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Biomedical subjects

E Matutes

Publications and source records attributed to E Matutes.

At least 19 recordsLinked to original sources
PubMed1996

Analysis of clonal rearrangements of the Ig heavy chain locus in acute leukemia.

Clonal rearrangements of the Ig heavy chain (IGH) locus occur in nearly all cases of B-cell precursor acute leukemia (BCP-ALL). Some of these rearrangements may be detected by polymerase chain reaction (PCR) using VH gene framework III (FRIII) and JH consensus primers. However, about 20% of BCP-ALLs fail to amplify with this technique. To determine the causes of these PCR failures and to investigate any possible association with specific subgroups of disease, we analyzed 72 acute leukemias of defined immunophenotype and cytogenetics, comparing FRIII with VH-family leader-specific PCR methods and Southern blotting. Of 37 BCP-ALL cases, 6 (16.2%) failed totally to amplify with FRIII and JH primers. None of these cases amplified with VH leader primers. Additionally, all cases retained germline VH6 genes and 5 of 11 rearranged alleles amplified with a consensus DH primer, indicating that these rearrangements represented biallelic DH-JH recombinations. Among the 6 FRIII and VH leader PCR-negative BCP-ALL cases, there was no common immunophenotype or consistent cytogenetic abnormality, although all showed structural chromosomal abnormalities and 3 of 5 successfully karyotyped had abnormalities of chromosome 12p. 13 cases with t(9;22)(q34;q11) Philadelphia chromosome-positive [Ph+]) and IGH rearrangements (9 BCP-ALL and 4 biphenotypic cases) were also analyzed. Of 23 rearranged IGH alleles, 19 (82%) were positive by FRIII PCR, and all 4 remaining alleles were amplified by VH leader primers. Use of the leader primers in these Ph+ cases also detected 3 additional clonal rearrangements that were not anticipated from Southern blotting; such unexpected bands were not observed in 21 other Ph- cases. The additional bands represented "new" and unrelated VH rearrangements rather than VH-VH replacement events. We conclude that biallelic DHJH rearrangements occur in a subgroup of BCP-ALL; in these cases, the activation of the full VHDHJH recombination mechanism had not occurred. Therefore, these cases of BCP-ALL were arrested at an early stage of B-cell differentiation. In contrast, all Ph+ BCP-ALLs and biphenotypic acute leukemias, which may represent the transformation of multipotent hemopoietic stem cells, had undergone VHDHJH recombination. Of 9 Ph+ BCP-ALL cases, 3 also showed ongoing VHDHJH rearrangement, reflecting the persistent expression of the VHDHJH recombinase. Finally, sequence analysis of 33 rearranged VHDHJH genes showed that only 3 including 2 Ph+ BCP-ALL maintained an intact open-reading frame. Loss of the open-reading frame occurred not only because of out-of-frame VHDH and DHJH joining, but also because of VH gene mutation and deletion. These data show that most BCP-ALLs may represent the neoplastic transformation of BCPs destined to die in the bone marrow.

Acute Disease↗
PubMed1996

C-kit receptor (CD117) expression in acute leukemia.

The murine monoclonal antibody YB5.B8 (CD117) identifies a transmembrane tyrosine kinase receptor encoded by the human c-kit proto-oncogene. In this study we investigated the expression of c-kit on different types of acute leukemia to determine the degree of specificity and sensitivity of this marker for the myeloid and lymphoid lineages. C-kit was positive in over half of the 115 cases of acute leukemia studied. Overall, two thirds of AML cases expressed c-kit, whereas only one of 23 ALL patients was c-kit positive. C-kit was also positive in 16 of 19 cases of myeloid blast crisis of myeloproliferative disorders and negative in four with a lymphoid phenotype. There was no correlation between c-kit expression and the degree of myeloid differentiation by FAB subtypes or other markers. We conclude that c-kit is a specific marker for the myeloid lineage, which is expressed early during hematopoietic differentiation and can aid the diagnosis of AML in difficult cases. More patients need to be tested to establish whether the expression of c-kit may define AML subgroups of prognostic significance.

Acute Disease↗
PubMed1996

2-Chlorodeoxyadenosine in the treatment of hairy cell leukaemia: differences in response in patients with and without abdominal lymphadenopathy.

We treated 26 patients with hairy cell leukaemia (HCL) with 2-chlorodeoxyadenosine, including nine with abdominal lymphadenopathy and of whom two had HCL-variant; 18 were previously treated. The overall response in 23 evaluable HCL patients was 100% with 87% complete remission (CR). The CR rate was 57% in patients with abdominal lymphadenopathy. Neither of the patients with HCL-variant achieved CR. Of four patients who had become refractory to 2'-deoxycoformycin, two achieved CR and one partial remission (PR). The lower response in HCL with abdominal lymphadenopathy supports the view that this represents a more resistant form of the disease.

Adult↗
PubMed1996

Trisomy 12 defines a group of CLL with atypical morphology: correlation between cytogenetic, clinical and laboratory features in 544 patients.

We have analysed the clinical and laboratory features in 544 patients with chronic lymphocytic leukaemia (CLL) with available cytogenetics and fluorescence in-situ hybridization (FISH) analysis for trisomy 12 in half of them, to examine the correlation between chromosome abnormalities and clinical or laboratory parameters. Five chromosome groups were defined: (1) trisomy 12 (18%), detected as the sole abnormality or associated with other changes; (2) del(13)(q12-14) (7%); (3) other abnormal karyotypes (20%); (4) normal karyotype (41%); and (5) no divisions (14%). There were no differences in the age distribution between the five groups. Clinical stages (Binet) were: A (74%), B (12%) and C (14%). Stage A was common in cases with del(13q)(82%), normal (84%) and other abnormal karyotypes (74%), whereas it was less common in trisomy 12 cases (64%) and those with no divisions (48%). Typical CLL morphology was found in 83% of cases; 10% had more than 10% prolymphocytes (CLL/PL) and 7% had other atypical features. CLL with trisomy 12 was the only group with a high frequency of either CLL/PL (31%) or atypical morphology (24%). Atypical morphology and CLL/PL were even more frequent when trisomy 12 was associated with other chromosomal abnormalities (70% v 46%). The incidence of cases with CLL/PL and other atypical morphology was significantly lower in the other chromosome groups (P < 0.001). There were no differences in immunophenotype among the various groups except for a higher frequency of stronger Smlg and FMC7 expression in cases with trisomy 12, particularly those with CLL/PL and other atypical morphology. Our findings confirm that trisomy 12 defines a subgroup of CLL with more frequent atypical morphology, including CLL/PL, stronger SmIg and FMC7 expression, more advanced stages (B and C in 18%) and possibly worse prognosis.

Aged↗
PubMed1996

Differential expression of CD3 and CD7 in T-cell malignancies: a quantitative study by flow cytometry.

Most T-cell antigens are expressed on normal and neoplastic T lymphocytes and for this reason it is not easy to distinguish between the immunophenotype of normal and malignant T cells. We have addressed this problem by comparing the levels of expression of CD3 and CD7 on T lymphocytes from 18 healthy donors with those of 61 cases of T-cell leukaemia using quantitative flow cytometry with a method that converts fluorescence intensity into number of antigen molecules per cell. Normal T lymphocytes expressed 124 +/- 25 CD3 and 20 +/- 3 x 10(3) CD7 molecules per cell. The mean CD3 values were significantly lower in all types of T-cell leukaemia than in normal T cells (P < 0.05), with the exception of Sezary syndrome. The lowest CD3 values were found in T-lymphoblastic leukaemia (T-ALL), 30 +/- 21 x 10(3), and adult T-cell leukaemia/lymphoma (ATLL), 38 +/- 31 x 10(3), followed by T-prolymphocytic leukaemia (T-PLL), 92 +/- 47 x 10(3), and granular lymphocyte leukaemia (GLL), 95 +/- 21 x 10(3). In contrast, the number of CD7 molecules was significantly higher in T-All, 35 +/- 7 x 10(3) (P < 0.01), and T-PLL, 29 +/- 12 x 10(3), than the normal controls (P < 0.01), whereas ATLL and GLL showed a low CD7 expression, 13 +/- 3 and 12 +/- 3 x 10(3), respectively. Our results show that the quantitative analysis of CD3 and CD7 and their combined evaluation may enable a distinction between normal and leukaemic T cells and could facilitate the monitoring of minimal residual disease. This study has also defined the T prolymphocyte as a cell of intermediate maturity between thymic derived and peripheral T lymphocytes.

Antigens, CD7↗
PubMed1996

The ultrastructure of mantle cell lymphoma and other B-cell disorders with translocation t(11;14)(q13;q32).

We conducted an ultrastructural study in 22 cases of B-lymphoproliferative disorders in leukaemic phase bearing the t(11;14) translocation. The features of peripheral blood leukaemic cells in nine cases of mantle cell lymphoma (MCL) were compared to those diagnosed as B-prolymphocytic leukaemia (B-PLL) (five cases), splenic lymphoma with villous lymphocytes (SLVL) (four cases), lymphoplasmocytic lymphoma (LPL) (one case), chronic lymphocytic leukaemia with > 10% prolymphocytes (CLL/ PL) (one case) and unclassified B-non Hodgkin's lymphoma (B-NHL) (two cases). The ultrastructural characteristics were also compared to those present in B-NHL without t(11;14), including cases of follicular centre lymphoma (FCL). This study shows that MCL has distinct ultrastructural features including a cleaved or indented nucleus with an even heterochromatin distribution, an absent or inconspicuous nucleolus, low N/C ratio, abundant mitochondria, a well developed Golgi zone, profiles of endoplasmic reticulum and centrioles. This pattern clearly differs from that found in FCL cells. The nuclear pattern of MCL cells also differed from the cells in the other disorders with t(11;14), but shared an organelle-rich cytoplasm, and features which were not apparent in cases without t(11;14). The cytoplasmic changes observed in cells bearing t(11;14) suggest increased cellular activity which may relate to the chromosome translocation and the resulting over-expression of bcl-1.

B-Lymphocytes↗
PubMed1996

Expression of TIA-1 and TIA-2 in T cell malignancies and T cell lymphocytosis.

OBJECTIVE: To investigate the reactivity with TIA-1 and TIA-2, two monoclonal antibodies that recognise, respectively, granular structures in T lymphocytes and the T cell receptor chain in cells from a variety of T cell disorders. METHODS: Cytoplasmic staining with TIA-1 and TIA-2 was carried out by the immunoalkaline phosphatase anti-alkaline phosphatase technique in 67 cases with a T cell disorder: 31 large granular lymphocyte (LGL) leukaemia, nine T-prolymphocytic leukaemia (T-PLL), five Sezary syndrome, four peripheral T cell lymphoma (PTCL), 13 T cell lymphocytosis, and five T-acute lymphoblastic leukaemia (T-ALL). All had over 75% abnormal T cells which were CD2+, CD3+, CD5+, CD7+, and negative with B cell markers. RESULTS: TIA-1 was positive in 77% cases of LGL leukaemia and half of the PTCL and T-ALL, whereas it was negative in all Sezary syndrome and most T-PLL (8/9) and reactive T-lymphocytosis (10/13). In LGL leukaemia, TIA-1 was positive irrespective of the membrane phenotype, whether CD8+, CD4- or CD4+, CD8-, and was more often positive in cases where cells were CD16+, CD56+, or CD57+. TIA-2 was positive in 60% of cases encompassing all diagnostic types of T cell disorder. There was no correlation between TIA-2 expression and that of other T cell markers, activation antigens, and natural killer markers. CONCLUSIONS: The pattern of TIA-1 expression in T cell malignancies may help in the differential diagnosis among LGL leukaemia (high expression), T cell lymphocytosis and other T cell diseases (low expression). As TIA-2 is expressed in over 95% mature T lymphocytes and thymic cells, its assessment may be useful to demonstrate aberrant phenotypes which can be exploited for detecting minimal residual disease.

Antibodies, Monoclonal↗
PubMed1996

Association between sarcoidosis and lymphoma revisited.

AIMS: To investigate the relation between sarcoidosis and lymphoma. METHODS: The hospital notes of five patients with sarcoidosis and a lymphoproliferative disorder were reviewed. Histological material on which the diagnoses of sarcoidosis and lymphoma were made was re-analysed. RESULTS: Four of the five patients had well documented sarcoidosis preceding the development of lymphoma by 18 months to 28 years; the fifth patient had lymphoplasmacytic lymphoma with a reactive granuloma reaction. Two patients had chronic sarcoidosis and three were treated with prednisolone. The types of lymphoma were: Hodgkin's disease (n = 1), B cell lymphoma (n = 2) (mantle cell and lymphoplasmacytic/local plasmacytoma) and large granular lymphocyte leukaemia (T cell) (n = 1). CONCLUSIONS: The association between sarcoidosis and lymphoma is confirmed, suggesting that sarcoidosis may be a predisposing factor for the development of a lymphoid malignancy due to disturbance of the immune system. All types of lymphoma may develop. The first case of T cell granular lymphocytic leukaemia in a patient with sarcoidosis has been documented.

Adult↗
PubMed1996

Differential expression of T cell antigens in normal peripheral blood lymphocytes: a quantitative analysis by flow cytometry.

AIMS: To obtain reference values of the level of expression of T cell antigens on normal lymphocyte subsets in order to disclose differences which could reflect their function or maturation stages, or both. METHODS: Peripheral blood from 15 healthy donors was processed by flow cytometry with triple colour analysis. For each sample phycoerythrin (PE) conjugated CD2, CD4, CD5, CD8, and CD56 monoclonal antibodies were combined with Cy5-R-phycoerythrin (TC) conjugated CD3 and fluorescein isothiocyanate (FITC) conjugated CD7; CD2- and CD7-PE were also combined with CD3-TC and CD4-FITC. Standard microbeads with different capacities to bind mouse immunoglobulins were used to convert the mean fluorescence intensity (MFI) values of the lymphocyte subsets identified by multiparametric flow cytometry into the number of antigen molecules per cell, measured as antibody binding capacity (ABC). RESULTS: CD4+ (helper/inducer) T cells exhibit a higher CD3 antigen expression compared with CD8+ (suppressor/ cytotoxic) T lymphocytes. Within the CD4+ T cells, the CD4+CD7- subset expressed a lower level of CD3 compared with CD4+CD7+ and CD8+CD7+ cells, and higher CD2 and CD5 expression than the main CD3+CD7+ subset. Major differences in antigen expression were also detected between CD3+ T cells and CD3-CD56+ natural killer (NK) cells: NK cells exhibited higher levels of CD7 and CD56 and lower levels of CD2 and CD5 than T cells. Significantly lower CD5 expression was also detected in the small CD5+ B lymphocyte subset compared with T cells. CONCLUSIONS: Quantitative flow cytometry with triple colour analysis may be used to detect antigen modulations in disease states and to increase the accuracy of diagnosis by comparison with findings in normal counterparts.

Adult↗
PubMed1996

Zeta chain and CD28 are poorly expressed on T lymphocytes from chronic lymphocytic leukemia.

We have analyzed the expression of the zeta chain of the T cell receptor/CD3 complex and the co-stimulatory molecule CD28 by dual colour immunofluorescence on T lymphocytes from patients with B cell chronic lymphocytic leukemia (CLL). Zeta chain was significantly reduced on CD3-positive lymphocytes from 33 patients compared with normal controls (P<0.0001). The values were lower in stages B and C than in stage A. In five patients tested in partial remission the values were normal. CD28, investigated in CD3, CD4 and CD8 positive T cells from 18 CLL patients appeared to be reduced in the three subsets but more marked in CD8-positive lymphocytes. The loss of zeta chain and CD28 in a proportion of circulating T lymphocytes from CLL may underlie some of the known functional abnormalities of these cells and the immunodeficiency associated with the disease.

CD28 Antigens↗
PubMed1996

Differential expression of B29 (CD79b) and mb-1 (CD79a) proteins in acute lymphoblastic leukaemia.

CD79 is a heterodimeric molecule comprising two polypeptide chains, B29 (CD79b) and mb-1 (CD79a). It is physically linked in the surface of B cells to membrane immunoglobulin, forming the B cell antigen receptor complex. Expression of the mb-1 (CD79a) chain has been studied in leukaemias and shown to be present in most B lineage acute lymphoblastic leukaemias (ALL). In contrast, little is known about the expression of B29 (CD79b) in this condition. Two monoclonal antibodies (MoAb) were used in this study by immunocytochemistry and flow cytometry: HM57, against an intracellular epitope of the mb-1(CD79a) chain, and SN8, reacting with an extracellular epitope of B29 (CD79b). Our aim was to investigate the expression of B29 (CD79b) in the various immunological subtypes of B lineage ALL and compare its cytoplasmic and membrane expression. Seventy-nine cases were studied, including 13 chronic myeloid leukaemia in B lymphoid blast crisis (CML-BC) and 66 ALL, subclassified as early B (two), common (28), pre-B (23), mature (five) and biphenotypic with B lymphoid commitment (eight). Most cases expressed mb-1 (CD79a) in the cytoplasm. B29 (CD79b) was expressed in the cytoplasm in 65% (15/23) of pre-B-ALL and in 14% (4/28) common-ALL but it was detected in the cell membrane in only three cases of mature B-ALL, being negative in all other B lineage subtypes ALL. Three of the biphenotypic leukaemias coexpressed cytoplasmic B29 (CD79b) and mu-chain. This was also seen in two cases of CML-BC, while four cases expressed only cytoplasmic B29 (CD79b) without mu-chain. Our results suggest that during B cell differentiation, B29 (CD79b) is expressed later than mb-1 (CD79a) in the cytoplasm and parallels the cytoplasmic expression of mu-chain. B29 (CD79b) is present in the membrane at a later stage compared to its cytoplasmic expression and found in mature B blasts (B-ALL) that express membrane Ig as it is in normal and leukaemic B lymphocytes.

Antibodies, Monoclonal↗
PubMed1996

Cytogenetic findings in acute biphenotypic leukaemia.

We have studied the immunological and cytogenetic features of 26 patients with acute leukaemia classified as biphenotypic according to a scoring system based on the number and lineage specificity of antigens expressed on the blast cells. The series included 19 adults (age >15 years) and seven children. The cases were distributed in four immunophenotypic groups: (1)coexpression of myeloid and B antigens, 18 cases (69 percent);(2)myeloid and T cell antigens, six (23 percent); (3) one case with trilineage differentiation; and (4) one case with coexpression of both B and T cell antigens. Cytogenetic analysis revealed a normal karyotype in four cases (15 percent) and abnormal clones in 22 (85 percent). Eight patients had the Philadelphia (Ph) translocation, t(9;22)(q34;q11), (31 percent), three cases had structural aberrations of 6q and two had 11q23 rearrangements, one with t(11;19) and a second with t(4;11); the other eight cases had different alterations including t(9;12)(q1;q1), t(8;21)(q22;q22), t(2;7) (p1?3;q3?4), t(7;12)(q11;p11), hyperdiploidy and other structural abnormalities. The chromosomal rearrangements in children were characterised by abnormalities of 11q23 in two cases and the Ph translocation in three. Our data indicate that biphenotypic features are common in cases presenting with t(9;22) as the eight cases included here represent 47 percent of all cases of Ph+ve acute leukaemia studied in our Institution. Biphenotypic acute leukaemias comprise a heterogeneous group of leukaemias involving pluripotent stem cells. Cytogenetic studies are essential in characterising these cases as they will disclose several poor prognosis chromosome aberrations of which the Ph chromosome is the most frequent.

Adolescent↗
PubMed1996

Correlation of trisomy 12 with proliferating cells by combined immunocytochemistry and fluorescence in situ hybridization in chronic lymphocytic leukemia.

Conventional G-banding and fluorescence in situ hybridization (FISH) were performed on peripheral blood samples of 340 consecutive untreated cases of chronic lymphocytic leukemia (CLL) for the detection of trisomy 12 and other chromosome abnormalities. These findings were correlated with the proliferative activity of CLL lymphocytes assessed by the monoclonal antibody Ki-67. Cytogenetic analysis displayed a normal karyotype in 131 (38.5%) cases, trisomy 12 in 68 (20%), 31 by G-banding and an additional 37 cases by FISH, other clonal abnormalities in 47 (14%), and no metaphases in 94 (27.5%). The percentage of Ki-67-positive cells was significantly higher in cases with trisomy 12 (4.1 +/- 4.48) than in cases with a normal karyotype (1.5 +/- 2.0), those with other clonal abnormalities (1.35 +/- 1.37) and cases with no metaphases (1.14 +/- 1.6) (P< 0.0001). Cases with trisomy 12 were associated with more advanced clinical stage, atypical morphology and a higher percentage of Ki-67+ve cells than cases lacking trisomy 12 (P< 0.0001). Although there was no direct correlation between the percentage of trisomic and proliferating cells, the combination of immunocytochemistry and FISH showed that most Ki-67-positive cells were trisomic for chromosome 12. Our results suggest that the association of trisomy 12 with a higher proliferative activity supports the view that this abnormality is a secondary event associated with disease progression in CLL.

Adult↗