Rads Of Total Body Irradiation Research Articles

Blockade of Programmed Death Ligand 1 Enhances the Therapeutic Efficacy of Combination Immunotherapy against Melanoma

Inhibition of antitumor T cell responses can be mediated by the productive interaction between the programmed death-1 (PD-1) receptor on T cells and its ligand PD-L1. PD-L1 is highly expressed on both murine bone marrow-derived dendritic cells (DCs) and B16 melanoma. In this study, in vitro blockade of PD-L1 interaction on DCs led to enhanced IFN-gamma production and cytotoxicity by Ag-specific T cells. In vivo, the systemic administration of anti-PD-L1 Ab plus melanoma peptide-pulsed DCs resulted in a higher number of melanoma peptide-specific CD8(+) T cells, but this combination was insufficient to delay the growth of established B16 melanoma. Although the addition of 600 rad of total body irradiation delayed tumor growth, further adoptive transfer of Ag-specific CD8(+) T cells was needed to achieve tumor regression and long-term survival of the treated mice. Lymphopenic mice treated with anti-PD-L1 Ab demonstrated increased activation and persistence of adoptively transferred T cells, including a higher number of CD8(+) T cells infiltrating the tumor mass. Together, these studies support the blocking of PD-L1 signaling as a means to enhance combined immunotherapy approaches against melanoma.

Up-Regulated Expression of the CXCR2 Ligand KC/GRO-α in Atherosclerotic Lesions Plays a Central Role in Macrophage Accumulation and Lesion Progression

Macrophage-mediated inflammation is central to atherogenesis. We have determined previously that the CXC chemokine receptor CXCR2 is involved in advanced atherosclerosis. We sought to determine whether one of the ligands of CXCR2, KC/GRO-alpha, can also modulate atherogenesis. KC/GRO-alpha(-/-) mice were generated and mated with the atherosclerosis-prone LDLR(-/-) mice. There was a significant reduction in atherosclerosis in mice lacking KC/GRO-alpha; however, this reduction was only approximately half that seen previously in mice lacking CXCR2 in the leukocyte. To determine whether CXCR2 is involved in the early formation of atherosclerosis, leukocyte-specific CXCR2(-/-) chimeric mice on LDLR(-/-) background were generated. Early fatty streak lesion formation in these mice was not affected by leukocyte CXCR2 deficiency whereas lesions were less developed in mice lacking leukocyte CXCR2 when atherosclerosis was allowed to progress to the intermediate stage. Macrophages were relatively sparse in the lesions of leukocyte CXCR2(-/-) mice despite robust MCP-1 expression. These studies indicate that KC/GRO-alpha/CXCR2 does not play a critical role in recruitment of macrophages into early atherosclerotic lesions but both arterial KC/GRO-alpha and leukocyte-specific CXCR2 expression are central to macrophage accumulation in established fatty streak lesions.

Expression of cathepsin G-like and α 1-antichymotrypsin-like proteins in reactive astrocytes

The central nervous system (CNS) of many different species responds to diverse neurologic injuries with an activation of astrocytes. Yet, the exact function of this reactive astrocytosis is unknown. In this report, mouse astrocytes were activated in vivo by focal penetrating brain injury. Reactive astrocytes were stained with antibodies raised against the serine protease cathepsin G (cat.G), the serine protease inhibitor α 1-antichymotrypsin (ACT), or the astrocytic marker glial fibrillary acidic protein (GFAP). Reactive astrocytes expressing both cat.G-like and ACT-like antigens were found around cerebral would margins between 18 h and 13 days after neural lesions. The injury-induced immunostaining was unaltered by 900 rads of total body irradiation, suggesting that the astroglial reaction was relatively independent of bone marrow-derived cells. The in vivo immunostaining was complemented with biochemical assays on cultured primary astrocytes. A synthetic peptide was used as a substrate in combination with specific inhibitors to identify a proteolytic activity within astroglial lysates and culture supernatants that closely resembles cat.G. This activity increased substantially upon stimulation of astrocytes with dibutyryl cyclic AMP and was neutralized by antibodies raised against cat.G. In a separate report, it was shown that astrocytes also contain an ACT-like inhibitory activity 38. The production of ACT- and cat.G-like antigens and activities by activated astrocytes should allow these cells to participate in a number of important biologic processes. Many of these processes may benefit the CNS by assisting in early wound repair. However, astroglial proteases and their inhibitors could also contribute to the pathogenesis of certain neurologic diseases.

Response to total body irradiation in dermatomyositis

There were two patients with severe dermatomyositis refractory to immunosuppressive therapy treated with 150 rad of total body irradiation given over a period of 5 weeks. Both patients responded promptly with minimal side-effects and remain in partial remission 42 and 18 months after completion of the treatment. Total body irradiation is effective in some patients with dermatomyositis who are refractory to standard therapy.

Effects of feeding lactobacillus GG on lethal irradiation in mice.

Mice exposed to 1400 rads of total body irradiation experienced 80%-100% mortality in 2 wk. Bacteremia was demonstrated in all dead animals. Feeding Lactobacillus GG strain reduced Pseudomonas bacteremia and prolonged survival time in animals colonized with this organism. In animals not colonized with Pseudomonas, feeding Lactobacillus GG also produced some reduction in early deaths, and there was less Gram-negative bacteremia in these animals compared with controls.

Administration of Fat Emulsions

In our bone marrow transplant unit, a 20-bed facility, we treat patients with leukemia and aplastic anemia. In preparation for a bone marrow transplant all patients receive 1,000 rads of total body irradiation and/or high-dose chemotherapy. This treatment destroys the patient's bone marrow function to prepare for the acceptance of a bone marrow transplant from a sibling or from one of the parents. Both the chemotherapy and the irradiation result in loss of appetite, nausea, vomiting, and severe mucositis leading to reduced oral intake. In addition, the body's nutritional requirements are increased by such occurrences as fever, infection, and graft-versus-host disease. Thus, at the same time oral intake is diminished, nutritional requirements are increased, leading to the possibility of a severely compromised nutritional state. One of the prominent nutritional deficiencies has been essential fatty acid deficiency (EFAD). This occurs both because of reduced oral intake of fats and because stored body fat is being used to meet energy requirements(1). The occurrence of EFAD can intensify the potential adverse effects of total body irradiation and chemotherapy. The common signs of EFAD

Allogeneic marrow transplantation for acute leukemia in relapse

The results of allogeneic marrow transplantation in 75 patients with acute lymphoblastic leukemia and 63 patients with acute non-lymphoblastic leukemia in relapse are reviewed. The effects of various chemotherapeutic regimens added to the basic regimen of cyclophosphamide (Cy) 60 mg/kg given on each of two successive days followed by 1000 rad of total body irradiation (TBI) were evaluated. The regimens tested were dimethylbusulphan (DMB), 1.3-bis(2-chlorethyl)-1-nitrosourea (BCNU) and daunorubicin. Seventeen of 138 patients are alive between three and nine and a half years from transplantation. The addition of other chemotherapeutic agents to the basic Cy and TBI regimen did not decrease relapse frequency or prolong survival.

Allogeneic marrow transplantation for acute lymphoblastic leukemia in remission using fractionated total body irradiation

Twelve patients with acute lymphoblastic leukemia in second to fourth remission received allogeneic marrow transplants following preparation with cyclophosphamide, 120 mg/kg and 1400 rad of fractionated total body irradiation. Two patients died of interstitial pneumonitis 32 and 62 days post-transplantation. Six patients relapsed between days 59 and 659 and four died of leukemia-related problems. Two patients who relapsed are currently alive, one in remission and one in relapse. Four patients are alive and free of disease 657 to 991 days following transplantation. This disease-free survival was not significantly better than the six of 22 disease-free survivors previously observed following cyclophosphamide and 1000 rad of total body irradiation given in a single exposure.

Result of Attempted Hematopoietic Reconstitution Using Isologous, Peripheral Blood Mononuclear Cells: A Case Report

Abstract In order to test the effect of peripheral blood mononuclear cell infusions on hematopoietic recovery in man we intensively leukapheresed a normal identical twin and obtained 9.8 x 10(10) peripheral blood mononuclear cells containing 4 x 10(5) CFU-C. These isologous cells were infused into his identical twin brother who had received 150 rad of total body irradiation and intensive combination chemotherapy as adjuvant therapy for Ewing's sarcoma. When compared to other patients receiving similar treatment, leukocyte recovery was accelerated by 3–4 wk, and occurred at a rate comparable to that induced by infusion of autologous cryopreserved marrow. Recovery of granulocytes, monocytes and platelets was not accelerated. The low number of CFU-C present in the preparation used ((one-eighth the number of CFU-C we usually obtain from bone marrow autograft collections) may have led to the pattern of hematopoietic recovery we observed in this patient.

Result of attempted hematopoietic reconstitution using isologous, peripheral blood mononuclear cells: a case report

In order to test the effect of peripheral blood mononuclear cell infusions on hematopoietic recovery in man we intensively leukapheresed a normal identical twin and obtained 9.8 x 10(10) peripheral blood mononuclear cells containing 4 x 10(5) CFU-C. These isologous cells were infused into his identical twin brother who had received 150 rad of total body irradiation and intensive combination chemotherapy as adjuvant therapy for Ewing's sarcoma. When compared to other patients receiving similar treatment, leukocyte recovery was accelerated by 3-4 wk, and occurred at a rate comparable to that induced by infusion of autologous cryopreserved marrow. Recovery of granulocytes, monocytes and platelets was not accelerated. The low number of CFU-C present in the preparation used ((one-eighth the number of CFU-C we usually obtain from bone marrow autograft collections) may have led to the pattern of hematopoietic recovery we observed in this patient.

Autologous Bone Marrow Transplantation Following Chemotherapy and Irradiation in Dogs With Spontaneous Lymphomas<xref ref-type="fn" rid="FN2">2</xref><xref ref-type="fn" rid="FN3">3</xref>

Thirty dogs with spontaneous lymphomas were administered two to six cycles of chemotherapy and were randomized into 3 groups to receive 800 rads of total body irradiation and autologous bone marrow transplantation. Of 10 dogs irradiated after chemotherapy-induced remission and infused with remission marrow (group 1), 8 (80%) had successful grafts and experienced remissions lasting 62 to 1024 days. Of 9 dogs irradiated during remission and infused with remission marrow mixed with autologous tumor cells (group 2), 6 (66%) had remission lasting 15 to 45 days. Eleven dogs with progressive tumor growth (relapse) following chemotherapy were irradiated and infused with remission marrow (group 3). Tumor remission lasting 39 to 350 days was observed in 5 dogs (45%) in this group, and 6 dogs died in less than 30 days. Dogs in groups 1 to 3 had median survival times of 216, 60, and 45 days, respectively. The prolonged survival times for dogs in group 1 compared to dogs in groups 2 and 3 suggest that protocols involving irradiation and autologous marrow grafting in this model would be most effective when these protocols are applied to animals having a minimum tumor burden at the time of irradiation and when the graftingmore » is done with tumor-free autologous marrow.« less

HÆMOPOIETIC RECOVERY IN EWING'S SARCOMA AFTER INTENSIVE COMBINATION THERAPY AND AUTOLOGOUS MARROW INFUSION

After the completion of combination therapy designed to achieve local control of Ewing's sarcoma, 13 patients with either truncal primary lesions or proven metastases were given 150 rad of total body irradiation over 5 weeks followed by cyclophosphamide, doxorubicin, imidazole carboxamide, and vincristine. 11 patients received autologous cryopreserved marrow infusions. In 2 patients marrow collections were not attempted. Two patterns of hæmopoietic recovery were observed: 8 patients, who had received marrow infusions, showed leucocyte, granulocyte, and platelet recovery by 27, 28, and 30 days. 5 patients, 3 of whom had also received marrow, showed more delayed recovery with leucocyte, granulocyte, and platelet recovery at 45, 53, and 77+ days. Delayed recovery in patients receiving marrow seemed to correlate with aberrations in marrow freezing-rate during phase change, and these aberrations could be shown to diminish post-freeze recovery of marrow granulocytemonocyte precursor cells.

Origin of human bone marrow fibroblasts.

We investigated the origin of bone marrow fibroblasts in three bone marrow transplant recipients with aplastic anaemia and leukaemia who received grafts from HLA-identical siblings of opposite sex. The patients were conditioned for transplantation with high doses of cytotoxic drugs and 300--1000 rads total body irradiation. After transplantation, bone marrow cells were cultured in T flasks for 3 weeks and the adherent cells were then trypsinized and passaged weekly. After several passages the cells had the typical morphology and growth pattern of fibroblasts. Metaphases from these cells were all of recipient sex type. In contrast, haematopoietic cells and lymphocytes obtained at the same time were of donor sex type. Our findings indicate that the human bone marrow fibroblast is not derived from a precursor common to haemotopoietic cells or lymphocytes. The bone marrow fibroblast appears to be a mesenchymal cell, unrelated to haematopoietic stem cells, which is capable of in vitro proliferation after as much as 1000 rads of total body irradiation.

1,3 Bis (2-chloroethyl)-1-nitrosourea-induced host resistance to syngeneic LSA mouse leukemia

A syngeneic tumor, the LSA lymphoma of C57BL mice, is considered non-antigenic or only weakly antigenic because no host reaction was elicited after repeated exposure of mice to killed tumor. However, easily detectable complete tumor resistance was elicited after 1 or 2 exposures to BCNU and tumor cure. Marked amplification of this host immunity in the BCNU treatment-cured mouse was shown by serial graded exposures to viable tumor grafts, and it was possible to establish virtually complete resistance to further tumor challenge by this method. A lymphoma spleen colony forming assay detected the antigenic properties of the LSA lymphoma in syngeneic C57BL mice. The host resistance state was easily prevented by sublethal irradiation of the mouse with 300 to 400 rad before tumor implantation. Once resistance was established, it was not altered by 450 rad of total body irradiation. These differences indicate a radiosensitive cell for the adaptive immune response and a less radiosensitive (and more complex) mechanism for the established tumor resistant state.

672 SUCCESSFUL BONE MARROW TRANSPLANTATION (BMT) FOR APLASTIC ANEMIA WITH AN HLA-MLC NON-IDENTICAL PARENT DONOR

Successful BMT was performed in a 9 yr. old black female with severe idiopathic aplastic anemia from her HLA-MLC non-identical father. Initial Hgb.=3.8 gm.%, retic. count=0.1%, WBC=700 cmm., platelets=10,000 cmm.; bone marrow biopsy was markedly hypocellular with 85% non-hematopoietic elements. Tissue typing showed the patient to be HLA A-1 B-8, A-26 B-7; the father was HLA A-1 B-8, A-3 B-7. In mixed leukocyte culture (MLC), there was low but significant bidirectional (10%; 16%) stimulation. Cell mediated lympholysis demonstrated no significant killing. Pre-transplant preparation included Procarbazine 15 mg/kg/day × 3 days, anti-thymocyte globulin (ATG) 15 mg/kg/day x 3 days, and 750 rads of total body irradiation by linear accelerator. Cells from the donor (6.9 × 108/kg) were infused intravenously. Graft versus host (GVH) prophylaxis following BMT consisted of IV Methotrexate weekly × 100 days and ATG and corticosteroids on days 7-21. By day 28 post BMT, cells of donor HLA type were present and marrow chromosomes were 100% XY. On days 29-31 mild GVH developed with fever, rash and slight liver enzyme elevations. Skin biopsy was consistent with GVH. Skin GVH resolved with Prednisone treatment. At 7 months post BMT peripheral counts are normal, however liver enzyme elevations persist. Marrow engraftment with minimal GVH in this patient should encourage further trials of BMT in patients without HLA-MLC identical siblings.

Neoplastic and life-span effects of chronic exposure to tritium. II. Rats exposed in utero.

A study was conducted to determine the effects on neoplasia incidence and life-span of exposure in utero to a major environmental radionuclide. Sprague-Dawley rats were continuously exposed to tritiated water (HTO) from conception through birth in doses of 0, 1, 10, 50, and 100 muCi HTO/ml body water. HTO administration was terminated at birth. Calculated cumulative doses during gestation were approximately 0, 6.6, 66, 330, and 660 rads of total body irradiation. Under these exposure conditions, the two highest doses resulted in sterile offspring. Animals surviving through 30 days postnatally were defined as the study population and observed until their deaths. Intrauterine exposures to doses up to 66 rads had no significant effects on either sex with respect to life-span, overall neoplasia incidence, incidence rate, or onset of mammary fibroadenomas. Females exposed to 330 or 660 rads were sterile and had lower incidence rates of mammary fibroadenomass than did controls; at 660 rads females had a lower incidence of overall neoplasia and reduced mean life-spans. Sterile male offspring had reduced mean longevity after irradiation at 660 rads. Regardless of dose group, females had significantly higher incidences of neoplasia and longer life-spans than males.

Studies of immunological reactivity following syngeneic or allogeneic marrow grafts in man.

This report describes studies of immunological reactivity in 13 marrow transplant recipients prepared for engraftment with 1,000 rads of total body irradiation, cyclophosphamide, or both, and observed for 5–30 months after engraftment. Three patients with hematological malignancy had identical twin donors; five patients with hematological malignancy and five with aplastic anemia had HL-A-matched sibling donors. Serum IgG, IgA, and IgM levels declined in all recipients and then returned to normal by day 100. Absolute lymphocyte counts were above 1,000/mm3 in all patients within 1–3 months after grafting. Two of the three twin recipients showed normal antibody responses to bacteriophage by days 39 and 365 postgrafting and cutaneous reactivity to dinitrochlorobenzene by days 393 and 678. These three patients had no significant infections. Allogeneic graft recipients showed a markedly decreased antibody response to bacteriophage. In contrast, several of these recipients developed cytomegalovirus or herpes zoster infections, from which they recovered, and they developed good antibody titers against these agents. Testing in mixed leukocyte culture showed that cells from recipients of syngeneic grafts displayed normal responses to allogeneic cells and phytohemagglutinin. Recipients of allogeneic grafts showed a wide range of responsiveness but clear stimulation was observed on nearly all occasions, even within the 1st month after grafting. Despite this in vitro reactivity, the allogeneic recipients, with one exception, could not be sensitized to dinitrochlorobenzene when tested between days 28 and 365. We conclude that recipients of syngeneic grafts show a satisfactory return of immunological reactivity. Recipients of allogeneic grafts show a long lasting immunological deficiency which points out the necessity for vigilance in early detection and treatment of infection and the need for additional measures to restore immunological competence in these patients.