Nucleotide sequence and polymorphism analysis of canine DRA cDNA clones.
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Amino Acid Sequence↗
Biomedical subjects
R Storb
Publications and source records attributed to R Storb.
Factors that influence collection and engraftment of autologous peripheral-blood stem cells.
PURPOSE: To analyze factors that affect the collection of peripheral-blood stem cells (PBSC) before transplant and the tempo of engraftment after transplant. PATIENTS AND METHODS: A consecutive series of 243 patients with breast cancer (n = 87), malignant lymphoma (n = 90), multiple myeloma (n = 32), or other malignancies (n = 34) had PBSC collected following stimulation with colony-stimulating factors (CSFs) or after chemotherapy followed by CSF. Infusion of PBSC was performed following myeloablative chemotherapy with chemotherapy with or without total-body irradiation (TBI). Postinfusion CSFs were administered to 72 patients. An analysis of factors that influence CD34+ cell yield was performed by linear regression. Cox regression analysis was used to determine factors that affect the kinetics of granulocyte and platelet recovery following infusion of PBSC. RESULTS: Mobilization with chemotherapy followed by CSF, a diagnosis of breast cancer, absence of marrow disease, no prior history of radiation therapy, and fewer cycles of conventional-dose chemotherapy were associated with a higher average daily yield of CD34+ cells. In the multivariate analysis, the CD34 content of infused cells and the use of a posttransplant CSF influenced neutrophil recovery after infusion of PBSC. CD34 content was also important for predicting platelet recovery. The use of postinfusion CSF was associated with a significant delay in platelet recovery in patients who received less than 5.0 x 10(6) CD34+ cells/kg, but there was no discernable effect in patients who received greater than 5.0 x 10(6) CD34+ cells/kg. CONCLUSION: Disease status and prior treatment influence the ability to mobilize PBSC. CD34 cell dose is an important predictor of engraftment kinetics after PBSC transplant, regardless of disease or mobilization technique. The use of postinfusion CSF improves neutrophil recovery, but at low CD34 doses can delay platelet recovery.
Adolescent↗
Peripheral-blood stem-cell collections after paclitaxel, cyclophosphamide, and recombinant human granulocyte colony-stimulating factor in patients with breast and ovarian cancer.
PURPOSE: Here we evaluate Taxol (paclitaxel; Bristol-Myers Squibb, Princeton, NJ) and cyclophosphamide (CY) with recombinant human granulocyte colony-stimulating factor (rhG-CSF) for mobilization of peripheral-blood stem cells (PBSCs) for autologous stem-cell transplantation in patients with breast and ovarian cancer. PATIENTS AND METHODS: PBSCs were collected from 17 patients with breast (n = 11), ovarian (n = 5), and gastric (n = 1) cancer after administration of Taxol (170 mg/m2 x 1) and CY (4 g/m2 x 1) followed by rhG-CSF (10 micrograms/kg/d). PBSC collections after Taxol and CY were compared with PBSC collections from nine patients with stage IV breast (n = 8) or stage III ovarian (n = 1) cancer who had received CY (4 g/m2 x 1) followed by rhG-CSF (16 micrograms/kg/d) for mobilization. RESULTS: Mean WBC and platelet counts on the first day of apheresis were 6.3 x 10(9)/L (range, 1.9 to 22.1) and 35 x 10(9)/L (range, 19 to 77), respectively. The median numbers of CD34+ cells, peripheral-blood mononuclear cells (PBMNC), and peripheral-blood total nucleated cells (PBTNC) collected were 13.02 x 10(6)/kg (range, 5.4 to 57.8; mean, 16.02), 6.86 x 10(8)/kg (range, 1.9 to 51.2), and 17.41 x 10(8)/kg (range, 2.4 to 106.6), respectively. In the comparison group, the median yield of CD34+ cells was 6.39 x 10(6)/kg (range, 0.2 to 28; mean, 10.01; P = .01). The mean daily yield of CD34+ cells/kg/collection was 3.5 (range, 0.8 to 28.9) after Taxol and CY, as compared with 1.3 (range, 0.1 to 7.0) for patients who received CY alone (P = .01). All patients who received CY and Taxol reached a target level of 5 x 10(6) CD34+ cells/kg, as compared with five of nine patients (55.5%) who received CY alone (P = .03). CONCLUSION: These data suggest that Taxol and CY followed by rhG-CSF mobilizes PBSCs in patients with advanced breast and ovarian cancer more effectively than this regimen without Taxol.
Adult↗
An update on allogeneic marrow transplantation for myelodysplastic syndrome.
Ninety-three patients with myelodysplastic syndrome with either excess blasts or life-threatening cytopenia received cyclophosphamide and total body irradiation (88 patients) or busulfan (5 patients) followed by allogeneic (90 patients) or syngeneic (3 patients) marrow infusion. Thirty-eight patients are disease-free survivors between 1.8 and 11.3 years (median 6.1 years) after transplantation. The median Karnofsky performance status of the survivors is 100%, and only 2 have a performance status of < 80%. Eighteen patients relapsed and the remaining 37 died without evidence for relapse. The 5-year actuarial disease-free survival, relapse, and non-relapse mortality rates are 40%, 29%, and 44%, respectively. The actuarial relapse rate was higher among patients with excess blasts at the time of transplantation compared to those without excess blasts (49% vs. 4%, p = 0.0001), resulting in a lower disease-free survival (31% vs. 54%, p = 0.07). Increasing age was associated with a lower disease-free survival (p = 0.02). In a multivariate analysis, younger age, shorter disease duration, and absence of excess blasts were associated with improved outcome. We recommend that patients with myelodysplasia with appropriate marrow donors, especially patients less than age 40, undergo marrow transplantation early during the disease course, before the disease progresses or life-threatening complications develop.
Adolescent↗
Bone marrow transplantation.
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Anemia, Aplastic↗
Prevention of graft-versus-host disease after bone marrow transplantation.
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Acute Disease↗
Ultrastructural localization of stem cell factor in canine marrow-derived stromal cells.
Stromal cell lines derived from canine long-term bone marrow cultures (LTBMC) were characterized regarding the expression of growth factors and especially the localization of stem cell factor (SCF) (c-kit ligand). One cell line (DO64) was immortalized by transformation with a retroviral vector containing the open reading frames (ORFs) E6 and E7 of the human papilloma virus type 16 (HPV-16). Transfection did not change cellular characteristics but rendered the cell line more independent from culture conditions. The transformed line DO64 consisted mainly of fibroblast-like cells. In addition, some cells showed endothelial and some smooth-muscle cell features. Stromal cells expressed a broad spectrum of surface markers, including low levels of major histocompatibility-complex (MHC) class-II antigens. A new murine monoclonal antibody (MAb), RG7.6 (IgG1), specific for canine SCF, recognized the majority of fibroblast-like stromal cells. The staining pattern for SCF showed perinuclear and intracytoplasmic dense areas. Immunoelectron microscopy revealed the localization of SCF in secretory vesicles, the perivesicular cytoplasm, and bound to the cytoplasmatic membrane. RNA analysis showed that stromal cells transcribed, in addition to SCF, messages for granulocyte colony-stimulating factor (G-CSF), granulocyte-monocyte CSF (GM-CSF), interleukin-6 (IL-6), and transforming growth factor-beta (TGF-beta). In summary, we have established and characterized canine marrow-derived stromal cell lines, and using the new MAb RG7.6, we have localized SCF to cytoplasmatic vesicles as well as the membrane of stromal cells.
Animals↗
Major histocompatibility complex class II-mediated inhibition of hemopoiesis in vitro and in vivo is abrogated by c-kit ligand.
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Animals↗
Influence of total nucleated cell dose from marrow harvests on outcome in patients with acute myelogenous leukemia undergoing autologous transplantation.
This retrospective study was conducted to determine whether the total number of nucleated cells (TNC)/kg collected at marrow harvest was associated with outcome in 151 patients with acute myelogenous leukemia (AML) who received an autologous purged (n = 67) or non-purged (n = 84) marrow transplant. At the time of transplant 33 patients were in first complete remission (CR), 47 in second CR, 54 in first relapse and 17 beyond second CR. Ninety patients received busulfan (BU) 16 mg/kg and cyclophosphamide (CY) 120 or 200 mg/kg, 51 patients received CY 120 mg/kg and total body irradiation (TBI) 12-15.75 Gy and 10 patients received BU 8 mg/kg, CY 60 mg/kg and TBI 12 Gy as conditioning regimens. Patients whose marrow harvest yielded < 2 x 10(8) TNC/kg did not undergo purging with 4-hydroperoxycyclophosphamide (4HC). This group of patients (n = 28) had a 100 day mortality of 50% and only 54% achieved a granulocyte levels of > 0.5 x 10(9)/l and 29% achieved platelet transfusion independence. Patients whose marrow harvest yielded 2-4 x 10(8) TNC/kg and did not undergo marrow purging had a 20% mortality by day 100, 91% recovered granulocytes to > 0.5 x 10(9)/l and 61% became platelet independent. Patients whose marrow harvest yielded 2-4 x 10(8) TNC/kg and underwent marrow purging with 4HC had a 50% mortality by day 100 and 58% achieved a granulocyte levels of > 0.5 x 10(9)/l and 42% became platelet transfusion independent.(ABSTRACT TRUNCATED AT 250 WORDS)
Acute Disease↗
Rapid engraftment after autologous transplantation utilizing marrow and recombinant granulocyte colony-stimulating factor-mobilized peripheral blood stem cells in patients with acute myelogenous leukemia.
This study was performed to determine whether peripheral blood stem cells (PBSCs) mobilized with recombinant granulocyte-colony stimulating factor (rhG-CSF) increase the tempo of granulocyte and platelet recovery when added to marrow in patients with acute myelogenous leukemia (AML) undergoing autologous bone marrow transplantation (BMT). Twenty six patients with AML had bone marrow harvested in first (n = 16) or second (n = 10) complete remission (CR) and cryopreserved. Patients received rhG-CSF alone (n = 20) or rhG-CSF following chemotherapy (n = 6). PBSCs were collected from 24 of the 26 patients a median of 7 (range 3-2130) days after marrow harvest. Two patients presumed to be in second CR did not have PBSCs collected because of early relapse. Fourteen patients in first CR (n = 3), second CR (n = 8) or first relapse (n = 3) proceeded to autologous BMT utilizing marrow + rhG-CSF-mobilized PBSCs. Engraftment parameters were compared with a historical group of 158 patients with AML who had received purged (n = 67) or unpurged (n = 91) autologous BMT without PBSCs. The median number of peripheral blood total nucleated and CD34+ cells collected from 24 patients was 19.55 x 10(8)/kg (range 1.83-54.83) and 5.59 x 10(6)/kg (1.23-34.79), respectively. All patients transplanted achieved a granulocyte level of > 0.5 x 10(9)/l with a median of 13 days (range 11-27 days) and platelets to 20 x 10(9)/l median 14 days (range 9-83 days).(ABSTRACT TRUNCATED AT 250 WORDS)
Acute Disease↗
Inhibition of hematopoiesis in long-term marrow cultures established on adherent layers from AcSDKP-treated dogs.
The tetrapeptide Acetyl-N-Ser-Asp-Lys-Pro (AcSDKP) interferes with G1/S phase progression in hematopoietic precursors. We investigated the effect of AcSDKP on in vitro and in vivo hematopoiesis in a canine model. AcSDKP, added daily for 2 weeks to long-term marrow culture (LTMC) at concentrations > 10(-8)M, reversibly inhibited colony-forming unit granulocyte/macrophage (CFU-GM) formation (p < 0.001 and p < 0.05 for 10(-6) and 10(-7)M, respectively). Inhibition was more profound when AcSDKP addition was begun at the initiation rather than at the time of recharging the cultures. Next, seven dogs were given AcSDKP in vivo at 50 (n = 2), 250 (n = 2), or 500 micrograms/kg/day (n = 3) via continuous infusion for 7 days. No adverse effects were observed. LTMCs were established on days -9, -2, +7, and +28 of AcSDKP. One week later (days -2, +5, +14, and +35), adherent layers were recharged with fresh autologous marrow, and CFU-GM in nonadherent cells was assayed weekly beginning 1 week after recharging. The cumulative number of CFU-GM harvested from LTMCs was dependent upon the time of initiation of LTMC. The difference between day -2 (adherent layer pre-AcSDKP; recharge on AcSDKP) and day +7 culture (adherent layer on AcSDKP; recharge after discontinuation of AcSDKP, p < 0.001) suggested an effect of AcSDKP on the adherent stromal layer. Ex vivo hematopoiesis partially recovered following discontinuation of AcSDKP, although CFU-GMs were still reduced in LTMCs established on day +28. Normal nonadherent cells recharged onto allogeneic adherent/layers obtained during AcSDKP treatment grew significantly fewer CFU-GM than cultures on adherent cells obtained before AcSDKP treatment (p < 0.05). Therefore, these data suggest that AcSDKP affects not only hematopoietic cells but also cells of the adherent layer.
Animals↗
Low IgG production by mononuclear cells from marrow transplant survivors and from normal neonates is due to a defect of B cells.
Low IgG production is a characteristic feature of both the immunodeficiency of recipients of bone marrow transplant (BMT) and the physiologic immunodeficiency of newborns. We tried to determine whether this is due to a B cell defect and whether this can be corrected in vitro by IgG production-promoting cytokines CD40 ligand (CD40L) and interleukin 10 (IL-10). Highly purified circulating B cells from patients at 1 year after transplant, normal neonates and normal adults were cultured with and without anti-mu plus CD40L plus IL-10. Proliferation was measured on day 4 and IgM and IgG production were measured on day 9. Proliferation and IgM production of B cells from patients and from neonates were somewhat low (43-67% of normal). In contrast, IgG production by B cells from patients and from neonates was markedly low (< or = 13% of normal). Decreased IgG production correlated with decreased percentage of B cells negative for membrane IgD (mIgD). We conclude that the low IgG production of normal neonates and patients at 1 year after transplant is, at least in part, caused by a defect of circulating B cells and that this defect cannot be corrected by CD40L and IL-10. Quantitative deficiency of switched mIgD-) B cells probably accounts for this defect.
Adult↗
Effects of rhIL-11 on normal dogs and after sublethal radiation.
The effects of recombinant human interleukin-11 (rhIL-11) were studied in normal dogs and dogs given otherwise sublethal total-body irradiation (TBI) without marrow transplantation. Ten normal dogs were given rhIL-11 subcutaneously, twice daily for 14 days at varying doses, two dogs at 30 micrograms/kg/day, four dogs at 60 micrograms/kg/day, two dogs at 120 micrograms/kg/day, and two dogs at 240 micrograms/kg/day. Peripheral blood platelet counts increased in all dogs. The increase in platelet counts ranged from 1.4 to 3.1 times the pre-treatment level. The greater increases of platelets were associated with higher doses (p = 0.01). No change in platelet size was evident except at the dose of 240 micrograms/kg/day. There were no changes in the total white blood cell (WBC) count or differential. A higher proportion of megakaryocytes with a DNA content of 32N/64N was observed in dogs treated with rhIL-11 at day 7 (n = 6) than for control dogs that did not receive rhIL-11 (n = 7; p = 0.01). In both peripheral blood and marrow, significantly increased hematopoietic progenitors (i.e, colony-forming unit granulocyte/macrophage [CFU-GM]) were present 7 and 14 days after the start of treatment. Concentrations of serum fibrinogen increased by a median of 155 mg/dL at day 7 of rhIL-11 (p < 0.01). Cholesterol also increased by a median of 52 mg/dL at day 14 (p < 0.01). There was a single death of a non-irradiated dog from pneumonitis on day 15 after the start of rhIL-11 administration at a dose of 120 micrograms/kg/day. All other non-irradiated dogs tolerated rhIL-11 without any significant adverse effects. Five dogs were given 200 cGy TBI without marrow grafting, followed by 240 micrograms/kg/day rhIL-11 subcutaneously in two divided doses for 28 days starting within 2 hours of TBI. The results in this group were compared with 10 dogs that had previously or concurrently been given 200 cGy without marrow grafting or hematopoietic growth factors. Two of the five treatment dogs died of pneumonitis on day 13 compared to one death among 10 control dogs on day 24. Among dogs that survived to hematologic recovery, the rhIL-11 dogs had decreased platelet counts (< 150,000) for a median of 24 days (range = 24 to 41) compared to a median of 28 days (range = 21-40) for the control group. Treatment with rhIL-11 increased platelet counts, platelet size, ploidy number of megakaryocytes, and marrow and peripheral blood CFU-GM in normal dogs.(ABSTRACT TRUNCATED AT 400 WORDS)
Animals↗
Phase II study of busulfan, cyclophosphamide and fractionated total body irradiation as a preparatory regimen for allogeneic bone marrow transplantation in patients with advanced myeloid malignancies.
A previous phase I dose escalation study determined that the maximum tolerated doses of busulfan and cyclophosphamide that could be combined with 12.0 Gy of total body irradiation were 7 mg/kg and 50 mg/kg, respectively. A phase II study of these three agents was carried out in 56 patients with advanced myeloid malignancies receiving allogeneic bone marrow transplants from HLA-identical donors. Cyclosporine with methotrexate or with prednisone was administered for prophylaxis against graft-versus-host disease. Grade 3 (n = 8) and 4 (n = 3) regimen-related toxicity occurred in 20% of patients, which was the maximum predicted from the phase I study. The 2-year actuarial probabilities of non-relapse mortality and relapse were 0.52 and 0.55, respectively. Fourteen patients survive, 12 in remission, 581-1761 days post-transplant. The actuarial probabilities of disease-free survival for patients with recurrent acute myeloid leukemia and advanced chronic myeloid leukemia at 2 years were 20% and 23%, respectively. When compared with our historical experience in patients receiving other treatment regimens, there was no apparent improvement in disease-free survival.
Adolescent↗
Engraftment of patients with lymphoid malignancies transplanted with autologous bone marrow, peripheral blood stem cells or both.
Forty six patients with lymphoid malignancies receiving autologous transplants using three different sources of hematopoietic stem cells were compared for engraftment parameters. Thirteen patients (five with multiple myeloma, seven with non-Hodgkin's lymphoma and one with Hodgkin's lymphoma) received autologous marrow with post-transplant growth factors (group 1). During the same time interval, 14 patients (five with multiple myeloma, six with non-Hodgkin's lymphoma and three with Hodgkin's lymphoma) were transplanted with autologous marrow plus recombinant granulocyte colony-stimulating factor (rhG-CSF)-mobilized peripheral blood stem cells (PBSC) and post-transplant growth factors (group 2). Nineteen patients (seven with multiple myeloma and 12 with non-Hodgkin's lymphoma) received rhG-CSF mobilized PBSC and post-transplant growth factors (group 3). All PBSC were collected after G-CSF mobilization (16 micrograms/kg/day s.c. for 6 days) without prior chemotherapy. After high-dose myeloablative chemotherapy or chemoradiotherapy, the median days to recovery of neutrophils to levels of 0.5 and 1.0 x 10(9)/l were 12 vs. 9 vs. 9 days (P = 0.0003 (group 1 vs. group 2) and P = 0.53 (group 2 vs. group 3)) and 13 vs. 10 vs. 10 days (P = 0.0003 (group 1 vs. group 2) and 0.92 (group 2 vs. group 3)) for groups 1, 2 and 3, respectively. The median day to platelet transfusion independence was 22 vs. 11 vs. 11 days (P = 0.001 (group 1 vs. group 2) and P = 0.50 (group 2 vs. group 3)) for groups 1, 2 and 3, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)
Adult↗
T cell reconstitution after bone marrow transplantation into adult patients does not resemble T cell development in early life.
Immune reconstitution after marrow transplantation has some characteristics of immune development in early life. Here we provide phenotypic data suggesting this may not be true for T cells (particularly CD4+ T cells) in the case of marrow transplantation into adults. T cells from 35 adult patients at 1 year after transplant, 14 normal neonates and 22 normal adults were studied by 3-color flow cytometry. Marked disparity between the phenotype of neonatal vs post-transplant T cells was found. Of the total CD4+ T cells, the neonates had supranormal percentages of CD45RAhigh, L-selectin+, CD29low/-, CD11alow/- and CD28+ cells whereas most patients at 1 year after transplant had subnormal percentages of these CD4+ T cell subpopulations. (sub/supra-normal denotes below/above normal adult values). Absolute blood counts of naive (CD45RAhigh, L-selectin+, CD29low/- and CD11alow/-) CD4+ T cells correlated inversely with patient age. Neonates had also supranormal percentages of CD45RAhigh, L-selectin+, CD29low/-, CD11alow/- and CD28+ CD8+ T cells whereas the patients (particularly those with chronic GVHD) tended to have subnormal percentages of these CD8+ T cell subpopulations. Contrary to the CD4+ T cells, there was no correlation between the absolute counts of CD45RAhigh, L-selectin+, CD29low/- or CD11alow/- CD8+ T cells and patient age. Whereas the vast majority of neonatal T cells were CD38high, most patient and normal adult T cells were CD38- or CD38intermediate (both CD4+ and CD8+ T cells). We conclude that, in contrast to early life, the production of naive CD4+ T cells is deficient in adult (and particularly elderly) transplant recipients.
Adolescent↗
Transplantation of hematopoietic stem cells.
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Anemia, Hemolytic, Congenital↗
Major histocompatibility complex class II expression is required for posttransplant immunological but not hemopoietic reconstitution in mice.
We had previously shown in a canine model that the administration of anti-MHC class II monoclonal antibody (MAB) immediately after autologous marrow transplantation prevented hemopoietic reconstitution. Since MHC class II expression in mice differs from that in dogs we were interested in determining the effect of MHC class II manipulation on posttransplant hemopoietic and immunological recovery in mice. Three murine models including MHC class II knock-out mice were studied. BALB/c mice (I-E+, I-A+) given anti-MHC class II MAB H81.9 (anti-I-E; 1 mg/kg/day, days 0-4) after TBI and infusion of syngeneic marrow or infused with anti-I-E purged marrow both showed normal hemopoietic reconstitution. Similarly, C57B1/6 mice (I-E-, I-A+) transplanted with M5/114 (anti-I-A) purged marrow recovered normal hemopoiesis. MHC class II knock-out (C2D) mice, which lack class II completely, also recovered normal hemopoiesis after TBI and transplantation with either normal or class II-deficient (C2D) marrow, although the kinetics of platelet recovery as determined by megakaryocytopoiesis and platelet counts on day 14 were slightly delayed. C57B1/6 mice transplanted with C2D marrow recovered normally. Immunologic recovery, however, was abnormal both in C2D recipients and in normal mice transplanted with C2D marrow: While CD8+ T lymphocytes recovered normally, no (or only very few) CD4+ T cells were identified posttransplant. Treatment of normal mice with anti-MHC class II MAB in vivo or transplantation of MHC class II-purged marrow, however, did not interfere with complete immunological recovery, although T cell maturation was slightly delayed. Thus, complete immunological reconstitution requires the expression of MHC class II on marrow-derived precursor cells, while the expression of MHC class II antigens is not a requirement for hemopoietic reconstitution in mice.
Animals↗