Biomedical subjects

G M Rigolin

Publications and source records attributed to G M Rigolin.

33 records · Page 2Linked to original sources

PubMed1996

Richter's syndrome in a case of atypical chronic lymphocytic leukaemia with the t(11;14)(q13;q32): role for a p53 exon 7 gene mutation.

Clinicobiological, histological, cytogenetic and molecular genetic studies were performed in a case of atypical B-cell chronic lymphocytic leukaemia (B-CLL) with the t(11;14)(q13;q32) evolving into Richter's syndrome (RS) in order (a) to determine the clonal relationship between the cell of origin for B-CLL and RS, and (b) to analyse genetic events underlying the disease progression in this patient. After 4 years following diagnosis, a rapid deterioration of the clinical picture occurred, concomitant with the appearance of large lymphoid blasts in peripheral blood (PB), bone marrow (BM) and ascites samples. A diagnosis of RS was made and cytogenetic analysis revealed karyotype evolution with trisomy 7 and del(17p) in addition to t(11;14). Fluorescence in situ hybridization showed 78% lymphoid blast cells obtained from ascites sample to be trisomic using a chromosome-7-specific pericentromeric probe. Whereas no rearrangement of the c-myc proto-oncogene was detected at disease progression, direct sequencing of p53 gene exon 5-9 revealed an exon 7 missense point mutation. This abnormality was not present in the CLL phase. Immunological staining with the monoclonal antibody PAb-1801, detecting the p53 protein product, revealed a negative pattern in the CLL phase, whereas 24% positivity was documented in representative samples obtained at RS. It is concluded that RS was cytogenetically related with B-CLL in this patient, suggesting the occurrence of a bona fide transformation and that the mutation of p53 exon 7, in association with the development of 17p deletion, possibly played a role in the development of RS.

Aged↗

PubMed1995

Photomultiplier voltage setting: possible important source of variability in molecular equivalents of soluble fluorochrome (MESF) calculation?

We evaluated the effect of flow cytometer photomultiplier (PMT) voltage setting on molecular equivalents of soluble fluorochrome (MESF) calculation by using two different quantitative microbead calibrating kits as calibrating standards and a third one as an unknown testing sample of stable intensity of fluorescence. Based on the analysis of residual values derived from linear regression and on the determinations of the resolution index, we demonstrated that, on a FACScan instrument, the window of photomultiplier setting for fluorescein isothiocyanate (FITC) fluorescence determinations is 590-670 V. However, the best region was in the range of 610-650 V. Within the 590-670 PMT voltage region, the evaluation of the same testing sample at different PMT voltages, at 590 V, gave MESF percentage differences of 15% and 13% on the two lower unknown standards of intensity compared to the values obtained at the nearby 630 V setting. In the analyzed PMT region, logarithmic amplifier response was globally maintained, whereas evaluation of the same unknown testing sample in both linear (Lin) and logarithmic (Log) amplification demonstrated (at the higher intensities of fluorescence) Log-Lin minimal differences within the 610-650 PMT voltage range. According to our data, it can be stated that PMT voltages between 610 and 650 provide the best instrument performances and that PMT setting may be proposed as a source of variability in MESF calculation that deserves a careful evaluation in quality-control trials.

Calibration↗

PubMed1995

Heterogeneity of lineage involvement by trisomy 8 in myelodysplastic syndrome. A multiparameter analysis combining conventional cytogenetics, DNA in situ hybridization, and bone marrow culture studies.

To better understand the role of trisomy 8 in myelodysplastic syndrome (MDS), we performed a multiparameter analysis combining conventional chromosome studies (CCS), fluorescence in situ hybridization (FISH), and bone marrow (BM) culture studies in two patients with MDS evolving into acute myeloid leukemia (AML). A mosaicism of a cytogenetically normal clone and a clone with trisomy 8 was detected in both patients throughout the course of the disease, a finding confirmed by FISH on BM cells. The relative size of the trisomic clone increased from 52% to 71% (p < 0.0001) and from 53% to 69% (p = 0.001) of all BM cells at the time of the leukemic switch in patients 1 and 2, respectively. Combined FISH and immunophenotyping of BM cells showed involvement of the granulomonocytic lineage in patient 1 and involvement of erythroid cells as well as of the granulomonocytic lineage in patient 2. Only disomic lymphocytes were detected in both patients. FISH on single hemopoietic colonies grown in semisolid media detected trisomic CFU-GM and disomic BFU-E in patient 1, whereas a proportion of CFU-GM and BFU-E deriving from the trisomic clone was detected in patient 2. However, the percent of trisomic colonies was lower than the percent of involved granulomonocyte precursors and involved erythroblasts, as detected by combined FISH and immunophenotyping on fresh BM samples. We have thus shown heterogeneity of lineage involvement by trisomy 8 in MDS undergoing transformation into AML. Although preferential growth of disomic clones may occur in vitro, the finding of an increased size of the trisomic clone at the time of leukemic switch suggests that these cells had proliferative advantage in vivo over cells without trisomy 8.

Aged↗

PubMed1995

Atypical chronic lymphocytic leukaemia with t(11;14)(q13;q32): karyotype evolution and prolymphocytic transformation.

In order to define better the cytological and clinical features of atypical B-cell chronic lymphocytic leukaemia (B-CLL) with t(11:14)(q13;q32), sequential morphologic immunological and cytogenetic studies were performed in seven patients belonging to a series of 72 consecutive cases presenting with a diagnosis of CLL or atypical CLL according to the FAB criteria. Cytologic diagnosis in these seven patients with t(11;14) was typical CLL in two cases presenting with < 10% large lymphocytes (LL) and prolymphocytes (PL) and atypical CLL in five cases in which LL and PL comprised between 10% and 55%. The diagnosis was supported by histologic findings on bone marrow biopsy (five cases) or splenectomy specimens (two cases). A progressive increase of peripheral LL and PL was observed, resulting in a switch of FAB diagnosis over a 6-60-month period from typical CLL into atypical CLL in two cases and from atypical CLL into prolymphocytic leukaemia in five cases. Immunophenotyping showed a mature B-cell phenotype with CD19, CD22, CD24 positivity and CD10 negativity in all patients. A bright-staining pattern for surface immunoglobulins (SIg) was detected in 6/7 cases, CD5 positivity in 6/7 cases, and CD23 positivity in 1/7 cases. The FMC-7 monoclonal antibody was positive in > 40% cells in 5/6 cases. Chromosome changes in addition to t(11;14) were seen in five cases; in two cases unbalanced translocations involving the 3q21 chromosome region, resulting in partial trisomy for the long arm of chromosome 3, were detected early in the course of the disease. Karyotype evolution that was associated with disease progression occurred in 3/6 assessable patients. Comparison of these findings with similar data from 65 B-CLL patients without t(11:14) showed that atypical morphology, switch of FAB diagnosis during the course of the disease, and karyotype evolution were more frequently seen in cases with t(11;14) (5/7 v 15/65 cases, P = 0.015, 7/7 v 7/65 cases, P < 0.0001, and 3/6 v 5/45 assessable cases, P = 0.04, respectively). The frequency of positivity for CD23 and bright SIg staining differed significantly in the two groups. It is concluded that t(11;14) identifies a cytologically atypical subset of B-CLL, characterized by frequent cytologic and cytogenetic evolution and by a distinct immunological profile, sharing some biological features with mantle cell lymphoma.

Antigens, CD↗

PubMed1995

CD34+ leukemic cells assessed by different CD34 monoclonal antibodies.

CD34 monoclonal antibodies (McAbs) are widely used to identify and isolate hemopoietic progenitors and to classify acute and chronic leukemias. We assessed the reactivity of 17 CD34 McAbs from the 5th International Workshop on Leukocyte Differentiation Antigens with a variety of cells types: normal bone marrow hemopoietic progenitors, 10 AML, 6 ALL, 11 CML. The reactivity for these McAbs was compared with that of reference CD34 McAbs (Q-Bend 10 and 8G12). For each cell population the % of McAb binding cells, the peak channel and the mean fluorescence intensity (MFI) of the positive cells was evaluated. The peak channel, the MFI and the number of positive cells varied significantly from case to case, depending on the McAb and the type of leukemia. According to the spectrum of reactivity three groups of McAbs were defined; however, 7 McAbs do not belong to any of these subgroups. These groups were not entirely in accordance with McAb classification based on enzyme cleavage that identified three epitopes of the CD34 molecule. Some reagents were found to be more specific for AML, other for ALL, CML or normal CD34+ cells. Normal bone marrow light density cells showed a significantly higher percentage of positive cells for 43A1 and MD34.2 McAbs compared to that documented for the remaining McAbs. AML cells showed the most variable pattern of expression for the CD34 McAbs. In leukemic samples, MESF (molecular equivalents of soluble fluorochrome) values ranged from 18,200 to 322,000 and the number of binding sites per cells was 5,000-81,000.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Monoclonal↗

PubMed1995

CD34+/CD33+ blast cells: correlation with FAB subtypes.

The reproducibility of FAB morpho-cytochemical classification in a multicenter trial was recently assessed as 78.1%. The relevance of immunophenotyping in AML is debated because of the lack of a clear prognostic impact and the necessity of uniform diagnostic criteria and standardised methodologies. Several authors, studying the morpho-immunological characteristics of AML blasts by flow cytometry, suggested the necessity for the definition of clusters of cells with similar morphological patterns and intensity of expression. In this respect we reviewed blast CD34 and CD33 intensity of expression in 65 AMLs classified according to the FAB criteria. Four morphological groups could be identified: 1) low side scatter (SSC) and forward scatter (FSC); 2) intermediate SSC and FSC; 3) high SSC and FSC; 4) combination of pattern 2 plus 3. Five immunological patterns were defined: a) high CD34 expression with negative or weak CD33; b) high CD34 and CD33 expression; c) weak to negative CD34 with high CD33 expression; d) coexistence of 2 different subpopulations; e) negative to weak CD34 and CD33 expression. Based on this morpho-immunological analysis we were able to subdivide AML patients into 5 homogeneous subgroups and a comparison with FAB classification showed a concordance of 73.9%. Regarding CD34 and CD33 intensity of expression, a correlation with prognosis was demonstrated among all M1-M2 patients and in the M2 and M4 subgroups. In conclusion even if immunophenotype cannot substitute the FAB approach to AML, we feel that a flow cytometry morpho-immunological analysis could be helpful in achieving a greater understanding and agreement between different Institutions and assist in the definition of more precise prognostic subgroups.

Antigens, CD↗

PubMed1994

Neutrophils from patients with myelodysplastic syndromes: relationship between impairment of granular contents, complement receptors, functional activities and disease status.

Myelodysplastic syndromes (MDS) are stem cell disorders of clonal origin in which infections and leukemic transformation are quite frequent. Neutrophils from 28 patients with MDS were analysed by flow cytometry for the expression of the two complement receptors CR1 and CR3, the antigenic reactivity of some granule constituents--myeloperoxidase, lysozyme, elastase, lactoferrin--and functional activities, such as locomotion, respiratory burst and cytotoxicity. The results were correlated with the FAB disease subtypes, grouped as low risk (RA) and high risk patients (RAEB, RAEB-t, CMML) and with 30 healthy subjects. A significant reduction in the percentage of neutrophil CR1, CR3 positivity and chemotaxis induced by endotoxin-activated serum was detected in the high risk group when compared with the low risk group and healthy controls. Furthermore, the high risk group also showed a low amount of myeloperoxidase, elastase, lysozyme and superoxide anion, but both low and high risk groups displayed reduced cellular cytotoxicity in comparison with the control. This work indicates that MDS patients belonging to the more advanced FAB categories frequently show multiple abnormalities in the expression of neutrophil complement receptors, and granular components (> 3), as well as in cell functions, suggesting the possibility of using these phenotypic abnormalities in the monitoring of disease progression.

Chemotaxis, Leukocyte↗

PubMed1994

Prognostic value of immunophenotypic characteristics of blast cells in acute myeloid leukemia.

In order to elucidate the prognostic role of cytofluorimetry analysis of leukemic cells in AML, the immunophenotypic characteristics of blast cells obtained from 66 AML patients belonging to M0-M2 and M4-M5 FAB subtypes have been investigated by flow cytometry using a large panel of monoclonal antibodies (McAbs) utilized in single, double, and triple fluorescence experiments. On a univariate analysis, four different immunophenotypic blast cell characteristics were found to be associated with a poor prognosis: expression of CD34 "bright" (ratio > 10 between fluorescence emission of positive cells and that of negative (isotypic) control-P/N ratio: mean MESF value: 265,000) in > 15% blast cells, co-expression of CD34 and CD33 in > 60% blast cells, expression of CD14 in > 30% leukemic cells, the MDR+ ("multiple drug resistant") phenotype. In contrast, the duration of remission, and overall survival of AML patients showing a "dim" CD34 expression (P/N ratio: 3-10: mean MESF value: 49,000) was similar to that of CD34- AML patients, irrespective of the percentage of positivity for CD34, which was, however, a predictive factor of survival in patients with higher CD34 fluorescence intensities in their blastic population. No correlation between FAB subtypes, prognosis and immunophenotype was found. The multivariate regression analysis showed that, besides age, only the combined expression of CD34 and CD33 had independent prognostic meaning. Indeed, in each FAB subtypes the CD34+/CD33+ phenotype was associated with a shorter survival and a lower mitotic rate. These data may contribute to the understanding of the discrepancies so far observed in the literature regarding the prognostic role played by the CD34 expression on leukemic AML blasts.

Antigens, CD↗

PubMed1993

Chromosome aberrations in CD34-positive acute myeloid leukemia. Correlation with clinicopathologic features.

Morphologic, immunologic, and cytogenetic features were studied in 30 newly diagnosed patients with CD34-positive (CD34+) de novo acute myeloid leukemia (AML) in comparison with 30 patients with CD34-negative (CD34-) AML. Karyotype at diagnosis was abnormal in 25/30 CD34+ AML patients, of which nine had major karyotype aberrations (MAKA). Clonal chromosome changes were detected in 9/30 patients with CD34- AML. The most frequent chromosome aberration in CD34+ patients was -5/5q-, an aberration showing a strong association with the M2 FAB subtype of AML. Other recurring chromosome changes involved chromosome 16q (four cases) and chromosome 17p (three cases). Total or partial monosomy 7q was detected in three cases. Among CD34- AML, two patients had the classical t(15;17) and two had structural aberrations of 6q. Among patients with CD34+ AML, nine had MAKA in association with trilineage myelodysplasia (TMDS). TMDS was infrequent in CD34+ AML without MAKA and in CD34- AML. Complete remission (CR) was achieved in 8/30 CD34+ AML (26%), as compared with 22/30 CD34- AML (73%), and median survival was 2 months in the former group and 8 months in the latter. No patient with CD34+ AML and MAKA achieved CR, whereas 8/21 CD34+ AML without complex chromosome changes or with normal karyotype achieved CR. In conclusion, a distinct cytogenetic profile may be associated with CD34+ AML. Cytogenetic findings in CD34+ AML may be clinically relevant in that they may disclose a subset of patients with MAKA with a low CR rate.

Adult↗

PubMed1993

[Immunophenotypic study of acute myeloid leukemia (AML): biological significance and prognostic implications].

The immunophenotype of 61 de novo AML referring to the Institute of Hematology of Ferrara between January 1988 and September 1992 was studied in order to evaluate the prognostic impact and the biological implications of this approach. In this study we confirm the possible aberrant expression of some antigens and the lack of a strict correlation between FAB classification and immunophenotype. The expression of the CD14 and CD34 was correlated with a worse prognosis, while the CD34+ patients further showed a lower probability to achieve a complete remission.

Aged↗

PubMedDate not supplied

Primary gastric lymphoma: distribution and clinical relevance of different epidemiological factors.

BACKGROUND: Over the last 10 years the incidence of primary gastric lymphomas (PGL), and in particular those of MALT origin, has significantly increased. Recent works correlated this epidemiological observation to Helicobacter pylori (HP) infection. On the other hand, new evidence demonstrating that occupational exposure to pesticides and solvents has played an important role in the pathogenesis of non-Hodgkin lymphomas (NHL) has emerged from studies involving large series of patients. METHODS: Thirty PGL patients, observed between 1986-1992, were subdivided according to HP infection, history of previous gastric disturbances (G) and exposure to pesticides and solvents (T). RESULTS: On the basis of these parameters we divided the patients into three groups: T+HP+ (8), T+HP- (7), T-HP+ (9). T+ patients had a positive history of gastric problems or a positive histological biopsy in 13.3% of cases, versus 66.7% in T- patients. The incidence of HP infection in the T+ group was 53%, which proved to be comparable to the statistics for northeastern Italy, while in the T- group the incidence of infection was 100%. CONCLUSIONS: On the whole these data suggest that HP infection could be considered a pathogenetic factor in 34% of patients, while occupational exposure to pesticides and solvents could have played a more important role in 66% of these cases.

Aged↗

PubMedDate not supplied

Study of prognosis in acute myeloid leukemias (AML) by cluster analysis.

BACKGROUND: Cluster analysis is particularly effective in detecting homogeneous subgroups among large series of observations. We applied this relatively uncommon approach to the study of prognosis in 137 patients affected by acute myeloid leukemia (AML). METHODS AND RESULTS: Employing simple presentation parameters (age, WBC, splenomegaly, hepatomegaly) we used cluster analysis to define 3 groups with different overall survival (p = 0.0019). This classification was obtained following a rescaling of the variables and principal component analysis. Validation was performed through random definition of a control group. With the same variables, univariate analysis demonstrated age was the only prognostic factor, while Cox's model was not significant. CONCLUSIONS: In our series cluster analysis allowed a better definition of prognosis than Cox's analysis. Since the 3 groups are well identifiable, each patient can be rapidly classified and his allocation confirmed by discriminant functions. For cluster 2 we were able to project a possible myelodysplastic evolution, while cluster 3 was more frequently associated with a monocytic blastic component. We think that cluster analysis deserves consideration as an alternative statistical approach in the analysis of large series of data; its usefulness lies in its power to define homogeneous prognostic or biologic subgroups and to elaborate further hypotheses for new studies.

Acute Disease↗

PubMedDate not supplied

Flow cytometry evaluation of urokinase-type plasminogen activator receptor (UPA-R) in acute myeloid leukemia cells.

The aim of this study was to investigate by flow cytometry the expression of the UPA-R (Urokinase type plasminogen activator receptor-CD87) on the blastic population of AML and ALL patients in order to evaluate whether the presence of this molecule could be associated with peculiar clinical and biologic features of leukemic cells. Five different monoclonal antibodies (MoAbs) (clones: 3B10#; VIM5*; 109#; 68#; 100#) were used in order to detect the distinct forms of this cellular receptor. Cell reactivity varied significantly from case to case, also depending on the MoAb used for the flow cytometry analysis. In brief, 3B10# and VIM5* MoAbs were found to be positive in more than 90% of monocytes and neutrophils from healthy subjects, while the number of positive cells was decreased (60%) using the 109# MoAb. However, either 68# and 100# MoAbs recognised only a low number of blood monocytes and neutrophils (8-20%), while lymphocytes were unreactive with all the five UPA-R MoAbs. ALL cells were found to be CD87 negative in all cases. Blasts from AML showed a heterogeneous pattern of expression for the UPA-R MoAbs, being the reactivity strictly dependent on the MoAb used, and, to a higher extent, on the degree and type of maturation of the blastic cells. The number of blasts recognising 3B10# and VIM5* MoAbs was significantly higher than that reacting with the remaining MoAbs irrespective of the FAB subtype. Since proteolytic enzymes, like UPA, play a key role in the dissolution of the extracellular matrix, and in facilitating the cell egress from the bone marrow, it is conceivable that the expression of the UPA-R could contribute to the invasive properties and, possibly, metastatic potential of leukemic cells.

Acute Disease↗

PubMedDate not supplied

Myelodysplastic syndromes with monocytic component: hematologic and cytogenetic characterization.

BACKGROUND AND OBJECTIVE: Patients with myelodysplastic syndromes (MDS) showing high numbers of abnormally localized immature precursors (ALIP) also display myelomonocytic antigens on immature cells, and it has been suggested that the presence of monocytosis could define a distinct subset of MDS characterized by poorer survival. The objective of this study was to analysis the incidence and significance of monocytosis among our series of patients with MDS and correlate the distributions of these elements with other hematologic features. METHODS: We evaluated the monocytic component in myelodysplastic syndromes in order to clarify the significance of monocytosis in MDS and its relationship with CMMoL and other MDS. Monocytosis was defined as a percentage of blood monocytes greater than 10%. RESULTS: Among a series of 139 consecutive MDS patients, we describe a group of 29 (20.8%) patients with monocytosis and dysplastic features involving multiple cell lineages which do not fulfill the criteria for diagnosis of CMMoL or aCML. These patients, who do not differ from MDS without monocytosis in the main clinical parameters, are characterized by relatively higher leukocyte (WBC 6.6 x 10(9)/L) and granulocyte counts (PMN 2.5 x 10(9)/L), hypercellular bone marrow and relatively poor prognosis. Among these patients, we observed a particularly high incidence of evolution to CMMoL (34.5%) and AML (17.2%) with monocytic component (FAB M4 and M5). Cytogenetic data demonstrated clonal chromosome changes in 11/13 patients with MDS and monocytosis, while only 19/41 patients without monocytosis showed clonal abnormalities. CONCLUSIONS AND PERSPECTIVES: The combination of hematologic and cytogenetic features in our study suggests that it is reasonable to consider myelodysplasia with monocytosis as a distinct disease subset of MDS, characterized by multilineage dysplasia along with a higher incidence of karyotype aberrations. The multi-step pathogenetic process in these patients may have reached a more advanced stage at which the relative or absolute increase in the number of monocytes may represent the first event in the subsequent progression of the disease towards acute leukemia.

Cell Lineage↗

PubMedDate not supplied

Potential clinical applications of rhGM-CSF in acute myeloid leukemia based on its biologic activity and receptor interaction.

BACKGROUND AND OBJECTIVE: Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a multilineage hemopoietic growth factor that stimulates proliferation, differentiation, and survival of progenitor cells, enhances the functional activities of mature myeloid effector cells, and plays a key role in host defense and the inflammatory process. Although the clinical use of rhGM-CSF in patients affected by lymphoid malignancies is widely accepted, its utility and safety in the management of acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS) is still controversial. The three main schedules adopted for clinical application of GM-CSF in AML are as follows: A) post-chemotherapy, in order to shorten the duration of neutropenia and/or monocytopenia; B) prechemotherapy to recruit blast cells into active cell cycle phases, and to increase their sensitivity to cell cycle-dependent cytotoxic drugs; C) as a mobilizing agent to induce the release of progenitor cells from bone marrow into circulation (peripheral blood progenitor cell transplantation-PBPC). The objective of this paper is to analyze the potential clinical applications of rhGM-CSF in AML. EVIDENCE AND INFORMATION SOURCES: The material examined in the present review includes several personal papers in this field and articles and abstracts published in journals covered by the Science Citation Index. STATE OF THE ART: Based on current knowledge, it may be argued that rhGM-CSF should be used only in a subset of AML patients at high risk of infection mortality, including elderly subjects, and/or in those AML patients who relapse or are resistant to induction treatment. However, the risk of stimulating the leukemic clone following GM-CSF therapy should be kept in mind when using this growth factor in the clinical setting, even though the great majority of the reported papers on this subject have shown that GM-CSF therapy does not affect relapse rates, frequency of remissions or patient life expectancy. PERSPECTIVES: It is likely that new data from controlled clinical trials will clarify the therapeutic role of GM-CSF in myeloid-derived malignancies, allowing the establishment of consensus guidelines for its use.

Acute Disease↗