M. Bregni - Academia.edu (original) (raw)

Papers by M. Bregni

Blood, 1991

Optimum methods of harvesting circulating hematopoietic progenitors for autologous transplantatio... more Optimum methods of harvesting circulating hematopoietic progenitors for autologous transplantation to support myeloablative cancer therapy are still uncertain, mostly because of the lack of an assay for marrow- repopulating stem cells. The CFU-GM assay, the commonly used indirect indicator of the quality of the graft, is poorly standardized and provides results evaluable only retrospectively. Based on the knowledge that hematopoietic progenitors express CD34 and CD33 differentiation antigens, we developed a dual-color direct immunofluorescence flow cytometry assay with the aim of replacing the CFU-GM assay advantageously. For this purpose, we applied both assays to 157 blood samples obtained daily throughout 20 different recoveries from pancytopenia induced by high-dose cyclophosphamide or etoposide cancer therapy with or without recombinant human GM colony-stimulating factor (rhGM-CSF). The appearance of CD34+ cells in the circulation indicated that hematopoietic progenitors had in...

Annals of Oncology, 2001

Background: This study evaluates the specificity of some reverse-transcriptase polymerase chain r... more Background: This study evaluates the specificity of some reverse-transcriptase polymerase chain reaction (RT-PCR) assays for the detection of residual tumor cells in breast cancer patients. The following markers have been analysed: carcinoembryonic antigen (CEA), cytokeratins (CK19 and CK20), polymorphic epithelial mucin (MUC-1), epidermal growth factor receptor (EGFR), maspin, and mammaglobin. RT-PCR was employed to detect breast cancer cells in peripheral blood (PB), bone marrow (BM), and stem cell leukoaphereses (PBPC). Patients and methods: We evaluated the specificity of our RT-PCR assays on a panel of breast cancer specimens (n = 30), on PBPC in patients undergoing high-dose chemotherapy (n = 38), on BM (n-7) and PB (n = 5) samples obtained from patients with breast cancer. Marrow cells, PB, and PBPC from normal subjects or hematological tumor patients were tested as negative controls. Results: Only maspin and mammaglobin met the criteria of sensitivity and specificity required for the detection of residual disease; they were expressed in 80% and 97% of breast cancer specimens, respectively, and not expressed in normal controls. CK19, CK20, EGFR, MUC-1, and CEA were sometimes expressed in normal blood cells and/or hematological tumors. Conclusions: Our data support the notion that maspin and mammaglobin are useful markers for RT-PCR detection of minimal residual disease (MRD) in breast cancer patients, and that perspective clinical studies are needed to determine wether RT-PCR assays will be useful in assessing prognosis, tailoring therapy, or developing new strategies fore.v vivo purging.

The International Journal of Cell Cloning, 1992

Motivations for developing a flow cytometry assay Cancer patients treated with intensive chemothe... more Motivations for developing a flow cytometry assay Cancer patients treated with intensive chemotherapy may or may not experience a n expansion of the circulating hematopoietic progenitor cell (CPC) compartment of sufficient magnitude to justify large-scale collection of these cells for subsequent autologous transplantation after further myeloablative cancer therapy. Lack of a clinical assay for realtime determination of the number of CPCs can expose patients with scarce or without overshoot of CPCs to insufficient or useless leukapheresis procedures (1). To overcome this problem, based on our initial observation that CPCs similarly to their marrow counterpart express the CD34 differentiation antigen (CD34+ cells) (2), we have developed a direct immunofluorescence flow cytometry assay to estimate CD34+ cells in the peripheral blood of cancer patients recovering from intensive chemotherapy-induced pancytopenia (3). This flow cytometry assay, replacing

International Journal of Radiation Oncology*Biology*Physics, 1990

Pulmonary function tests (standard vital capacity, WC, total lung capacity, TLC, forced expirator... more Pulmonary function tests (standard vital capacity, WC, total lung capacity, TLC, forced expiratory volume in 1 second-forced vital capacity ratio, FEVl/FVC, carbon monoxide transfer factor, DLCO) were prospectively evaluated in patients (median age 25 years, 13-52 years; median follow-up 20 months, 6-51 months) with Hodgkin's disease (15 patients), non-Hodgkin's lymphoma (9 patients), and inflammatory breast cancer (3 patients) treated with sequential high-dose therapy comprising the following phases over approximately 2 months: a) cyclophosphamide (7 g/m'); b) vincristine (1.4 mg/m*), methotrexate (8 g/m'), and cisplatinum (120 mg/m*) or etoposide (2 g/m*); c) total body irradiation (TBI; 12.5 gy, 5 fractions over 48 hours), intravenous melphalan (120-180 mg/m*), and transplantation of autologous peripheral blood and/or bone marrow hematopoietic stem cells. Within 2 months after transplantation, 12 patients also received 25 Gy radiotherapy boost to mediastinum and clavicular regions. In viva dosimetry evaluations of fractionated TBI treatments showed that mean radiation dose absorbed by lungs was 12.18 Gy (97.4% of TBI dose). Despite such a high radiation dose, we observed only transient and subclinical decrease of SVC, TLC, and DLCO. The decrease of SVC, TLC, and DLCO was more evident and prolonged in patients receiving radiotherapy boost. All parameters progressively recovered to normal values within 2 years after transplantation. In contrast, FEVl/FVC remained within normal limits in all patients, thus demonstrating the absence of obstructive ventilatory changes. In addition, no interstitial pneumonia was observed.

Bone Marrow Transplantation, 1997

Blood Cells, Molecules, and Diseases, 1996

Mobilized peripheral blood hematopoietic progenitor cells obtained from cancer patients treated w... more 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 &amp;quot;in vivo expansion&amp;quot; such that additional ex vivo expansion of primitive progenitor cells may not be possible.

Best Practice & Research Clinical Haematology, 1999

The mobilization of haematopoietic progenitor cells is a multifactorial process, still poorly und... more The mobilization of haematopoietic progenitor cells is a multifactorial process, still poorly understood at the molecular level. Mobilized haematopoietic progenitors, as defined by the expression of CD34 cell surface molecule, comprise heterogeneous subpopulations of cells committed to different haematopoietic lineages. Haematopoietic progenitors may be mobilized by chemotherapy alone, haematopoietic growth factors alone, or by chemotherapy plus haematopoietic growth factors. The choice of a mobilization regimen that allows an optimal yield of progenitors with a minimum number of leukaphereses should incorporate, in most patients, a disease-specific chemotherapeutic agent(s) plus a haematopoietic growth factor, to be continued until completion of harvest.

Acta Dermato Venereologica, 2013

Journal of Clinical Oncology

2525 Background: Genetically-modified-lymphocytes (GML) can induce a specific immune response aga... more 2525 Background: Genetically-modified-lymphocytes (GML) can induce a specific immune response against the transgene product (HSV-TK), when injected into immunocompetent patients (Science 276:1719, 1997). In tumor-bearing mice the infusion of tumor-associated-antigen (TAA)-transduced lymphocytes induced protective immunity and long-term memory. In this experimental setting, the T-cell effectors were generated by a cross-presentation pathway, mediated by host dendritic cells (DCs). Indeed, DCs isolated from lymph nodes of treated mice contained fragments of labeled GML and were able to elicit antigen-specific responses ex vivo. Methods: To validate this novel vaccination strategy in humans, we treated 10 patients affected by stage IIIc/IV MAGE-A3+ melanomas with escalating doses of autologous GML expressing MAGE-A3 and the tracer antigen HSV-TK. HSV-TK allows to monitor the immune status of the patients. Immune responses were monitored by DTH and in vitro T cell assays (semi-quantitat...

Blood

Reduced intensity conditioning (RIC) followed by allogeneic stem cell transplantation (SCT) is as... more Reduced intensity conditioning (RIC) followed by allogeneic stem cell transplantation (SCT) is associated with durable engraftment, low transplant-related mortality (TRM) and clinical remissions in hematologic malignancies. In addition, RIC followed by allograft is feasible also in patients who had failed previous autologous (auto) SCT. Here, we report the outcome of 118 lymphoma pts receiving RIC and allogeneic SCT from HLA-identical sibling donors. Histologic types included in the study were: chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL; n=26), low-grade non-Hodgkin lymphoma (LG-NHL; n=24, 21 follicular NHL), high-grade NHL (HG-NHL, B cell n=28, T cell n=17), Hodgkin disease (HD; n=23). Median age was 50 years (range: 20–69). 47% of pts had refractory disease (46% CLL/SLL; 33% LG-NHL; 44% HG-NHL; 56% HD) and 48% have failed an auto SCT (15% CLL/SLL; 33% LG-NHL; 55% HG-NHL; 82% HD). All pts received debulkying chemotherapy followed by the same RIC regimen with t...

La radiologia medica, 2011

The original version of this article, unfortunately, contained a mistake. The name of the first a... more The original version of this article, unfortunately, contained a mistake. The name of the first author should have been A. Bagliani instead of A. Baggiani Nella versione originale di questo articolo era contenuto un refuso accidentale. Il nome del primo autore avrebbe dovuto essere A. Bagliani anziché A. Baggiani.

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Tumori

Metastatic cancer of unknown primary site is characterized by a poor prognosis. Nevertheless, in ... more Metastatic cancer of unknown primary site is characterized by a poor prognosis. Nevertheless, in the case of suspected breast cancer, patients may have the same prognosis of stage II patient". Technetium-99m-hexakis-2-methoxy-isobuty1-isonitrile (sestamibi) is a lipophilic cation widely employed as a myocardial perfusion imaging agent. Latest diagnostic applications in oncology confirmed [99ffiTc]-sestamibi as an useful tracer for the visualization of a variety of primitive and secondary tumors2,4,7. 10. Nuclear medicine procedures using single photon emission tomography (SPET) are currently being assessed with the aim of improving lesion detection1. These techniques proved to have a higher specificity than planar acquisition. Moreover, new instrumentation has made it possible to acquire and reconstruct SPET images within a short time, making it a useful tool in clinical practice.

Bone marrow transplantation, 1994

With the aim of facilitating the ex vivo manipulation of peripheral blood hematopoietic progenito... more With the aim of facilitating the ex vivo manipulation of peripheral blood hematopoietic progenitors (CPCs = circulating progenitor cells) collected by leukapheresis, we removed polymorphonuclear cells and monocytes that naturally adhere to nylon wool fibers. Leukapheresed cells harvested at the time of hematopoietic recovery after cancer therapy with high-dose cyclophosphamide plus hematopoietic growth factors were incubated with nylon wool fibers for 1 h at 37 degrees C. Evaluation of the cells non-adherent to the nylon wool in all experiments (n = 14) showed that the median recovery of nucleated cells and CPCs detected as CD34+ cells, CFU-GM and BFU-E was 16.4% (range 4.8%-34.0%), 60.0% (range 30.8-80.8%), 60.9% (range 33.4-74.5%) and 65.5% (range 30.8-69.2%), respectively. Therefore exposure to the nylon wool determined a selective removal of mature cells and a complementary enrichment of CPCs. The wide range of results depended on the significantly different cell compositions of...

Pathologie-biologie, 1992

Cancer research, Jan 15, 1997

Peptide presentation by autologous dendritic cells (DCs) is a new tool to activate tumor antigen-... more Peptide presentation by autologous dendritic cells (DCs) is a new tool to activate tumor antigen-specific T cells in melanoma patients. However, it is not known whether autologous DCs, differentiated by two of the most efficient protocols (from CD34+ progenitors or from monocytes), are equally effective as professional antigen-presenting cells (APCs) when the patients have a low frequency of peptide-specific precursors. To this end, a limiting dilution assay was applied to evaluate the frequency of antigen-specific CTL precursors (CTLps) in peripheral blood of HLA-A*0201+ melanoma patients. Then, from two melanoma patients showing low frequency of CTLps to melanoma antigen-A/melanoma antigen recognized by T cell (Melan-A/Mart-1)(27-35) peptide, autologous DCs were differentiated from granulocyte colony-stimulating factor-mobilized CD34+ progenitors or from monocytes. CD34+- and monocyte-derived DCs were characterized by a similar proportion of CD1a+ cells expressing HLA class II ant...

Bone marrow transplantation, 1996

Contamination of autologous blood cell transplants with cells of follicular non-Hodgkin's lym... more Contamination of autologous blood cell transplants with cells of follicular non-Hodgkin's lymphoma (F-NHL) may contribute to relapse of the malignancy after potentially curative high doses of chemotherapy and radiotherapy. In an attempt to circumvent this limitation, we have evaluated various techniques of selection of CD34+ cells to eliminate malignant cells from blood cell transplants of five patients with F-NHL undergoing high-dose sequential therapy. The contamination of F-NHL cells was evaluated using a nested PCR assay for the detection of bcl-2-IgH rearrangement with a sensitivity of one F-NHL cell in 10(5) normal cells. In two experiments with blood cell transplant fractions of 0.5 x 10(9) nucleated cells, negative selection of CD34+ cells by removal of B cells and other mature cells that naturally adhere to nylon wool fibers decreased the number of CD19+ B cells detectable by flow cytometry but failed to eliminate bcl-2-IgH-positive F-NHL cells detectable by PCR. In con...

Haematologica

H uman gene therapy has progressed from theoretical to practical reality in a short time. Because... more H uman gene therapy has progressed from theoretical to practical reality in a short time. Because transduction of specific target cells is the critical step in gene therapy, improvement of gene transfer methods has been the principal aim of research. Retroviral-mediated gene transfer, which was developed the early 1980's in animal models, is the procedure most used today. Vectors need to be engineered that will target specific cell types, insert their genetic information into a safe site on the genome, and be regulated by normal physiological signals. When efficient vectors of this type are produced (retroviral, viral, synthetic, or a combination of the three), gene therapy will probably have a profound impact on medical practice. The first approved gene transfer (only a marker gene) protocol was begun in May 1989 and the first clinical gene therapy trial, which was for the correction of adenosine deaminase (ADA) deficiency, was initiated in September 1990. Many had assumed that hematopoietic stem cells would be the first target of gene therapy because they persist long term and involve numerous diseases of all lineages. The European Concerted Action (ECA) workshop on Gene Transfer into Hemopoietic Cells took place in Marseille on October 17 th and 18 th , 1995.

European journal of histochemistry : EJH, 1994

Journal of clinical oncology : official journal of the American Society of Clinical Oncology, 1994

To assess the toxicity, efficacy, and applicability of high-dose therapy with bone marrow and/or ... more To assess the toxicity, efficacy, and applicability of high-dose therapy with bone marrow and/or peripheral-blood autotransplantation in high-risk, previously untreated patients with multiple myeloma. Thirteen consecutive patients with high-labeling index (LI) multiple myeloma received a novel high-dose sequential (HDS) regimen consisting in the high-dose administration of cyclophosphamide (7 g/m2) followed by vincristine (1.4 mg/m2) plus methotrexate (8 g/m2 with leucovorin rescue), etoposide (2 g/m2) and, finally, total-body irradiation (TBI; 10 Gy) plus melphalan (120 mg/m2) with autografting of peripheral-blood hematopoietic progenitor cells. Recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF; 5 micrograms/kg/d) was continuously infused after cyclophosphamide and etoposide both to accelerate hematopoietic recovery and to expand/mobilize the hematopoietic progenitor-cell pool. Among 13 patients, 12 completed the program; 10 (or 77%) achieved a complete r...