sequence of each procedure decrease the incidence of unresponsiveness. Implantation of kidney allografts 20 hr before removal of marrow, followed by irradiation and marrow reconstitution produced unresponsiveness in only 3 of 21 dogs. Kidneys transplanted at the time of marrow replacement were tolerated in 4 of 16 irradiated recipients. When kidneys were placed into irradiated dogs, and an interval of 40 hr was provided before marrow reconstitution, only 5 of 19 animals became unresponsive. Twelve dogs unresponsive to renal allografts received skin grafts from the respective kidney donors. Five dogs were unresponsive to such skin allografts; the remaining seven dogs rejected the skin grafts within 115 days. Second-, third-, and fourth-set skin grafts from the kidney donor were rejected in a progressively more rapid fashion. Survival and function of the kidney allografts were not affected by the rejection of successive skin grafts from the kidney donor. Skin grafts from other DLA-identical donors and DLA-incompatible skin grafts were rejected by the same recipients in uniform fashion.