Search PubMed⌕ Search

Biomedical subjects

R Hoffman

Publications and source records attributed to R Hoffman.

At least 127 records · Page 7Linked to original sources
PubMed1996

A monoclonal antibody recognizing CD43 (leukosialin) initiates apoptosis of human hematopoietic progenitor cells but not stem cells.

CD43 (the major sialoglycoprotein of leukocytes) is an adhesion molecule broadly expressed on hematopoietic cells. A monoclonal antibody recognizing this molecule induces apoptosis of lineage marker-negative bone marrow hematopoietic progenitor cells (HPCs) that express CD34 at a high density (CD34hiLIN-). However, not all cells within this population undergo apoptosis on stimulation via CD43. Dividing progenitor cells are most highly affected, whereas more primitive quiescent cells survive anti-CD43 monoclonal antibody treatment. These surviving cells (1) are enriched for cobblestone area-forming cells, (2) repopulate fragments for human fetal bone implanted into C.B-17 scid/scid mice, (3) have a potential to differentiate in vivo to myeloid and lymphoid cells, and (4) have a high proliferative potential in long-term stromal cell-free liquid culture. These data indicate that cells with hematopoietic stem cell characteristics are relatively resistant to CD43-mediated apoptosis as compared with HPCs. Thus, CD43 may be specifically involved in the regulation of HPC proliferation.

Adult↗
PubMed1996

Differential mobilization of myeloma cells and normal hematopoietic stem cells in multiple myeloma after treatment with cyclophosphamide and granulocyte-macrophage colony-stimulating factor.

Peripheral blood stem cells (PBSCs) mobilized with high-dose chemotherapy and hematopoietic growth factors are now widely used to support myeloablative therapy of multiple myeloma and effect complete remissions in up to 50% of patients with apparent extension of event-free and overall survival. Because tumor cells are present not only in bone marrow, but also in virtually all PBSC harvests, it is conceivable that autografted myeloma cells contribute to relapse after autotransplants. In this study, the kinetics of mobilization of normal hematopoietic stem cells were compared with those of myeloma cells present in PBSC harvests of 12 patients after high-dose cyclophosphamide and granulocyte-macrophage colony-stimulating factor administration. CD34+ and CD34+Lin-Thy+ stem cell contents were measured by multiparameter flow cytometry, and myeloma cells were quantitated by immunostaining for the relevant Ig light chain and by a quantitative polymerase chain reaction for the myeloma-specific CDRIII sequence. Results indicated marked heterogeneity in the percentages of mobilized stem cells among different patients (0.1% to 22.2% for CD34+ cells and 0.1% to 7.5% for CD34+Lin-Thy+ cells, respectively). The highest proportions of hematopoietic progenitor cells were observed early during apheresis, with 9 of 12 patients mobilizing adequate amounts of CD34+ cells for 2 autotransplants (> 4 x 10(6)/kg) within the first 2 days, whereas peak levels (percent and absolute numbers) of myeloma cells were present on days 5 and 6 (0.5% to 22.0%). During the last days of collection, mobilized tumor cells exhibited more frequently high labeling index values (1% to 10%; median, 4.4%) and an immature phenotype (CD19+). The differential mobilization observed between normal hematopoietic stem cells and myeloma cells can be exploited to reduce tumor cell contamination in PBSC harvests.

Biomarkers↗
PubMed1996

Hierarchical structure of human megakaryocyte progenitor cells.

Megakaryocytopoiesis is a complex biological process involving a series of cellular events that begins with the pluripotent hematopoietic stem cell and ultimately results in the biogenesis of platelets. A hierarchy of megakaryocyte (MK) progenitor cells has been previously defined based upon studies of in vitro megakaryocytopoiesis. Ontogeny-related changes in MK progenitor cells were analyzed in order to further define this cellular hierarchy. Unifocal colony-forming unit-megakaryocyte (CFU-MK)-derived colonies cloned from fetal bone marrow (FBM) formed after fewer days of in vitro culture and were 2.6-fold larger than those colonies cloned from adult bone marrow (ABM). The frequency of CFU-MK-derived colonies cloned from ABM was significantly greater. MK colonies, however, cloned from FBM morphologically consisted of both pure MK colonies and mixed colonies containing MKs, in which a core of CD41- cells were surrounded by CD41+ MKs. Large colonies resembling the primitive BFU-MK also were assayed from both FBM and ABM. These BFU-MK-derived colonies appeared after fewer days of incubation when FBM was assayed, compared to ABM, but at a significantly lower frequency. In addition, large unifocal MK colonies consisting of >300 cells (300-1000) appeared from cells cloned from fetal, but not adult, marrow. This type of colony represents a unique type of MK progenitor cell, termed the high-proliferative-potential cell-MK. Such colonies represent the progeny of the most primitive human MK progenitor cell identified to date. We also attempted to investigate the process of commitment of stem cells to the MK lineage. We explored the actions of thrombopoietin (TPO) on primitive hematopoietic cells in order to gain an understanding of stem cell commitment. CD34+ Thy-1+ Lin- marrow cells, which are enriched for pluripotent hematopoietic stem cells, were shown to express c-Mpl by the polymerase chain reaction. In addition, TPO alone was capable of inducing CD34+ Thy-1+ Lin- cells after two to three weeks to produce progeny composed entirely of MKs. These studies indicate that TPO has a profound effect on hematopoietic stem cells, and that the hierarchy of MK progenitor cells begins with the pluripotent hematopoietic stem cell.

Adult↗
PubMed1996

Long-term hematopoietic culture-initiating cells are more abundant in mobilized peripheral blood grafts than in bone marrow but have a more limited ex vivo expansion potential.

Mobilized peripheral blood hematopoietic progenitor cells obtained from cancer patients treated with high-dose cyclophosphamide (7g/m2) followed by G-CSF, GM-CSF, IL-3, PIXY321, or combinations of these cytokines have been successfully used for autologous stem cell transplantation. We investigated the ability of hematopoietic progenitor cells (HPC) derived from mobilized peripheral blood (PB) to undergo ex vivo expansion in short term cultures by enumerating numbers of de novo generated CD34+ cells, assayable progenitor cells, and the frequency of long-term hematopoietic culture-initiating cells (LTHC-IC). These parameters were examined in CD34+ cells generated in culture through the use of cell tracking with the membrane dye PKH2. Fresh isolated mobilized CD34+ cells contained 0.49 +/- 0.36% LTHC-IC. However, due to the high number of total CD34+ cells in mobilized PB, the absolute number of LTHC-IC was higher than that contained in a bone marrow (BM) harvest. Mobilized CD34+ cells were stained with PKH2 and incubated with SCF, IL-3, and IL-6. After 5 to 6 days, numbers of total CD34+ cells and clonogenic progenitors increased 1.4- and 2.2-fold, respectively. Numbers of total progenitors continued to increase such that 10 to 12 days after the initiation of cultures a 6.4-fold increase was demonstrable. However, between days 5 and 7 of culture, the frequency of LTHC-IC in CD34+PKH2bright cells (cells which did not divide) was less than 50% of that determined for fresh cells, while the frequency among CD34+PKH2dim cells (cells that had divided) was very low or undetectable. However, moderately higher frequencies of LTHC-IC were detected following expansion for 48 hours only. In similar assays, both BM and cord blood cells were capable of generating LTHC-IC in CD34+PKH2dim cells but not to expand the overall number of these progenitors. These observations suggest that although mobilized PB CD34+ cells contain large numbers of LTHC-IC, these cells might not be capable of further ex vivo expansion and generation of additional LTHC-IC in vitro. Furthermore, these data indicate that mobilized PB CD34+ cells may have undergone maximal "in vivo expansion" such that additional ex vivo expansion of primitive progenitor cells may not be possible.

Antigens, CD34↗
PubMed1996

Effect of aminoguanidine and cyclosporine on lung allograft rejection.

BACKGROUND: Aminoguanidine, a nitric oxide synthase inhibitor, has been shown to reduce the inflammatory allogeneic response. Here we used it in combination with cyclosporine to evaluate its effect on a clinically relevant immunosuppressive protocol. METHODS: Orthotopic left lung transplantation was performed in 120 rats, of which 24 were syngeneic Lewis to Lewis controls, and allogeneic transplantations were performed across major histoincompatibility barriers (ACI to Lewis). We studied synchronous histologic changes accompanying cytokines and nitric oxide synthase messenger RNA by reverse transcriptase polymerase chain reaction in the grafted lungs. Nitrate/nitrite, oxidized degradation products of nitric oxide, were measured in the whole blood, as were concentrations of cyclosporine. Lung tissue was immunohistochemically stained for nitric oxide synthase protein. Rats receiving allografts were either untreated (24) or received low-dose cyclosporine (232 +/- 105 ng/mL blood by high-performance liquid chromatography), high-dose cyclosporine (2,046 +/- 664 ng/mL), aminoguanidine alone (800 mg. kg-1. day-1 intraperitoneally), or aminoguanidine plus low-dose cyclosporine. RESULTS: The results suggest that aminoguanidine combined with low doses of cyclosporine can reduce the allogeneic response across major histoincompatibilities in rodent lung transplantation. Its biologic effect may not exclusively depend on the inhibition of nitric oxide synthase and may, by other means, reduce proinflammatory cytokines. CONCLUSIONS: Aminoguanidine may be an effective adjuvant to conventional immunosuppression.

Animals↗
PubMed1996

Inhibition of angiogenesis and murine tumour growth by laminarin sulphate.

LAM S5 is a polysulphated derivative of the glucan laminarian that inhibits basic fibroblast growth factor (bFGF) binding and the bFGF-stimulated proliferation of fetal bovine heart endothelial (FBHE) cells. This report demonstrates that LAM S5 has anti-angiogenic activity, as shown by inhibition of tubule formation by endothelial cells cultured on Matrigel and inhibition of vascularisation of the chick chorioallantoic membrane. In addition, LAM S5 caused a tumour growth delay of the murine RIF-1 tumour of 2.6 days (P = 0.01).

Animals↗
PubMed1996

Managed care and the infectious diseases specialist.

There is growing demand to contain health care costs and to reassess the value of medical services. The traditional hospital, academic, and research roles of the infectious disease (ID) specialist are threatened, yet there is an increasing need for expertise because of growing antimicrobial resistance and emerging pathogens. Opportunities exist to develop and expand services for the care of patients infected with human immunodeficiency virus and in infection control, epidemiology, outcomes research, outpatient intravenous therapy, and resource management. It is important for ID physicians to appreciate the principles involved in managed care and the areas in which ID services can be valuable. To be effective, physicians need to know about tools such as practice guidelines, physician profiling, outcomes monitoring, computerized information management, risk sharing, networking, and marketing, as well as related legal issues. With a positive attitude toward learning, application, and leadership, ID physicians can redefine their role and expand their services through managed care.

Ambulatory Care↗
PubMed1996

Inhibition of nitric oxide synthesis does not improve interleukin-2-mediated antitumor effects in vivo.

Nitric oxide (NO) decreases cytotoxicity and proliferation of cytotoxic lymphocytes (CTLs) in vitro. Both can be prevented by inhibitors of the NO synthase (NOS). To elucidate whether inhibition of the IL-2-induced NOS could boost efficacy of IL-2-stimulated CTLs in vivo, we assessed lung metastases in mice injected with IL-2, the NOS inhibitor aminoguanidine (AG), their combination and the diluent. No improvement was observed for IL-2 + AG compared to IL-2 while NO production was normalized. Since NO causes one of the two major side effects of IL-2 treatment, hypotension, we further studied whether capillary leak could be attributed to NO, too. While IL-2-inducible NO was reduced to control levels by AG, pulmonary edema was unaffected. Thus a decrease in NO does not improve antitumor effects of IL-2-stimulated CTLs nor does it attenuate IL-2-associated capillary leak.

Animals↗
PubMed1996

Polymeric surface modifications of tantalum stents.

PURPOSE: To compare two kinds of polymer-coated tantalum stents with bare tantalum stents (control) to determine if the coatings can improve thromboresistance. METHODS: Twenty-seven Fontaine-Dake stents were balloon expanded in three 8-mm x 80-cm polytetrafluoroethylene (PTFE) grafts; 9 stents were bare tantalum (T); 9 were coated with polyetherurethane (PL); and 9 were coated with parylene (PA). There were 9 stents placed in each graft as follows: 3 tantalum, 3 polyetherurethane, and 3 parylene. In swine whose platelets had been radiolabeled with indium 111, the ends of each stented graft were connected to 14F femoral and venous sheaths to create an ex vivo fistula. Each graft was exposed to blood for 30, 60, and 120 minutes. At the end of each test period, the stented grafts were disconnected from the sheaths, flushed with saline until clear, and then flushed with formalin. The stents were removed from the grafts, and a radionuclide well counter recorded radionuclide counts from each stent type at each period of blood contact. These values were converted to platelet density per 1000 microns 2. Stents were then photographed and scanned with electron microscopy (EM) for qualitative analysis. Possible significant differences in platelet adhesion with the three types of stents (both between stent groups and within stent groups) were examined using a two-tailed Student's t-test. RESULTS: There were significantly fewer platelets adsorbed on PA versus T at all time periods (p < 0.005); on PL versus T at 60 and 120 minutes (p < 0.005); and on PA versus PL at 30 and 120 minutes (p < 0.0005). There was no significant difference in platelet density within each stent group (p = 0.1). Mean platelet density (number of platelets per 1000 microns 2 +/- SD) was as follows: at 30 minutes: T = 1891 +/- 965; PL = 373 +/- 193; and PA = 27 +/- 3; at 60 minutes: T = 6226 +/- 1621; PL = 1573 +/- 793; and PA = 1185 +/- 710; at 120 minutes: T = 5307 +/- 591; PL = 3164 +/- 318; and PA = 180 +/- 100. Gross inspection of the 120-minute groups demonstrated focal areas of thrombus on T, less on PL, and none on PA. Scanning EM demonstrated extensive platelet accumulation covering T at all time periods, less on PL, and even less on PA.

Animals↗
PubMed1996

CD34+Thy-1+Lin- stem cells from mobilized peripheral blood.

Over the last ten years there has been increasing use of mobilized peripheral blood (MPB) progenitor cells as grafts for autologous transplantation. Among the cells comprising these MPB autografts is a subpopulation of CD34+Thy-1+Lineage (Lin)- cells, which is enriched for hematopoietic stem cell (HSC) activity. The percentage of CD34+ cells which express Thy-1 is higher in some samples of MPB than in bone marrow (BM). Using myeloid and erythroid cell depletion prior to high speed cell sorting, it is possible to purify sufficient numbers of CD34+Thy-1+Lin-HSCs from a MPB leukapheresis sample for use as an autograft. CD34+Thy-1+Lin-cells will potentially provide a tumor-depleted autograft for cancer patients. This HSC population is also being studied as a potential target for gene transfer for the treatment of patients with HIV, cancer and a variety of genetic disorders.

Animals↗
PubMed1996

Detection of human cells in human/sheep chimeric lambs with in vitro human stroma-forming potential.

In utero transplantation of preimmune fetal sheep with human hematopoietic stem cells results in stable long-term hematopoietic chimerism. To clarify the mechanisms of support of human stem cells in chimeric sheep, we established stromas from bone marrow of 30 sheep transplanted in utero with human hematopoietic stem cells from adult bone marrow adult peripheral blood, or fetal liver. We examined the stromas for the presence or absence of human stromal elements in vitro. Human stromal elements were detected in 12 sheep as assessed by polymerase chain reaction (PCR) using human HLA-DQalpha-specific primers. The human origin of the PCR product was confirmed by Southern blotting using an HLA-DQalpha-specific probe. However, none of these stromal cells were positive for CD45 or CD14 as determined by fluorescence-activated cell sorting (FACS) analysis or by message expression using reverse transcriptase (RT)-PCR. In an attempt to further characterize these cells fibroblasts were isolated by panning, and DNA analysis confirmed the human origin of these cells in the same lambs. Of the fetuses injected with the highly enriched cells from adult human bone marrow 36% were found to harbor cells capable of forming human stromal elements in vitro in their marrow. Of those injected with human fetal liver and peripheral blood stem cells, 42 and 40%, respectively, exhibited in vitro human stromal cell-forming ability. These results indicate the long-term persistence of cells capable of giving rise to components of human marrow stroma in vitro in the human/sheep xenograft model.

Animals↗
PubMed1996

Detection of a primitive megakaryocyte progenitor cell in human fetal bone marrow.

Ontogeny-related changes in megakaryocyte (MK) progenitor cells were a analyzed to further define the cellular hierarchy occurring during human MK development. CD34+ cells were selected from human low-density adult bone marrow (ABM) or unfractionated fetal bone marrow (FBM) and assayed for MK colony formation in a serum-depleted fibrin clot assay system. At days 3, 7, 12, 16, 21, and 28 of incubation, MK colonies were analyzed for colony number, size and number of foci of development. Unifocal CFU-MK-derived colonies cloned from FBM formed after fewer days of in vitro culture and were 2.6-fold larger than those colonies cloned from ABM, However, the frequency of CFU-MK derived colonies cloned from ABM was significantly greater. the MK colonies cloned from FBM morphologically consisted of both pure MK colonies and mixed MK-containing colonies, in which a core of CD41- cells were surrounded by CD41+MKs. Large colonies resembling the primitive burst-forming unit-MK (BFU-MK) also were assayable from both FBM and ABM. These BFU-MK-derived colonies appeared after fewer days of incubation when FBM was assayed compared to ABM, but at a significantly lower frequency. In addition, large unifocal MK colonies consisting of >300 cells (300-1000) appeared from cells cloned from FBM but not ABM. This type of colony, which appears to represent a unique type of MK progenitor cell, was termed a high proliferative potential cell-MK (HPPC-MK). such colonies represent a relatively rare MK progenitor. We conclude that there are a significant ontologic changes in human MK progenitor cell development. The kinetics of MK colony formation from ABM differs significantly from that of FBM. Furthermore, the proliferative capacity of fetal MK progenitor cells appear to be far greater than that of ABM. In addition, a more primitive lineage-restricted MK progenitor cell with extensive proliferative capacity, the HPPC-MK, was assayable exclusively from FBM.

Adult↗
PubMed1996

Fetal bone marrow CD34+CD41+ cells are enriched for multipotent hematopoietic progenitors, but not for pluripotent stem cells.

We have investigated the expression of CD41a (gpIIbIIIa) on a subpopulation of human fetal bone marrow (FBM) CD34+ progenitor cells. Human FBM CD34+Lin- cells were subfractionated into CD41a+ and CD41a- subpopulations by flow cytometry. All the megakaryocyte colony-forming cells (CFU-MK) and almost all the burst-forming units-megakaryocyte (BFU-MK) were found within the CD41a+ subpopulation. In addition, a 14-fold greater number of granulocyte-macrophage colony-forming units (CFU-GM) and a five-fold greater number of mixed lineage progenitor cells (CFU-mix) were observed within the CD34+Lin-CD41a+ subpopulation compared to the CD34+Lin-CD41a- subpopulation. The high proliferative potential of CD34+Lin-CD41a+ cells was demonstrated by their capacity to expand in in vitro culture containing human plasma and recombinant Mpl ligand (thrombopoietin [Tpo]) with production of over 80% CD41b+ (gpIIb+) MKs. However, in long-term bone marrow cultures, the CD34+Lin-CD41a- population contained a significantly higher frequency of cobblestone area-forming cells (CAFC) than the CD34+Lin-CD41a+ population, indicating the presence of a primitive hematopoietic stem cell (HSC) population within the CD34+Lin-CD41a- subset. These data suggest that fetal CD34+Lin-CD41a+ cells are enriched for MK progenitor cells (CFU-MK and BFU-MK), myeloid progenitors, and CFU-mix but do not contain the more primitive CAFC.

Antigens, CD34↗
PubMed1996

Identification of CD34+ subsets after glycoprotease selection: engraftment of CD34+Thy-1+Lin- stem cells in fetal sheep.

Epitopes on the CD34 molecule detected by some CD34 antibodies can be cleaved by a unique glycoprotease from Pasteurella haemolytica, which cleaves only glycoproteins rich in O-linked glycans. A method to isolate CD34+ cells from adult bone marrow was developed subsequently, in which CD34+ cells were isolated in high purity and yield following immunomagnetic bead selection and detachment with the glycoprotease. Using a variety of other cell-surface markers shown here to be insensitive to glycoprotease, committed progenitors of T lymphoid, B lymphoid, monomyeloid, megakaryoblastic, or erythroid lineages could be identified. Significantly, candidate hematopoietic stem cells (HSC) that are contained within a CD34+Lin- (CD2-, CD14-, CD15-, CD16-, CD19-) (or CD34+CD38-) subset expressing the Thy-1 antigen (CDw90), c-kit receptor (CD117), and CDw109 but lacking expression of CD71 and HLA-DR antigens also were detected. Functionally distinct subsets of glycoprotease-selected CD34+ cells were identified and subfractionated using flow cytometry and fluorescence-activated cell sorting (FACS). These subsets included candidate HSCs expressing the CD34+Thy-1+Lin- phenotype, which were sorted from a CD34+ fraction of a mobilized peripheral blood (MPB) sample. In a fetal sheep model, when CD34+Thy-1+Lin- cells were injected intraperitoneally, they were capable of homing to the marrow, where they generated long-term multilineage hematopoiesis and maintained human CD34+ cells, indicating that candidate HSC subsets of CD34+ cells selected with this highly specific enzyme were capable of engraftment in vivo. The ability to identify and purify virtually any phenotypically defined subset of glycoprotease-selected CD34+ stem/progenitor cells should facilitate the study of hematopoiesis in vitro and in animal models in vivo as well as the development of novel genetic techniques for the correction of specific blood cell disorders in humans.

Adult↗
PubMed1996

Recombinant human interleukin-3 (rhIL-3) enhances the mobilization of peripheral blood progenitor cells by recombinant human granulocyte colony-stimulating factor (rhG-CSF) in normal volunteers.

To identify a precisely timed and safe protocol for progenitor cell mobilization, we studied the effects of rhIL-3 and rhG-CSF administration to normal volunteers. rhG-CSF 5 micrograms/kg/d was administered subcutaneously (s.c.) for 7 consecutive days either alone or preceded by rhIL-3 5 micrograms/kg/d s.c. for 4 consecutive days in sequential or partially overlapping schedules. The combined cytokines were well-tolerated--adverse effects were similar to those of the individual agents. Total white blood cell (WBC) and neutrophil counts rose briskly in response to rhG-CSF, and peak mean values were similar between treatment cohorts. Mean platelet counts were modestly elevated during rhG-CSF treatment only in the cohorts receiving rhIL-3 and rhG-CSF. Mean circulating CD34+ cells peaked on day 5 in the rhG-CSF group (38.9+/-14.3/microliter), day 6 in the sequential rhIL-3/rhG-CSF group (56.4+/-12.4/microliter), and day 6 in the partial overlap group (46.1+/-10.9/microliter). On day 3, mean CD34+ cell counts of the subjects who received sequential treatment were markedly higher than observed in the other groups (p<0.05) and were estimated to have been sufficient for collection of adequate grafts by single 10-L leukapheresis procedures in 60% of subjects. Circulating clonogenic cells (CFU-GM and/or BFU-E) were substantially higher in the sequential group than the rhG-CSF group on days 3-6 but were only minimally elevated above baseline in the partial overlap group. The numbers of circulating CD34+/Lin-/Thy-1+ cells (putative stem cells) were increased substantially, especially in the sequential group. On the basis of this pilot trial, we conclude that priming with rhIL-3 is a safe and well-tolerated method for enhancing the mobilization of human blood progenitors and stem cells by rhG-CSF.

Adult↗
PubMed1996

[clinical application of multiplanar and 3D reconstruction of spiral CT in diagnosis of acetabulum fractures].

This review describes recent visualizations of computed tomography for the diagnosis of acetabular fractures. The techniques of conventional and helical-CT for the imaging of the acetabulum are compared. Furthermore, the different methods of multiplaner and three-dimensional reconstructions e.g. shaded surface display, maximum intensity projection, and volume rendering are presented. Figures of each method are demonstrated and the diagnostic potential of multiplanar and three-dimensional imaging for fractures of the pelvis is discussed.

Acetabulum↗