Mobilization of peripheral blood stem cells in advanced ovarian cancer

High-dose chemotherapy with hematopoietic support has been expected to improve the survival of advanced ovarian cancer patients in recent years. An essential component of such treatment has been the ability to collect and reinfuse a large number of peripheral blood stem cells (PBSCs) following high dose therapy. This study was designed to determine which clinical and hematological factors would be better indicators to collect the proper volume of PBSCs. Thirteen patients received a total of 24 courses of induction chemotherapy and 69 of apheresis. We usually mobilized stem cells using CEP chemotherapy (cisplatin 50-70 mg/m2, epirubicin 50 mg/m2 and cyclophosphamide 1.5 g/m2) with G-CSF and CEE regimen (cyclophosphamide 2.0 g/m2, epirubicin 50 mg/m2, and etoposide 50 mg/m2) as a salvage for mobilization. We obtained an average 5 x 10(6)/kg of CD34+ cells for 3 days as one course. The number of CD34+ cells collected significantly depended on the platelets and reticulocytes on the first day of apheresis, but not a nadir of WBCs. It is concluded that apheresis should be started on recovery of WBCs to 5,000-10,000/microliters, of immature granulocytes to > or = 10% and of reticulocytes to > or = 20%. This study confirmed the feasibility of collecting enough PBSCs to use standard chemotherapy of ovarian cancer patients.     

Malignancy-associated remission of systemic lupus erythematosus maintained by autologous peripheral blood stem cell transplantation

Many investigators worldwide are currently exploring the role of peripheral blood stem cell transplantation (PBSCT) in managing autoimmune diseases. We report the case of a woman with systemic lupus erythematosus (SLE) with mucocutaneous and renal involvement, who underwent PBSCT for stage IVB Hodgkin's disease. Following the development of the lymphoma, she has had a prolonged clinical and serologic remission of the SLE. The potential effects of lymphoproliferative disorders and PBSCT on the course of SLE are considered.     

Mobilization of hematopoietic stem cells (CFU-C) into the peripheral blood of man by endotoxin

Pseudomonas endotoxin was administered to three normal subjects in order to assess possible effects on stem cell circulation. Increased numbers of granulopoietic stem cells (CFU-C) transiently appeared in the peripheral blood, reaching a maximum after the time of maximal leukopenia. There was a mean 4-fold increase in CFU-C per ml of blood following endotoxin. A high proportion of these CFU-C did not require an added source of colony-stimulating activity for growth in agar.     

Successful engraftment of haploidentical stem cell transplant for familial haemophagocytic lymphohistiocytosis using both bone marrow and peripheral blood stem cells

Familial haemophagocytic lymphohistiocytosis (HLH) is a disease with a very poor prognosis unless patients receive a bone marrow transplant. It is often difficult to find an HLA-matched donor and haploidentical familial donors may be considered. The main complication of this type of transplant is graft rejection. We describe a patient with familial HLH who received a haploidentical transplant using both mobilized peripheral blood and bone marrow stem cells in an attempt to overcome graft rejection by increasing the stem cell dose. The peripheral blood stem cell inoculum was CD34 enriched using a Cellpro column and T-cell depleted by Campath-1M, the patient received conditioning for a matched sibling donor transplant with the addition of Campath 1G. There was rapid and full engraftment and the patient remains disease free at 5 months. This technique may be applicable for other fatal inborn errors in the absence of an HLA-matched donor.     

Autologous peripheral blood stem cell transplantation and adoptive immunotherapy with activated natural killer cells in the immediate posttransplant period

Relapse after high-dose chemotherapy supported by peripheral blood stem cell transplantation (HDC-PBSCT) is the main cause of therapeutic failure in patients with lymphoma and breast cancer. Adoptive immunotherapy with activated natural killer (A-NK) cells and interleukin 2 might eliminate surviving residual tumor without adding to toxicity. Eleven patients with relapsed lymphoma and one with metastatic breast cancer were entered on a pilot clinical trial of HDC-PBSCT followed on day 2 after transplant by infusion of cultured autologous A-NK cells. Simultaneously, recombinant human interleukin 2 (rhIL-2) was initiated as a 4-day continuous i.v. infusion at 2 x 10(6) IU/m2/day, referred to as high-dose rhIL-2. Therapy with high-dose rhIL-2 was followed by a 90-day continuous i. v. infusion at 3 x 10(5) IU/m2/day, referred to as low-dose rhIL-2. All patients engrafted and nine completed treatment. Posttransplant days to a neutrophil count of 500/microliter and to a platelet count of 50,000/microliter were similar to comparable patients treated with HDC-PBSCT alone. Generation of A-NK cells for therapy was feasible in all patients except the three patients with Hodgkin's disease, whose cells did not proliferate in culture. Overall toxicity associated with early posttransplant transfer of A-NK cells and interleukin 2 did not differ from that observed with peripheral blood stem cell transplantation alone in comparable patients. There was early amplification of natural killer cell activity in the peripheral blood of four patients that appeared to result from the transfused A-NK cells. Adoptive transfer of A-NK cells and rhIL-2 during the pancytopenic phase after HDC-PBSCT was feasible and well tolerated, did not adversely affect engraftment, and resulted in amplified natural killer activity in the peripheral blood during the immediate posttransplantation period.     

A significant percentage of normal T lymphocytes express HLA-DP in the peripheral blood

HLA-DP expression has been widely investigated on T lymphocytes activated under different conditions. In the present study, a double staining procedure was used in flow cytometric experiments to define DP expression on normal peripheral blood T lymphocytes. In about two-thirds of the case analyzed, DP was expressed on a higher percentage of normal peripheral T lymphocytes than DR was. This was particularly true for 1 of the 16 cases investigated in which the percentage of T lymphocytes expressing DP was 46% and in which DP expression was mainly the prerogative of CD8+ and CD56+ lymphocytes.     

Collection of allogeneic peripheral blood progenitor cells by two protocols on an apheresis system

BACKGROUND: Granulocyte-colony stimulating factor-mobilized allogeneic peripheral blood progenitor cells (PBPCs) are replacing bone marrow in transplantation for the treatment of several hematologic malignancies. The advantages of PBPCs are offset by the donor-associated disadvantages of granulocyte-colony stimulating factor side effects and the risk of apheresis-like platelet loss. STUDY DESIGN AND METHODS: For each individual, the first donation of allogeneic PBPCs by apheresis on the Spectra, using either the standard protocol Version 4.7 (45 donors, [Version 4.7]) or the AutoPBSC (60 donors, [AutoPBSC]) was compared. Between July 1995 and May 1996, all donors enrolled underwent Version 4.7 apheresis. Since May 1996, the majority of donors underwent AutoPBSC apheresis. For statistical analysis, only data from the first apheresis for each individual donor was considered for independent values. RESULTS: These results indicate a similar collection efficiency for CD34+ cells in the first apheresis of each donor (54% Version 4.7 vs. 53% AutoPBSC, p = 0.8). The apheresis time was longer with the AutoPBSC (233 min vs. 251 min, p = 0.005), whereas the loss of platelets was significantly lower (p < 0.001) with the AutoPBSC (28% vs. 19%). The mean number of CD34+ cells collected in the first apheresis component was 4.0 x 10(8) (Version 4.7) versus 3.8 x 10(8) (AutoPBSC). CONCLUSION: Both apheresis protocols collect sufficient numbers of PBSCs for allogeneic transplantation. The AutoPBSC operates in a fully automatic fashion, avoiding manual adjustment and interindividual variations. The loss of platelets is lower with AutoPBSC than with Version 4.7, but the apheresis time is slightly longer.     

The role of blood stem cells in hematopoietic cell renewal

It has been the purpose of this keynote address to review available evidence for the notion that the stem and progenitor cells circulating in the peripheral blood play a decisive role in the homeostasis of blood cell formation distributed throughout dozens of bone marrow units in the skeleton. Furthermore, if this notion is correct, one could speculate that the quantity and quality of stem and progenitor cells in the blood should reflect the functional state of the hematopoietic stem cell system throughout the skeletal bone marrow and provide a new tool for the evaluation of alteration in blood cell production. On this basis, the following questions are considered: A) What do we know about the quality and quantity of blood stem cells in steady-state conditions? B) In what way do blood stem cells respond to perturbations of the "steady-state" of blood cell formation? C) Which role do blood stem cells play during hemopoietic development assuming that the establishment of bone marrow hemopoiesis requires the "seeding" of blood stem cells into an appropriate cellular environment? D) What is the role of blood stem cells in hemopoietic regeneration after partial body irradiation with a small volume of marrow (and hence stem cells) protected? and E) What are the mechanisms and/or kinetics of hemopoietic recovery if stem cells introduced into the circulation were collected from exogenous (autologous or allogeneic) sources? In this review presentation, experimental work of our group and of other members of the scientific community is summarized. It becomes obvious that blood stem and progenitor cells play a key role in hematopoietic homeostasis. Furthermore, their physiology and pathophysiology deserve rigorous experimental studies in order to develop a novel tool in the diagnosis and prognosis of neoplastic and non-neoplastic disorders of blood cell formation.     

Dendritic cell-based vaccines in the setting of peripheral blood stem cell transplantation: CD34+ cell-depleted mobilized peripheral blood can serve as a source of potent dendritic cells

We are investigating the use of tumor-pulsed dendritic cell (DC)-based vaccines in the treatment of patients with advanced cancer. In the current study, we evaluated the feasibility of obtaining both CD34+ hematopoietic stem/ progenitor cells (HSCs) and functional DCs from the same leukapheresis collection in adequate numbers for both peripheral blood stem cell transplantation (PBSCT) and immunization purposes, respectively. Leukapheresis collections of mobilized peripheral blood mononuclear cells (PBMCs) were obtained from normal donors receiving granulocyte colony-stimulating factor (G-CSF) (for allogeneic PBSCT) and from intermediate grade non-Hodgkin's lymphoma or multiple myeloma patients receiving cyclophosphamide plus G-CSF (for autologous PBSCT). High enrichment of CD34+ HSCs was obtained using an immunomagnetic bead cell separation device. After separation, the negative fraction of mobilized PBMCs from normal donors and cancer patients contained undetectable levels of CD34+ HSCs by flow cytometry. This fraction of cells was then subjected to plastic adherence, and the adherent cells were cultured for 7 days in GM-CSF (100 ng/ml) and interleukin 4 (50 ng/ml) followed by an additional 7 days in GM-CSF, interleukin 4, and tumor necrosis factor alpha (10 ng/ml) to generate DCs. Harvested DCs represented yields of 4.1+/-1.4 and 5.8+/-5.4% of the initial cells plated from the CD34+ cell-depleted mobilized PBMCs of normal donors and cancer patients, respectively, and displayed a high level expression of CD80, CD86, HLA-DR, and CD11c but not CD14. This phenotypic profile was similar to that of DCs derived from non-CD34+ cell-depleted mobilized PBMCs. DCs generated from CD34+ cell-depleted mobilized PBMCs elicited potent antitetanus as well as primary allogeneic T-cell proliferative responses in vitro, which were equivalent to DCs derived from non-CD34+ cell-depleted mobilized PBMCs. Collectively, these results demonstrate the feasibility of obtaining both DCs and CD34+ HSCs from the same leukapheresis collection from G-CSF-primed normal donors and cancer patients in sufficient numbers for the purpose of combined PBSCT and immunization strategies.     

Factors affecting the efficiency of collection of CD34-positive peripheral blood cells by a blood cell separator

BACKGROUND: The yield of CD34-positive cells obtained from an apheresis procedure is determined, in part, by the efficiency of collection. Optimization of the efficiency of CD34-positive peripheral blood cell collection requires identification of predictive factors. STUDY DESIGN AND METHODS: Demographic, stem cell collection, mobilization, and disease-related measures from autologous and allogeneic donors undergoing 252 progenitor cell apheresis procedures were retrospectively reviewed. Statistical relationships between CD34 collection efficiency and the various measures were determined by correlation and multiple linear regression analysis. RESULTS: CD34 collection efficiency inversely correlated with the peripheral white cell count, hematocrit, and serum albumin concentration (R2 = 0.29). White cell count was the single best predictor of CD34 efficiency (R2 = 0.19). Donor groups with cytopenias (patients vs. normal donors; increased cycles of prior chemotherapy; bone marrow involvement; chemotherapy plus growth factor mobilization) had higher collection efficiencies. Only 29 percent of the variability in the data could be attributed to white cell count, hematocrit, and albumin concentration. The majority of the remaining variability was due to unexplained differences between donors. CONCLUSION: CD34 collection efficiencies show considerable variation. Higher peripheral white cell counts, hematocrits, and/or albumin concentrations result in decreased CD34 collection efficiency, but most of the variability in the data is not accounted for by these three factors.     

Second attempts at mobilization of peripheral blood stem cells in patients with initial low CD34+ cell yields

The abuse of alcohol causes health and social problems, such as sickness, death, injury, pain, suffering and crime. These harms impose an economic burden on society. Resources are used or foregone as a consequence of alcohol abuse. This article provides an estimate of the economic cost of alcohol abuse in Ontario in 1992. It uses the cost-of-illness method, in particular, the human-capital approach to estimate the prevalence-based economic costs of alcohol abuse. This methodology is consistent with international guidelines formulated at the 1994 International Symposium on Economic and Social Costs of Substance Abuse. The direct and indirect economic costs of alcohol abuse are identified and estimated. The total economic cost of alcohol abuse, from a societal perspective, is estimated to be US$2261.10 million in Ontario in 1992.     

碳硫分析仪长期稳定性测量方法研究

碳硫分析仪一般使用标准样品绘制校准曲线后直接测定未知样品中碳、硫元素含量,通过标准样品或控制样品的实时核查实现对仪器及校准曲线的长时间监控使用,因此,测量出仪器的长期稳定性时间上限(T_MAX)十分必要。长期稳定性测量即是监控测量结果的准确度,包括精密度和正确度。针对本实验室内的碳硫分析仪设计长期稳定性试验,利用相关标准中的重复性及实验室内再现性精密度数据以及χ²统计量,对测量数据进行各时段内精密度、各时段内正确度、时段内重复性、时段间总精密度以及总均值正确度的检验,给出了碳硫分析仪的长期稳定性时间上限7h。在此时间内,仪器无需任何校正,节省了时间和成本。     

Total parenteral nutrition on energy metabolism in children undergoing autologous peripheral blood stem cell transplantation

The resting energy expenditure (REE) and the respiratory quotient (RQ) were measured longitudinally using indirect calorimetry to examine the effects of total parenteral nutrition (TPN) on energy metabolism in children undergoing autologous peripheral blood stem cell transplantation (PBSCT). There were six children (two males and four females) and the age ranged from five to 13 years (median, eight yrs). The diagnosis included acute lymphocytic leukemia (ALL; 4), neuroblastoma (NBL; 1) and primitive neuroectodermal tumor (PNET; 1). TPN was started after the patients were stabilized following PBSCT (group A; n = 3) or before the initiation of high-dose cytoreductive chemotherapy (HCC) (group B; n = 3). Duration of HCC before PBSCT was identical between the two groups (six to eight days). Average total calorie and protein intake during HCC was significantly higher for group B than for group A. The %REE, the percentage of REE to the predicted basal energy expenditure (BEE), in group A showed 133 +/- 19%, 129 +/- 14% and 146 +/- 11% during three periods of HCC (days -8 to -1 of PBSCT), bone marrow suppression (days 0 to 11 of PBSCT) and bone marrow recovery (days 12 to 22 of PBSCT), respectively. In contrast, those in group B were 10% to 20% lower than those in group A at all periods. Carbohydrate oxidation rates during HCC in group A were significantly lower than those in group B, and those were not different between both groups during post-PBSCT periods. Fat oxidation rates in both groups were similar at all stages of periods. In contrast, protein degradation rates in group A were significantly higher than those in group B at all stages of the period. From these results, we concluded that commencement of TPN administration prior to HCC in the patients undergoing PBSCT provides beneficial effects to maintain better energy metabolic and nutritional status.     

Luminol-enhanced, whole blood chemiluminescence of human neutrophils evaluated by means of an automated, computer-assisted, and high-sensitivity luminescence analyzer

To evaluate the usefulness of two standardized commercially available amplification assays for the detection of Mycobacterium tuberculosis: Amplicor test (Roche) and MTD-Amplified direct test (Gen-Probe) a total of 281 respiratory specimens from 198 patients with symptoms of pulmonary diseases were examined and compared with conventional methods. Fifty-seven specimens were positive and 218 negative by both amplification assays. Three specimens were reactive by Amplicor only, and three by MTD only. In comparison with culture, the sensitivity, specificity, positive predictive value and negative predictive value were 96.0, 94.8, 80.0, and 99.1%, respectively, for the Amplicor test; the corresponding values were 94.0, 94.4, 78.3, and 98.6%, respectively, for the MTD. However, when 28 specimens from 14 patients on antituberculous therapy were excluded the improvement in PPV and specificity of both assays was obtained. In conclusion, both commercially available amplification tests are almost equally sensitive and specific and are suitable for the implementation in daily routine work in the specialized clinical laboratories.     

High-dose chemotherapy with autologous peripheral blood stem cell transfusion in the treatment of advanced testis cancer

Four patients with advanced testis cancer were treated by high-dose chemotherapy supporting by autologous peripheral blood stem cell transplantation. High-dose chemotherapy (carboplatin 250 mg/m2 or nedaplatin 200 mg/m2, etoposide 1,500 mg/m2, ifosphamide 7.5 g/m2 was given and peripheral blood stem cell transfusion was performed 72 hours after the last dose of chemotherapy. High-dose chemotherapy. was given followed by 1 or 2 cycles of pre high-dose therapy consisting of cisplatin 100 mg/m2 or carboplatin 500 mg/m2, etoposide 450 mg/m2, ifosphamide 6 g/m2. All 4 patients were evaluable. Three patients obtained a complete response and one showed a partial response. The partial responder was given RPLND. The RPLND specimen showed necrotic tissue.     

High-dose etoposide enables the collection of peripheral blood stem cells in patients who failed cyclophosphamide-induced mobilization

A case of an intracranial cavernous angioma, which presented with headaches and seizures in a pregnant patient, is described. Diagnosis was established with magnetic resonance imaging. A computer-assisted literature search uncovered no previously reported case of intracranial cavernous angioma initially presenting during pregnancy.     

Two cases of adenosquamous cell carcinoma of advanced cervical cancer treated by carboplatin-based chemotherapy with peripheral blood stem cell autotransplant

In two cases with adenosquamous cell carcinoma of advanced cervical cancer, carboplatin-based chemotherapy was given intraarterially from the internal iliac artery as neoadjuvant chemotherapy, and peripheral blood stem cells (PBSCs) were harvested. After the operation, conventional intravenous chemotherapy with PBSC autotransplant was performed. PBSCs were mobilized by neoadjuvant chemotherapy and G-CSF administration. By the apheresis procedures, 0.7-2.6 x 10(6)/kg CD34 positive cells were obtained. They had no severe side effects from intravenous chemotherapy with PBSCT, and they were free of disease 20 months. Neoadjuvant chemotherapy and G-CSF administration may be capable of mobilization of PBSCs, and chemotherapy with PBSCT may be useful in radioresistant advanced adenosquamous carcinoma of the cervical cancer.     

High-dose therapy with peripheral blood stem cell transplantation results in a significant reduction of the haemopoietic progenitor cell compartment

In order to study the effect of high-dose therapy with peripheral blood stem cell transplantation (PBSCT) on the haemopoietic reserve in man, the number and composition of bone marrow (BM) and peripheral blood (PB)-derived progenitor cells were examined in 137 cancer patients. In 45 patients, paired samples from BM and PB were obtained before PBSC mobilization and 6-27 months after transplantation. Following PBSCT. the proportion of CD34+ cells was significantly smaller than before mobilization (BM 1.99 +/- 0.24 versus 0.8 +/- 0.09, P < 0.001), and no change was observed at several follow-up visits thereafter. The reduction was most pronounced for the primitive BM progenitor subsets such as the CD34+/DR- and CD34+/ Thy-1+ cells. The impairment of hematopoiesis was also reflected by a significant reduction in the plating efficiency of BM and PB samples. No relationship was found between the decrease in the proportion of CD34+ cells and any particular patient characteristics, kind of high-dose therapy or the CD34+ cell content in the autograft. In conclusion, high-dose therapy with PBSC transplantation is associated with a long-term impairment of the haemopoietic system. The reduction in the number of haemopoietic progenitor cells is not associated with a functional deficit, as peripheral blood counts post-transplantation were normal in the majority of patients.