Search PubMed⌕ Search

Biomedical subjects

S Rutella

Publications and source records attributed to S Rutella.

At least 55 records · Page 3Linked to original sources
PubMed1999

Expression of thrombospondin receptor (CD36) in B-cell chronic lymphocytic leukemia as an indicator of tumor cell dissemination.

BACKGROUND AND OBJECTIVE: The expression of CD36 antigen has not been conclusively associated with human B-lymphocytes although CD36 was recently detected in a human B-cell angiotropic lymphoma where it might be involved in lymphoblast-endothelial cell adhesion. We investigated the expression of CD36 in B-cell chronic lymphocytic leukemia (CLL) by multiparameter flow cytometry; results were correlated with clinical features. DESIGN AND METHODS: CD36 expression was evaluated on peripheral blood and bone marrow samples from 24 patients affected by CD5+ B-CLL. Mononuclear cells were isolated by Ficoll-Hypaque density gradient centrifugation, were labeled with fluorochrome-conjugated monoclonal antibodies under standard experimental conditions and were analyzed by flow cytometry. CD36 expression was quantified both in terms of frequency of CD19+CD36+ cells and of mean fluorescence intensity (MFI-R) of CD36+ cell populations. The intensity of CD36 expression was arbitrarily classified as weak (MFI-R ranging from 3 to 6; score 0), moderate (MFI-R ranging from 6 to 9; score 1), intermediate (MFI-R ranging from 9 to 11; score 2) or strong (MFI-R ranging from 11 to 17; score 3). RESULTS: CD36 could be detected on 3% (range 2-5) of normal CD19+ B-lymphocytes and on 45% (range 30-75) of neoplastic CD19+ B-cells. When CLL patients were stratified according to CD36 staining intensity, higher hemoglobin levels (Hb) were recorded in patients assigned to score 0 (Hb = 14.3 g/dL; range 13.9-15.1) compared to patients scoring 1-2 (Hb = 11.2; range 10.3-12.2) or 3 (Hb = 9.8; range 9.6-11.6; p=0.0053). Similarly, higher platelet counts (Plt) were found in patients scoring 0 (Plt = 282x10(3)/microL; range 244-319), compared to patients with intermediate (Plt = 175x10(3)/microL; range 144-238) and high scores (Plt = 149x10(3)/microL; range 103-230; p=0.044); lymphocyte count (Ly) was significantly higher in patients assigned to score 3-4 (Ly = 23.3x10(3)/microL, range 13-30) compared to score 0-2 (Ly = 9.8x10(3)/microL, range 8.5-10.8; p=0.045). CLL patients expressing CD36 at intermediate-to-strong intensity (MFI-R = 14, range 9-16) were more frequently assigned to Rai stages III-IV than stages I-II (CD36 MFI-R = 9, range 6.5-11; p=0.005) and stage 0 (CD36 MFI-R = 6, range 4-7.3; p<0.001). Interestingly, bone marrow diffuse histology was strongly associated with higher CD36 expression (MFI-R = 8.7; range 4.7-13.9) compared to non-diffuse patterns of bone marrow infiltration (MFI-R = 6.7; range 5.2-9.3; p=0.0019). In multivariate regression analysis, CD36 staining intensity significantly and independently correlated with diffuse BM histology (p=0.033). INTERPRETATION AND CONCLUSIONS: The present report provides the first evidence of CD36 expression on CD19+ B-cells from CLL; the correlations with clinical parameters strongly support the view that CD36 might favor tumor cell spreading. Whether high CD36 expression levels on CLL CD19+ B-cells identify an aggressive disease subset remains to be further confirmed in larger series of patients.

Aged↗
PubMed1999

Atypical Sweet's syndrome in a neutropenic patient with acute myeloid leukemia, secondary to a RAEB-T, simulating thrombophlebitis.

We report a rare case of a patient with acute myeloid leukemia following refractory anemia with excess of blasts transformed (RAEB-T) who presented a clinical picture suggestive of thrombophlebitis. The ultrasonographic procedure and the response to corticosteroid treatment suggest that this condition was compatible with an atypical Sweet's syndrome.

Acute Disease↗
PubMed1999

Long-term immune recovery after CD34+ immunoselected and unselected peripheral blood progenitor cell transplantation: a case-control study.

BACKGROUND AND OBJECTIVE: CD34+ stem cell selection induces extensive T-cell depletion as a consequence of ex vivo manipulation. The impact of T-cell depletion on long-term immunologic recovery after autologous CD34+ peripheral blood progenitor cell transplantation (CD34+ PBPCT) is not well characterized. We compared the long term immunologic recovery in two groups of patients submitted to CD34+ PBPCT or unselected autologous peripheral blood progenitor cell transplantation (uPBPCT). DESIGN AND METHODS: Eight patients in both groups were closely matched for diagnosis, age, disease status at transplantation and conditioning regimen and lymphocyte phenotype was prospectively evaluated during long-term post-transplantation follow-up. RESULTS: At a median of 18 months after transplantation, CD3+ lymphocyte subset remained below the normal range in both groups. CD19+ B lymphocytes subset after CD34+ PBPCT was within the normal range in both groups. CD4+ lymphocytes were depressed while the CD8+ lymphocyte subset was increased in group A and in the normal range in group B. As a result, inversion of CD4/CD8 ratio was documented in both groups. T-activated lymphocytes (CD3DR+) and natural killer (CD16/56+) cells were increased in both groups. INTERPRETATION AND CONCLUSIONS: Long-term immune recovery appears to be unaffected by extensive ex vivo manipulation in this adult population when compared to recovery after unmanipulated PBPCT. CD34+ selection, although causes an extensive depletion of T lymphocytes in the graft does not represent a risk factor for delayed CD4+ recovery late after transplantation. Elevated numbers of NK cells and activated T-cells, which have antineoplastic activity, are maintained late after autologous CD34+ transplantation.

Adult↗
PubMed1998

Flow cytometric detection of perforin in normal human lymphocyte subpopulations defined by expression of activation/differentiation antigens.

We investigated with three-color flow cytometry the expression of perforin (Pf) in normal human lymphocyte subpopulations identified by means of activation and differentiation-related antigens. Interestingly, Pf could be detected in a substantial subset (13 +/- 2%) of memory CD4+ CD45RO+ cells, on relevant percentages of memory (CD45RO+) and naive (CD45RA+) CD8+ cells and on virtually all CD3- CD16+, CD3- CD56+ and NKB1+ natural killer cells, as expected. The analysis of fluorescence intensity showed higher levels of Pf expression on CD8dim and NK cells compared to CD8bright and CD4+ lymphocytes, Pf and CD69, HLA-DR, CD95 and CD25 activation/differentiation-related antigens were never co-expressed. On average, 15 +/- 3% of CD3+ CD28+ cells were found to be Pf+, in line with a previously activated or memory cell type. Comparable percentages of CD8+ CD11b- (cytotoxic) and CD8+ CD11b+ (suppressor) T cells were Pf+. Multiparameter flow cytometry is a powerful tool to detect minute fractions of Pf-expressing cells in heterogeneous populations.

Biomarkers↗
PubMed1998

Functional, phenotypic and molecular characterization of cytokine low-responding circulating CD34+ haemopoietic progenitors.

Circulating CD34+ cell populations characterized by a low rate (up to five) or high rate (more than five) of cell divisions were isolated from 8 d cultures in the presence of stem cell factor (SCF), interleukin-3 (IL-3), granulocyte-macrophage colony stimulating factor (GM-CSF), granulocyte colony stimulating factor (G-CSF), erythropoietin (EPO), Flt3 ligand and Peg-rHu megakaryocyte growth and development factor (Peg-rHuMGDF) using the fluorescent dye 5,6-carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) and flow cytometric cell sorting. Phenotypic characterization of cells which had experienced up to five divisions (CFDA-SEbright) showed a similar surface antigen expression to starting, freshly isolated CD34+ cells. Conversely, cells which experienced more than five divisions (CFDA-SEdim) showed a differentiating behaviour, down-regulating CD34 antigen and acquiring differentiation markers. CFDA-SEbright cells were significantly enriched in CD105 (endoglin) positive precursors as compared to both freshly isolated CD34+ and CFDA-SEdim cells. Functional analysis indicated that CFDA-SEbright had a 3-fold and 10-fold greater cumulative cloning efficiency as compared to freshly isolated CD34+ cells and CFDA-SEdim cells, respectively. CFDA-SEbright cells retained the vast majority of LTC-IC and showed a LTC-IC frequency 2.8-fold higher than that found in freshly isolated CD34+ cells. RT-PCR and Western blot analyses showed significantly higher bcl-2 RNA and protein levels in CFDA-SEbright cells as compared to freshly isolated CD34+ and CFDA-SEdim cells. This study indicates that cytokine low-responding circulating CD34+ cells (CFDA-SEbright cells) represent a functionally, phenotypically and molecularly distinct multipotent progenitor population with biological properties associated with primitive precursors.

Antigens, CD↗
PubMed1998

Immunophenotypic profile of peripheral blood eosinophils in acute graft-vs.-host disease.

We used a flow cytometry technique, the "FOG" method (formaldehyde and octylglucopyranoside), to investigate the expression of activation antigens, i.e., CD4, CD23, CD25, HLA-DR, and the EG2 epitope of eosinophilic cationic protein, on peripheral blood eosinophils (PBEs) in leukemic patients who had developed acute graft-vs.-host disease (aGVHD) with eosinophilia after allogeneic bone marrow transplantation (alloBMT) or leukocyte buffy coat infusion. A comparative analysis was performed in transplanted patients not affected by aGVHD and in other conditions commonly associated with peripheral eosinophilia, i.e., interleukin (IL)-2 immunotherapy and allergy. CD25, recognizing the p55 subunit of IL-2 receptor, was detected in all patients with aGVHD except two who, at the onset of eosinophilia, were already receiving methylprednisolone intravenously. The specificity of our findings is confirmed by the absence of reactivity with anti-CD25 mAb in PBEs from transplanted patients not affected by aGVHD. Interestingly, the expression of CD25 progressively declined after steroid therapy. CD25 was also expressed after rhIL-2 administration, probably reflecting analogous mechanisms of eosinophil activation. No aGVHD or rhIL-2-treated patient showed reactivity with anti-CD4, CD23, or HLA-DR. CD25 and CD23 antigens were detected in 29% and 36% of allergic patients only. The accessibility of the EG2 epitope was significantly enhanced in all study groups compared with controls. In vitro activation of normal eosinophils with phorbol 12-myristate 13-acetate upregulated CD9 and EG2 expression but failed to induce the CD25 antigen, suggesting that selective activating stimuli may be required. The functional significance of in vivo CD25 expression and the role of activated PBEs in the development of cellular and cytokine-mediated tissue destructive processes in aGVHD remain to be clarified.

Adolescent↗
PubMed1998

Assessment of hematological and immunological function during long-term follow-up after peripheral blood stem cell transplantation.

BACKGROUND AND OBJECTIVE: Long-term hemopoietic and immunological profile after autologous peripheral blood progenitor cells transplantation (PBPCT), in patients affected by hematological malignancies is largely unknown. The aim of this work was to detect the impact of high dose chemotherapy and PBPCT on hemopoietic and immunological function compared to conventional chemotherapy. DESIGN AND METHODS: Patients had to fulfill the following criteria: continuous complete remission after PBPCT, follow-up longer than 12 months, no chemo or radiotherapy or biological response modifiers after PBPCT. Twenty-five patients were considered eligible for this analysis. Stable and complete hemopoietic reconstitution (Hb > 12 g/dL, WB > 4.0 x 10(9)/L, ANC > 1.5 x 10(9)/L and Pits count > 150 x 10(9)/L), morphological examination of peripheral blood and bone marrow, cytogenetic analysis and immunological profile were evaluated at 12 months and yearly thereafter. RESULTS: Immunological reconstitution showed a persistent reduction of CD4/CD8 ratio up to five years after PBPCT. This reduction was related to a persistent increase of CD8+ lymphocytes and a constant reduction of CD4+ lymphocytes. INTERPRETATION AND CONCLUSIONS: Defects observed in PBPCT patients are induced by the procedure itself, by the conditioning regimen or both. The different behavior in the immune reconstitution of CD8+ subset after PBPCT may be favored by an extrathymic origin of these cells while CD4+ subset recovery which is thymus-dependent is impaired after PBPCT in adult population. Long-term hemopoietic reconstitution after PBPCT is rapidly obtained and is stable over the years, long-term immunological function seems to be abnormal in these patients and these abnormalities are long-lasting.

Adult↗
PubMed1998

Serum of healthy donors receiving granulocyte colony-stimulating factor induces T cell unresponsiveness.

The effects of serum from healthy donors receiving recombinant human granulocyte colony-stimulating factor (rhG-CSF) (G-serum) on blast transformation, expression of activation-related antigens, secretion of interleukin (IL)-2, and proliferation were evaluated in allogeneic lymphocytes stimulated with phytohemagglutinin. Escalating concentrations of G-serum induced 27%, 47%, and 70% suppression of lymphocyte proliferation; interestingly, CD4+ and CD8+ cells underwent blast transformation and up regulated early (CD69) and late (CD25, HLA-DR, and CD71) activation-related antigens. Negligible fractions of apoptotic cells were found after mitogenic challenge, suggesting that the strongly diminished proliferation was not attributable to extensive activation-induced programmed cell death of responding T cells. The levels of IL-2 in cultures containing G-serum were comparable to those in cultures performed without G-serum; however, high concentrations of exogenous IL-2 restored lymphocyte mitogenesis regardless of G-serum concentration. These findings--cell enlargement, upregulation of activation-related antigens, inability to proliferate after mitogenic stimulus, and restoration of cell division by exogenous IL-2--resembled those associated with "partial activation" of lymphocytes, a fundamental control mechanism of tolerance induction in T cell clones. Soluble immunoregulatory mediators infused with allogeneic hematopoietic progenitor products collected after rhG-CSF administration could induce T cell unresponsiveness in vivo, thus preventing clonal expansion and amplification of immune responses, and could account for the unexpectedly reduced incidence and severity of graft vs. host disease compared with allogeneic marrow infusion.

Adult↗
PubMed1998

Expression of p15INK4B in normal hematopoiesis.

The cyclin-dependent kinase inhibitor (CDKI) p15INK4B (p15) induces cell cycle arrest in G0/G1 phase. Several studies report deletion or transcriptional loss of the p15 gene in myeloid and lymphoid hematological malignancies, and a possible role as a tumor suppressor gene has been proposed for this CDKI. In this study we evaluated the expression of p15 by cytofluorometric, immunohistochemical, and reverse transcriptase-polymerase chain reaction (RT-PCR) methods in CD34+ progenitors (both during steady state and after chemotherapy and/or granulocyte-colony stimulating factor [G-CSF] administration) and in cells belonging to different hematopoietic differentiative lineages. We found that p15 is not expressed in normal G0/G1-arrested peripheral blood (PB)- or bone marrow (BM)-CD34+ cells. Moreover, p15 is expressed in G0/G1-blocked CD34+ cells mobilized by chemotherapy and G-CSF but not in CD34+ cells mobilized by G-CSF alone. To clarify the role of p15 in normal hematopoiesis, we used flow cytometry to investigate its expression in normal differentiating BM and PB cells. We found that p15 was expressed in cells belonging to the granulocyte-monocyte lineage and in B and T lymphocytes, whereas erythroid and megakaryocytic cells were p15 negative. These findings, which were confirmed both by immunohistochemical and RT-PCR analysis, definitely establish a linkage between p15 expression and granulocyte-monocyte differentiation.

Antigens, CD34↗
PubMed1997

Differential sensitivity of leukemic and normal hematopoietic progenitors to the killing effect of hyperthermia and quercetin used in combination: role of heat-shock protein-70.

Autologous bone-marrow transplantation (ABMT) is widely used in the treatment of acute leukemias where a matched sibling donor is not available for allogeneic transplantation. However, a major problem in ABMT is relapse, and ex vivo purging may be very important in preventing it. We show here that quercetin enhances the growth-inhibitory effect of hyperthermia (HT) in AML (19 cases) and ALL (6 cases) leukemic blasts. Furthermore, the inhibitory effect of this combined treatment resulted in leukemic-cell apoptosis. On the contrary, normal hematopoietic progenitors were neither growth-inhibited nor induced to apoptosis by HT-plus-quercetin treatment. To explain this difference in sensitivity of leukemic and normal hematopoietic progenitors, we analyzed the effect of quercetin on heat-induced expression of heat-shock protein-70 (HSP-70), which has been shown to be important in regulating thermosensitivity. We found that quercetin inhibits heat-induced HSP-70 expression both at protein and at mRNA levels in AML and ALL blasts. In normal CD34+ progenitors, the combined treatment with HT and quercetin did not reduce HSP-70 expression and did not induce cell apoptosis. Considering the difference in heat sensitivity of normal CD34+ and leukemic progenitors in the presence of quercetin, the combined use of HT and quercetin could constitute a purging protocol for ABMT.

Adult↗
PubMed1997

Inhibition of lymphocyte blastogenic response in healthy donors treated with recombinant human granulocyte colony-stimulating factor (rhG-CSF): possible role of lactoferrin and interleukin-1 receptor antagonist.

The effects of rhG-CSF on lymphocyte blastogenesis were evaluated in six healthy donors, submitted to progenitor cell mobilization for allogeneic transplantation. Neutrophil, monocyte and lymphocyte count increased 6.7-fold, 5.3-fold and 2.0-fold on day +4 of rhG-CSF as compared with baseline. The DNA stimulation index (DNA SI) of 72 h phytohemagglutinin (PHA)-treated cultures decreased from 20% (15-35.5) prior to rhG-CSF to 6.7% (1.5-11.9; P = 0.0026), 8% (4-12; P = 0.0091) and 15% (9-22; P = 0.0091) on days +2, +4 and +6; similarly, reactivity to concanavalin A decreased from 18% (12-20) to 1.8% (0.5-7; P < 0.01), 3% (2-8; P < 0.01) and 5% (2-11; P = 0.009). No changes of lymphocyte response to pokeweed mitogen were observed. DNA SI of PHA-treated cultures inversely correlated with neutrophil and monocyte count. IL-1 receptor antagonist (IL-1ra) and lactoferrin (Lf) plasma levels sharply increased and correlated with neutrophil and monocyte count. IL-10 increased five-fold on day +2, returned to pretreatment values thereafter and did not show any correlation with DNA SI, suggesting that it was not responsible for the observed phenomena. Interestingly, DNA SI of PHA-treated cultures inversely correlated with IL-1ra and Lf levels. CD3+ and CD19+ lymphocyte activation status, ie CD23, CD25, CD30 and HLA-DR coexpression, was not affected by rhG-CSF administration. Pharmacological doses of rhG-CSF in healthy donors inhibit lymphocyte blastogenesis via an increased production and/or release of immunoregulatory soluble mediators, ie IL-1ra and Lf, by primed neutrophils and monocytes.

Adult↗
PubMed1997

Autologous stem cell transplantation: evaluation of erythropoietic reconstitution by highly fluorescent reticulocyte counts, erythropoietin, soluble transferrin receptors, ferritin, TIBC and iron dosages.

The plasma concentrations of erythropoietin (Ep), soluble transferrin receptors (sTfRs), iron, total iron binding capacity (TIBC) and ferritin were monitored in five leukaemia patients undergoing autologous bone marrow stem cell transplantation (BMSCT) and in 10 lymphoma and 21 ovarian cancer patients undergoing autologous peripheral blood SCT (PBSCT); 9/21 ovarian cancer patients received recombinant human G-CSF and Ep and six recombinant human GM-CSF and Ep following SCT. All parameters were evaluated in relation to the kinetics of erythroid reconstitution as evaluated by haemoglobin (Hb) and reticulocyte levels [including the fraction of immature reticulocytes, also called highly fluorescent reticulocytes (HFR)]. Leukaemia patients undergoing BMSCT showed only a delayed (occurring at days 35-50 after SCT) and partial RBC, neutrophil and platelet recovery, whereas all patients undergoing PBSCT exhibited a rapid (occurring at days 10-15 after SCT) and sustained haemopoietic recovery. The various levels of erythroid rescue observed among these patients markedly influenced the kinetics of the different parameters investigated: (i) in leukaemia BMSCT patients sTfRs declined following SCT and remained at low levels thereafter, whereas Ep, iron. TIBC and ferritin showed a progressive and significant increase; (ii) in the different groups of patients undergoing PBSCT: (a) sTfR levels first declined following SCT and then returned to pre-therapy values at days 12-16, this response preceded erythropoietic recovery; (b) Ep, total iron, TIBC and ferritin showed an initial increase in the first days following SCT and then returned to pre-therapy values. Altogether, these observations indicate that: (i) both sTfR levels and reticulocyte counts are predictive parameters of erythropoietic recovery; (ii) coordinated changes of biochemical parameters underlying iron metabolism (iron, TIBC and ferritin) accompany erythroid rescue following SCT.

Adolescent↗
PubMed1997

RhG-CSF-mobilized CD34+ peripheral blood progenitors are myeloperoxidase-negative and noncycling irrespective of CD33 or CD13 coexpression.

Cycling status, myeloperoxidase expression, informative surface markers, and proliferative potential of peripheral blood hemopoietic progenitors (PBHP) were evaluated by flow cytometry in 10 patients affected by resistant lymphoma, and submitted to stem cell mobilization with combination chemotherapy (MiCMA) followed by recombinant human granulocyte colony-stimulating factor (rhG-CSF, 5 micrograms/kg/day). CD34+ PBHP coexpressed myeloid-associated and activation antigens, i.e., CD33 (96%, range 85-99), CD13 (99.5%, range 99.4-99.8), HLA-DR (99%, range 96.5-99.8), and CD38 (98%, range 90-100), lacked intracytoplasmic myeloperoxidase (MPO, < 3%), and resided in the Gzero/G1 phase of the cell cycle (96.5%, range 81-99.5, compared with 70%, range 49-78 bone marrow HP; p = 0.0007), independently of surface membrane phenotype; S-phase percentages of sorted CD34+ CD33(+)/-, CD34+CD38(+)/-, CD34+HLA-DR(+)/-, CD34+CD45RA(+)/-, CD34+CD45RO(+)/-, and CD34+CD41a(+)/- subpopulations were always negligible. In colony assays, 5-week-old long-term cultures seeded with CD34+CD33- cells yielded as many colonies as did CD34+CD33+ cells. In conclusion, rhG-CSF-mobilized CD34+ PBHPs contain noncycling, highly immature progenitors in which the expression of myeloid-associated antigens, i.e., CD33 or CD13, might not be indicative of myeloid commitment.

Antigens, CD↗